Role of histidine residues in EcoP15I DNA methyltransferase activity as probed by chemical modification and site-directed mutagenesis

Towards understanding the catalytic mechanism of M.EcoP15I [EcoP15I MTase (DNA methyltransferase); an adenine methyltransferase], we investigated the role of histidine residues in catalysis. M.EcoP15I, when incubated with DEPC (diethyl pyrocarbonate), a histidine-specific reagent, shows a time- and concentration-dependent inactivation of methylation of DNA containing its recognition sequence of 5'-CAGCAG-3'. The loss of enzyme activity was accompanied by an increase in absorbance at 240 nm. A difference spectrum of modified versus native enzyme shows the formation of N-carbethoxyhistidine that is diminished by hydroxylamine. This, along with other experiments, strongly suggests that the inactivation of the enzyme by DEPC was specific for histidine residues. Substrate protection experiments show that pre-incubating the methylase with DNA was able to protect the enzyme from DEPC inactivation. Site-directed mutagenesis experiments in which the 15 histidine residues in the enzyme were replaced individually with alanine corroborated the chemical modification studies and established the importance of His-335 in the methylase activity. No gross structural differences were detected between the native and H335A mutant MTases, as evident from CD spectra, native PAGE pattern or on gel filtration chromatography. Replacement of histidine with alanine residue at position 335 results in a mutant enzyme that is catalytically inactive and binds to DNA more tightly than the wild-type enzyme. Thus we have shown in the present study, through a combination of chemical modification and site-directed mutagenesis experiments, that His-335 plays an essential role in DNA methylation catalysed by M.EcoP15I.

Growth and Modification of Cluster-Assembled Thin Films

Thin film applications have become increasingly important in our search for multifunctional and economically viable technological solutions of the future. Thin film coatings can be used for a multitude of purposes, ranging from a basic enhancement of aesthetic attributes to the addition of a complex surface functionality. Anything from electronic or optical properties, to an increased catalytic or biological activity, can be added or enhanced by the deposition of a thin film, with a thickness of only a few atomic layers at the best, on an already existing surface. Thin films offer both a means of saving in materials and the possibility for improving properties without a critical enlargement of devices. Nanocluster deposition is a promising new method for the growth of structured thin films. Nanoclusters are small aggregates of atoms or molecules, ranging in sizes from only a few nanometers up to several hundreds of nanometers in diameter. Due to their large surface to volume ratio, and the confinement of atoms and electrons in all three dimensions, nanoclusters exhibit a wide variety of exotic properties that differ notably from those of both single atoms and bulk materials. Nanoclusters are a completely new type of building block for thin film deposition. As preformed entities, clusters provide a new means of tailoring the properties of thin films before their growth, simply by changing the size or composition of the clusters that are to be deposited. Contrary to contemporary methods of thin film growth, which mainly rely on the deposition of single atoms, cluster deposition also allows for a more precise assembly of thin films, as the configuration of single atoms with respect to each other is already predetermined in clusters. Nanocluster deposition offers a possibility for the coating of virtually any material with a nanostructured thin film, and therein the enhancement of already existing physical or chemical properties, or the addition of some exciting new feature. A clearer understanding of cluster-surface interactions, and the growth of thin films by cluster deposition, must, however, be achieved, if clusters are to be successfully used in thin film technologies. Using a combination of experimental techniques and molecular dynamics simulations, both the deposition of nanoclusters, and the growth and modification of cluster-assembled thin films, are studied in this thesis. Emphasis is laid on an understanding of the interaction between metal clusters and surfaces, and therein the behaviour of these clusters during deposition and thin film growth. The behaviour of single metal clusters, as they impact on clean metal surfaces, is analysed in detail, from which it is shown that there exists a cluster size and deposition energy dependent limit, below which epitaxial alignment occurs. If larger clusters are deposited at low energies, or cluster-surface interactions are weaker, non-epitaxial deposition will take place, resulting in the formation of nanocrystalline structures. The effect of cluster size and deposition energy on the morphology of cluster-assembled thin films is also determined, from which it is shown that nanocrystalline cluster-assembled films will be porous. Modification of these thin films, with the purpose of enhancing their mechanical properties and durability, without destroying their nanostructure, is presented. Irradiation with heavy ions is introduced as a feasible method for increasing the density, and therein the mechanical stability, of cluster-assembled thin films, without critically destroying their nanocrystalline properties. The results of this thesis demonstrate that nanocluster deposition is a suitable technique for the growth of nanostructured thin films. The interactions between nanoclusters and their supporting surfaces must, however, be carefully considered, if a controlled growth of cluster-assembled thin films, with precisely tailored properties, is to be achieved.

La norme du corps hybride : Une éthique de la reconstruction et de l'amélioration du corps humain en chirurgie

Aujourd'hui, techniques et technologies interagissent avec le corps humain et donnent aux personnes la possibilité de reconstruire leur corps, mais aussi de l'améliorer et de l'augmenter. L'hybridation est un processus technologique visant à compenser les défaillances humaines. L'augmentation de la puissance d'être est exaltée (santé, sexualité, performance, jeunesse), pourtant son accès n'est pas pour tous. Ce livre propose de démêler les différentes représentations du corps hybride et les projets qui les sous-tendent.

Conformational restrictions of peptides via backbone modification: Solution and crystal-state analysis of Boc-L-Pro-DZPhe-Gly-NH2

An N-alpha-protected model tripeptide amide containing, in the central position, an alpha,beta-dehydrophenylalanine (Z-configurational isomer), Boc-L-Pro-DELTA-Z-Phe-Gly-NH2 (Boc, tert-butyloxycarbonyl), has been synthesized by solution methods and fully characterized. IR absorption and H-1 NMR studies provided evidence for the occurrence of a significant population of a conformer containing two consecutive, intramolecularly H-bonded (type II-III') beta-bends in solution. However, an X-ray diffraction analysis clearly indicates that only the type-II beta-bend structure survives in the crystal state.

Chemical modification of sheep plasma glycoprotein

Glycoprotein isolated from sheep plasma was chemically modified, and the effect of chemical modification on biological activities and immunological cross reactions has been studied. The removal of sialic acid resulted in a change in the “overall conformation” of the glycoprotein as evidenced by a decrease in viscosity of the glycoprotein solution and an increased susceptibility of the glycoprotein to proteolytic enzymes. Sialic acid-free glycoprotein no longer inhibited the tryptic activity or prolonged the clotting time of plasma. However, it could react with the antiserum to sheep plasma glycoprotein. The periodate oxidation of sheep plasma glycoprotein resulted in a complete loss of inhibition of trypsin activity, prolongation of plasma clotting time, and the ability to cross-react with the rabbit antiserum. The significance of periodate oxidation in relation to the possible sequence of sugars in the carbohydrate prosthetic group in the glycoprotein is discussed. Iodination and heating in buffers of acid and alkaline pH values of sheep plasma glycoprotein resulted in complete loss of trypsin activity and ability to prolong plasma clotting time. Iodination of the glycoprotein did not affect the immunological cross-reactivity.

The Role of Interface Modification on Thermal Degradation and Crystallization Behavior of Composites from Commingled Polypropylene Fiber and Banana Fiber

The thermal degradation behavior of banana fiber and polypropylene/banana fiber composites has been studied by thermogravimetric analysis. Banana fiber was found to be decomposing in two stages, first one around 320 degrees C and the second one around 450 degrees C. For chemically treated banana fiber, the decomposition process has been at a higher temperature, indicating thermal stability for the treated fiber. Activation energies for thermal degradation were estimated using Coats and Redfern method. Calorific value of the banana fiber was measured using a constant volume isothermal bomb calorimeter. rystallization studies exhibited an increase in the crystallization temperature and crystallinity of polypropylene upon the addition of banana fiber. POLYM. COMPOS., 31:1113-1123, 2010. (C) 2009 Society of Plastics Engineers.

¿Feminismo indígena? : Un análisis crítico del discurso sobre los textos de la mujer en el movimiento zapatista 1994-2009

This work analyses texts on indigenous women´s participation in the Mexican Zapatista Army, Ejército Zapatista de Liberación Nacional. The EZLN came to public attention after ten years of clandestine organization in 1994 in Chiapas, a southern state of Mexico neighboring Guatemala. Along the invasion of various municipalities in Chiapas, the Zapatista Army published their own Revolutionary Laws, directed to the Mexican government that included a section on women´s own laws. The indigenous women´s participation in a guerrilla movement in the economically poorest area of Mexico raised many questions among Mexican feminists and some of them fiercely criticized the laws for not being liberating or feminist at all. The question is, did the indigenous women want the laws to be feminist? To answer the main research question How is the position of women constructed in the Zapatista discourse? I analyze texts by various actors in the discourse within the theoretical framework of critical discourse analysis and the feminist theories of intersectionality. The connecting point in this interdisciplinary framework is the question of power and hegemony. The actors in the discourse are the women commanders themselves, the men commanders, the Zapatista spokesperson, subcomandante Marcos and the Mexican feminists. The texts analyzed are the letters of the EZLN to the media and discourses in public reunions, first published in Mexican newspapers and international discussion lists on the Internet and after 2005, on the Zapatista´s own webpage. The results show that instead of discussing whether the Zapatista women´s participation is feminist or not, the action itself provoked such wide discussion of the diversity within the feminist movement that it is a contribution itself. The work also shows that the use of language can be one tool in the quite recent paradigm of intersectionality in feminist theories.

Crystal And Molecular Structure Of A Dimethyl Sulphoxide Complex With Lanthanum Nitrate, La(No3)34.(Ch3)2so

The crystal structure of the complex La(NO3)3.4(CH3)2SO has been solved by the heavy-atom method. The complex crystallizes in the monoclinic space group C2/e with four formula units in a unit cell of dimensions a= 14.94, b= 11.04, c= 15.54 A and fl= 109 ° 10'. The parameters have been refined by threedimensional least-squares procedures with anisotropic thermal parameters for all atoms except hydrogen. The final R index for 1257 observed reflexions is 0.094. The La 3 + ion is coordinated by ten oxygen atoms with La-O distances varying from 2.47 to 2.71 A. The geometry of the coordination polyhedron is described.

Le Grand Tour Du reste de l’Europe jusqu’à la Laponie – et retour

Rencontre entre le Sud et le Nord Depuis des siècles, les pays méditerranéens ont entretenu des relations actives avec le Nord de l’Europe. Les regions du Nord ont été conquises et converties, manipulées et colonisées, mais aussi intégrées dans les réseaux commerciaux, scientifi ques, culturels et touristiques. Les voyageurs du Sud de l’Europe ont peu à peu découvert les regions plurielles du Nord de la Finlande, de la Suède, de la Norvège et de la presqu’île de Kola. L’attraction focntionnait aussi dans l’autre sens. Pour les gens du Nord, scientifi ques ou artistes, le Sud de l’Europe constituait une destination aussi exotique aue la Laponie pour ceux du Sud. Dans ce recueil, nous proposons dix points de vue sur ces échanges et réseaux entre la Sud et le Nord. Le premier chapitre présente Tornio, connue très tôt comme la porte de la Laponie, les premières relations scientifi ques entre le Nord et le Sud, ainsi que les exhibitions de Samis dans les métropoles européennes. Le deuxième chapitre propose une analyse des experiences et des impressions de voyageurs italiens, espagnols et français. Le dernier chapitre est consacré aux voyages d’artistes et écrivains fi nlandais en France, principalement à Paris. Ces rencontres entre le Sud et le Nord qui participèrent à la remise en cause, la relativisation et la construction des identities nationals et ethniques, permirent aussi de créer une conscience européenne commune.

New family of thallium cuprate superconductors not containing calcium or barium: TlSrn+1−xLnxCunO2n+3+δ (Ln=La, Pr, or Nd)

The first two members of the new TlSrn+1−xLnxCunO2n+3+δ (Ln=La, Pr, or Nd) series of superconducting cuprates possessing 1021 and 1122 type structures are described. The n=1 (1021) members with Tcs around 40 K have electrons or holes as the majority charge carriers depending on x. The n=2 (1122) cuprate (Ln=Pr or Nd) shows a Tc in the 80–90 K range.

Rare-earth chromium citrates as precursors for rare-earth chromites: lanthanum biscitrato chromium(III) dihydrate, La[Cr(C6H5O7)2]·2H

The thermal decomposition of lanthanum biscitrato chromium(III) dihydrate has been studied in static air and dynamic argon atmospheres. The complex decomposes in four steps: dehydration, decomposition of the citrate to an intermediate oxycarbonate, formation of LaCrO4(V) from oxycarbonate, and finally decomposition of LaCrO4(V) to LaCrO3. Formation of LaCrCrO4(V) requires the presence of oxygen The decomposition behaviour of a mechanical mixture of lanthanum citrate hydrate and chromium citrate hydrate was compared with that of the citrato complex. Both the starting material and the intermediates were characterized by X-ray diffraction, IR electronic and ESR spectroscopy, surface area and magnetic susceptibility measurements, as well as by chemical analysis. A scheme is proposed for the decomposition of lanthanum biscitrato chromium(III) dihydrate in air. LaCrO3 can be obtained at temperatures as low as 875 K by isothermal decomposition of the complex.

La música en la fiesta del dios Xipe Totec

In Aztec mythology and religion, Xipe Totec ("our lord the flayed one") was a life-death-rebirth deity, god of agriculture, vegetation, the east, disease, spring, goldsmiths, silversmiths and the seasons. The music used for his worship has been described by a diversity of documents (carved stones, depicted codices, and chronicles made by the first European friars arrived to Mexico). This article explores the symbolic content of these descriptions, especially in association with Xochipilli Macuilxochitl, master of music and poetry.

El teponaztli en la tradición musical mexica

Lecture about the teponaztli (a specific type of slit drum), traditionally used by the Aztec (or Mexica) people, and its associated repertoire. Special attention is paid to the usage of this instrument in the Cantares Mexicanos (a song compilation from the 16th century) accompaniment.