598 research outputs found

    An antibody raised against a pathogenic serpin variant induces mutant-like behaviour in the wild-type protein

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    A monoclonal antibody (mAb) that binds to a transient intermediate may act as a catalyst for the corresponding reaction; here we show this principle can extend on a macro molecular scale to the induction of mutant-like oligomerization in a wild-type protein. Using the common pathogenic E342K (Z) variant of α1-antitrypsin as antigen-whose native state is susceptible to the formation of a proto-oligomeric intermediate-we have produced a mAb (5E3) that increases the rate of oligomerization of the wild-type (M) variant. Employing ELISA, gel shift, thermal stability and FRET time-course experiments, we show that mAb5E3 does not bind to the native state of α1-antitrypsin, but recognizes a cryptic epitope in the vicinity of the post-helix A loop and strand 4C that is revealed upon transition to the polymerization intermediate, and which persists in the ensuing oligomer. This epitope is not shared by loop-inserted monomeric conformations. We show the increased amenity to polymerization by either the pathogenic E342K mutation or the binding of mAb5E3 occurs without affecting the energetic barrier to polymerization. As mAb5E3 also does not alter the relative stability of the monomer to intermediate, it acts in a manner similar to the E342K mutant, by facilitating the conformational interchange between these two states

    Peer Review—The Newcomers' Perspective

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    The World Academy of Young Scientists argue that double blind peer-review will generate a better perception of fairness and equality in global scientific funding and publishin

    Tidal breathing parameters measured using structured light plethysmography in healthy children and those with asthma before and after bronchodilator

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    Structured light plethysmography (SLP) is a light‐based, noncontact technique that measures tidal breathing by monitoring displacements of the thoracoabdominal (TA) wall. We used SLP to measure tidal breathing parameters and their within‐subject variability (v) in 30 children aged 7–16 years with asthma and abnormal spirometry (forced expiratory volume in 1 sec [FEV1] <80% predicted) during a routine clinic appointment. As part of standard care, the reversibility of airway obstruction was assessed by repeating spirometry after administration of an inhaled bronchodilator. In this study, SLP was performed before and after bronchodilator administration, and also once in 41 age‐matched controls. In the asthma group, there was a significant increase in spirometry‐assessed mean FEV1 after administration of bronchodilator. Of all measured tidal breathing parameters, the most informative was the inspiratory to expiratory TA displacement ratio (IE50SLP, calculated as TIF50SLP/TEF50SLP, where TIF50SLP is tidal inspiratory TA displacement rate at 50% of inspiratory displacement and TEF50SLP is tidal expiratory TA displacement rate at 50% of expiratory displacement). Median (m) IE50SLP and its variability (vIE50SLP) were both higher in children with asthma (prebronchodilator) compared with healthy children (mIE50SLP: 1.53 vs. 1.22, P < 0.001; vIE50SLP: 0.63 vs. 0.47, P < 0.001). After administration of bronchodilators to the asthma group, mIE50SLP decreased from 1.53 to 1.45 (P = 0.01) and vIE50SLP decreased from 0.63 to 0.60 (P = 0.04). SLP‐measured tidal breathing parameters could differentiate between children with and without asthma and indicate a response to bronchodilator

    LayeredCast -A Hybrid Peer-to-Peer Live Layered Video Streaming Protocol

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    Abstract-Peer-to-Peer overlay networks are an attractive foundation for video streaming. However, live Peer-to-Peer media streaming systems face many challenges such as bandwidth heterogeneity, node churn, and selfish nodes. Although many tree based and mesh based streaming protocols have been proposed, each has its own drawbacks such as unreliability and unfairness in tree based and long startup delay and complex scheduling in mesh based protocols. In this paper, we propose a new video streaming protocol called LayeredCast main features of which are: 1) Hybrid: Drawbacks of the simple approaches are compensated using a hybrid of mesh and tree overlays. 2) Layered Video: Provides an adaptive scheme to enhance the video quality using a layered video codec for heterogeneous clients. 3) QoS: LayeredCast scheduling aims at moving complexity of Multi-Service network core to the network clients application layer, thus providing better QoS over simple regular networks. LayeredCast&apos;s tree network pushes the base layer to all peers while the enhancement layers and missing base layer segments are pulled over a mesh network by peers with extra bandwidth using a new data-driven scheduling scheme. We have evaluated the performance of LayeredCast on an innovative simulation framework. Simulation results verify better performance of LayeredCast in term of decodable video frames over CoolStreaming, especially when network resources are limited

    Ultrastructural and molecular studies on fat and thin macrocysts of Sarcocystis spp. isolated from naturally infected goats

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    ABSTRACT There is considerable confusion concerning the structure and synonymity of macroscopic sarcocysts in goats. In the present work, ultrastructural and molecular study on fat and thin macrocysts of Sarcocystis isolated from naturally infected goats were investigated. Two forms of macrocystic sarcocysts were collected from fresh oesophagus, diaphragm, skeletal muscles and heart of slaughtered goats in abattoirs of Tehran and Ghazvin provinces of Iran. Then ultrastructural and molecular studies were carried out contemporaneous using PCR technique and transmission electron microscope respectively. Based on morphological examinations both cysts differ in shape with naked eye but they had the same wall structure with electron microscopy study. The 18S rRNA amplification by PCR showed a 637-bp band for both forms. The results of sequencing of PCR product of the cysts indicated these two kinds of cysts were identical to each other and also to Sarcocystis moulei. The present result, suggests that fat and thin cysts are the same and identical species

    The fungus Neurospora crassa displays telomeric silencing mediated by multiple sirtuins and by methylation of histone H3 lysine 9

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    <p>Abstract</p> <p>Background</p> <p>Silencing of genes inserted near telomeres provides a model to investigate the function of heterochromatin. We initiated a study of telomeric silencing in <it>Neurospora crassa</it>, a fungus that sports DNA methylation, unlike most other organisms in which telomeric silencing has been characterized.</p> <p>Results</p> <p>The selectable marker, <it>hph</it>, was inserted at the subtelomere of Linkage Group VR in an <it>nst-1 </it>(n<it>eurospora </it>s<it>ir </it>t<it>wo</it>-1) mutant and was silenced when <it>nst-1 </it>function was restored. We show that NST-1 is an H4-specific histone deacetylase. A second marker, <it>bar</it>, tested at two other subtelomeres, was similarly sensitive to <it>nst-1 </it>function. Mutation of three additional SIR2 homologues, <it>nst-2</it>, <it>nst-3 </it>and <it>nst-5</it>, partially relieved silencing. Two genes showed stronger effects: <it>dim-5</it>, which encodes a histone H3 K9 methyltransferase and <it>hpo</it>, which encodes heterochromatin protein-1. Subtelomeres showed variable, but generally low, levels of DNA methylation. Elimination of DNA methylation caused partial derepression of one telomeric marker. Characterization of histone modifications at subtelomeric regions revealed H3 trimethyl-K9, H3 trimethyl-K27, and H4 trimethyl-K20 enrichment. These modifications were slightly reduced when telomeric silencing was compromised. In contrast, acetylation of histones H3 and H4 increased.</p> <p>Conclusion</p> <p>We demonstrate the presence of telomeric silencing in Neurospora and show a dependence on histone deacetylases and methylation of histone H3 lysine 9. Our studies also reveal silencing functions for DIM-5 and HP1 that appear independent of their role in <it>de novo </it>DNA methylation.</p

    Broadband Antireflection and Light Extraction Enhancement in Fluorescent SiC with Nanodome Structures

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    We demonstrate a time-efficient and low-cost approach to fabricate Si3N4 coated nanodome structures in fluorescent SiC. Nanosphere lithography is used as the nanopatterning method and SiC nanodome structures with Si3N4 coating are formed via dry etching and thin film deposition process. By using this method, a significant broadband surface antireflection and a considerable omnidirectional luminescence enhancement are obtained. The experimental observations are then supported by numerical simulations. It is believed that our fabrication method will be well suitable for large-scale production in the future

    Synthetic heterochromatin bypasses RNAi and centromeric repeats to establish functional centromeres

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    In the central domain of fission yeast centromeres, the kinetochore is assembled on CENP-A cnp1 nucleosomes. Normally, small interfering RNAs generated from flanking outer repeat transcripts direct histone H3 lysine 9 methyltransferase Clr4 to homologous loci to form heterochromatin. Outer repeats, RNA interference (RNAi), and centromeric heterochromatin are required to establish CENP-A Cnpl chromatin. We demonstrated that tethering Clr4 via DNA-binding sites at euchromatic loci induces heterochromatin assembly, with or without active RNAi. This synthetic heterochromatin completely substitutes for outer repeats on plasmid-based minichromosomes, promoting de novo CENP-A Cnpl and kinetochore assembly, to allow their mitotic segregation, even with RNAi inactive. Thus, the role of outer repeats in centromere establishment is simply the provision of RNAi substrates to direct heterochromatin formation; H3K9 methylation-dependent heterochromatin is alone sufficient to form functional centromeres.SCOPUS: ar.jinfo:eu-repo/semantics/publishe
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