President’s Prize 2022
President’s Prize 2022 – via Zoom Webinar, Friday, 29th April, 11:00 – 12.30pm.
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The Academy continues with it’s award scheme for graduates in the degree courses in Biomedical Science from TU Dublin, GMIT and MTU/UCC. Each year the colleges are requested to nominate their two students who have excelled in the research project component of their final exams. This year, the nominated students will present their research findings live via Zoom webinar. The President’s Prize is awarded to the student who presents a project judged to be the winner by a panel of selected members and other professionals. The winner goes forward to compete for the Martin Nicholson Award at the EPBS (European Association for Professions in Biomedical Sciences) annual conference.
Booking and Registration:
Format:
11:00 Presidents address – Bernadette Jackson
11:05 Contestants will have 10 minutes to present and 5 minutes for Q&A.
List order as below, alphabetically by surname.
Judges will return after the presentations
A study of Methicillin Resistant Staphylococcus aureus Isolates from an Irish Hospital: assessing the geographical spread and clonality of spa types
Nicola Blake,
Supervisors: Dolores Crowley, Julie Twomey, Dr Helen O’Shea, Munster Technological University
Abstract:
Introduction: Staphylococcus aureus can progress from commensal status to cause life-threatening illness. This is dictated by its arsenal of virulence factors, and potentiated by acquisition of antibiotic resistance traits. Methicillin-resistant S. aureus (MRSA) has therefore been established as a persistent and formidable pathogen within our hospital and community settings. Rapid and precise detection of these organisms allows earlier intervention, which reduces admission length, mortality and healthcare costs. This requires the knowledge of trends emerging in the national and international context which may carry the risk of further antibiotic resistance or virulence factor acquisition. Therefore surveillance of isolates from our hospitals serves as a valuable tool for future planning of testing, treatment and prevention of MRSA. This study seeks to conduct surveillance of 241 MRSA isolates from Cork University Hospital, Ireland (2016) within both national and international contexts. These isolates had been typed using agr, SCCmec and spa typing, as well as restriction modification testing. This allows the lineage and associated methicillin and fusidic acid resistance patterns of these isolates to be investigated in this study. Due to the hypervariability of the spa typing region, a large number of spa types is expected. This project sought to show the geographical spread of particular spa types, and to investigate whether alignment of spa type short sequence repeats could allow related strains to be grouped together, easing interpretation.
Materials and Methods: Data manipulation was conducted using R. Zeemaps was used to map spa types geographically. Clonality of spa types was investigated using alignment of constituent Short Sequence Repeats. This was then cross-referenced with the lineage determined by restriction modification testing, and in the literature.
Results: All isolates were community-acquired, with the ST22-MRSA-IVh clone predominating, at 92%. ST22-MRSA-IVh was associated with spa t032, t022 and t2945. Mapping showed spa types were previously isolated in Western Europe. Short Simple Repeat alignment showed 38 of 54 spa types showed similarity to t032. There was concordance between sequence type and Groupings I, IV, VI, while II, III, V and VII required repeating of restriction modification testing. It was also shown that 95.1% of fusidic acid resistant isolates had the L461K fusA mutation, which is vertically transmitted.
Conclusion: The MRSA isolates of CUH were highly clonal, dominated by the endemic clone. Mapping of spa types showed localised clustering to common travel areas. SSR alignment of spa types, the most diverse parameter, showed the majority were related to EMRSA15-associated t032. The Short Simple Repeat alignment system was largely reliable at correctly assigning spa types of the same sequence type.
Biography:
Nicola Blake graduated in Biomedical Science
from Munster Technological University/University
College Cork and is currently undertaking her
clinical laboratory placement in
University Hospital Waterford.
Validation of the Biomic V3 Automated Zone Reader for the Comparison of Direct Susceptibility Testing to a Standardised Method for Antibiotic Susceptibility Testing on Urine Samples
Zara Brady, BSc. (Hons) in Medical Science, Galway-Mayo Institute of Technology
Supervisors: Niamh Carpenter & Larry O’Neill, Microbiology Department, Cavan General Hospital,
Dr Debbie Corcoran, Lecturer, Galway-Mayo Institute of Technology.
Abstract
Introduction: Direct Susceptibility Testing (DST) is a method whereby susceptibility testing is performed directly on the patient’s sample using disk diffusion. DST is extremely advantageous as the susceptibility profiles are obtained 24-hours earlier than standardised antibiotic susceptibility testing (AST), enabling a rapid turnaround time. This aids in the prompt administration of tailored antibiotic therapy which helps to prevent the emergence of antibiotic resistant strains. This is especially important for critically ill patients.
This study aimed to validate the Biomic V3 automated disk diffusion reader for use on midstream urine (MSU) samples where Escherichia coli (E. coli) has been identified as the primary pathogen. The rationale for this study was to determine if performing DST on E. coli MSU samples agrees with the standardised method for AST in Cavan General Hospital, which is the Becton Dickinson (BD) Phoenix 100 analyser, based on EUCAST guidelines.
Methods: Only MSU samples where greater than 105 organisms/mL of E. coli were isolated in pure culture on chromogenic agar were included in the study. Out of the 295 samples that were set up for DST, only 111 samples could be utilised for the study.
Results: When utilised on samples where E. coli was identified as the primary pathogen, the overall agreement obtained between standardised AST on the Phoenix analyser and DST using the Biomic V3 was 97.9%. A total of 16 (1.3%) very major errors, 2 (0.2%) major errors, and 8 (0.7%) minor errors were seen. All antibiotics except Amoxicillin-clavulanate achieved an agreement of over 95%. A Cohen’s kappa of 0.902 was achieved overall when comparing both methodologies.
Conclusion: DST can be confidently used as a method of AST for MSU samples where E. coli has been identified as the primary pathogen. However, Amoxicillin-clavulanate should not be reported on patient samples when using DST. Due to the high number of non-interpretable samples set up for DST, the methodology could be reserved for patients who would benefit most from the decreased turnaround time.
Biography
Zara graduated from Galway-Mayo Institute of Technology (GMIT)
in Spring 2021. She is currently working as a medical scientist in the
Biochemistry department in Cavan General Hospital, where she completed
her clinical laboratory placement. She is currently studying for her
MSc. in medical science at GMIT.
The incorporation of point-of-care analyser ROTEM®, to enhance the treatment of major haemorrhage.
Ashley Hendy, National Maternity Hospital in the Blood Transfusion Laboratory
Abstract
The incorporation of point-of-care analysers, like ROTEM®, to enhance the treatment of major haemorrhage is being recognised internationally. ROTEM® analyses whole blood samples to allow rapid analysis of coagulation parameters.
In 2018, ROTEM® was introduced to enhance the treatment of post-partum haemorrhage in the National Maternity Hospital (NMH). However, the implementation of ROTEM® in this setting required evaluation. The British Society of Haematology provide guidelines for the use of ROTEM® in treatment of major haemorrhage. The use of ROTEM® in NMH was evaluated in comparison to these guidelines and a quality improvement initiative was designed to address the downfalls identified.
This study revealed under-utilisation of ROTEM® analysis in NMH. In 2019-2020, 79.6% of patients eligible for ROTEM® analysis were not tested on this platform. When incorporating ROTEM® analysis in NMH, the massive haemorrhage pathway must be activated, as per the locally verified ROTEM® algorithm. This study identified the massive haemorrhage pathway is potentially under-activated in this setting. In 2019-2020, 60.9% of patients analysed on ROTEM® in NMH satisfied the specified blood loss trigger for ROTEM® analysis, yet only 14.7% were treated alongside an active massive haemorrhage pathway. Review of ROTEM® records found no documented training records of staff in ROTEM® analysis and significant issues with identifying IQC failures.
A survey of ROTEM®-users revealed training was not standardised for ROTEM® in NMH. To address these findings, a ROTEM®-User Training Programme was drafted and participation in an accredited external quality assurance scheme was initiated.
Biography: Ashley, graduated with First-Class Honours
in Biomedical Science from Technological University Dublin,
2021. In final year she majored in Biochemistry, Transfusion
and Transplant Immunology. She is currently working in the
National Maternity Hospital in the Blood Transfusion Laboratory.
To determine a novel role for G protein-coupled receptors in Pancreatic ductal adenocarcinoma pathogenesis.
Fiachra Nolan, Medical Scientist in the Galway Clinic
Supervisors: Dr Brian Moran, Karen Finn, Ryan McGowan and Áine Sally
Abstract
Pancreatic ductal adenocarcinoma (PDAC) accounts for 90% of all pancreatic carcinomas and is the 11th most common cancer in Ireland. PDAC has a mortality of 10.8 deaths per 100,000 population. The aim of this investigation is to determine a novel role for G protein-coupled receptors in PDAC pathogenesis. Alongside this aim the role of GPCRs may be used to enhance patient treatment through their modulation, increasing cancerous cell death while retaining living non-cancerous cells. This aims to reduce mortality and extend the patient life expectancy.
G protein- coupled receptors (GPCRs) are seven-transmembrane receptors and control downstream signalling which may promote or inhibit ion channels or enzyme production. GPCRs may have varied expression between normal human pancreatic ductal epithelium cell lines (HPDE) and PANC-1 cells which are a cancerous cell line obtained from a 56-year-old Caucasian male PDAC patient. Both cell lines are cultured in vitro to allow investigation.
TaqMan qPCR probes were used to determine the relative expression of specific G protein- coupled receptors known as GPCR-X and GPCR-Y in PANC-1 and HPDE cells. These receptors have been renamed due to further research being undertaken. GPCR-X shows 7.2-fold upregulation in PANC-1 cells compared to normal cells (p<0.001), while GPCR-Y has similar expression in normal and PANC-1 cells. Indicating that GPCR-X may play a role in the pathogenesis of PDAC while the involvement of GPCR-Y is unlikely.
MTT and NR cell viability assays using PANC-1 cells cultured in vitro, assessed the effect of agonising and antagonising GPCR-X in combination and comparison to gemcitabine. The conventional pancreatic cancer treatment gemcitabine, appears ineffective treating PDAC, producing a maximal 22% cytotoxicity, as expected due to the highly resistant nature of PDAC. Agonising GPCR-X with specific agonists creates enhanced in vitro cell growth in comparison to untreated cells. Antagonising GPCR-X with specific antagonists alone creates up to 43% cytotoxicity (p<0.0001). A combination of antagonising GPCR-X and treatment with gemcitabine creates a maximal 69% cytotoxicity (p<0.0001). This indicates that antagonising GPCR-X has greater therapeutic potential than gemcitabine alone and is enhanced in combination with gemcitabine, as supported by statistical p-values in comparison with gemcitabine alone. This creates potential for use of GPCR-X as a novel therapeutic target for PDAC.
Biography:
Fiachra graduated from GMIT in 2021
and is currently working as a
Medical Scientist in the Galway Clinic.
Does the Level of Expression of Cellular mRNA Encoding gC1qR Change with Alphavirus Infection?
Madeleine Quinn, clinical placement in University Hospital Limerick
Supervisor: Dr. Martina Scallan, Microbiology Department in UCC.
Abstract:
gC1qR is a well-studied pathogen recognition receptor (PRR) which interacts with many viruses. Viruses such as Human Immunodeficiency Virus (HIV), Hepatitis C Virus (HCV), Epstein Barr Virus (EBV), Rubella Virus (RV), and Herpes Simplex Virus (HSV) are known to interact with cellular gC1qR in a manner which favours their own viral replication. Semliki Forest Virus (SFV) capsid protein has been reported to co-immunoprecipitate with gC1qR leading to speculation that SFV may also manipulate gC1qR to favour its own viral replication. In order to evaluate the relationship between gC1qR and SFV, confluent and sub- confluent baby hamster kidney (BHK) cell monolayers were infected with either the L10 or SFV A7(74) strain of SFV, with parallel mock infected controls. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) analysis was carried out and the cycle threshold (C#) values obtained were used to calculate the level of expression of the gene of interest using ∆∆C#. In confluent BHK monolayers infection increased the expression of gC1qR while expression decreased upon infection of sub-confluent monolayers. The expression of gC1qR in uninfected BHK cells was highest in cells that were most confluent and lowest in the least confluent monolayers. The affect confluency has on the accumulation of SFV9 RNA was found to differ between virus strains. 𝛽-Actin and GAPDH reference genes displayed divergent expression patterns when compared (as reference genes) across different confluences of uninfected cells and between uninfected and virus infected cells.
Biography:
Madeleine graduated in Biomedical Science from
UCC/MTU. and is currently on clinical placement
in University Hospital Limerick (UHL-2022).
Molecular and phenotypic investigations to assess risk to human health associated with a collection of Pseudomonas aeruginosa of clinical origin.
Fionn Sheridan, Medical Scientist, Microbiology laboratory, St. Vincent’s University Hospital.
Abstract:
Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that is increasingly implicated in outbreaks of a variety of healthcare-associated infections with high mortality. P. aeruginosa demonstrates high antimicrobial resistance, with the ability to form biofilm that further enhances antimicrobial resistance, establishes chronic infection and allows persistence in the healthcare environment.
This study aimed to assess the risk of clinically isolated P. aeruginosa to human health through phenotypic and genotypic characterisation methods. A total of 52 clinical P. aeruginosa isolated from various anatomical sites were investigated for antimicrobial susceptibility, biofilm formation, biofilm metabolism and Congo red morphotype. Whole genome sequencing was then applied to perform in-silico virulence and antimicrobial resistance genotyping, MLST, SNP analysis and pan-genome alignment. High level resistance was seen against all beta-lactams, ciprofloxacin and gentamicin, with high susceptibility to amikacin and colistin. All clinical isolates were biofilm formers, while most were strong biofilm formers and metabolically inactive. Highly resistant and virulent genotypes were identified in our collection. Congo red morphotypes were diverse and could not be correlated to a virulence genotype. MLST showed the presence of international high-risk clones, while SNP analysis and pan-genome alignment showed multiple closely-related isolates. AST and biofilm studies highlight the need for new antimicrobial agents and anti-biofilm therapies to combat these infections.
Furthermore, the discovery of multiple international high-risk clones, and associated horizontally transferred AMR genes, highlights the importance of surveillance programmes that monitor their dissemination. Moreover, the application of WGS in identifying potentially persistent strains in the healthcare environment can inform infection control teams to identify and eradicate these strains.
Biography:
Fionn graduated with a First Class Honours in
Biomedical Science, from the
Technological University Dublin.
He is currently work as a Medical Scientist
in the Microbiology laboratory of
St. Vincent’s University Hospital.