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Home / Books / Modification and Editing of RNA
Modification and Editing of RNA
Editors: Henri Grosjean1, Rob Benne2
Affiliations: 1: Laboratoire d'Enzymologie et Biochimie Structurales, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France; 2: Department of Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
Content Type: Monograph
Category: Microbial Genetics and Molecular Biology
e-ISBN : 9781555818296
DOI: 10.1128/9781555818296
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This volume examines the various aspects of post-transcriptional RNA processing, including biosynthetic, genetic, and functional aspects of nucleoside modification and editing by nucleoside conversion as well as by nucleoside insertion/deletion.
Written by leading researchers in the field, it includes descriptions of the different types of modification/editing, identification of the correspondending enzymes, discussion of cis- and trans- acting sequences, machineries, mechanisms, cellular locations, the biological role of individual modified/edited bases, involvement in diseases like cancer and AIDS, and possible evolutionaly connections between different processes.
Modification and Editing of RNA is a valuable reference for researchers, teachers and students, and will be a useful tool for the planning of future research to solve many questions that remain.
Electronic Only, 610 pages, illustrations, appendixes, index.
Chapter 1 : Historical Perspectives on RNA Nucleoside Modifications
Author: Byron G. Lane
This chapter provides the historical roots of current avenues of research on RNA nucleoside modifications by describing some functional sites, structural environments, and biosynthetic strategies with which the modifications are associated. It describes selected examples of nucleoside modifications in ribosomal, transfer, messenger, and small nuclear RNAs. Emphasis is placed on conspicuous association of the most abundant nucleoside modifications in eukaryotic rRNA with the peptidyl transfer site in the ribosome. The chapter also recounts the historical and chronological development of general knowledge about RNA structure and function. The early identifications of modified nucleosides relied heavily on application of the same "new" approaches (anion-exchange and filter-paper chromatography and UV spectrophotometry) that were described in considerable detail in the first volume of The Nucleic Acids. In Ross Hall's monograph (1971), progress could already be reported about the biogenesis of base-and sugar-methylated nucleosides. When 2'- and 3'-O-aminoacyladenosines were prepared from the aminoacylated termini of tRNAs, they were among the first modified nucleosides to be identified as RNA constituents. Aminoacyladenosines, corresponding to the 20 (unmodified) amino acids found in proteins, have never previously been classified as modified nucleosides. The first hypermodified nucleoside to be reported and later correctly identified was described in a report in 1965. It seems fitting to remark that the direct functional consequence (frameshifting) of the first reported RNA editing event was instantly transparent, whereas the functional consequence of the first reported, and by far the most abundant, RNA modification (Ψ), has for more than 40 years remained all but opaque.
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Chapter 2 : RNA-Modifying and RNA-Editing Enzymes: Methods for Their Identification
Authors: Henri Grosjean, Yuri Motorin, Annie Morin
This chapter reviews the methods that have been successfully used to date for the identification of RNA modification and editing enzymes. The development of recombinant DNA and RNA techniques and techniques for chemical synthesis of DNA and RNA offers adequate alternatives for obtaining synthetic or semisynthetic unmodified or partially modified RNA substrates suitable for analyzing enzymatic posttranscriptional RNA modifications both in vivo and in vitro. Stepwise automated chemical synthesis of DNA oligonucleotides is widely used for producing starting material for gene construction, primers for PCR, DNA splints, or DNA probes for hybridization and deoxyribozymes. Two distinct approaches are generally used to gain the information on the precise location of modification or editing sites in RNA molecules. The first is based on direct sequencing of pure RNA and is applied mostly for the analysis of rather short RNA molecules, while the other is based on the use of reverse transcription with unfractionated RNA mixtures and specific oligonucleotide primers. Microinjection of antisense RNA, with the aim of blocking translation of selected mRNAs in the Xenopus laevis oocyte or fertilized frog eggs, revealed the existence of what was first identified as an unwinding RNA protein. With the recent access to complete genome sequences, it has become possible to search for homologous genes in the different genomes, to clone them into suitable vectors after amplification by PCR, and to express them in appropriate host cells.
Chapter 3 : Detection and Structure Analysis of Modified Nucleosides in RNA by Mass Spectrometry
Author: Pamela F. Crain
This chapter reviews applications of electrospray ionization (ESI) to the analysis of RNA and its constituents. Some examples of modified nucleosides discovered by liquid chromatography (LC)/mass chromatography (MS) and characterized include four sugar-methylated nucleosides in tRNA from hyperthermophilic archaea, 5-methylcarboxymethyl-2'-O-methyluridine in rat liver tRNASer[Sec] *, and two sugar-methylated nucleosides in the cap4 subunit of trypanosomal mRNA. A method has been implemented for mapping modified nucleosides in RNA, based on electrospray ionization mass spectrometry of RNase T1-derived oligonucleotides. The ability to mass measure the small peaks suggests that oligonucleotides can be analyzed at levels lower than the 20 pmol of tRNA utilized in the present example. To examine levels of modified nucleosides in tRNA as a function of growth temperature in the thermophilic archaeon Pyrococcus furiosus, it was grown at 70, 85, and 100°C. The strategies and examples of nucleic acid analysis using mass spectrometry presented in this chapter utilize ESI because of its successful implementation for the analysis of nucleic acids and their constituents from the monomer level in digests all the way up to intact tRNA and 5S rRNA.
Chapter 4 : Incorporation of Modified Nucleotides into RNA for Studies on RNA Structure, Function and Intermolecular Interactions
Authors: Robert A. Zimmermann, Michael J. Gait, Melissa J. Moore
This chapter focuses on techniques for intentionally incorporating both natural and unnatural modified nucleotides into RNA molecules. The incorporation of modified nucleotides can also facilitate the structural analysis of RNAs and RNA-containing complexes. The chapter provides a broad overview of the types of experiments that can be performed with chemically modified RNAs. It describes the properties of particular modified nucleotides, and discusses techniques for their incorporation into polynucleotide chains. It presents a number of examples of the ways in which synthetic approaches have been used to investigate a variety of problems. It also illustrates the methods that pertain to individual areas of expertise. Many of the modified nucleotides retain the structural properties of their unmodified counterparts and can be randomly incorporated into polynucleotide chains by enzymatic synthesis. RNA molecules with modified nucleotides at predetermined sites can also be constructed by semisynthetic methods in which the modified residue is introduced at the 5' end of the 3' RNA fragment by transcription and then ligated to the 5' RNA fragment. A novel strategy for the site-specific incorporation of modified nucleotides makes use of circularly permuted RNAs (cpRNAs). The approaches described in this chapter will prove useful in the future in the many exciting areas of research involving naturally modified and edited RNAs.
Chapter 5 : Biophysical and Conformational Properties of Modified Nucleosides in RNA (Nuclear Magnetic Resonance Studies)
Author: Darrell R. Davis
The location and identity of a large number of the modified nucleosides found in tRNA are highly conserved across diverse species and between tRNA isoacceptors. Conserved modification patterns in rRNAs and small nuclear RNAs also suggest that particular modified nucleosides have critical roles, although these functions are not well described compared to modifications found in tRNA. The nuclear magnetic resonance (NMR) determined biophysical properties have also provided an explanation for changes in important functions of tRNA during protein synthesis such as codon-anticodon recognition. This chapter talks about NMR determination of nucleoside conformation. The use of the percentage of 3'-endo sugar conformation as a measure of base stacking originated from NMR investigations on ribonucleotide dimers and trimers. Nucleoside modification in either the base or sugar can have significant effects on the sugar conformation, the glycosyl conformation, and base stacking. The 3'-gauche effect is mechanistically related to the anomeric effect in that it arises due to a geometrically dependent overlap between σ* and n lone pair molecular orbitals. Nucleoside bases that are in a stacked geometry generally have aromatic NMR proton shifts that are upfield of the positions seen in the absence of significant stacking. The chapter also focuses on conformational and thermodynamic effects of specific nucleoside modifications. Our knowledge of the fundamental biophysical properties of modified nucleosides will be critical to understanding how modification has been used throughout biology to optimize the function of biochemically critical processes involving RNA.
Chapter 6 : Effects of Pseudouridylation on tRNA Hydration and Dynamics: a Theoretical Approach
Authors: Pascal Auffinger, Eric Westhof
This chapter focuses on the structural implications resulting from the occurrence of this residue in tRNA and also presents an overview of the tRNA pseudouridylation sites. The principle of molecular dynamics (MD) resides in the numerical integration of the Newtonian equations of motion. The potential energy function contains several terms that account for covalent bond stretching, bond angle bending, harmonic dihedral bending, and nonbonded interactions including van der Waals and Coulombic terms. The residence time of this water molecule, which is consistently observed in several MD simulations, was estimated to be significantly longer than 500 ps. This water molecule forms an important structural link between the nucleotide backbone and the modified base and, thus, reduces the conformational mobility of the RNA close to the pseudouridylation site. Summarizing the preceding information suggests strongly that the main function of pseudouridylation close to the anticodon is to stabilize the structure of the loop by reducing its intrinsic dynamics, likely to avoid codon misreading. Pseudouridines are also important in ribosomal RNA and small nuclear RNA. It would not be surprising that in most structural contexts pseudouridines would stabilize rRNA and snRNA in a similar manner as in tRNA. In snRNA, the occurrence of pseudouridines in single-stranded regions may be required to improve their recognition features. Recent MD studies on the conformational behavior of 2'-OH groups in tRNA may be considered as a first approach toward the investigation of the roles of 2'-O-methyl groups.
Chapter 7 : Modulatory Role of Modified Nucleotides in RNA Loop-Loop Interaction
Authors: Henri Grosjean, Claude Houssier, Pascale Romby, Roland Marquet
The anticodon stem loop of tRNA is one of the most remarkable hairpins in RNAs that present several modified nucleotides arising by posttranscriptional processes. Many different modifications have been identified at position 34. These modifications are considered to be important for the specificity, fidelity, and efficiency of the tRNA functions, mainly in decoding the genetic message during translation on the ribosome. This chapter summarizes the information available about the role of modified nucleotides present at positions 34 and 37 in modulating tRNA anticodon-anticodon interaction. Implications for codon-anticodon interaction and comparison with other types of RNA-RNA interacting systems are also discussed in this chapter. One way to evaluate the effect of modified nucleotides on the stability and dynamic properties of anticodon-anticodon complexes is the temperature-jump relaxation technique. The chapter describes two RNA loop-loop interacting systems that do not involve modified nucleotides. Their comparison with the loop-loop interacting systems involving hairpins of tRNA molecules described in this chapter are interesting to better understand how modified nucleotides can restrict the interaction to only a few selected nucleotides within the interacting RNA loops. Comparison to other types of loop-loop complexes involving RNAs other than tRNAs, the so-called "kissing" complexes, is also made. The main difference between these different types of loop-loop complexes is the extent of base pairing between the nucleotides of the interacting loops.
Chapter 8 : Mechanisms of RNA-Modifying and -Editing Enzymes
Authors: George A. Garcia, DeeAnne M. Goodenough-Lashua
This chapter focuses on mechanistic issues involved in RNA-modifying enzymes. From a chemical/structural viewpoint, modified nucleosides can be divided into two groups. The first group consists of relatively "simple" modifications (e.g., methylation, thiolation, deamination, and isomerization). The second group of modified nucleosides consists of more "complex" modifications (e.g., multiple modifications and hypermodifications), involving a multi-enzyme pathway or, as is the case with queuine and archaeosine, involving biosynthetic precursors synthesized by other enzymes for this purpose alone. The X-ray crystal structure of a nucleoside adenosine deaminase has been determined, and a zinc ion and an ordered water molecule have been located in the active site. The X-ray crystal structure of cytidine deaminase complexed with uridine has also recently been determined. The miaA enzyme utilizes Δ2-isopentenyl pyrophosphate (IPP, or dimethylallyl diphosphate) as the isopentenyl group donor. Δ2-IsopentenyI pyrophosphate is utilized by a number of isopentenyl transferases leading to various isoprenoids and ultimately to steroids. Two reaction mechanisms have been postulated for the isopentenyl transferases, an associative mechanism and a dissociative mechanism. Technological advances in RNA generation (both by in vitro transcription and by chemical synthesis) along with advances in molecular biological approaches (to identify, clone and express modifying enzyme genes) have been predominantly responsible for the renewed activity in RNA modification and editing research.
Chapter 9 : Structural Basis of Base Exchange by tRNA-Guanine Transglycosylases
Authors: Christophe Romier, Dietrich Suck, Ralf Ficner
Contrary to most other modified bases, queuine and archaeosine biosyntheses start outside the tRNA and require a base exchange at the tRNA level, a reaction catalyzed by enzymes known as tRNA-guanine transglycosylases (TGTs). Several groups have reported the isolation of the eukaryotic TGT, but these reports do not agree on its oligomeric state and the size of its subunits. An archaebacterial TGT has been isolated very recently. This enzyme is a 78-kDa protein that has been shown through partial sequencing to be sequence-related to the prokaryotic TGT. TGT is certainly one of the most interesting enzymes of the queuine biosynthesis pathway because it catalyzes a reaction (a base exchange) which is also observed in many other cellular processes involving DNA and RNA. The structure of Zymomonas mobilisTGT was solved by the well-known technique of multiple isomorphous replacement (MIR). The soaking approach employed with the small preQ1, molecule cannot be used with a tRNA macromolecule and the only way to get the structure of a prokaryotic TGT-tRNA complex is by cocrystallization of the components. The three-dimensional structure of queuosine monophosphate shows that the β-configuration of the ribose is preserved. The recent occurrence in sequence databases of eukaryotic and archaebacterial proteins highly homologous to the prokaryotic TGTs defines a clear phylogenetic link and suggests that TGTs from all three kingdoms have a common fold and a common catalytic mechanism.
Chapter 10 : Biosynthesis and Functions of Modified Nucleosides in Eukaryotic mRNA
Authors: Joseph A. Bokar, Fritz M. Rottman
This chapter focuses on the sequence-specific methylation of internal adenosine residues to form N6-methyladenosine. It talks about nucleosides modifications present in cellular mRNA; the nucleoside modifications present in virion RNA and viral mRNA are also be discussed because of the nearly complete overlap of the spectrum of modifications present. Lower eukaryotes such as yeast and Neurospora contain only cap 0. Collectively, these cap methylation events account for the presence of most of the nucleoside modifications in eukaryotic mRNA, namely, 7-methylguanine (m7G), all four 2'-O-methylnucleosides (Nm), and N6,2'-Odimethyl-adenosine (m6Am). Three modified nucleosides have been reported within internal regions of eukaryotic mRNA: N6methyladenosine (m6A), 5-methylcytidine (m5C), and inosine. A study examined short (20 nucleotide) synthetic RNA substrates and confirmed that the in vitro specificity closely paralleled the frequency with which sequences are methylated in vivo as originally described by Schibler. This study was extended by evaluating an additional two m6A sites in a similar fashion. Camper and his coworkers utilized an inhibitor, S-tubericidinylhomocysteine (STH), which is a structural analog of S-adenosylmethionine (SAM) and a potent inhibitor of SAM-dependent methyltransferases. In this study, the effects of the inhibitor on cap methylation and internal methylation were closely monitored. Posttranscriptional modification of nucleosides within mRNA molecules is an integral aspect of premRNA processing. These modifications are characteristic of eukaryotic mRNA only; no equivalent nucleotide modifications have been found in any of the prokaryotic mRNAs characterized so far.
Chapter 11 : Posttranscriptional Modifications in the U Small Nuclear RNAs
Authors: Séverine Massenet, Annie Mougin, Christiane Branlant
This chapter deals with the posttranscriptional modifications of metabolically stable small nuclear RNAs (snRNAs) and the possible link to the function of these molecules. A study of the posttranscriptional modifications in the AT-AC specific spliceosomal snRNAs was started, and the results obtained are described in this chapter. The identity and location of all the detected posttranscriptional modifications are shown in the chapter. Interestingly, the sequence involved in where this pseudoknot structure is located contains posttranscriptional modifications: two 2'-O methylations and three pseudouridines. Posttranscriptional modifications may be required for stabilization of the pseudoknot structure. The importance of the posttranscriptional modifications of the human U2 snRNA for spliceosome activity was clearly demonstrated by reconstitution of the spliceosome with HeLa cell nuclear extracts depleted of U2 snRNA and complemented with in vitro transcribed U2 snRNA. It has now been clearly demonstrated that 5'-terminal posttranscriptional modifications play an essential role for the biogenesis of the spliceosomal U snRNPs in eukaryotes, and this is probably also the case for the U snRNAs involved in the recently discovered minor splicing machinery of vertebrates. The various recent experiments that are previously summarized in the chapter are strongly in favor of a direct functional role of U snRNA posttranscriptional modifications in spliceosome assembly and splicing.
Chapter 12 : The Pseudouridine Residues of rRNA: Number, Location, Biosynthesis, and Function
Authors: James Ofengand, Maurille J. Fournier
This chapter focuses on the pseudouridine (5-β-D-ribofuranosyluracil; Ψ) residues in rRNA. This subject has been reviewed previously both in a detailed analysis of the work on eukaryotic rRNA up to circa 1990, which also includes methylated nucleosides, and in a review of more recent work. The chapter talks about number and locations of Ψ in small-subunit (SSU) and large-subunit (LSU) rRNAs. Synthesis of cytoplasmic rRNAs in eukaryotic cells involves the action of a large population of small nucleolar RNAs (snoRNAs). Site selection in each case involves base pairing of a guide snoRNA with the rRNA segment to be modified, and selection of a nucleotide located at a constant distance from an additional determinant(s) in the snoRNA. The two types of guide function are provided by snoRNAs in separate families known as the box C/D and H/ACA box families, respectively. Each family contains snoRNAs required for rRNA processing, but the main function of these RNAs is the modification of rRNA nucleotides. There is no firm evidence so far for an essential role for any in the cell, and a number of cases are known where deletion of a single Ψ has no obvious effect. It is likely that additional pseudouridine synthases will be identified and characterized and that they will come from new and familiar sources.
Chapter 13 : Small Nucleolar RNAs Guide the Ribose Methylations of Eukaryotic rRNAs
Authors: Jean-Pierre Bachellerie, Jérôme Cavaillé
This chapter provides a progress report on developments that should set the stage for dissecting the rRNA-ribose methylation machinery and for deciphering the role of these modifications in ribosome assembly and function. Interestingly, ribose methylations are far from being uniformly distributed within the universally conserved domains of eukaryotic rRNAs. The finding that small RNA guides specify the sites of ribose methylation in eukaryotic rRNAs raises the issue that RNA cofactors also could be involved in prokaryotes. Prokaryotic rRNAs have considerably fewer ribose methylations than eukaryotic rRNAs, with only 4 sites in E. coli. Two prokaryotic rRNA ribose methylases have been characterized so far that are each specific for a single site in rRNA. The presence of a complex repertoire of site-specific, trans-acting snoRNA guides sufficient to precisely target the modification through the formation of a common canonical duplex structure with the rRNA substrate strongly suggests that the enzymology of the reaction is quite simple, at least as to the variety of protein enzymes involved. snoRNA-guided ribose methylation of rRNA involves the formation of scores of 10-21 bp long intermolecular RNA duplexes along the elongating pre-rRNA transcript in eukaryotes. Analyzing the structure of the methylase(s) and pseudouridine synthase(s) involved in the modification of eukaryotic rRNAs, with particular reference to RNA editing enzymes, also recognizing double-stranded RNA structures could be illuminating.
Chapter 14 : Functional Aspects of the Three Modified Nucleotides in Yeast Mitochondrial Large-Subunit rRNA
Author: Thomas L. Mason
This chapter focuses on the possible functional significance of the three highly conserved modified nucleotides in the otherwise minimally modified yeast mitochondrial large-subunit rRNA (LSU rRNA). The three modified nucleotides in yeast mitochondrial LSU rRNA, Gm2270, Um2791, and pseudouridine 2819, are equivalent to the Gm2251, Um2552, and pseudouridine 2580, respectively, in peptidyl transferase center (PTC) of Escherichia coli LSU rRna. The PET56 nuclear gene encodes an rRNA ribose methyltransferase (Pet56p) required for the formation of 2'-O-methylguanosine at G2270 in yeast mitochondrial LSU rRNA (G2251 in E. coli numbering), and PET56 function is essential for the formation of functional mitochondrial ribosomes. While PET56 is normally essential for the formation of functional yeast mitochondrial ribosomes, a dominant extragenic mutation has been isolated (SRM1-1) that suppresses, albeit very weakly, pet56 loss-of-function mutations without restoring methylation at G2270 in yeast mitochondrial LSU rRNA. The presence of only three modified nucleotides at highly conserved positions in the PTC of yeast mitochondrial LSU rRNA has fueled speculation that these particular modification might have special functional significance. The recent progress in the identification of genes responsible for specific modifications in E. coli, yeast mitochondrial, and yeast cytoplasmic rRNAs will accelerate the genetic analysis of rRNA modification in these systems. It will be particularly informative to compare the in vivo effects of depleting pseudouridine 2580 in E. coli and yeast mitochondria and Gm2251 and Um2552 in all three ribosomal systems.
Chapter 15 : Regulatory Aspects of rRNA Modification and Pre-rRNA Processing
Authors: Denis L. J. Lafontaine, David Tollervey
The major types of posttranscriptional modification are the isomerization of uracil to pseudouridine, 2′-O-methylation of the ribose moieties and base methylation. In addition to their roles in pre-rRNA modification, both Dim1p and Cbf5p are required for pre-rRNA processing, and Pet56p is required for synthesis of mitochondrial large ribosomal subunits. This chapter discusses data on Dim1p and Cbf5p that provide insights on the type of systems that may act to coordinate the many steps in ribosome synthesis and the origins of the snoRNA-directed rRNA modification systems present in eukaryotic cells. Uncoupling of the dimethylation and prerRNA processing defects showed that Dimlp rather than the dimethylation activity is required for prerRNA processing. This observation did not, however, determine whether Dim1p is itself directly required for pre-rRNA processing. The binding of Dim1p to the pre-rRNA may therefore be monitored to ensure that it fulfills its functions in both modification and assembly. Ribosome synthesis is a highly dynamic process during which a vast number of processing, modification, and assembly reactions occur simultaneously. The requirement for Cbf5p in pre-rRNA processing appears to be quite different from the requirement for Dim1p. Eukaryotic ribosome synthesis involves very complex pre-rRNA processing and assembly pathways. These include numerous steps occurring in different cellular compartments and requiring a plethora of RNA and proteins, many of them in the form of snoRNPs that only transiently associate with the preribosomal particles. The archaea may hold the final clues to understanding the origins of the small nucleolar ribonucleoproteins (snoRNPs)-directed systems of rRNA modification.
Chapter 16 : Editing of tRNA
Authors: David H. Price, Michael W. Gray
The term "RNA editing" was first coined more than a decade ago to describe the phenomenon of uridine insertion into trypanosomatid mitochondrial transcripts. Unlike mRNAs, transcripts of tRNA and rRNA genes are themselves converted into the functional entities they encode. RNA editing events represent additional steps in posttranscriptional processing and, like nucleoside modifications, they may occur at different stages in the pathway. To quantify RNA editing one usually compares the intensity of the bands on a sequencing (or primer extension) ladder that correspond to the edited and unedited versions of the RNA. The first example of tRNA editing in a mitochondrial (mt) system is that reported to occur in the ameboid protozoon Acantbamoeba castellanii. The observed nucleotide substitutions consisted of both purine-to-purine and pyrimidine- to-purine changes, suggesting a mechanism involving base or nucleotide replacement. More recently, it has been shown that nascent mRNAs present in stalled RNA polymerase complexes are substrates for editing by both cytidine and dinucleotide insertion. In a particular study the isolated RNAs were found to be edited to within 14 to 22 nucleotides of the stalled polymerase, suggesting that the insertional editing activity in Physarum is able to act quite close to the site of RNA synthesis. The most recently described mode of tRNA editing is that found in the mitochondria of metazoa.
Chapter 17 : RNA Editing by Base Conversion in Plant Organellar RNAs
Authors: Anita Marchfelder, Stefan Binder, Axel Brennicke, Volker Knoop
Awareness of the RNA editing process in plants grew gradually. The first discrepancies in nucleotide identities between mRNA and DNA were noted as the appearance of thymidylates in the sequence ladders of cDNAs at positions of cytidylates in the genomic sequence. The existence of RNA editing in plant mitochondria resolved initial problems with nonconserved amino acid codons found in the plant gene sequences. A schematic view of RNA editing in plant organelles is discussed in this chapter. In analogy to the analysis of RNA editing in other systems, the chapter also presents a separate discussion of the process in plant organelles, that is, the biochemistry of the modification of nucleotide identity and the specificity determinants, which decide where editing should occur. Evolutionary selection and streamlining should have eliminated RNA editing, unless there is some advantage to be gained for the genetic system. RNA editing significantly increases the efforts required to analyze gene expression in plant organelles. RNA editing in plastids and mitochondria also complicates in vitro manipulation of organellar genes and expression. The similarity of the RNA editing processes in plastids and mitochondria of a given plant cell suggests a common evolution of the respective editing processes and very similar mechanistic features.
Chapter 18 : Apolipoprotein Β mRNA Editing
Authors: Benny Hung-Junn Chang, Paul P. Lau, Lawrence Chan
Apolipoprotein B (apoB) mRNA editing was the first instance of RNA editing described in vertebrates. The subcellular location for apoB mRNA editing has been examined in rat liver. The situation in vivo, however, is much more complex, because the presence of the mooring sequence is not necessarily associated with editing of its upstream C. When researchers in 1997 overexpressed apobec-1 in mice, a novel mRNA, NAT1, was found to be edited. The evolutionary relationship of apobec-1 with the cytidine/cytidylate deaminases has been noted by a number of investigators. This chapter outlines a few salient points concerning the evolution of the apobec-1-related deaminase gene family, taking into consideration some novel homologous sequences published earlier. Based on the current knowledge of the area, a model of the apoB mRNA editing enzyme complex is presented. The presence of a lipoprotein phenotypic effect in animals with only partial inhibition of editing activity in the liver contrasts sharply with the relative lack of an effect in apobec-1 knockout mice that do not edit apoB mRNA either in the liver or the small intestine.
Chapter 19 : Adenosine-to-Inosine Conversion in mRNA
Authors: Susan M. Rueter, Ronald B. Emeson
The conversion of adenosine to inosine (A-to-I) by RNA editing, at specific positions within RNAs, represents an increasingly common posttranscriptional modification for generating diversity and flexibility in eukaryotic gene expression. This chapter describes a number of mRNA substrates that undergo A-to-I editing events, the effects that such alterations in coding potential have upon protein function, and the candidate enzymes responsible for such posttranscriptional modifications. While nucleotide sequence analyses have revealed the presence of a CGG codon encoding the critical regulatory arginine residue within the TM2 region of GluR-B cDNAs, a glutamine (CAG) codon was found in GluR-B genomic DNA at this position Q/R site. The editing of RNAs encoding glutamate receptor subunits results from the conversion of genomically encoded adenosine residues to the guanosine-like nucleotides found in cDNAs generated from mature GluR mRNA transcripts. The hepatitis delta virus (HDV) is a subviral human pathogen whose packaging and propagation are dependent on concurrent infection with the hepatitis B virus. Recent studies of RNA editing in polyomavirus have suggested that this posttranscriptional modification represents a mechanism by which RNA transcripts expressed early after viral infection are inactivated subsequent to viral DNA replication (the late phase). While future work will further define the cellular and molecular basis for dsRNA adenosine deamination, the functional consequences of subtle A-to-I changes in mRNAs and the alterations in amino acid sequence for resultant proteins are bound to be profound.
Chapter 20 : Nucleoside Deaminases for Cytidine and Adenosine: Comparison with Deaminases Acting on RNA
Author: Charles W. Carter, Jr.
This chapter reviews the structure of Escherichia coli cytidine deaminase and implications for evolutionary relationships between members of the cytidine deaminase superfamily. It then summarizes details of the E. coli cytidine deaminase (ECCDA) reaction mechanism, with particular reference to the role of conserved residues and aspects of substrate specificity relating to the nucleoside modifying and editing enzymes. The chapter concludes with a discussion of models for RNA editing deaminases derived from sequence homologies and structural studies. APOBEC-1, the sole representative of what will likely be a family of editing cytidine deaminases acting on RNA substrates, shares this signature and, is outlined in detail, it likely has significant tertiary and quaternary homology to the E. coli cytidine deaminase, ECCDA. An attempt has been made to rationalize the naming of editing adenosine deaminases, calling them adenosine deaminases acting on RNA (ADARs). Conversion of cytidine to uridine involves two successive steps: hydration of the 3-4 double bond, and subsequent elimination of the leaving ammonia molecule with formation of the keto tautomer of the pyrimidine. A broad, representative sample of APOBEC-1 mutants was examined by biochemical assays for homodimerization, RNA binding, and RNA editing. The X-ray structure has been determined for murine adenosine deaminase, which catalyzes deamination of adenosine to inosine. The secondary and tertiary structure motifs of the two nucleoside deaminases are completely unrelated. The outlines described in the chapter of how C-to-U and A-to-I editing occurs provide coherent hypotheses based on known structures of related enzymes.
Chapter 21 : Mitochondrial mRNA Editing in Kinetoplastid Protozoa
Authors: Stephen L. Hajduk, Robert S. Sabatini
The mRNAs in the mitochondrion of kinetoplastid protozoa are often modified by the precise addition or deletion of uridylates (U's) by a process termed RNA editing. This chapter focuses on the molecular basis of RNA editing, and the integration of biochemical mechanism with the structure of the editing machinery. The mitochondrial mRNAs are translated on mitochondrial ribosomes and produce functional proteins. It is clear that while minicircles do not encode proteins directly, they play an important role in RNA maturation by encoding most of the guide RNAs (gRNAs) necessary for the editing of mitochondrial mRNAs. The secondary structures determined for the four gRNAs, while similar, have Gibbs free energies well below those predicted for the most stable structures for these RNAs. In a series of UV cross-linking studies, gRNAs binding proteins have also been identified from Trypanosoma brucei mitochondrial extracts. When different gRNAs specific to different mRNA editing sites are incubated with mitochondrial extracts, the same eight proteins are able to bind. Based entirely on analysis of genomic and cDNA sequences, we can predict with some certainty that RNA editing is obligatory for the formation of mitochondrial mRNAs with correct initiation and termination codons and continuous open reading frames to encode these mitochondrial proteins.
Chapter 22 : RNA Editing in Physarum Mitochondria
Authors: Jonatha M. Gott, Linda M. Visomirski-Robic
In vitro approaches have facilitated mechanistic studies designed to look at the timing, accuracy and efficiency of insertional editing of newly synthesized RNAs. Because different assays are required for these distinct RNA populations, the two approaches are discussed separately. Importantly, upon incubation under editing conditions, nucleotide insertion into previously synthesized, unedited RNA was not observed, even though newly transcribed RNA made during the ''chase'' reaction was fully edited . The lack of unedited regions in RNAs labeled in isolated mitochondria under standard, non-limiting nucleotide concentrations and the proximity of RNA editing to the site of transcription, coupled with the inability of the editing activity to insert nucleotides into previously synthesized RNAs, argue that there is a limited ''window of opportunity'' for editing in Physarum mitochondria. Given the proximity of nucleotide insertion to the active site of transcription, it is likely that the Physarum mitochondrial RNA polymerase plays some role, direct or indirect, in the editing process. Nonencoded nucleotides are known to be added very close to the growing end of the RNA chain in Physarum mitochondria , and it is possible that they are added concurrently with RNA synthesis. Although transcription normally is templated, many RNA polymerases are capable of adding nucleotides in a nontemplated or pseudotemplated manner. Much of the mitochondrial genome of Physarum has yet to be sequenced, and it is possible that additional types of editing events may yet be discovered upon characterization of the remaining genes and their RNA products.
Chapter 23 : Cotranscriptional Paramyxovirus mRNA Editing: a Contradiction in Terms?
Authors: Daniel Kolakofsky, Stéphane Hausmann
The Paramyxovirinae currently are organized in three genera: the paramyxovirus genus (including Sendai virus [SeV], human parainfluenza virus type 1 [hPIV1], and human and bovine PIV3); morbilliviruses [e.g., measles virus and the distemper viruses]; and rubulaviruses (e.g., mumps virus and simian virus 5 {SV5}]). The first rubulavirus P gene sequence determined correctly was that of SV5. This yielded the surprising result that there was no single open reading frame (ORF) large enough to code for the SV5 P protein. mRNA transcribed from cDNAs that were exact copies of the SV5 P gene could be translated into the shorter V protein encoded by the 5' proximal ORF, but not into the longer P protein. Paramyxovirus RNA synthesis is restricted to the cytoplasm, and these viruses fend for themselves in all aspects of mRNA synthesis. Given that they formed their poly (A) tails by the unusual mechanism of polymerase stuttering, this mechanism was immediately suggested as one that could also account for the unusual G insertions. Experimental evidence in favor of a stuttering mechanism came from work with SeV, a member of the remaining paramyxovirus genus. SeV mRNAs made in vitro with purified virions contained the same pattern of G insertions as those found intracellularly, indicating that the process was carried out by viral gene products.
Chapter 24 : Intracellular Locations of RNA-Modifying Enzymes
Author: B. Edward H. Maden
This chapter talks about the subcellular locations in which the modifications to eukaryotic RNA occur, and focuses mainly on tRNA and rRNA. Most tRNA molecules are built to a common general design, giving rise to the well-known cloverleaf secondary structure and "inverted L" tertiary structure. Availability of the genes and the means of manipulating their expression is of great importance, as stressed for several examples in the chapter, including PUS1 in the section on cytoplasmic tRNA, TRM1 and MODS in the section on mitochondrial tRNA and DIM1 in the section on cytoplasmic rRNA. Although the chapter has concentrated on eukaryotes, in which the questions of intracellular locations and organization are fairly obvious, it should not be excluded that some level of intracellular organization of RNA-modifying enzymes could also occur in prokaryotes, for example by multienzyme interactions between RNA-modifying enzymes and other enzymes of RNA function, such as aminoacyl-tRNA synthases.
Chapter 25 : Genetics and Regulation of Base Modification in the tRNA and rRNA of Prokaryotes and Eukaryotes
Author: Malcolm E. Winkler
This chapter describes some of the better-understood systems, primarily in bacteria and yeast, that exhibit regulation of modification enzyme activity or gene expression. It attempts to bring out the common themes about regulation that are emerging from ongoing studies of the modification process. The efficiency of nonsense codon suppression has been exploited to analyze the effects of tRNA modification on decoding and codon context in bacteria and eukaryotes. New genetic approaches combined with high-pressure liquid chromatography (HPLC) screening and reverse genetics will likely lead to the identification of the remaining tRNA and rRNA modification genes in bacteria and yeast. A recent determination by quantitative Western immunoblotting showed that the MiaA (i6A37) prenyltransferase is also a moderately abundant enzyme at about 650 monomers per cell (≈ 1 μM) in bacteria growing exponentially in enriched minimal-glucose medium. MiaA may need to be present in comparatively high cellular amounts, because its activity is strongly competitively inhibited for its prenyl substrate, dimethylallyl diphosphate (alternatively called Δ2-isopentenyl pyrophosphate), by nucleotide di- and triphosphates. The genes whose expression is affected by these undermodifications need to be identified by two-dimensional gel analyses or genetic methods using random lacZ fusions. After these targets are identified, it will be possible to access whether the magnitude of the effects of undermodification on gene expression is sufficiently large to readily indicate physiological significance.
Chapter 26 : Links between tRNA Modification and Metabolism and Modified Nucleosides as Tumor Markers
Authors: Glenn R. Björk, Torgny Rasmuson
tRNA is a key component in translation. The various tRNA species decode at different speeds and fidelity that depend on the availability of the tRNA species, of the codon choice, and of the codon context. Some tRNA may act as a cofactor in enzymatic reactions. The first precursor to porphyrins in many bacteria and organelles is glutamate-1-semialdehyde (GSA), which is formed after the ligation of Glu to tRNAGlu. In bacteria, an unexpected link between tRNA modification and intermediary metabolism is the requirement of chorismic acid or its derivative in a previously unknown metabolic pathway to synthesize uridine-5-oxyacetic acid (cmo5U34) and its methyl ester (mcmo5U34). Thus, whereas links between tRNA modification and metabolism are well documented in bacteria, such links also exist in eukaryotes. In a study of human immunodeficiency virus (HIV)-positive intravenous heroin addicts, researchers found that serum ψ was an independent predictor of progression to AIDS in Centers for Disease Control and Prevention (CDC) stage A2 HIV-infected patients. Interestingly, queuine in higher organisms may have such a role, since it influences the physiology of the cell in a tRNA independent way. In multicellular organisms, there is turnover of stable RNA, and modified nucleosides are found intact in rather large amounts in serum and in the urine. Therefore, urinary excretion of modified nucleosides reflects the turnover of RNA. Most of the RNA, which contains modified nucleosides in the multicellular eukaryotes, is stable and participates in translation (rRNA and tRNA) or in splicing (the various U RNAs).
Chapter 27 : Modified Nucleosides in Translation
Author: James F. Curran
Cellular physiology is fundamentally dependent on the functions of translational apparatus, and these functions are dependent on modified nucleosides. This chapter examines the translational functions of modified nucleosides in the anticodon arm of tRNA. There are modifications at other positions within tRNA, but our knowledge of translational effects is limited to the modifications in the anticodon region. Emphasis is placed on the effects that the loss of specific modifications has on the activities of tRNA. Before considering the effects of modifications on translation, it is helpful to review certain aspects of the decoding process. Further work on the translational mechanism is needed to fully understand the roles of modified nucleosides in the important cellular process. The chapter discusses the effects of modified nucleosides at various positions in the anticodon arm. There are data on the translational effects of a subset of the modified nucleosides that occur within the anticodon arm. The chapter talks about a discussion of unmodified U34 because models for decoding by the modified forms are extended from those for unmodified U. Virtually all tRNAs contain modified nucleosides within the anticodon region, and it has become abundantly clear that they contribute to translation in a number of ways. The future looks very promising for the continued study and understanding of the roles that modified nucleosides play in the fundamentally important translational process.
Chapter 28 : Importance of Modified Nucleotides in Replication of Retroviruses, Plant Pararetroviruses, and Retrotransposons
Author: Roland Marquet
Retrotransposons are widespread transposable elements that are closely related to retroviruses. As for all retroids, the replication cycle of retroviruses, retrotransposons, and pararetroviruses includes a reverse transcription step. Reverse transcription of the retroviral genome takes place in the cytoplasm of the infected cell. In retroviruses and retrotransposons, the viral enzymes, including reverse transcriptase (RT), are synthesized as Gag-Pol precursors, while the pol gene of cauliflower mosaic virus (CaMV; the caulimovirus prototype) is expressed separately from the gag gene. It is not only the modified nucleotides of tRNAs, but also those present in the genomic RNA itself might affect the replication of retroviruses and retrotransposons. The identification of such modifications and their possible roles are described in this chapter. Modified nucleotides might be critical for a number of steps in the replication life cycle of retroviruses, plant pararetroviruses, and retrotransposons. The importance of modified nucleotides of the primer tRNA for encapsidation and reverse transcription is well established. It is important to realize that no general conclusion that would be valid for all retroviruses, plant pararetroviruses, and retrotransposons can be drawn regarding the importance of modified nucleotides. Human retroviruses cause several fatal diseases, including AIDS (HIV-1 and HIV-2), leukemia (HTLV-I and HTLV-II), and possibly multiple sclerosis. Obviously, defining the exact roles of modified nucleotides in the replication life cycles of plant pararetroviruses, retrotransposons, and especially retroviruses deserves more attention.
Chapter 29 : Modified Nucleosides Always Were: an Evolutionary Model
Authors: Nicolas Cermakian, Robert Cedergren
This chapter evaluates the distribution of modified nucleosides in certain RNA species of the three domains of life (archaea, bacteria, and eukaryotes). Fourteen modifications are present in all three domains and of these several are found at comparable positions in tRNAs . First, most of the conserved modified nucleosides are in or near the anticodon loop. There is strong phylogenetic evidence for the early appearance of modified nucleosides. The incorporation of modified nucleosides into nucleic acid polymers is quite plausible, and the use of a particular nucleoside may depend only on its concentration in the primordial soup. Again, considering the chemistry of nucleosides, it would seem that the moderately modified nucleosides would be present in the highest concentrations in the primordial soup along with the standard nucleosides. In parallel, the phylogenetic evidence suggests that some simple modified nucleosides existed at the point when the major organismal groups diverged. In the case of tRNAs, there is no evidence for present-day RNA guiding, although such a system could be imagined early in the evolution of replicating cells. The current evidence of the enzymes involved in RNA-guided modification leaves the door open to speculation that RNA has a greater role in this process than simply guiding modification.
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For the sake of projects, the newly mixed mortar should have good workability, and the hardened mortar should have the required strength, the bending power to the bottom face, little deformation and durability.
The workability of fresh mortar refers to the comprehensive properties of mortar easy for construction and good for quality, including mobility and water retention. The mortar with good mobility is easy to be paved thinly and evenly on bricks and bonded with floors well.
The mobility of mortar is the property that mortar can flow under the role of dead weight and exterior force. Fluidity is expressed by "sinking degree", usually determined by the consistency of mortar. The bigger the sinking degree is, the better the mobility will be.
The selection of mortar mobility should be determined by masonry types, construction conditions, and weather. The consistency of masonry mortar should be selected according to project requirement.
The water retention of mortar refers to the property of mortar to maintain moisture. When the mortar with good water retention is used in transportation, standing and pavement, water will not escape from mortar soon and the necessary consistency can be maintained. To keep a certain amount of water in mortar is easy to manipulate and also guarantees the normal hydration of cement to maintain the strength of masonry.
The water retention of mortar is expressed by layering degree, measured by mortar layering degree instrument. The mortar with good water retention has the layering degree of 10-30mm, and if it is more than 30mm, the water retention will be bad and easy to segregate; the mortar whose layering degree is less than 10 mm is not good for construction. Based on many experiments, the layering degree of cement mortar should be no more than 30mm, and the layering degree of cement mix mortar should be no more than 20mm.
The hardened mortar should have big strength which is expressed by strength grade. Compressive strength is the main basis for mortar strength. The strength grade of mortar, expressed by fm, is determined by compressive strength average (MPa) measured through curing a group of six cube specimens with side length of 70.7mm for 28d, the standard test method.
The strength grades of mortar include: M20, M15, M10, M7.5, M5 and M2.5, the six grades.
The strength of mortar is connected with its surface material. For the ordinary cement mortar, the following equation can be used to calculate its compressive strength.
Brick and stone masonry is a solid entity composed of many blocks that are bonded by mortar as a whole. Thus, it is required that mortar must have a certain adhesion stress over bricks and stones. Generally, the more the compressive strength of mortar is, the bigger its adhesion stress will be. In addition, its adhesion stress is related to the surface, cleanness, and humidity of bricks and stones, as well as the construction and curing conditions. For example, bricklaying needs watering, and the surface without clay will improve the adhesion stress and ensure the quality of masonry.
It is easy for mortar to get deformed when it bears loads or the temperature changes. If it deforms greatly or unevenly, the quality of masonry and surface will decrease and cause shrinkage and crack. When fine aggregates are used to mix the mortar, its deformation is bigger than the ordinary mortar. In order to prevent the cracks caused by uneven shrinkage deformation, hemp cut, paper strip and others fabric materials can be mixed in the surface mortar.
The durability of mortar refers to the property to withstand wear and tear in the long-term use. The hydraulic masonry that usually contacts with water should be impermeable and frost-resistant, so the impermeability and frost resistance of hydraulic masonry should be considered.
The frost resistance of mortar refers to the property to resist freeze-thaw cycle. Mortar is frozen and damaged because the water in its pores expands due to freeze and breaks the pores. Thus, dense mortar and the mortar with closed pores have good frost resistance. In addition, the factors influencing the frost resistance of mortar also include cement types, strength grades, and water-cement ratio.
The frost resistance of mortar is the property to resist the infiltration of pressure water. It is mainly related to density and size and structure of the inner pores. The connecting pores inside mortar and the cellular structures and, pores formed when it is moulded, all of which can lead to water seepage of mortar.
WHAT ARE THE EFFECTS ON CONCRETE DUE TO SEGREGATION?
WHAT IS LAITANCE IN CONCRETE & HOW IT AFFECTS CONCRETE?
WHAT ARE THE PROPERTIES OF FRESH CONCRETE?
I am diploma student, I was finished my study but i am not have job so please help me.
I love this site as an undergraduate student offering Bsc Construction.


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Wangari Muta Maathai (Nyeri, Kenya, 1940. április 1. – Nairobi, 2011. szeptember 25.) kenyai környezetvédő és politikus. 2004-ben Nobel-békedíjat kapott "a fenntartható fejlődéshez, demokráciához és békéhez való hozzájárulása" miatt. Ő volt az első afrikai nő, aki megkapta a díjat. Maathai egyúttal választott parlamenti képviselő és 2003. január – 2005. november között környezetvédelmi és természeti erőforrás miniszterhelyettes volt Mwai Kibaki kormányában.

Tanulmányai
Miután Kenyában elvégezte az iskolát, Maathai biológiát tanult az Egyesült Államokban és Németországban. Visszatérve Nairobiba, az ottani egyetemen ő volt az első afrikai nő, aki PhD címet szerzett állatorvosi szakon. 1971-ben az állatorvosi anatómia professzora lett, majd a kar dékánja.

Aktivista és politikus
1977-ben Maathai megalapította a Zöld Öv Mozgalmat demokratikus, környezetvédő civil szervezetet, amely azóta több mint 30 millió fát ültetett szerte az országban a talaj eróziója megelőzése érdekében. Innen ered a "Fák Asszonya" beceneve. Ezután egyre tevékenyebb szerepet vállalt környezetvédelmi és női jogi kérdésekben.

Maathai volt az előző elnöke a Kenyai Nők Nemzeti Tanácsának (Maendeleo Ya Wanawake). Az 1980-as években a férje elvált tőle, mondván, hogy túl erős egyéniség és nem tudja irányítani. A válóperes bíróság egyetértett a férjjel.

Daniel Arap Moi elnöksége alatt többször ült börtönben és erősen támadták amiatt, hogy többpárti választásokat követelt, valamint a korrupt politika és a törzsi politika felszámolását. 1989-ben Maathai majdnem teljesen egyedül megmentette a nairobi Uhuru Parkot azzal, hogy leállíttatta a Moi üzlettársainak az építkezését.

2002-ben Maathait képviselővé választották. 2003-tól ő volt a miniszterhelyettes a Környezet, Természeti Erőforrások és Vadvilág Minisztériumában. 2003-ban megalapította Kenyai Mazingira Zöld Pártot.

1997-ben, a második kenyai többpárti választáson indult az elnökségért, de a pártja visszavonta a jelölést. 2005 márciusában őt választották az Afrikai Unió Gazdasági, Szociális és Kulturális Tanácsának első elnökévé.

Egyike volt a nyolc zászlóvivőnek a 2006. évi téli olimpiai játékok nyitó ünnepségén.

2007-ben a The 11th Hour című filmben szerepelt, melyben a globális felmelegedési válságról beszélt.

Nobel-békedíj

Maathai az első afrikai nő és az első környezetvédő, aki Nobel-békedíjat kapott.

"Maathai bátran fellépett a korábbi elnyomó rendszer ellen Kenyában" – így jelentette be a norvég Nobel Békedíj Bizottság a 2004-es díj odaítélését. "Kivételes tetteivel felhívta a figyelmet a politikai elnyomásra mind otthon, mind külföldön. Munkássága sokaknak jelentett ösztönzést a demokratikus jogaikért folytatott harcukban, különösen a nők nyertek bátorítást belőle helyzetük javításához."

Kritikák
A Nobel-békedíj odaítélése után Maathai AIDS-szel kapcsolatos nyilatkozatai körül vita alakult ki a kenyai és nemzetközi sajtóban.

A kenyai East African Standard napilap augusztus 31-én megjelent cikkében idézte Maathainak egy szülővárosában, Nyeriben tartott szemináriumon elhangzott véleményét, miszerint az AIDS kórokozóját, a HIV-et tudósok állították elő azzal a céllal, hogy megtizedeljék Afrika népességét. "Ne legyünk naivak. Az AIDS nem Isten afrikaiakra vagy feketékre mért büntetése, hanem az elnyomás eszköze, amelyet gonosz tudósok hoztak létre, bár hogy pontosan kik, azt talán sosem fogjuk megtudni." A cikk megjelenése után Maathai hangsúlyozta, hogy ő sosem gyanúsított meg egyetlen országot sem a vírus előállításával, de a vírus eredetével kapcsolatos titkolózást gyanúsnak tartja.

A Nobel-békedíj elnyerését követő napon tartott sajtókonferencián Nairobiban Maathai megismételte HIV-vel kapcsolatos véleményét: "Egyesek azt mondják, az AIDS a majmoktól származik, de én kételkedem ebben, mivel mi időtlen idők óta a majmok mellett éltünk; mások azt mondják, Isten átka, de szerintem ez lehetetlen. Valójában tudósok hozták létre biológiai hadviselés céljából. Miért övezte mindig is annyi titok az AIDS-et? Amikor az eredete iránt tudakozódik az ember, sokan felkapják a fejüket. Ez gyanakvóvá tesz."

Maathai a Time magazinnak nyilatkozva így tisztázta pozícióját: "Fogalmam sincs, hogy ki hozta létre az AIDS-et, és azt sem tudom, hogy az biológiai fegyver-e. De azt tudom, hogy az ilyesmik nem a Holdról jönnek. Mindig is úgy gondoltam, hogy fontos elmondani az embereknek az igazságot, de azt gyanítom, van olyan igazság, amelyet nem szabad nagyon hangoztatni. (…) Az AIDS-re gondolok. Biztos vagyok benne, hogy az emberek tudják, honnan jött. És egészen biztos vagyok benne, hogy nem a majmoktól ered. Miért ne lehetne minket arra ösztönözni, hogy feltegyük magunknak ezeket a kérdéseket?"

Később, 2004 decemberében egy a Nobel-bizottság által kiadott nyilatkozatában Maathai azt írta, a HIV körüli összeesküvés-elméletek "gonoszak és rombolóak".

Jegyzetek

Fordítás

További információk
 A Zöld Öv Mozgalom honlapja
 Nobel díj

1940-ben született személyek
2011-ben elhunyt személyek
Nobel-békedíjasok
Nobel-díjas nők
Kenyai politikusok
Kenyai nők

redpajama_set_name: RedPajamaWikipedia


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Motherwell 09/10 Canterbury Away Kit
Motherwell unveiled the new 2009/2010 Away Kit, in association with PSL and Canterbury.
Record sales of the new home kit has created a buzz around the change strip but with Canterbury facing much publicized difficulties recently, the launch was unfortunately delayed. This has also caused problems confirming an on-sale date but that should be confirmed soon.
The strip is available to view, in person, at the 'Well shop.
QPR 09/10 Lotto Third Kit
Queens Park Rangers unveiled the new 09/10 Third kit made by Lotto. QPR will wear our brand new third kit in Tuesday night's Carling Cup first round tie against Exeter City.
The R's lock horns with the Grecians at St James' Park in a 19.45pm kick-off - and they'll be donning the new navy blue number for the very first time.
Only 1500 limited edition shirts have been produced and R's Kit Manager, Gary Doyle, has already given it the thumbs up.
"It's a lovely new kit and the players are all really looking forward to wearing it for the first time," he told
"They really like the look and the feel of it and it looks great on."
Middlesbrough 09/10 Adidas Away Kit
Middlesbrough unveiled their 09/10 Adidas away Kit, which the team will wear in action for the first time in the Carling Cup tie at Nottingham Forest later this month.
Modelled here by new signing Mark Yeates and young defender Jonathan Grounds, the 2009-10 away strip features a cyan shirt, shorts and socks, with the unmistakeable adidas stripes.
The away strip is being launched on the back of record home kit sales. Like the home shirt, the away top is available in short and long-sleeved versions.
The away shirt also features adidas Climacool fabric, an integrated system of technologies to regulate the wearer's body temperature, take sweat away from the body and allow air to circulate close to the skin.
Reebok 1995-96 Fiorentina Home Shirt
Fiorentina 1995-96 home shirt by Reebok as worn during the 95-96 season when Batigol's incredible scoring record helped the side to their first Coppa Italia title in 21 years with goals in both legs of the Final against Atalanta.
Adidas 1988-90 West Germany Home Shirt
Perhaps the most iconic football shirt of all time, as worn at the Italia '90 World Cup when Franz Beckenbauer's outfit memorably defeated a Gascoigne-inspired England in the Semi-Finals before an Andy Brehme penalty in the Final saw off Maradona's Argentina to crown the side World Champions for the third time.
Umbro 1991 Scotland Match Worn Home Full Kit
Extremely rare player kit believed to have been worn by Durie during the 1992 European Championship Group 2 qualifying match against San Marino played at Hampden park, Glasgow on 13th November 1991.
Adidas 1992 USA Home Shirt
Classic design home shirt as worn in the build up to the 1994 World Cup which the USA hosted.
Meyba 1984-89 Barcelona Home Shirt
FC Barcelona home shirt as worn when Terry Venables guided Barça to their only La Liga title of the 80s.
Hamilton Academical 09/10 Nike Away Kit
Hamilton Academical unveiled the new 09/10 away Kit made by Nike with the logo of their new sponsor -Shoreline Energy International, who have agreed to sponsor the club's two away strips for 2009/10.
Manager Billy Reid is pictured with senior players Simon Mensing & Derek Lyle with the kit which will be worn for the first time at Rugby Park, Kilmarnock in the opening fixture of the Clydesdale Bank Scottish Premier League season.
Also pictured with the players is Accies Chief Executive George Paterson and Godfrey Mitchell one of the executive directors of Shoreline International.
Northern Ireland 09/10 Umbro Away Kit
Northern Ireland unveiled the new 09/10 away kit made by Umbro.
The full kit is available to buy from Saturday 22nd August onwards.
Thanks to Daniel
Velez Sarsfield 09/10 Penalty Kits
Club Atlético Vélez Sársfield unveiled the new 09/10 kits made by Penalty.
Velez foundation dates back to 1910. It was on a rainy day when three men sheltered from a summer storm in Vélez Sársfield Railway Station. They were Julio Guglielmone, Martin Portillo and Nicolas Martín Moreno.
Together, the friends established the Argentines of Velez Sarsfield Athletic Club(Club Atlético Argentinos de Velez Sarsfield) and appointed Luis Barredo as their first chairman. Their first home was the piece of land located at United Provincies St.-Provincias Unidas-, currently Juan Bautista Alberdi, Convention St, currently Jose Bonifacio, and Mariano Acosta and Ensenada. During its infancy, the team sported white shirts but soon changed to navy blue shirts with white shorts.
Hansa Rostock 09/10 Masita kits
Hansa Rostock unveiled the new 09/10 kits made by Masita.
Hansa also presented its new main sponsor WINDSTÄRKE11. "WINDSTÄRKE11" is an initiative of several companies involved in the wind energy sector in Germany.
Deportivo La Coruna 09/10 lotto kits
Manuel Pablo, Juca, Aranzubia, Juan Carlos Valerón and youth Alvaro Queijeiro revealed the new deportivo la coruna 09/10 kits made by Lotto to the public and media.
After the previous contract with Canterbury of New Zealand was cancelled due to causes attributable to the same company, Depor contacted Italian firm Lotto Sport and decided to sign a new deal that's converting this sports apparel manufacturer in Depor's supplier of uniforms for the next four seasons.
Shrewsbury Town 09/10 Prostar Third Shirt
Shrewsbury Town unveiled the new 09/10 third kit for when they travel away from the Prostar Stadium this coming season.
The existing blue and black kits have caused a few colour clashes on occasions and Town have commissioned a white change kit option.
This now gives Town the three options of kit over the upcoming Coca Cola League 2 campaign with the white shirt being classed as the 3rd kit.
Fans have asked if it would be possible to get hold of the new shirt and we have arranged with Prostar for the shirt to be stocked in the club shop.
Thanks to Danny
Charlton Athletic 09/10 Joma Away Shirt
Charlton Athletic have revealed their new blue and black striped away kit for the 2009/10 campaign, which went on sale for the first time before the start of the season on last Saturday.
And with stocks of the new red home kit again running low following a high number of sales, supporters were be able to purchase the new away jerseys for the first time last weekend.
Charlton retail manager Michael Rea said: "Given how late in the summer we finalised the new sponsorship deal with Kent Reliance Building Society, we're really pleased to be able to have launched both new kits before the start of the season.
"Despite the delivery problems, sales have been really good for the red home shirts, and we're expecting the away kit to be just as popular."
The away kit, which features black shorts and black socks with a blue band around the top, is reminiscent of the seahawk and blue denim away kit that the Addicks wore two seasons ago.
"The seahawk and denim kit was one of the most sought after kits in recent years, so we're urging supporters who want to get their hands on the away kit to purchase it as early as possible," added Rea.
Fans should note that the Valley Superstore will be closed from 1pm on Friday to enable staff to prepare the store for the kit launch the following morning.

redpajama_set_name: RedPajamaCommonCrawl


text:
Category Archives: Art Rant
Wicked Inspiration
Posted by kittycalash in Art Rant, History, Living History, personal, Philosophy, Reenacting
anarchist guide to historic house museums, Hamilton, historical reenactors, history, interpretation, literature, living history, passion, progressive reenacting, Reenacting
You know that musical? The one you can't get tickets to unless they were willed to you by your grandmother because she was lucky enough to stumble out of the Tardis right at the box office on opening day, but had to buy them for, like, six years from now, and they're actually for the Kansas City production? The one in which the founding fathers are, you know, brown? My friends and family assumed I'd hate the whole idea, but I don't. Like so many people, I love it, and not just for the music, though my preferred method of psyching myself up for GeeDubs1790 or What Cheer Day is listening to the Stones or the Beastie Boys all the way up.
So, what then, interests me in "Hamilton," and why do I think it relates to interpretation? This: the way that the show filters the concerns of the late eighteenth century through the lens of the twenty-first could make for enlightening listening.
My Shot gives me goosebumps: why? Is it because history and civics are finally sexy?
Maybe. But "Hamilton" as a whole, per Rebecca Mead in the New Yorker, "is a hymn to the allure that America promises the immigrant who aspires to reach its shores; it is also an argument for the invigorating power that this nation's porous borders, and porous identity, have always offered."
Porous identity. That's part of why I'm fascinated. But just as "Sleep No More" and Occupy Providence (really really) were partial inspirations for What Cheer Day, "Hamilton" strikes me as the kind of production worth paying attention to. No, I am not suggesting that the paunchy reenactors start channeling their inner Biggie, though I might well pay to see that.
No, what I'm suggesting is that we reconsider what it is we're doing out there on the field and in the historic houses, and not just what but how and why. Hamilton takes an unexpected approach to history and it's going gangbusters, while Amazing Grace tanked. So it's not about the costuming authenticity, though I implore you not to give that up. It's about the passion. It's about coming at things sideways.
What makes you love doing laundry, drilling with precision, telling local gossip, making soap, or whatever it is that you love best about your place in the past? Why does all this matter to you so much? What is it that we can learn about today as you teach us about the past? Let loose that love and passion, share those insights, and ten to one you'll have more fun and excite more visitors.
Maintain an Even Strain
Posted by kittycalash in Art Rant, Events, History, Living History, personal
common people, feminism, first person interpretation, interpretation, living history, women's history, women's work
Dude: I am conflicted. There are folks out there doing excellent work, but after reading some recent posts around the interwebs, I kept thinking, "Stockholm Syndrome much?"
I'm as much of a narcissist as the next person, and I think I recognize some of the folks being called out in various places for being critical of women's roles in living history events. So, organize my own events? Come up with my own things to do?
Cool: challenge accepted.
I am, in fact, throwing down for the pleasure and pain of running a farm in late June. No, I didn't organize it, but I was asked to take on a challenge and I have accepted, roping my favorite tailor into the effort as well. It'll mean a bunch of studying, but in a pinch, I can always clean the house. We can rake, make refugees stay in the yard, and try as hard as we can to keep Quakers from putting radical ideas in the slaves' heads. I think it will be hard, unpleasant, and uncomfortable—and that's what I don't like about the suggestions in the otherwise honestly well-intended and meant-to-inspire posts.
Playing at Quadrille. Oil on canvas by Francis Hayman. Birmingham Museums and Art Gallery
They're so nice. They reinforce women's subordinate roles in the past and present. Children's activities? I might die, really, I might. If that's your bag, go for it, please! We need it. It's simply not something I can do.
No more can I talk about What People Wore. It's not that I don't care (y'all know I do) but that I want to move past the surface.
It's not enough just to look great.
The Heir: Tom Finds New Wealth. William Hogarth.
Dive deep: find the dirt. Find the hard stuff. You don't have to be nice. That's my personal problem with what I've been reading: between the lines I keep hearing a voice suggesting that we be nice girls, that we simmer down. No, I'm sorry. I can't. Reader, if you can, go for it.
But if you can't, I want to tell you: Keep pushing. Keep asking. Keep speaking up. Challenge the status quo. Our Girl History did a great post on Well Behaved Women, and I fully support the work people are doing to represent the Well Behaved and the marginalized (shout out to the veteran with the knife-grinding cart: well imagined, sir!).
John Russell. The Blind Beggar and His Granddaughter, oil on canvas, 18th century. The Bowes Museum UK.
Bring it. Bring the ordinary.
But if you can't be ordinary or run the children's games or be subservient or show how women dressed, that's okay. For the love of god, someone, be desperate.
Be hungry, be angry, be resentful, be religious.
Whatever you do, don't be afraid to speak your mind.
That Belongs in a Museum
Posted by kittycalash in Art Rant, Collecting, Museums, personal
art history, collections, historic house museums, museum collections, museum practice, personal, philosophy
Sotheby's Sale 8446, Lot 20. Easy Chair, Philadelphia ca 1770
We're taking a brief break from this week's nightlife programming to bring you this special report. Regular snark resumes with the next post.
It's not a secret that I work in a museum. I operate in a world of objects that cannot be touched, sat upon, slept in, worn– you get the idea. Well, sometimes things that I believe belong in a museum don't end up in one. Sometimes beautiful objects with great stories and deep resonance with the museum I think they belong in don't make it there. Sometimes museum professionals take phone calls from irate family members who are incensed that you're even talking to someone in the family about how to organize their materials. Sometimes objects do make it, but then, before the paperwork is executed, the gift is rescinded. People get weird about stuff.
Sometimes they get weird about stuff because of what it's worth. Eleven years ago, a family chose not to fight over objects, but instead sold their family furniture. They grossed nearly $13 million. Thirteen Million Dollars: Enough for a baker's dozen of Dr. Evils. With chairs that sell for $204,000 (Sale 8278, lot 586), it's no wonder people get weird about stuff because of money.
Lot 586. It's a pity the catalog is no longer available online, but even Sotheby's has to conserve server space. It was a beautiful chair: a 1763 upholstered easy chair– upholstered by Plunket Fleeson of Philadelphia.* I pored over that catalog page in the Important Americana sale catalog. It would have come in just before Christmas, or just after, slick clay-coated pages printed with fine ink. When the auction catalogs arrive at work, we stand in the kitchen-mail room and bury our noses deep in the gutter: smells like money.**
Staircase Group (Portrait of Raphaelle Peale and Titian Ramsay Peale I)
Charles Willson Peale. Philadelphia Museum of Art, E1945-1-1
I wanted that chair, Lot 586, that walnut easy chair made in Philadelphia. Wrong town, you say? Mais non, Philadelphia was the place to buy fancy goods– especially upholstered goods– in the 18th century. Providence merchants were trading with Philadelphia, the town that set the style for the colonies.*** It was sophisticated, urbane, refined. So, when Sarah Brown was pregnant with their first child, John Brown sent to Philadelphia for an easy chair. It would be the best.
He'd already ordered a tea table and roundabout chairs from Newport at the time of his marriage in 1760. These objects were about more than function: they were signifiers of taste and sensibility as much as wealth. So, as the time came closer for Sarah's confinement, John Brown became increasingly agitated with Plunket Fleeson, who was delaying the delivery of the chair. John Brown was concerned for Sarah's comfort postpartum, and said so in a letter. I can't quote or link to it, because it's in private hands, quoted in the catalog entry for lot 586.****
So what about that chair? Well… I got permission from the Authorities (a Board-level committee, with the support of the Executive Director) and we bid on the chair by phone. You know already we were not the winning bid. We were willing to bid a lot– really, a lot of money for us– for this chair.
Not because it's worth so much. Not because of Plunket Fleeson, or the quality of the carvings, or the craftsmanship, really.
Because Sarah Brown sat in that chair cradling her son, James Brown, in 1763. She sat in that chair with the children who came after him, the babies who lived, and the babies who died.
Because that chair told a story about a family, about a relationship between a husband and wife, a man and a woman, at its most basic level.
That chair told a story about love.
Even I am a sucker for love.
Love is why people get weird about stuff, about the chairs, the family photos, the workbaskets, heck– the drill presses. We imbue objects with meaning, with memories, that substitute for the people we love when they're gone. Sometimes it's a t-shirt that smells of a lover. Sometimes it's sewing basket used by three generations of women. And sometimes it's a chair.
In the case of the easy chair, imagining the cradling comfort of the chair and the memories it recalls is simple. In the case of, say, shield back side chairs, the leap is a little harder. But perhaps– just perhaps– arranged around a festively set table, those chairs conjure memories of holiday meals, birthday dinners, graduation parties. Maybe those chairs take you back to the people and times when you felt loved.
Or maybe they're worth $100,000, and you consign them to auction.
But if you value the story as much as the object, here's the funny thing: you can keep that memory forever, and share the story with everyone, if you give that object to a museum.*****
When people really love an object and are fighting with their families over who gets to keep the things, I don't play Solomon. I tell them a story about memory, and preservation, and about endurance. Sometimes I can convince them. Sometimes I can't.
When I can't, I tell them I understand, and that my priority– and my institution's priority– is not things, but people. They can keep their object: we'll be here when they're ready. It's all true, and I do mean it.
But inside, I feel like Indiana Jones, soaking wet on the deck of a pitching ship.
That belongs in a museum.
*If money were no object, I'd buy it for that name alone.
**The quality decreased in 2008, when the financial crisis hit everybody hard, but the paper weight and ink have been creeping back up in quality of late.
***I know, it sounds crazy now, but it was true. At the time of the American Revolution, it was the second largest English speaking city in the world, after London.
****You are correct, sir: I'd like the letter book it's in, too.
*****Subject to acceptance. Some rules and regulations apply. Leaning is touching. Don't lick it.
Posted by kittycalash in Art Rant, Clothing, History, Living History, Research
18th century, 18th century clothes, 18th century clothing, dress, engravings, fashion, gowns, Research, style, women
I'm of an age (none of your beeswax, thanks for asking) where most of the clothes in the shops are simply not for me. Not only am I picky about labor standards, fabric and construction quality, the styles aren't for me. I'm too old. I create enough havoc walking (falling) down the street as it is that I do not need to look like the mid-life crisis I'm having involves a desperate search to recapture my youth through H & M or Hot Topic or whatever-the-kids-are-wearing clothing. I've achieved "a certain age," and as with my everyday closet, my living history wardrobe has to reflect my age as well. Pity, really.
Following the Fashion. Hand colored etching by James Gillray, published by Hannah Humphrey, 1794. British Museum 1851,0901.706
But more's the pity here: I don't have a clear idea, really, how age was perceived in the various decades I interpret. I have a clearer idea of how different body shapes and class levels were perceived and taunted, though sometimes body shape is an analogue for age.
Is this my daughter Ann? Pen and ink print study by Samuel Hieronymus Grimm, 1774. British Museum 2011,7084.56
There's a classic image that juxtaposes age and youth. Here's how the British Museum describes this image:
'Is this my Daughter Ann?'; satire on fashion. A street where, before a house [on the right], over the door of which the name 'Love Joy' is written, a sedan chair has been brought, in order to carry away a young lady, who, in a towering toupée, and other articles of fashionable attire of this period, is leaving the house in company with a young soldier, who caresses her as they go; she looks fondly at him. An old woman, in what was then an 'old-fashioned' costume is interposing to prevent the departure of the damsel. 1774
Okay… "An old woman, in what was then an 'old-fashioned' costume." You will note that the old woman is wearing what we typically wear here in New England to represent everyday clothing for middling and lower sort American colonists for most of the 1770s. I don't want to put too much stock in the cataloger's description, because I'm not sure that print curators or curatorial assistants always know as much about material culture as they think they do. And we know that six weeks is not six years when it comes to transmitting fashion changes and updates across an ocean. I think "old fashioned" might not be exactly or entirely right as a descriptive phrase.
What does the old woman's clothing really signify? That she's rural and not urban? That's she's poor? That she's old? How 'old fashioned' is that costume by 1774… in a context other than London courtesan couture? And how do we translate the clues we have trouble deciphering into a dress code for living history?
Is this my Daughter Ann, mezzotint by James Watson after S. H. Grimm, published by Sarah Sledge, 1774. British Museum J,5.104
Well, happily, there's a verse under the image in this print.
Is this my Daughter Ann
The Matron thus Surprised exclaims,
And the deluded Fair One Blames
But had the Mother been as Charming
She had Thought the Mutual sport no harm.
This Moral's an undoubted Truth
Age envies Still the Joys of Youth
So this print is not about fashion. It's about sex. (Well, duh. You were wondering when I'd bring that up.) It's also, in a way, about hypocrisy, isn't it? But the verse gives us the clue that the mother's clothes are meant to be matronly. "Conservative because of her age" might be a better descriptive phrase than "old fashioned" in that catalog record.
But does that mean that those of us who have achieved "a certain age" might also consider whether we, too, should be dressing in a more "conservative because of her age" style? I don't really know.
But look here: Zoffany, ca. 1762.
David Garrick and Mary Bradshaw in David Garrick's "The Farmer's Return". Johann Zoffany, ca.1762. YCBA B1981.25.731
Here we're looking at a rural woman ten to twelve years before "My daughter Ann." The costumes are very similar; maybe the mother in Ann really is just old fashioned.
Here's another version of "My daughter Ann," this time more clearly fashion focused, without the sexual overtones.
Print made by Francis E. Adams, active ca.1760–1775, British, Heyday! Is This My Daughter Anne!, 1773, Mezzotint and etching on medium, moderately textured, cream laid paper, YCBA, B1970.3.820
Yale helpfully provides a transcription of the verse at the bottom:
HEYDAY! Is this my DAUGHTER ANNE! | Heyday! the country Matron in surprize, | Is this my Daughter thus bedizell'd? cries. | To Town she lately went a Damsel plain: | But scarcely now is to be known again. | That City to its Vanities has brought her, | And banish'd the good Housewifery I taught her. | Why, Child you'll frighten here our honest People: | They'll say you've on your Head a London Steeple.
My best guess is that this print is skewering both Anne and her mother: Anne, for being so outlandishly fashion-forward, and her mother, for being so far behind. But again, that's only a guess.
Fashionable Furniture, or, The Glories of the Past
Posted by kittycalash in Art Rant, History, Research
19th century, Ackermann's Repository, fashion, Federal style, furniture, historic interiors, style
include window drapes.
Ackermans's Repository of Arts., etc. April, 1817.
Every now and then, someone argues with me that the historic house where I work would not have had window curtains or drapes. Sometimes they like to expand that argument to "there were no curtains at all" in early Federal America. The reasoning is usually that textiles were too expensive to "waste" on window dressing. If you know me, you know this kind of argument is a Bad Idea. The public fight (I was angrily accosted by a now-former docent during a public presentation) is known as The Great Curtain Kerfuffle, and resulted in my reply that the owner of the house could very well afford anything he pleased. Fabric is Money.*
There's another iteration of my argument: Color is Money.
Ackermans's Repository of Arts., etc. April, 1817. page 244
Those of us who shop at IKEA are not going to have "the most fashionable style of decoration" in our 1817 homes. The Willings and Binghams of Philadelphia were models for the most fashionable families of Providence, and while well before this 1817 plate, the Binghams were recorded draping their chairs with orange and red silks. In early 19th century Providence, the John Innis Clark family had silk covers on their sofas and chairs in 1808, and plenty of carpets and curtains in their Benefit Street home from the 1790s on.
"Crimson is very rich, but blue is handsomer," wrote Eliza Ward to her sister, Mrs John Innis Clark, in the 1790s. Curtains and covers were fringed (Mrs Hazard Gibbes was blue and yellow). Windows were dressed, and younger, less affluent relatives received hand-me-down curtains. In 1803, Elizabeth Watters in Wilmington, North Carolina was having a carpet "wove in true Scotch taste in imitation of Highland plaid."
John Phillips (1719-1795) Oil on canvas by Joseph Steward,1794-1796. Sack Gallery, Hood Museum of Art, Dartmouth College.
Some carpets, no? Maybe the new mantra is Carpets are Money.
But quite aside from an obvious display of wealth, what we have to realize about these images and letters is that they are depicting a world that looks very different from our own. Color sensibilities, tolerance for pattern mixing, non-matchy-matchy sewing and dressing. We have to abandon our 21st century aesthetic sensibilities when we dress ourselves or our spaces for the past, and really embrace the vivacity of that world. Sensory overload, perhaps, but getting closer to what the world of the past looked like will help us see– in every sense– the way the people of the past did.
*I may or may not have made additional statements afterwards to the effect that of course wealthy Americans squatted naked in the corners of their well-appointed mansions gnawing raw meat until Benjamin Franklin invented fire and fabric. I should be sorry about that, but I don't seem to be.


mvpreed2 started a topic BDC's FIRST 2017-18 Predictions Thread!
BDC's FIRST 2017-18 Predictions Thread!
Combining a bit of our past (PGU), with a bit of the future (BDC), I'd like to bring the very first PREDICTIONS thread.


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