Sensitive and specific detection of viable Mycobacterium avium subsp. paratuberculosis in raw milk by the Peptide-mediated magnetic separation (PMS)-phage assay

This study describes further validation of a previously described Peptide-mediated magnetic separation (PMS)-Phage assay, and its application to test raw cows’ milk for presence of viable Mycobacterium avium subsp. paratuberculosis (MAP). The inclusivity and exclusivity of the PMS-phage assay were initially assessed, before the 50% limit of detection (LOD50) was determined and compared with those of PMS-qPCR (targeting both IS900 and f57) and PMS-culture. These methods were then applied in parallel to test 146 individual milk samples and 22 bulk tank milk samples from Johne’s affected herds. Viable MAP were detected by the PMS-phage assay in 31 (21.2%) of 146 individual milk samples (mean plaque count of 228.1 PFU/50 ml, range 6-948 PFU/50 ml), and 13 (59.1%) of 22 bulk tank milks (mean plaque count of 136.83 PFU/50 ml, range 18-695 PFU/50 ml). In contrast, only 7 (9.1%) of 77 individual milks and 10 (45.4%) of 22 bulk tank milks tested PMS-qPCR positive, and 17 (11.6%) of 146 individual milks and 11 (50%) of 22 bulk tank milks tested PMS-culture positive. The mean 50% limits of detection (LOD50) of the PMS-phage, PMS-IS900 qPCR and PMS-f57 qPCR assays, determined by testing MAP-spiked milk, were 0.93, 135.63 and 297.35 MAP CFU/50 ml milk, respectively. Collectively, these results demonstrate that, in our laboratory, the PMS-phage assay is a sensitive and specific method to quickly detect the presence of viable MAP cells in milk. However, due to its complicated, multi-step nature, the method would not be a suitable MAP screening method for the dairy industry.

The temperate Burkholderia phage AP3 of the Peduovirinae shows efficient antimicrobial activity against B. cenocepacia of the IIIA lineage

Burkholderia phage AP3 (vB_BceM_AP3) is a temperate virus of the Myoviridae and the Peduovirinae subfamily (P2likevirus genus). This phage specifically infects multidrug-resistant clinical Burkholderia cenocepacia lineage IIIA strains commonly isolated from cystic fibrosis patients. AP3 exhibits high pairwise nucleotide identity (61.7%) to Burkholderia phage KS5, specific to the same B. cenocepacia host, and has 46.7% - 49.5% identity to phages infecting other species of Burkholderia. The lysis cassette of these related phages has a similar organization (putative antiholin, putative holin, endolysin and spanins) and shows 29-98% homology between specific lysis genes, in contrast to Enterobacteria phage P2, the hallmark phage of this genus. The AP3 and KS5 lysis genes have conserved locations and high amino acid sequence similarity. The AP3 bacteriophage particles remain infective up to 5 h at pH 4-10 and are stable at 60°C for 30 min, but are sensitive to chloroform, with no remaining infective particles after 24 h of treatment. AP3 lysogeny can occur by stable genomic integration and by pseudo-lysogeny. The lysogenic bacterial mutants did not exhibit any significant changes in virulence compared to wild-type host strain when tested in the Galleria mellonella moth wax model. Moreover, AP3 treatment of larvae infected with B. cenocepacia revealed a significant increase (P < 0.0001) in larvae survival in comparison to AP3-untreated infected larvae. AP3 showed robust lytic activity, as evidenced by its broad host range, the absence of increased virulence in lysogenic isolates, the lack of bacterial gene disruption conditioned by bacterial tRNA downstream integration site, and the absence of detected toxin sequences. These data suggest the AP3 phage is a promising potent agent against bacteria belonging to most common B. cenocepacia IIIA lineage strains.

Novel inhibitors of the tRNA-dependent amidotransferase of "Helicobacter pylori": Peptides generated by phage display and dipeptide-like compounds

Cette thèse présente la découverte de nouveaux inhibiteurs de l’amidotransférase ARNt-dépendante (AdT), et résume les connaissances récentes sur la biosynthèse du Gln-ARNtGln et de l’Asn-ARNtAsn par la voie indirecte chez la bactérie Helicobacter pylori. Dans le cytoplasme des eucaryotes, vingt acides aminés sont liés à leur ARNt correspondant par vingt aminoacyl-ARNt synthétases (aaRSs). Ces enzymes sont très spécifiques, et leur fonction est importante pour le décodage correct du code génétique. Cependant, la plupart des bactéries, dont H. pylori, sont dépourvues d’asparaginyl-ARNt synthétase et/ou de glutaminyl-ARNt synthétase. Pour former le Gln-ARNtGln, H. pylori utilise une GluRS noncanonique nommée GluRS2 qui glutamyle spécifiquement l’ARNtGln ; ensuite, une AdT trimérique, la GatCAB corrige le Glu-ARNtGln mésapparié en le transamidant pour former le Gln-ARNtGln, qui lira correctement les codons glutamine pendant la biosynthèse des protéines sur les ribosomes. La formation de l’Asn-ARNtAsn est similaire à celle du Gln-ARNtGln, et utilise la même GatCAB et une AspRS non-discriminatrice. Depuis des années 2000, la GatCAB est considérée comme une cible prometteuse pour le développement de nouveaux antibiotiques, puisqu’elle est absente du cytoplasme de l’être humain, et qu’elle est encodée dans le génome de plusieurs bactéries pathogènes. Dans le chapitre 3, nous présentons la découverte par la technique du « phage display » de peptides cycliques riches en tryptophane et en proline, et qui inhibent l’activité de la GatCAB de H. pylori. Les peptides P10 (CMPVWKPDC) et P9 (CSAHNWPNC) inhibent cette enzyme de façon compétitive par rapport au substrat Glu-ARNtGln. Leur constante d’inhibition (Ki) est 126 μM pour P10, et 392 μM pour P9. Des modèles moléculaires ont montré qu’ils lient le site actif de la réaction de transmidation catalysée par la GatCAB, grâce à la formation d’une interaction π-π entre le résidu Trp de ces peptides et le résidu Tyr81 de la sous-unité GatB, comme fait le A76 3’-terminal de l’ARNt. Dans une autre étude concernant des petits composés contenant un groupe sulfone, et qui mimiquent l’intermédiaire de la réaction de transamidation, nous avons identifié des composés qui inhibent la GatCAB de H. pylori de façon compétitive par rapport au substrat Glu-ARNtGln. Cinq fois plus petits que les peptides cycliques mentionnés plus haut, ces composés inhibent l’activité de la GatCAB avec des Ki de 139 μM pour le composé 7, et de 214 μM pour le composé 4. Ces inhibiteurs de GatCAB pourraient être utiles pour des études mécanistiques, et pourraient être des molécules de base pour le développement de nouvelles classes d’antibiotiques contre des infections causées par H. pylori.

Suggestions to public libraries and local historical societies relative to collecting and preserving materials of local history, 1904

Public libraries and historical societies should work together in collecting and preserving materials of local history. This brochure is a guide for their collaboration.

A Multimedia Video Player Based of FFmpeg

With the rapid development of Internet technologies, video and audio processing are among the most important parts due to the constant requirements of high quality media contents. Along with the improvement of network environment and the hardware equipment, this demand is becoming more and more imperious, people prefer high quality videos and audios as well as the net streaming media resources. FFmpeg is a set of open source program about the A/V decoding. Many commercial players use FFmpeg as their displaying cores. This paper designed a simple and easy-to-use video player based on FFmpeg. The first part is about the basic theories and related knowledge of video displaying, including some concepts like data formats, streaming media data, video coding and decoding. In a word, the realization of the video player depend on the a set of video decoding process. The general idea about the process is to get the video packets from the Internet, to read the related protocols and de-encapsulate the protocols, to de-encapsulate the packaging data and to get encoded formats data, to decode them to pixel data that can be displayed directly through graphics cards. During the coding and decoding process, there could be different degrees of data losing, which is called lossy compression, but it usually does not influence the quality of user experiences. The second part is about the principle of the FFmpeg decoding process, that is one of the key point of the paper. In this project, FFmpeg is used for the main decoding task, by call some main functions and structures from FFmpeg class libraries, packaging video formats could be transfer to pixel data, after getting the pixel data, SDL is used for the displaying process. The third part is about the SDL displaying flow. Similarly, it would invoke some important displaying functions from SDL class libraries to realize the function, though SDL is able to do not only displaying task, but also many other game playing process. After that, a independent video displayer is completed, it is provided with all the key function of a player. The fourth part make a simple users interface for the player based on the MFC program, it enable the player could be used by most people. At last, in consideration of the mobile Internet’s blossom, people nowadays can hardly ever drop their mobile phones, there is a brief introduction about how to transplant the video player to Android platform which is one of the most used mobile systems.

Metrics that compare academic libraries, and how we stack up within CIC

This was presented during the 2nd annual Library Research and Innovation Practices at the University of Maryland Libraries, McKeldin Library, on June 8, 2016.

The changing role of subject librarians in academic libraries

Discusses the roles that subject librarians (or 'subject specialists') play in contemporary UK academic libraries. Argues that subject librarians, who still form a significant grouping of senior staff in most UK academic libraries, continue to have a significant role to play in the delivery of library services and that applies to both traditional and electronic library services. Discusses the traditional role of subject librarians and analyzes the way in which this role is changing. Those areas where the changing responsibilities are extensions of traditional roles into new areas are pinpointed, together with examples of where subject librarians are performing new roles and adopting new ways of working. Areas where the changing role of subject librarians can be specifically identified include: greater emphasis on liaison with users; advocacy of the collections; adopting new roles; dealing with user enquiries in new ways; working with technical staff; selecting electronic library materials; carrying out more information skills training; having a greater involvement in the implementation of educational technology; team working and project working. Presents practical examples based on experiences at Nottingham university and other UK research libraries. The redesign and relaunch of Nottingham University Library Web site is described to illustrate many of these points.

Exploring protein-RNA interactions with site-directed mutagenesis and phage display

The importance of RNA as a mediator of genetic information is widely appreciated. RNA molecules also participate in the regulation of various post-transcriptional activities, such as mRNA splicing, editing, RNA stability and transport. Their regulatory roles for these activities are highly dependent on finely tuned associations with cognate proteins. The RNA recognition motif (RRM) is an ancient RNA binding module that participates in hundreds of essential activities where specific RNA recognition is required. We have applied phage display and site-directed mutagenesis to dissect principles of RRM-controlled RNA recognition. The model systems we are investigating are U1A and CUG-BP1. In this dissertation, the molecular basis of the binding affinity of U1A-RNA beyond individual contacts was investigated. We have identified and evaluated the contributions of the local cooperativity formed by three neighboring residues (Asn15, Asn16 and Glu19) to the stability of the U1A-RNA complex. The localized cooperative network was mapped by double-mutant cycles and explored using phage display. We also showed that a cluster of these residues forms a “hot spot” on the surface of U1A; a single substitution at position 19 with Gln or His can alter the binding properties of U1A to recognize a non-cognate G4U RNA. Finally, we applied a deletion analysis of CUG-BP1 to define the contributions of individual RRMs and RRM combinations to the stability of the complex formed between CUG-BP1 and the GRE sequence. The preliminary results showed RRM3 of CUG-BP1 is a key domain for RNA binding. It possibly binds to the GRE sequence cooperatively with RRM2 of CUG-BP1. RRM1 of CUG-BP1 is not required for GRE recognition, but may be important for maintaining the stability of the full-length CUG-BP1.

The Lights are on but Nobody's Home: the Future of Academic Libraries?

This viewpoint article looks at the challenges facing academic libraries and argues that collaboration and engagement are vital to their survival. Cet article d’opinion examine des défis auxquels des bibliothèques académiques font face, et soutient que la collaboration et l’engagement sont indispensables pour leur survie.

A Grand Tour: A Catalogue of Eighteenth-Century Print Works at Memorial University Libraries

This catalogue highlights forty-seven of the 1,180 eighteenth-century imprints held by Memorial University Libraries. Intended as a general introduction to eighteenth-century literature in its original formats, the work is aimed at students and teachers of book history and bibliography, as well as at the general reader. Consequently, the focus is broad, highlighting the emerging free press, imaginative literature—particularly the novel—travel literature, street literature, illustration, as well as works of religion, philosophy, science, and medicine. The introduction discusses each of the works presented in the catalogue and makes a case for the collection as a whole as representing a range of developments both in eighteenth-century literature and in the book trade. Catalogue entries highlight the physical artifact, offering both description and photographic evidence. Each entry contains information about the author and the content of the work, and attempts to place the work in its literary context.

More : the newsletter for South Carolina libraries 

This June 2016 newsletter from the South Carolina State Library, volume 48, issue 6, features news and updates for South Carolina libraries.

INFORMATION, LIBRARIES AND DEVELOPMENT Address at the Inauguration of-Manifests Central American Seminar on IFLA / Unesco Public Libraries, School Libraries and the Internet ‖. 26 to 29 March 2007

It is a matter of pride that Costa Rica and the National University, both with extensive experience in the development of library we are the headquarters for the first time, an event that brings together three different activities on this theme: the Central American Seminar, Workshops and Standing Committee of the International Federation of Library Associations and Institutions, which will strengthen the monitoring and Manifestos on Public Libraries, Schools and Internet IFLA-UNESCO. Thank you very much to all of you for allowing us this honor and a double thanks to the comrades of the UNA university who took the initiative and made this opportunity possible.

Libraries and open access: the implications of open-access publishing and dissemination for libraries in higher education institutions

This chapter discusses the consequences of open-access (OA) publishing and dissemination for libraries in higher education institutions (HEIs). Key questions (which are addressed in this chapter) include: 1. How might OA help information provision? 2. What changes to library services will arise from OA developments (particularly if OA becomes widespread)? 3. How do these changes fit in with wider changes affecting the future role of libraries? 4. How can libraries and librarians help to address key practical issues associated with the implementation of OA (particularly transition issues)? This chapter will look at OA from the perspective of HE libraries and will make four key points: 1. Open access has the potential to bring benefits to the research community in particular and society in general by improving information provision. 2. If there is widespread open access to research content, there will be less need for library-based activity at the institution level, and more need for information management activity at the supra-institutional or national level. 3. Institutional libraries will, however, continue to have an important role to play in areas such as managing purchased or licensed content, curating institutional digital assets, and providing support in the use of content for teaching and research. 4. Libraries are well-placed to work with stakeholders within their institutions and beyond to help resolve current challenges associated with the implementation of OA policies and practices.

More : the newsletter for South Carolina libraries 

This July 2016 newsletter from the South Carolina State Library, volume 48, issue 7, features news and updates for South Carolina libraries.