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Search Results: 1 - 10 of 5469 matches for " TIMOTHY MAHONY "
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Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates
Antonino S Cavallaro, Donna Mahony, Margaret Commins, Timothy J Mahony, Neena Mitter
Microbial Cell Factories , 2011, DOI: 10.1186/1475-2859-10-57
Abstract: The expression of a truncated form of BVDV-E2 protein (E2-T1) in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0) containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies.We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit vaccine. The optimised E. coli expression system for E2-T1 combined with methodologies for solubilisation, refolding and integrated endotoxin removal presented in this study should prove useful for other vaccine applications.Bovine viral diarrhoea virus (BVDV) infection of cattle is linked to economically important diseases with losses in the USA being estimated to US$10-40 million per million calves [1] and US$6 million per million calves in the UK [2]. BVDV is a member of the Pestivirus genus within the Flavivirus family. The BVDV genome is a positive sense RNA molecule with one open reading frame (ORF) encoding for a polyprotein which is cleaved into the structural and
Back to BAC: The Use of Infectious Clone Technologies for Viral Mutagenesis
Robyn N. Hall,Joanne Meers,Elizabeth Fowler,Timothy Mahony
Viruses , 2012, DOI: 10.3390/v4020211
Abstract: Bacterial artificial chromosome (BAC) vectors were first developed to facilitate the propagation and manipulation of large DNA fragments in molecular biology studies for uses such as genome sequencing projects and genetic disease models. To facilitate these studies, methodologies have been developed to introduce specific mutations that can be directly applied to the mutagenesis of infectious clones (icBAC) using BAC technologies. This has resulted in rapid identification of gene function and expression at unprecedented rates. Here we review the major developments in BAC mutagenesis in vitro. This review summarises the technologies used to construct and introduce mutations into herpesvirus icBAC. It also explores developing technologies likely to provide the next leap in understanding these important viruses.
Repertoire of Bovine miRNA and miRNA-Like Small Regulatory RNAs Expressed upon Viral Infection
Evgeny A. Glazov, Kritaya Kongsuwan, Wanchai Assavalapsakul, Paul F. Horwood, Neena Mitter, Timothy J. Mahony
PLOS ONE , 2009, DOI: 10.1371/journal.pone.0006349
Abstract: MicroRNA (miRNA) and other types of small regulatory RNAs play a crucial role in the regulation of gene expression in eukaryotes. Several distinct classes of small regulatory RNAs have been discovered in recent years. To extend the repertoire of small RNAs characterized in mammals and to examine relationship between host miRNA expression and viral infection we used Illumina's ultrahigh throughput sequencing approach. We sequenced three small RNA libraries prepared from cell line derived from the adult bovine kidney under normal conditions and upon infection of the cell line with Bovine herpesvirus 1. We used a bioinformatics approach to distinguish authentic mature miRNA sequences from other classes of small RNAs and short RNA fragments represented in the sequencing data. Using this approach we detected 219 out of 356 known bovine miRNAs and 115 respective miRNA* sequences. In addition we identified five new bovine orthologs of known mammalian miRNAs and discovered 268 new cow miRNAs many of which are not identifiable in other mammalian genomes and thus might be specific to the ruminant lineage. In addition we found seven new bovine mirtron candidates. We also discovered 10 small nucleolar RNA (snoRNA) loci that give rise to small RNA with possible miRNA-like function. Results presented in this study extend our knowledge of the biology and evolution of small regulatory RNAs in mammals and illuminate mechanisms of small RNA biogenesis and function. New miRNA sequences and the original sequencing data have been submitted to miRNA repository (miRBase) and NCBI GEO archive respectively. We envisage that these resources will facilitate functional annotation of the bovine genome and promote further functional and comparative genomics studies of small regulatory RNA in mammals.
Expression of Two N1 Clones with Single Amino Acid Dissimilarity of Avian Influenza H5N1 Virus
HAYATI Journal of Biosciences , 2012,
Abstract: Two clones of N1 gene derived from isolate A/Dk/Tangerang/Bbalitvet-ACIAR-TE11/2007 (H5N1) exhibit single mismatch of amino acid sequence at position 242 that is threonine and methionine for the clone #3 and #5, respectively. In order to evaluate the effect of the amino acid substitution, these clones were inserted into two different expression vectors that are pEGFP-C1 and pcDNA-3.3 TOPO TA cloning. Subsequently, the respective recombinant clones were transfected into eukaryotic cells, including CEF, RK13 and VERO using Lipofectamine ‘plus’ reagent. As a result, the clone #3 retaining atypical sequence showed lower expression level rather than the clone #15 in both vectors and all type of cells. The 3D conformational modelling revealed that the mutation occurs in the inner part of glycoprotein embedded within envelope or matrix. Therefore, the missense mutation seems has no effect on the antigenic properties of neuraminidase but this substitution by any means causes lethal mutagenesis in the individual gene expression by reducing level of protein transcript.
A Successful Pregnancy Outcome after Surgical Decompression of Type I Arnold-Chiari Malformation  [PDF]
Patricia Ip, Susmita Pankaja, Fidelma O’Mahony
Open Journal of Obstetrics and Gynecology (OJOG) , 2015, DOI: 10.4236/ojog.2015.51007
Abstract: Type I Arnold-Chiari malformation (ACM) usually presents in adulthood and consists of a downward displacement of the cerebellar tonsils through the foramen magnum. A 25-year-old woman presented with a 5-month history of headache associated with blurred vision, tinnitus and sickness. Imaging recognised the need for surgical intervention, but whilst awaiting for surgery she fell pregnant. Considering the risks of neurological deterioration, the woman underwent surgical decompression of type I ACM at 15 weeks gestation. She subsequently presented with progressively worsening headaches during late pregnancy from 35 weeks. The obstetric plan was initially induction of labour at term but since the onset of worsening symptoms, this date was brought forward to 39 + 1 weeks gestation. She proceeded to have a normal delivery with no neonatal complications and an uneventful puerperium followed. Since the delivery, the patient reported fewer symptoms, showed no signs of neurological deficit and a repeat magnetic resonance imaging of the head showed good relief of neural compression. This case illustrates how judicious selection of the appropriate mode of delivery of women following surgically corrected ACM and a multidisciplinary approach is critical in the successful management of the antepartum period and labour.
Toll-Like Receptor mRNA Expression Is Selectively Increased in the Colonic Mucosa of Two Animal Models Relevant to Irritable Bowel Syndrome
Declan P. McKernan,Aoife Nolan,Elizabeth K. Brint,Siobhain M. O'Mahony,Niall P. Hyland,John F. Cryan,Timothy G. Dinan
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0008226
Abstract: Irritable bowel syndrome (IBS) is largely viewed as a stress-related disorder caused by aberrant brain-gut–immune communication and altered gastrointestinal (GI) homeostasis. Accumulating evidence demonstrates that stress modulates innate immune responses; however, very little is known on the immunological effects of stress on the GI tract. Toll-like receptors (TLRs) are critical pattern recognition molecules of the innate immune system. Activation of TLRs by bacterial and viral molecules leads to activation of NF-kB and an increase in inflammatory cytokine expression. It was our hypothesis that innate immune receptor expression may be changed in the gastrointestinal tract of animals with stress-induced IBS-like symptoms.
Infection control in general practices in Buffalo City and OR Tambo District Municipalities, South Africa
Don O'Mahony
African Journal of Primary Health Care & Family Medicine , 2012, DOI: 10.4102/phcfm.v4i1.268
Abstract: Background: Good infection control practices are effective in reducing rates of infection in health care settings. Studies in primary care in developed countries indicate that many general practitioners (GPs) do not comply with optimal infection control practices. There are no published studies from developing countries in Southern Africa. Objectives: The aim of this study was to describe infection control practices in private GP surgeries in the Buffalo City and OR Tambo District Municipalities in the Eastern Cape Province, South Africa. Method: A literature review was conducted to appraise current best practice with respect to Standard Infection Control and Transmission Based Precautions. A questionnaire, inquiring into GPs’ actual practices, was posted to each surgery. Results: The valid response rate was 34% (47/140). Methods used to sterilise instruments in 40 practices were: ultraviolet sterilisation (23), chemical disinfection (14), boiling water (7), and steam autoclave (2). Compounds used for chemical disinfection included organotin quaternary, chlorhexidine and benzyl ammonium chloride with a quaternary complex. Twenty-two (47%) used a hand rub. Sixteen (35%) GPs stated that they had a policy to promptly triage patients who are coughing, and 23 (50%) had a policy for airflow movement in the surgery. All practices appropriately disposed of sharps. Thirty-seven (80%) expressed interest in a seminar on infection control. Conclusions: Overall, GPs were aware of infection control precautions. Ultraviolet sterilisers and chlorhexidine are not recommended, however, for sterilisation or high level disinfection of medical instruments, and their use should be discontinued. Hand rubs are underutilised. GPs should implement Transmission Based Precautions to prevent airborne and droplet infections. Contr le des infections dans les cabinets de médecine générale dans les municipalités de Buffalo City et du district d’OR Tambo, en Afrique du Sud Contexte: De bonnes pratiques de contr le des infections sont efficaces pour réduire les taux d’infection dans les lieux administrant des soins de santé. Des études sur les soins primaires dans les pays développés indiquent que de nombreux médecins généralistes ne respectent pas les pratiques de contr le des infections optimales. Aucune étude émanant des pays en voie de développement en Afrique australe n’a jamais été publiée. Objectifs: L’objectif de cette étude consistait à décrire les pratiques de contr le des infections dans les cabinets de consultation de généralistes privés dans les municipalités de Buffalo City
Microneedle Array Design Determines the Induction of Protective Memory CD8+ T Cell Responses Induced by a Recombinant Live Malaria Vaccine in Mice
John B. Carey,Frances E. Pearson,Anto Vrdoljak,Marie G. McGrath,Abina M. Crean,Patrick T. Walsh,Timothy Doody,Conor O'Mahony,Adrian V. S. Hill,Anne C. Moore
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0022442
Abstract: Vaccine delivery into the skin has received renewed interest due to ease of access to the immune system and microvasculature, however the stratum corneum (SC), must be breached for successful vaccination. This has been achieved by removing the SC by abrasion or scarification or by delivering the vaccine intradermally (ID) with traditional needle-and-syringes or with long microneedle devices. Microneedle patch-based transdermal vaccine studies have predominantly focused on antibody induction by inactivated or subunit vaccines. Here, our principal aim is to determine if the design of a microneedle patch affects the CD8+ T cell responses to a malaria antigen induced by a live vaccine.
Health equity in the New Zealand health care system: a national survey
Nicolette F Sheridan, Timothy W Kenealy, Martin J Connolly, Faith Mahony, P Alan Barber, Mary Anne Boyd, Peter Carswell, Janet Clinton, Gerard Devlin, Robert Doughty, Lorna Dyall, Ngaire Kerse, John Kolbe, Ross Lawrenson, Allan Moffitt
International Journal for Equity in Health , 2011, DOI: 10.1186/1475-9276-10-45
Abstract: A national survey of district health boards (DHBs) was undertaken on macro approaches to chronic condition management with detail on cardiovascular disease, chronic obstructive pulmonary disease, congestive heart failure, stroke and diabetes. Additional data from expert informant interviews on program reach and the cultural needs of Māori and Pacific peoples was sought. Survey data were analyzed on dimensions of health equity relevant to strategic planning and program delivery. Results are presented as descriptive statistics and free text. Interviews were transcribed and NVivo 8 software supported a general inductive approach to identify common themes.Survey responses were received from the majority of DHBs (15/21), some PHOs (21/84) and 31 expert informants. Measuring, monitoring and targeting equity is not systematically undertaken. The Health Equity Assessment Tool is used in strategic planning but not in decisions about implementing or monitoring disease programs. Variable implementation of evidence-based practices in disease management and multiple funding streams made program implementation difficult. Equity for Māori is embedded in policy, this is not so for other ethnic groups or by geography. Populations that conventional practitioners find hard to reach, despite recognized needs, are often underserved. Nurses and community health workers carried a disproportionate burden of care. Cultural and diversity training is not a condition of employment.There is a struggle to put equity principles into practice, indicating will without enactment. Equity is not addressed systematically below strategic levels and equity does not shape funding decisions, program development, implementation and monitoring. Equity is not incentivized although examples of exceptional practice, driven by individuals, are evident across New Zealand.Inequalities preventable by reasonable means are unfair, and in health are indicators of distributional differences in the health status of the
Structural Aspects of the Interaction of Dairy Phages with Their Host Bacteria
Jennifer Mahony,Douwe van Sinderen
Viruses , 2012, DOI: 10.3390/v4091410
Abstract: Knowledge of phage-host interactions at a fundamental level is central to the design of rational strategies for the development of phage-resistant strains that may be applied in industrial settings. Phages infecting lactic acid bacteria, in particular Lactococcus lactis and Streptococcus thermophilus, negatively impact on dairy fermentation processes with serious economic implications. In recent years a wealth of information on structural protein assembly and topology has become available relating to phages infecting Escherichia coli, Bacillus subtilis and Lactococcus lactis, which act as models for structural analyses of dairy phages. In this review, we explore the role of model tailed phages, such as T4 and SPP1, in advancing our knowledge regarding interactions between dairy phages and their hosts. Furthermore, the potential of currently investigated dairy phages to in turn serve as model systems for this particular group of phages is discussed.
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