2,295 research outputs found

    A Single-stranded DNA-binding Protein of Bacteriophage T7 Defective in DNA Annealing

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    The annealing of complementary strands of DNA is a vital step during the process of DNA replication, recombination, and repair. In bacteriophage T7-infected cells, the product of viral gene 2.5, a single-stranded DNA-binding protein, performs this function. We have identified a single amino acid residue in gene 2.5 protein, arginine 82, that is critical for its DNA annealing activity. Expression of gene 2.5 harboring this mutation does not complement the growth of a T7 bacteriophage lacking gene 2.5. Purified gene 2.5 protein-R82C binds single-stranded DNA with a greater affinity than the wild-type protein but does not mediate annealing of complementary strands of DNA. A carboxyl-terminal-deleted protein, gene 2.5 protein-Delta26C, binds even more tightly to single-stranded DNA than does gene 2.5 protein-R82C, but it anneals homologous strands of DNA as well as does the wild-type protein. The altered protein forms dimers and interacts with T7 DNA polymerase comparable with the wild-type protein. Gene 2.5 protein-R82C condenses single-stranded M13 DNA in a manner similar to wild-type protein when viewed by electron microscopy

    Initial Registration of 3D Parcels

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    Registering the rights of a 3D parcel provides certainty of ownership, protection of rights and unambiguous spatial location. While not all cadastral jurisdictions in the world maintain a digital cadastral database, the concepts of such registration hold true regardless of whether it is a paper-based cadastre or a digital one. Similarly, the motivations and purpose for the creation of a 2D cadastre for individual jurisdictions applies for 3D cadastres as well. It provides a security of ownership of 3D parcel, protects the rights of the owners, and provides valuable financial instruments such as mortgage, collateral, valuation and taxation. The current life cycle of the development of a land parcel includes processes beginning from outside the cadastral registration sphere, such as zoning plans and permits, but has a direct impact on how a certain development application is processed. Thus, in considering the changes required to allow a jurisdiction to register 3D, it is important to note the sphere of influence that could have an impact on 3D registration. These include planners, notaries, surveyors, data managers and registrars; however for the purpose of this paper, the research is focused on the core 3D aspects that are institutional, legal and technical. This paper explores approaches and solutions towards the implementation of initial 3D cadastral registration, as derive by current procedures of registration of 3D parcels in various countries worldwide. To this purpose, the paper analyses the categorisations and approaches to 3D spatial units and examines the validation requirements (constraints) on a cadastral database, at various levels of maturity. In this view, 3D data storage and visualization issues are examined in relation to the level of complexity of various jurisdictions, as provided by the results of the country inventory combined with a worldwide survey in 2010 and updated in 2014 (Van Oosterom, et al, 2014). It seems that significant progress has been achieved in providing legal provisions for the registration of 3D cadastres in many countries and several have started to show 3D information on cadastral plans such as isometric views, vertical profiles or text environment to facilitate such data capture and registration. Moreover, as jurisdictions progress towards an implementation of 3D cadastres, much 3D data collected in other areas (BIM, IFC CityGML files, IndoorGML, InfraGML and LandXML) open up the possibility of creating 3D cadastral database combining the existing datasets. The usability, compatibility and portability of these datasets is a low cost solution to one of the costliest phases of the implementation of 3D cadastres, which is the initial 3D data capture

    Formation of a DNA Loop at the Replication Fork Generated by Bacteriophage T7 Replication Proteins

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    Intermediates in the replication of circular and linear M13 double-stranded DNA by bacteriophage T7 proteins have been examined by electron microscopy. Synthesis generated double-stranded DNA molecules containing a single replication fork with a linear duplex tail. A complex presumably consisting of T7 DNA polymerase and gene 4 helicase/primase molecules was present at the fork together with a variable amount of single-stranded DNA sequestered by gene 2.5 single-stranded DNA binding protein. Analysis of the length distribution of Okazaki fragments formed at different helicase/primase concentrations was consistent with coupling of leading and lagging strand replication. Fifteen to forty percent of the templates engaged in replication have a DNA loop at the replication fork. The loops are fully double-stranded with an average length of approximately 1 kilobase. Labeling with biotinylated dCTP showed that the loops consist of newly synthesized DNA, and synchronization experiments using a linear template with a G-less cassette demonstrated that the loops are formed by active displacement of the lagging strand. A long standing feature of models for coupled leading/lagging strand replication has been the presence of a DNA loop at the replication fork. This study provides the first direct demonstration of such loops

    Detection of Central Retinal Artery Occlusion by Point-of-Care Ultrasound in the Emergency Department: A Case Series.

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    Central retinal artery occlusion (CRAO) is a rare, but serious, diagnosis that can lead to blindness, most often due to thromboembolic disease. In the emergency department (ED), CRAO can present as acute, painless loss of vision. Physicians need quick ways to rule in this diagnosis due to the time-sensitive nature of the event. We describe two patients in this cases series who present to the same ED with unilateral painless vision loss and histories that include notable risk factors such as thromboembolic and atherosclerotic disease. Upon arrival, point-of-care ultrasound (POCUS) done at the bedside allowed for quick determination of CRAO. The importance of this case series is to emphasize the efficacy of POCUS in evaluating patients with painless vision loss in the ED setting

    Three-Dimensional Microscopy Characterization of Death Receptor 5 Expression by Over-Activated Human Primary CD4+ T Cells and Apoptosis

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    Activation-induced cell death is a natural process that prevents tissue damages from over-activated immune cells. TNF-Related apoptosis ligand (TRAIL), a TNF family member, induces apoptosis of infected and tumor cells by binding to one of its two death receptors, DR4 or DR5. TRAIL was reported to be secreted by phytohemagglutinin (PHA)-stimulated CD4+ T cells in microvesicles

    A Study of the Diverse T Dwarf Population Revealed by WISE

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    We report the discovery of 87 new T dwarfs uncovered with the Wide-field Infrared Survey Explorer (WISE) and three brown dwarfs with extremely red near-infrared colors that exhibit characteristics of both L and T dwarfs. Two of the new T dwarfs are likely binaries with L7+/-1 primaries and mid-type T secondaries. In addition, our follow-up program has confirmed 10 previously identified T dwarfs and four photometrically-selected L and T dwarf candidates in the literature. This sample, along with the previous WISE discoveries, triples the number of known brown dwarfs with spectral types later than T5. Using the WISE All-Sky Source Catalog we present updated color-color and color-type diagrams for all the WISE-discovered T and Y dwarfs. Near-infrared spectra of the new discoveries are presented, along with spectral classifications. To accommodate later T dwarfs we have modified the integrated flux method of determining spectral indices to instead use the median flux. Furthermore, a newly defined J-narrow index differentiates the early-type Y dwarfs from late-type T dwarfs based on the J-band continuum slope. The K/J indices for this expanded sample show that 32% of late-type T dwarfs have suppressed K-band flux and are blue relative to the spectral standards, while only 11% are redder than the standards. Comparison of the Y/J and K/J index to models suggests diverse atmospheric conditions and supports the possible re-emergence of clouds after the L/T transition. We also discuss peculiar brown dwarfs and candidates that were found not to be substellar, including two Young Stellar Objects and two Active Galactic Nuclei. The coolest WISE-discovered brown dwarfs are the closest of their type and will remain the only sample of their kind for many years to come.Comment: Accepted to ApJS on 15 January 2013; 99 pages in preprint format, 30 figures, 12 table

    Brain-Specific Phosphorylation of MeCP2 Regulates Activity-Dependent Bdnf Transcription, Dendritic Growth, and Spine Maturation

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    Mutations or duplications in MECP2 cause Rett and Rett-like syndromes, neurodevelopmental disorders characterized by mental retardation, motor dysfunction, and autistic behaviors. MeCP2 is expressed in many mammalian tissues and functions as a global repressor of transcription; however, the molecular mechanisms by which MeCP2 dysfunction leads to the neural-specific phenotypes of RTT remain poorly understood. Here, we show that neuronal activity and subsequent calcium influx trigger the de novo phosphorylation of MeCP2 at serine 421 (S421) by a CaMKII-dependent mechanism. MeCP2 S421 phosphorylation is induced selectively in the brain in response to physiological stimuli. Significantly, we find that S421 phosphorylation controls the ability of MeCP2 to regulate dendritic patterning, spine morphogenesis, and the activity-dependent induction of Bdnf transcription. These findings suggest that, by triggering MeCP2 phosphorylation, neuronal activity regulates a program of gene expression that mediates nervous system maturation and that disruption of this process in individuals with mutations in MeCP2 may underlie the neural-specific pathology of RTT

    Multisystem diseases and infections

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    Differential diagnosis of fevers?, Fever without localizing features?, Sepsis?, Cancer?, General rules of cancer management?, Rheumatoid arthritis?, Osteoarthritis?, Systemic lupus erythematosus?, Typhoid and paratyphoid fevers?, Rickettsioses?, Bartonella?, Ehrlichia?, Coxiella?, Relapsing fevers?, Leptospirosis?, Brucellosis?, Plague?, Melioidosis?, Anthrax?, African trypanosomiasis?, American trypanosomiasis?, Visceral leishmaniasis (kala-azar)?, Infectious mononucleosis?, Measles?, Arboviruses and zoonotic haemorrhagic fever viruses , Ebola and Marburg virus diseases, Crimean-Congo haemorrhagic fever, Rift Valley fever, Lassa fever, Hantavirus infections, Severe fever and thrombocytopenia, Zika virus, Japanese encephalitis , Dengue virus, Yellow fever, West Nile virus , Kyasanur Forest Disease, Chikungunya, Ross River fever, O'nyong nyon
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