Identification and characterization of a single-stranded DNA-binding protein from the archaeon Methanococcus jannaschii

Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria and eukarya. We report here the identification and characterization of the SSB of an archaeon, Methanococcus jannaschii. The M. jannaschii SSB (mjaSSB) has significant amino acid sequence similarity to the eukaryotic SSB, replication protein A (RPA), and contains four tandem repeats of the core single-stranded DNA (ssDNA) binding domain originally defined by structural studies of RPA. Homologous SSBs are encoded by the genomes of other archaeal species, including Methanobacterium thermoautotrophicum and Archaeoglobus fulgidus. The purified mjaSSB binds to ssDNA with high affinity and selectivity. The apparent association constant for binding to ssDNA is similar to that of RPA under comparable experimental conditions, and the affinity for ssDNA exceeds that for double-stranded DNA by at least two orders of magnitude. The binding site size for mjaSSB is ≈20 nucleotides. Given that RPA is related to mjaSSB at the sequence level and to Escherichia coli SSB at the structural level, we conclude that the SSBs of archaea, eukarya, and bacteria share a common core ssDNA-binding domain. This ssDNA-binding domain was presumably present in the common ancestor to all three major branches of life.

The inhibition of antigen-presenting activity of dendritic cells resulting from UV irradiation of murine skin is restored by in vitro photorepair of cyclobutane pyrimidine dimers

Exposing skin to UVB (280–320 nm) radiation suppresses contact hypersensitivity by a mechanism that involves an alteration in the activity of cutaneous antigen-presenting cells (APC). UV-induced DNA damage appears to be an important molecular trigger for this effect. The specific target cells in the skin that sustain DNA damage relevant to the immunosuppressive effect have yet to be identified. We tested the hypothesis that UV-induced DNA damage in the cutaneous APC was responsible for their impaired ability to present antigen after in vivo UV irradiation. Cutaneous APC were collected from the draining lymph nodes of UVB-irradiated, hapten-sensitized mice and incubated in vitro with liposomes containing a photolyase (Photosomes; Applied Genetics, Freeport, NY), which, upon absorption of photoreactivating light, splits UV-induced cyclobutane pyrimidine dimers. Photosome treatment followed by photoreactivating light reduced the number of dimer-containing APC, restored the in vivo antigen-presenting activity of the draining lymph node cells, and blocked the induction of suppressor T cells. Neither Photosomes nor photoreactivating light alone, nor photoreactivating light given before Photosomes, restored APC activity, and Photosome treatment did not reverse the impairment of APC function when isopsoralen plus UVA (320–400 nm) radiation was used instead of UVB. These controls indicate that the restoration of APC function matched the requirements of Photosome-mediated DNA repair for dimers and post-treatment photoreactivating light. These results provide compelling evidence that it is UV-induced DNA damage in cutaneous APC that leads to reduced immune function.

DNA double-strand break repair proteins are required to cap the ends of mammalian chromosomes

Recent findings intriguingly place DNA double-strand break repair proteins at chromosome ends in yeast, where they help maintain normal telomere length and structure. In the present study, an essential telomere function, the ability to cap and thereby protect chromosomes from end-to-end fusions, was assessed in repair-deficient mouse cell lines. By using fluorescence in situ hybridization with a probe to telomeric DNA, spontaneously occurring chromosome aberrations were examined for telomere signal at the points of fusion, a clear indication of impaired end-capping. Telomeric fusions were not observed in any of the repair-proficient controls and occurred only rarely in a p53 null mutant. In striking contrast, chromosomal end fusions that retained telomeric sequence were observed in nontransformed DNA-PKcs-deficient cells, where they were a major source of chromosomal instability. Metacentric chromosomes created by telomeric fusion became even more abundant in these cells after spontaneous immortalization. Restoration of repair proficiency through transfection with a functional cDNA copy of the human DNA-PKcs gene reduced the number of fusions compared with a negative transfection control. Virally transformed cells derived from Ku70 and Ku80 knockout mice also displayed end-to-end fusions. These studies demonstrate that DNA double-strand break repair genes play a dual role in maintaining chromosomal stability in mammalian cells, the known role in repairing incidental DNA damage, as well as a new protective role in telomeric end-capping.

Neurotransplantation of magnetically labeled oligodendrocyte progenitors: Magnetic resonance tracking of cell migration and myelination

Demyelination is a common pathological finding in human neurological diseases and frequently persists as a result of failure of endogenous repair. Transplanted oligodendrocytes and their precursor cells can (re)myelinate axons, raising the possibility of therapeutic intervention. The migratory capacity of transplanted cells is of key importance in determining the extent of (re)myelination and can, at present, be evaluated only by using invasive and irreversible procedures. We have exploited the transferrin receptor as an efficient intracellular delivery device for magnetic nanoparticles, and transplanted tagged oligodendrocyte progenitor cells into the spinal cord of myelin-deficient rats. Cell migration could be easily detected by using three-dimensional magnetic resonance microscopy, with a close correlation between the areas of contrast enhancement and the achieved extent of myelination. The present results demonstrate that magnetic resonance tracking of transplanted oligodendrocyte progenitors is feasible; this technique has the potential to be easily extended to other neurotransplantation studies involving different precursor cell types.

On the acidity and reactivity of HNO in aqueous solution and biological systems

The gas phase and aqueous thermochemistry and reactivity of nitroxyl (nitrosyl hydride, HNO) were elucidated with multiconfigurational self-consistent field and hybrid density functional theory calculations and continuum solvation methods. The pKa of HNO is predicted to be 7.2 ± 1.0, considerably different from the value of 4.7 reported from pulse radiolysis experiments. The ground-state triplet nature of NO− affects the rates of acid-base chemistry of the HNO/NO− couple. HNO is highly reactive toward dimerization and addition of soft nucleophiles but is predicted to undergo negligible hydration (Keq = 6.9 × 10−5). HNO is predicted to exist as a discrete species in solution and is a viable participant in the chemical biology of nitric oxide and derivatives.

Interaction of the β sliding clamp with MutS, ligase, and DNA polymerase I

The β and proliferating cell nuclear antigen (PCNA) sliding clamps were first identified as components of their respective replicases, and thus were assigned a role in chromosome replication. Further studies have shown that the eukaryotic clamp, PCNA, interacts with several other proteins that are involved in excision repair, mismatch repair, cellular regulation, and DNA processing, indicating a much wider role than replication alone. Indeed, the Escherichia coli β clamp is known to function with DNA polymerases II and V, indicating that β also interacts with more than just the chromosomal replicase, DNA polymerase III. This report demonstrates three previously undetected protein–protein interactions with the β clamp. Thus, β interacts with MutS, DNA ligase, and DNA polymerase I. Given the diverse use of these proteins in repair and other DNA transactions, this expanded list of β interactive proteins suggests that the prokaryotic β ring participates in a wide variety of reactions beyond its role in chromosomal replication.

Transcription factor EGR-1 suppresses the growth and transformation of human HT-1080 fibrosarcoma cells by induction of transforming growth factor beta 1.

The early growth response 1 (EGR-1) gene product is a transcription factor with role in differentiation and growth. We have previously shown that expression of exogenous EGR-1 in various human tumor cells unexpectedly and markedly reduces growth and tumorigenicity and, conversely, that suppression of endogenous Egr-1 expression by antisense RNA eliminates protein expression, enhances growth, and promotes phenotypic transformation. However, the mechanism of these effects remained unknown. The promoter of human transforming growth factor beta 1 (TGF-beta 1) contains two GC-rich EGR-1 binding sites. We show that expression of EGR-1 in human HT-1080 fibrosarcoma cells uses increased secretion of biologically active TGF-beta 1 in direct proportion (rPearson = 0.96) to the amount of EGR-1 expressed and addition of recombinant human TGF-beta 1 is strongly growth-suppressive for these cells. Addition of monoclonal anti-TGF-beta 1 antibodies to EGR-1-expressing HT-1080 cells completely reverses the growth inhibitory effects of EGR-1. Reporter constructs bearing the EGR-1 binding segment of the TGF-beta 1 promoter was activated 4- to 6-fold relative to a control reporter in either HT-1080 cells that stably expressed or parental cells cotransfected with an EGR-1 expression vector. Expression of delta EGR-1, a mutant that cannot interact with the corepressors, nerve growth factor-activated factor binding proteins NAB1 and NAB2, due to deletion of the repressor domain, exhibited enhanced transactivation of 2- to 3.5-fold over that of wild-type EGR-1 showing that the reporter construct reflected the appropriate in vivo regulatory context. The EGR-1-stimulated transactivation was inhibited by expression of the Wilms tumor suppressor, a known specific DNA-binding competitor. These results indicate that EGR-1 suppresses growth of human HT-1080 fibrosarcoma cells by induction of TGF-beta 1.

Stabilization of diverged tandem repeats by mismatch repair: evidence for deletion formation via a misaligned replication intermediate.

A functional methyl-directed mismatch repair pathway in Escherichia coli prevents the formation of deletions between 101-bp tandem repeats with 4% sequence divergence. Deletions between perfectly homologous repeats are unaffected. Deletion in both cases occurs independently of the homologous recombination gene, recA. Because the methyl-directed mismatch repair pathway detects and excises one strand of a mispaired duplex, an intermediate for RecA-independent deletion of tandem repeats must therefore be a heteroduplex formed between strands of each repeat. We find that MutH endonuclease, which in vivo incises specifically the newly replicated strand of DNA, and the Dam methylase, the source of this strand-discrimination, are required absolutely for the exclusion of "homeologous" (imperfectly homologous) tandem deletion. This supports the idea that the heteroduplex intermediate for deletion occurs during or shortly after DNA replication in the context of hemi-methylation. Our findings confirm a "replication slippage" model for deletion formation whereby the displacement and misalignment of the nascent strand relative to the repeated sequence in the template strand accomplishes the deletion.

Da tradução ou não de pronomes e partículas pronominais do Italiano ao Português: algumas escolhas feitas por dois tradutores

Este estudo é movido pela curiosidade quanto a como se resolvem, nas traduções do italiano para o português, questões de colocação pronominal. Encontramos, normal e frequentemente, na língua italiana, principalmente na língua falada, pronomes cujos equivalentes em português existem em gramáticas normativas da língua portuguesa, mas que, na prática, não são utilizados pelos falantes e escritores brasileiros. Encontramos, também, na língua italiana, um significativo número de verbos pronominais (como esserci, volerci, averne etc.) e um considerável número de verbos pronominais múltiplos (como andarsene, farcela, fregarsene etc.) que, juntamente com esses pronomes, constituem, para os professores brasileiros de italiano língua estrangeira (LE), elementos difíceis de trabalhar na sala de aula. Além disso, tais elementos também podem dificultar o trabalho dos tradutores, que devem fazer determinadas escolhas ao traduzi-los para o português. Como são traduzidos os pronomes combinados do italiano nas versões brasileiras? Será que os portugueses, que possuem, por exemplo, tais pronomes utilizam-nos em todos os casos em que os encontramos nos textos de partida? E as partículas pronominais são simplesmente eliminadas no texto de chegada ou são substituídas? Tais aspectos, se observados e organizados, podem levar a uma melhor compreensão das duas línguas em contato e dar subsídios a estudantes, professores e tradutores. Pensando nessa dificuldade, esta pesquisa buscou e listou alguns autores e obras disponíveis para consulta e analisou um corpus com cento e sessenta e três ocorrências de pronomes no italiano, mais sete acréscimos de pronomes no português brasileiro (PB) e/ou português europeu (PE), partindo do romance Uno, nessuno e centomila de Luigi Pirandello e suas respectivas traduções em PB e PE. Nosso objetivo consiste em encontrar respostas úteis à diminuição do estranhamento, por parte de um italiano, que escuta, de um brasileiro, frases sem pronomes (ainda que o italiano as entenda) e/ou a sensação de inadequação e, até mesmo, de desconforto, por parte de um brasileiro, ao produzir frases com todos os pronomes. No corpus analisado, temos uma amostra das escolhas e respectivas traduções propostas pelos tradutores para casos de pronomes reflexivos, de pronomes pessoais do caso reto, de pronomes pessoais do caso oblíquo, de pronomes combinados e de partículas pronominais ne, ci e vi, com manutenções, omissões, trocas por outros pronomes (possessivos, retos, oblíquos, demonstrativos) e, até mesmo, uma espécie de compensação numérica com a inclusão de palavra inexistente no texto de partida.

Compactação e elevação da vida útil de transformadores de distribuição empregando óleo vegetal isolante.

A busca no aumento da vida útil dos transformadores de distribuição, redução dos custos de manutenção e mitigação de falhas, leva ao desenvolvimento de novos materiais e critérios de exploração diferenciados desses ativos. Esta pesquisa apresenta o desenvolvimento de um novo transformador de distribuição compacto e mais eficiente utilizando óleo vegetal isolante totalmente biodegradável. Além de biodegradável, o óleo vegetal utilizado possui menor agressividade ambiental e maior capacidade térmica aumentando, a capacidade de carregamento do transformador sem comprometer a sua vida útil. A fim de atestar essa menor agressividade em relação ao óleo mineral, ensaios foram efetuados em um equipamento que permaneceu 12 anos em operação. O equipamento foi totalmente desmontado para análise e coleta de amostras de papel e óleo vegetal isolante. As análises visam comprovar a menor agressividade em relação ao óleo mineral, apresentadas na revisão bibliográfica. A menor agressividade torna possível a proposição de uma nova filosofia de planejamento de redes de distribuição utilizando uma quantidade menor dos novos transformadores para uma mesma carga, tornando-a mais compacta e eficiente.

Surréalisme Sous-L’Eau : science and surrealism in the early films and writings of Jean Painlevé

A French biologist who moved in Surrealist circles, Jean Painlevé began making films about underwater creatures in 1927, and by 1982 had created over two hundred films on a broad range of natural, scientific, and political subjects. His underwater films remain the most ethereal and poetic works in his oeuvre, and he specifically used cinema to capture the mystery and wonder of nature.

Perceived emotional intelligence, general intelligence and early professional success: predictive and incremental validity

Although the study of factors affecting career success has shown connections between biographical and other aspects related to ability, knowledge and personality, few studies have examined the relationship be-tween emotional intelligence and professional success at the initial career stage. When these studies were carried out, the results showed significant relationships between the dimensions of emotional intelligence (emotional self-awareness, self-regulation, social awareness or social skills) and the level of professional competence. In this paper, we analyze the relationship between perceived emotional intelligence, measured by the Trait Meta-Mood Scale (TMMS-24) questionnaire, general intelligence assessed by the Cattell factor "g" test, scale 3, and extrinsic indicators of career success, in a sample of 130 graduates at the beginning of their careers. Results from hierarchical regression analysis indicate that emotional intelligence makes a specific contribution to the prediction of salary, after controlling the general intelligence effect. The perceived emotional intelligence dimensions of TMMS repair, TMMS attention and sex show a higher correlation and make a greater contribution to professional success than general intelligence. The implications of these results for the development of socio-emotional skills among University graduates are discussed.

Restoration of religious temples: intervention in the church “Nuestra Señora de Belén” in Crevillente (Alicante)

Heritage conservation has raised historical problems usually centered in defects resulting from water leaks. Thus, any intervention is presented as a difficult task, both due to building techniques to be used and the lack of economic resources in many cases. In relation to the temples existing in Alicante (Spain), water drainage is solved with pitched roofs on slope formation (in vaulted naves) or directly supported on the vaulted elements (in the domes). Since those construction systems are composed by brick and plaster, the presence of moisture is problematic, and represents a risk of losing the strength capacity and therefore the stability of the dome. An example of this problem is the dome of the church “Nuestra Señora de Belén” in Crevillente, built with solid bricks, it has the highest diameter of the province (18th century). This historic building has been restored on several occasions in the recent years due to moisture, cracks or fissures. The study of these works give an idea of the difficulties of maintenance, conservation and proper restoration of such kind of buildings as unique and valued constructions in our heritage.

Ngara region, Tanzania and Burundi, 1994, Defense Mapping Agency (DMA) Series Y745, Sheet 4976-III (Raster Image)

This layer is a georeferenced raster image of the United States Defense Mapping Agency (DMA) Series Y745, Tanzania, 1:50,000 Topographic Line Map (TLM) Series sheet map entitled: Ngara. Printed in: 1994. Covers portions of Ngara region, Tanzania and a portion of Burundi. Sheet: 4976-III. Edition statement: Ed. 1 - DMA. The image inside the map neatline is georeferenced to the surface of the earth and fit to World Geodetic System (1984) coordinates. All map collar information is also available as part of the raster image. Tanzania 1:50:000 Series Y745 maps are in English. Each source map in the series is printed in color at a scale of 1:50,000. Series source sheets were published in 1994-1995 by the United States Defense Mapping Agency, Hydrographic/Topographic Center. The source map was scanned and georeferenced for Harvard University's Center for Geographic Analysis' AfricaMap project by East View Cartographic. Individual TLM sheets covering Burundi (40 sheets in total) were selected from the TLM worldwide series. DMA Topographic Line Map series maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 50 feet, with some sheets having supplemental meter contours, form lines, hachures, shading, and/or spot heights. Depths shown by bathymetric isolines. Please pay close attention to map collar information on projections, spheroid, compilation dates, legend information, and keys to grid numbering and other numbers which appear inside the neatline.

New York, N.Y. and vicinity, 1961 : Paterson, N.J.-N.Y. (8 Sheet Set) (Raster Image)

This layer is a georeferenced raster image of the United States Geological Survey 7.5 minute topographic sheet map entitled: New York and vicinity : Paterson, N.J.-N.Y., 1955. It is part of an 8 sheet map set covering the metropolitan New York City area. It was published in 1961. Scale 1:24,000. The source map was prepared by the Geological Survey from 1:24,000-scale maps of Hackensack, Paterson, Orange, and Weehawken 1955 7.5 minute quadrangles. The Orange quadrangle was previously compiled by the Army Map Service. Culture revised by the Geological Survey. Hydrography compiled from USC&GS charts 287 (1954), 745 (1956), and 746 (1956). The image inside the map neatline is georeferenced to the surface of the earth and fit to the Universal Transverse Mercator (UTM) Zone 18N NAD27 projection. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. USGS maps are typical topographic maps portraying both natural and manmade features. They show and name works of nature, such as mountains, valleys, lakes, rivers, vegetation, etc. They also identify the principal works of humans, such as roads, railroads, boundaries, transmission lines, major buildings, etc. Relief is shown with standard contour intervals of 10 and 20 feet; depths are shown with contours and soundings. Please pay close attention to map collar information on projections, spheroid, sources, dates, and keys to grid numbering and other numbers which appear inside the neatline. This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.