Cytoplasmic dynein is associated with slow axonal transport.

Neuronal function is dependent on the transport of materials from the cell body to the synapse via anterograde axonal transport. Anterograde axonal transport consists of several components that differ in both rate and protein composition. In fast transport, membranous organelles are moved along microtubules by the motor protein kinesin. The cytoskeleton and the cytomatrix proteins move in the two components of slow transport. While the mechanisms underlying slow transport are unknown, it has been hypothesized that the movement of microtubules in slow transport is generated by sliding. To determine whether dynein, a motor protein that causes microtubule sliding in flagella, may play a role in slow axonal transport, we identified the transport rate components with which cytoplasmic dynein is associated in rat optic nerve. Nearly 80% of the anterogradely moving dynein was associated with slow transport, whereas only approximately 15% of the dynein was associated with the membranous organelles of anterograde fast axonal transport. A segmental analysis of the transport of dynein through contiguous regions of the optic nerve and tract showed that dynein is associated with the microfilaments and other proteins of slow component b. Dynein from this transport component has the capacity to bind microtubules in vitro. These results are consistent with the hypothesis that cytoplasmic dynein generates the movement of microtubules in slow axonal transport. A model is presented to illustrate how dynein attached to the slow component b complex of proteins is appropriately positioned to generate force of the correct polarity to slide microtubules down the axon.

Distance determination in proteins using designed metal ion binding sites and site-directed spin labeling: application to the lactose permease of Escherichia coli.

As shown in the accompanying paper, the magnetic dipolar interaction between site-directed metal-nitroxide pairs can be exploited to measure distances in T4 lysozyme, a protein of known structure. To evaluate this potentially powerful method for general use, particularly with membrane proteins that are difficult to crystallize, both a paramagnetic metal ion binding site and a nitroxide side chain were introduced at selected positions in the lactose permease of Escherichia coli, a paradigm for polytopic membrane proteins. Thus, three individual cysteine residues were introduced into putative helix IV of a lactose permease mutant devoid of native cysteine residues containing a high-affinity divalent metal ion binding site in the form of six contiguous histidine residues in the periplasmic loop between helices III and IV. In addition, the construct contained a biotin acceptor domain in the middle cytoplasmic loop to facilitate purification. After purification and spin labeling, electron paramagnetic resonance spectra were obtained with the purified proteins in the absence and presence of Cu(II). The results demonstrate that positions 103, 111, and 121 are 8, 14, and > 23 A from the metal binding site. These data are consistent with an alpha-helical conformation of transmembrane domain IV of the permease. Application of the technique to determine helix packing in lactose permease is discussed.

Genetic rearrangements in the rfb regions of Vibrio cholerae O1 and O139.

The recent emergence of a pathogenic new non-O1 serotype (O139) of Vibrio cholerae has led to numerous studies in an attempt to identify the origins of this new strain. Our studies indicate that O139 strains have clear differences in the surface polysaccharides when compared with O1 strains: the lipopolysaccharide can be described as semi-rough. Southern hybridization with the O1 rfb region demonstrates that O139 strains no longer contain any of the rfb genes required for the synthesis of the O1 O-antigen or its modification and also lack at least 6 kb of additional contiguous DNA. However, O139 strains have retained rfaD and have a single open reading frame closely related to three small open reading frames of the O1 rfb region. This region is closely related to the H-repeat of Escherichia coli and to the transposases of a number of insertion sequence elements and has all the features of an insertion sequence element that has been designated VcIS1. Transposon insertion mutants defective in O139 O-antigen (and capsule) biosynthesis map to the same fragment as VcIS1. Preliminary sequence data of complementing clones indicate that this DNA encodes a galactosyl-transferase and other enzymes for the utilization of galactose in polysaccharide biosynthesis. We propose a mechanism by which both the Ogawa serotype of O1 strains and the O139 serotype strains may have evolved.

Illusory contours activate specific regions in human visual cortex: evidence from functional magnetic resonance imaging.

The neural basis for perceptual grouping operations in the human visual system, including the processes which generate illusory contours, is fundamental to understanding human vision. We have employed functional magnetic resonance imaging to investigate these processes noninvasively. Images were acquired on a GE Signa 1.5T scanner equipped for echo planar imaging with an in-plane resolution of 1.5 x 1.5 mm and slice thicknesses of 3.0 or 5.0 mm. Visual stimuli included nonaligned inducers (pacmen) that created no perceptual contours, similar inducers at the corners of a Kanizsa square that created illusory contours, and a real square formed by continuous contours. Multiple contiguous axial slices were acquired during baseline, visual stimulation, and poststimulation periods. Activated regions were identified by a multistage statistical analysis of the activation for each volume element sampled and were compared across conditions. Specific brain regions were activated in extrastriate cortex when the illusory contours were perceived but not during conditions when the illusory contours were absent. These unique regions were found primarily in the right hemisphere for all four subjects and demonstrate that specific brain regions are activated during the kind of perceptual grouping operations involved in illusory contour perception.

The antiproliferative activity of c-myb and c-myc antisense oligonucleotides in smooth muscle cells is caused by a nonantisense mechanism.

Smooth muscle cell (SMC) proliferation is thought to play a major role in vascular restenosis after angioplasty and is a serious complication of the procedure. Developing antisense (AS) oligonucleotides as therapeutics is attractive because of the potentially high specificity of binding to their targets, and several investigators have reported inhibition of SMC proliferation in vitro and in vivo by using AS strategies. We report here the results of our experiments on vascular SMCs using AS oligonucleotides directed toward c-myb and c-myc. We found that significant inhibition of SMC proliferation occurred with these specific AS sequences but that this inhibition was clearly not via a hybridization-dependent AS mechanism. Rather, inhibition was due to the presence of four contiguous guanosine residues in the oligonucleotide sequence. This was demonstrated in vitro in primary cultures of SMCs and in arteries ex vivo. The ex vivo model developed here provides a rapid and effective system in which to screen potential oligonucleotide drugs for restenosis. We have further explored the sequence requirements of this non-AS effect and determined that phosphorothioate oligonucleotides containing at least two sets of three or four consecutive guanosine residues inhibit SMC proliferation in vitro and ex vivo. These results suggest that previous AS data obtained using these and similar, contiguous guanosine-containing AS sequences be reevaluated and that there may be an additional class of nucleic acid compounds that have potential as antirestenosis therapeutics.

A constant radius of curvature model for the organization of DNA in toroidal condensates.

Toroidal DNA condensates have received considerable attention for their possible relationship to the packaging of DNA in viruses and in general as a model of ordered DNA condensation. A spool-like model has primarily been supported for DNA organization within toroids. However, our observations suggest that the actual organization may be considerably different. We present an alternate model in which DNA for a given toroid is organized within a series of equally sized contiguous loops that precess about the toroid axis. A related model for the toroid formation process is also presented. This kinetic model predicts a distribution of toroid sizes for DNA condensed from solution that is in good agreement with experimental data.

O português de herança em território fronteiriço: a LH em Olivença como arma para preservação de um grupo minoritário

Esta pesquisa tem como objetivo investigar a língua portuguesa nativa e de herança sobreviventes em território fronteiriço de língua oficial espanhola. Como locus de pesquisa selecionamos Olivença, uma cidade da Espanha em que a língua portuguesa se faz em situações específicas de uso. A relevância desta pesquisa traduz-se no fato de que espaços fronteiriços mantêm sobrepostas em espaços geográficos contíguos algumas realidades que se sobrepõem: a realidade da oficialidade linguística e a realidade do sentimento de pertença sociolinguística. Nesta tese, partimos da identificação, em trabalho de campo, da presença da língua portuguesa no território atualmente espanhol (mas historicamente português) e na constatação de que há uma flutuação de identificação-identidade linguística. No espaço geográfico em que fizemos incursão científica, duas cidades sobrepõem-se historicamente e duas geografias políticas, por outro lado, avizinham-se e roçam-se continuamente. Ao tomar contato com essa cidade, hipotetizamos que o sentimento de pertença linguística estaria presente entre os falantes mais velhos, que manteriam a herança de traços lusitanos em sua comunicação, mesmo ao falar o espanhol. A justificativa é que, logo de chegada, já avistáramos recintos comerciais com nomes portugueses e, contrariamente, não ouvíamos o som lusitano nas ruas. Sabemos que o domínio espanhol numa cidade outrora portuguesa tenderia a apagar vestígios portugueses. No entanto, em grupos íntimos pressupúnhamos o português como língua corrente. Durante o trabalho de campo, identificamos fortes valores culturais sendo empunhados como armas de resistência entre descendentes de portugueses, fazendo correr numa velocidade acentuada a reorganização dos valores lusitanos em redutos da cidade espanhola. Essa força e essa velocidade pareciam ser as molas propulsoras de uma mudança linguística muito sorrateira, que impactava o sentimento de unidade de um segmento social da comunidade sociolinguística. Isso nos inspirou a dar um passo investigativo seguinte em direção aos mais jovens, que tinham o espanhol como língua materna, mas tinham o português como língua de herança. À pergunta central sobre a força do português como língua de herança buscamos respostas por meio de duas outras questões mais indiretas feitas aos sujeitos entrevistados: será que os mais jovens percebiam-se como portugueses? será que os elementos culturais lusitanos presentes nas ruas eram reconhecidos como vinculados à língua de herança? Foi assim que passamos a recolher pistas sobre os traços de resiliência do português como língua incrustada na região espanhola de Olivença.

The accretion environment in Vela X-1 during a flaring period using XMM-Newton

We present analysis of 100 ks contiguous XMM-Newton data of the prototypical wind accretor Vela X-1. The observation covered eclipse egress between orbital phases 0.134 and 0.265, during which a giant flare took place, enabling us to study the spectral properties both outside and during the flare. This giant flare with a peak luminosity of 3.92+0.42-0.09 × 1037 erg s-1 allows estimates of the physical parameters of the accreted structure with a mass of ~1021 g. We have been able to model several contributions to the observed spectrum with a phenomenological model formed by three absorbed power laws plus three emission lines. After analysing the variations with orbital phase of the column density of each component, as well as those in the Fe and Ni fluorescence lines, we provide a physical interpretation for each spectral component. Meanwhile, the first two components are two aspects of the principal accretion component from the surface of the neutron star, the third component seems to be the X-ray light echo formed in the stellar wind of the companion.

World Map Showing Routes of Exploration and Discovery, 1790 (Image 8 of 8) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Chart of the world on Mercators projection : exhibiting all the new discoveries to the present time, with the tracks of the most distinguished navigators since the year 1700 carefully collected from the best charts, maps, voyages, &c. extant and regulated from the accurate astronomical observations made in three voyages performed under the command of Captn. James Cook in the years 1768, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 & 80, compiled and published by A. Arrowsmith, geographer; by permission of Simon McTavish Esq[r] is correctly delineated the discoveries of Mr. McKenzie laid down from his original journal in the year 1789. It was published by A. Arrowsmith, April 1, 1790. Scale [ca. 1:20,000,000]. This layer is image 8 of 8 total images of the seven sheet source map. Covers portions of eastern Asia. The original map sheet contains multiple maps on one sheet. Because the map images are non-contiguous and possibly at differing scales, each map image was georeferenced separately. The image inside the map neatline is georeferenced to the surface of the earth and fit to the 'Mercator' 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. This map shows features such as drainage, cities and other human settlements, territorial boundaries, shoreline features, and more. Relief shown by hachures. Depths shown by soundings. Includes routes, locations, and dates of James Cook's voyages. This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection and the Harvard University Library as part of the Open Collections Program at Harvard University project: Organizing Our World: Sponsored Exploration and Scientific Discovery in the Modern Age. Maps selected for the project correspond to various expeditions and represent a range of regions, originators, ground condition dates, scales, and purposes.

World Map Showing Routes of Exploration and Discovery, 1790 (Image 7 of 8) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Chart of the world on Mercators projection : exhibiting all the new discoveries to the present time, with the tracks of the most distinguished navigators since the year 1700 carefully collected from the best charts, maps, voyages, &c. extant and regulated from the accurate astronomical observations made in three voyages performed under the command of Captn. James Cook in the years 1768, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 & 80, compiled and published by A. Arrowsmith, geographer; by permission of Simon McTavish Esq[r] is correctly delineated the discoveries of Mr. McKenzie laid down from his original journal in the year 1789. It was published by A. Arrowsmith, April 1, 1790. Scale [ca. 1:20,000,000]. This layer is image 7 of 8 total images of the seven sheet source map. Covers portions of Australia and Southeast Asia. The original map sheet contains multiple maps on one sheet. Because the map images are non-contiguous and possibly at differing scales, each map image was georeferenced separately. The image inside the map neatline is georeferenced to the surface of the earth and fit to the 'Mercator' 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. This map shows features such as drainage, cities and other human settlements, territorial boundaries, shoreline features, and more. Relief shown by hachures. Depths shown by soundings. Includes routes, locations, and dates of James Cook's voyages. This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection and the Harvard University Library as part of the Open Collections Program at Harvard University project: Organizing Our World: Sponsored Exploration and Scientific Discovery in the Modern Age. Maps selected for the project correspond to various expeditions and represent a range of regions, originators, ground condition dates, scales, and purposes.

Massachusetts, Connecticut, and Rhode Island, 1825 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of Massachusetts, Connecticut and Rhode Island : constructed from the latest authorities, drawn by D.H. Vance ; engraved by J.H. Young. It was published by A. Finley in 1825. Scale [ca. 1:700,000]. Covers also portions of New York, New Jersey, Vermont, and New Hampshire. The image inside the map neatline is georeferenced to the surface of the earth and fit to the USA Contiguous Albers Equal Area Conic projection (Meters). 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, or other information associated with the principal map. This map shows features such as roads, drainage, state, county, and town boundaries, and more. Relief is shown pictorially. Includes statistical table. This layer is part of a selection of digitally scanned and georeferenced historic maps of New England from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

Massachusetts, Connecticut, and Rhode Island, 1819 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: A map of Massachusetts, Connecticut and Rhodeisland, by E. Ruggles; engraved by M.M. Peabody. It was published in 1819. Scale [ca. 1:424,000]. Covers Massachusetts, Connecticut, Rhode Island, and portions of Vermont, New Hampshire, Maine, New York, and New Jersey. The image inside the map neatline is georeferenced to the surface of the earth and fit to the USA Contiguous Albers Equal Area Conic projection (Meters). 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, or other information associated with the principal map. This map shows features such as roads, bridges, societies, drainage, lighthouses, coastal hazards, state, county, and town boundaries, and more. Relief shown by hachures. This layer is part of a selection of digitally scanned and georeferenced historic maps of New England from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New England, 1826 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: A map of the New England states : Maine, New Hampshire, Vermont, Massachusetts, Rhode Island & Connecticut, with the adjacent parts of New York & lower Canada, compiled and published by Nathan Hale ; engraved by J.V.N. Throop. It was published by in 1826. Scale [ca. 1:506,880]. Covers also a portion of the state of New York and the province of Quebec, Canada. The image inside the map neatline is georeferenced to the surface of the earth and fit to the USA Contiguous Albers Equal Area Conic projection (Meters). 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, or other information associated with the principal map. This map shows features such as roads, churches, drainage, state, county, and town boundaries, and more. Relief shown by hachures. Includes inset: Northern & eastern part of Maine and part of lower Canada and New Brunswick. Scale [ca. 1:1,267,200]. This layer is part of a selection of digitally scanned and georeferenced historic maps of New England from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

Maine, New Hampshire, and Vermont, 1845 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Map of Maine, New Hampshire, and Vermont : compiled from the latest authorities, [by] J.H. Young, sc. It was published by S. Augustus Mitchell in 1845. Scale [ca. 1:1,001,000]. Covers also portions of New York, Massachusetts, and the provinces of Quebec and New Brunswick, Canada. The image inside the map neatline is georeferenced to the surface of the earth and fit to the USA Contiguous Albers Equal Area Conic projection (Meters). 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, or other information associated with the principal map. This map shows features such as roads, railroads, canals, drainage, state, county, and town boundaries, and more. Relief shown by hachures. Includes inset: North part of Maine. Scale [ca. 1:2,505,000]. This layer is part of a selection of digitally scanned and georeferenced historic maps of New England from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

New England, 1862 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: G. Woolworth Colton's railroad, township & distance map of New England : with adjacent portions of New York, Canada & New Brunswick, drawn, engraved & published by G. Woolworth Colton. It was published in 1862. Scale 1:900,000. The image inside the map neatline is georeferenced to the surface of the earth and fit to the USA Contiguous Albers Equal Area Conic projection (Meters). 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, or other information associated with the principal map. This map shows features such as roads, railroads, drainage, state, county, and town boundaries, and more. Relief shown by hachures. Includes table of distances and inset: The eastern portion of Massachusetts on an enlarged scale. Scale 1:350,000. This layer is part of a selection of digitally scanned and georeferenced historic maps of New England from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.