2013 Projects (1-30)

1. Vascular tissue engineering: endothelial cell-mediated effects on extracellular matrix synthesis by arterial smooth muscle cells in culture

Brophy MJ, Flanagan TC1

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

The mechanical strength of tissue-engineered vascular grafts has been promoted by methods that enhance extracellular matrix (ECM) synthesis by seeded cells, including bioreactor conditioning/growth factor supplementation. Endothelial cells are an important component of such grafts due to their anti-thrombogenic properties, but there are conflicting data in the literature concerning their effect on ECM synthesis by arterial smooth muscle cells (SMCs). The present study aims to determine whether or not an overlying layer of endothelial cells, and/or their secretions, enhances or inhibits ECM production by co-cultured SMCs, which may determine an optimal time-point for graft endothelialisation.

For 2-D analysis, commercially-available human umbilical artery SMCs (HUASMCs) were cultured either alone, in direct contact within human umbilical artery endothelial cells (HUAECs), or exposed to secretions from HUAECs using co-culture well inserts. For preliminary 3-D analysis, HUASMCs were embedded within fibrin gels and were cultured either alone, with a surface monolayer of HUAECs, or in HUAEC-conditioned medium. All cultures were maintained for 12 days and ECM development (type-I collagen, elastin, fibronectin) was analysed using immunofluorescence microscopy. In addition, collagen and elastin production was quantified using spectrophotometric assay.

In 2-D culture, it was observed that HUASMCs and HUAECs in direct cell contact produced an increased amount of elastin (~7.6%) when compared to HUASMCs cultured alone, while collagen production was significantly reduced (~29%). These results were corroborated by immunofluorescence microscopy. Preliminary results from 3-D fibrin gel culture showed a 26.8% increase in elastin production in the group containing HUASMCs and a surface monolayer of HUAECs compared with HUASMCs cultured alone, while collagen production in 3-D gel constructs was also significantly reduced in this group (~39%), mirroring the 2-D results. Strikingly, a clear potent relationship between both cell types was observed, with significant contraction of gels in this group alone to a fraction of their size after 3 days in culture.

The results of the present study indicate that seeding of ECs onto fibrin-based vascular grafts should be completed toward the end of the in vitro conditioning period to allow for adequate deposition of collagen and elastin proteins by seeded SMCs and to prevent premature tissue contraction. 

Acknowledgement:

This project was funded by the National Children’s Research Centre. MJB also acknowledges funding through a HRB Summer Student Scholarship.

  • Presenting Author: Ms Megan Brophy
  • Supervisor: Dr Thomas Flanagan
  • Co-Supervisor: Dr Nobue Itasaki

2. A survey of compliance with the peripheral vascular cannula (pvc) care bundle as implemented by SVUH in order to reduce the incidence of blood stream infections by ensuring appropriate PVC care.

Anderson T,  FitzGerald S1, Schaffer K1, Flynn A.

1St. Vincent’s University Hospital, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Approximately one third of SVUH in-patients have a peripheral venous cannula (PVC) in-situ. Appropriate care of PVCs is vital to reduce the risk of local and/or bloodstream infection. The use of PVC care bundles can reduce the risk of infection. Current hospital guidelines recommend that PVCs that have been in situ for more than 72 hours may remain in place if still in use; however, PVCs should not remain in situ for more than 96 hours.

A single day point prevalence audit showed that PVC bundle forms were in use in 9 of 19 wards in SVUH. A subsequent prospective study of all patients with PVCs was carried out in these 9 wards over a four week period. Information was gathered during daily visits to all nine wards during which the PVC care bundle forms were reviewed and each  PVC was observed using the Visual Infusion Phlebitis (VIP) score.

A total of 222 PVCs were observed in 126 patients. For two thirds of PVCs (146/222; 66%), a PVC care bundle form was in use on the first day of review. Not all forms were completed in full each day. The date of insertion of PVCs was documented in 73% of cases (163/222). Approximately one fifth of PVCs (48/222; 22%) were removed prior to patient discharge, and patient notes were not available to review documentation of the date of PVC removal. Of the remainder, the date of removal was documented in two thirds (112/174; 64%).

Almost half of all PVCs observed (90/222; 41%) were removed within 72 hours; 41% remained in place for 72-96 hours and one fifth (39/222; 18%) were in for longer than 96 hours. Most of the PVCs in situ for more than 72 hours (122/129; 95%) were still in use.

Phlebitis/infection occurred in 10% of PVCs (22/222). There were no bloodstream infections related to PVC during the study period.  Most cases of phlebitis/infection occurred within 72 hours (18/22; 81%), and none occurred after 96 hours.

  • Presenting Author: Ms Toni Anderson
  • Supervisor: Dr Susan FitzGerald
  • Co-Supervisor: Dr Kirsten Schaffer

3. Uncovering the “sumo” in HIV-1 latency

Ho, MZG1, McCartin A2, Kulkarni A2, Gautier VM2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2UCD Centre for Research in Infectious Diseases, University College Dublin, Belfield, Dublin 4.

HIV-1 remains a threat with approximately 34-million people infected. Antiretroviral therapy, while efficacious in targeting replicating viruses, is innocuous against latent viral reservoirs, constituting a major obstacle towards a cure. Epigenetic mechanisms mediate HIV-1 repression. In this context, we recently identified SUMO-2/3 (Small Ubiquitin-like Modifier) post-translational modifications at the silenced HIV-1 promoter, suggesting that SUMO contributes to HIV-1 latency. Here, we characterize desumoylating enzyme Sentrin Specific Peptidase 1 (SENP1) as a novel modulator of HIV-1 silencing/transcription.

To determine SENP1 effects on HIV-1 latency, HeLa-LTR cells, containing the repressed HIV-1 promoter upstream of luciferase reporter gene, were transfected with SENP1 +/- Tat (HIV-1 trans-activator which drastically enhances HIV-1 gene transcription). Transfected cells were halted in G0 and G1 phases and analyzed using luciferase assay, protein assay, and Western Blot.

SENP1 overexpression reactivated the HIV-1 promoter by 10-fold. Interestingly, this effect was suppressed in G0 cells treated with SAHA, a Histone Deacetylase (HDAC) inhibitor, suggesting that sumoylated HDAC (known epigenetic silencer of HIV-1 promoter) is one potential target of SENP1.

Remarkably, HIV-1 Tat and SENP1 synergistically reactivated the promoter by 854-fold vis-à-vis a 153-fold reactivation by Tat. Western Blot analysis revealed that Tat seemingly increased SENP-1 expression, potentially explaining the synergism observed. Similarly, SAHA treatment partially diminished SENP1 effect in Tat co-transfected G0 cells.

This study highlights SUMO as a potential molecular switch controlling HIV-1 latency attesting to its value in translational medicine. Further research is needed to elucidate the mechanisms involved in SENP1-mediated HIV-1 reactivation.

Acknowledgment:

This work was supported by the Wellcome Trust vacation scholarship 

  • Presenting Author: Mr Matthew Ho-Zhi-Guang
  • Supervisor: Dr Virginie Gautier
  • Co-Supervisor: Dr Anurag Kulkarni

4. Paediatric presentation to primary care

O’Brien K1,Breen N2, Mac Guinness J3

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2, 3 Lecturers in General Practice, UCD School of Medicine and Medical Sciences, University College Dublin and Greystones Harbour family practice.

GPs are typically the first and most common point of contact for medical services for children. Patients often present early with poorly differentiated and nonspecific symptoms4. Most guidance on acute presentations originates from studies based in secondary care5.

This study documents presentation, process and outcome of children’s presentations to a general practice in a calendar year.  Data on all paediatric presentations in 2012 (total 3,088 presentations) to a GP training practice in Wicklow, Ireland was analysed with SPSS, following coding.  An open access urgent kid’s clinic (UKC) is run at the beginning of each week-day.

Ninety two per cent of illness presentations are dealt with exclusively in primary care. Referrals to OPD occurred in 5.2%, to A&E in 3.1%, with less than 0.1% admitted.  Twenty-five per cent of presentations were to the UKC; less than 1% to the out-of-hours deputizing service. Cough is the presenting complaint in 24.8% of cases; fever in 7.2%. Forty per cent of all diagnoses are respiratory/ENT.  Antibiotics are prescribed for approximately 30% of all presentations, although less than 10% of URTI diagnoses received an initial script for antibiotics. Consultation rates of GMS/non-GMS patients were similar.

Most paediatric illness is treated in the community and serious illness is rare. Ways to support doctors in dealing with less differentiated, non-specific presentations need to be addressed. The evidence-base for guidelines on clinical approach needs to be grounded in the health-care environment in which the guidelines will be used.  These data suggest that, in general practice, a focus on symptom interpretation is essential.

References:

1. Van den Bruel A, et al. Signs and symptoms for diagnosis of serious infections in children: a prospective study in primary care.Br J Gen Pract. 2007 July 1; 57(540): 538–546

2. Craig JC, Williams GJ, Jones M, Codarini M, Macaskill P, Hayen A, et al. The accuracy of clinical symptoms and signs for the diagnosis of serious bacterial infection in young febrile children: prospective cohort study of 15 781 febrile illnesses. BMJ2010;340:c1594

  • Presenting Author: Ms Kate O’Brien
  • Supervisor: Dr Nick Breen
  • Co-Supervisor: Dr Janette MacGuinness

5. Investigation into the coverage of cancer research in the irish press

Ryan S 1, O'Connor A 2, Rowland M 3, Walsh S 2

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2 Research Dept., Irish Cancer Society, 43-45 Northumberland Road, Dublin 4.

3UCDSchool of Medicine and Medical Science, Mater Misericordiae University Hospital, Eccles St. Dublin 7.

Since 1963 the Irish Cancer Society has invested over €30 million in cancer research. As the Society relies on public donations, promotion of cancer research in the media is vital. This study aimed to determine how cancer research is featured in the Irish press.

All newspaper articles, from national and local newspapers, that mentioned ‘cancer’ from 2010-2012 were collected using a media-monitoring service. A database was established and a formula developed, using factors including circulation, article placement and length, to analyse the prominence of research articles.

In total 35,216 cancer articles were collected, of which only 5.9% focussed on research. Of these, just 22% related to Irish research. Of the Irish research, research funded by the Irish Cancer Society featured highest (25% of articles), with the National Cancer Registry Ireland second at 14%. The Irish Independent had the highest relative prominence of research articles. However the Irish Times had the strongest focus on Irish research.  There was significantly higher traceability for Irish research (58%) than international research (47%, p<0.001). Overall, breast cancer research articles dominated (24%) followed by prostate (13%). Compared to disease burden, measured by disability-adjusted life years(1), breast, prostate and skin cancer were over-reported while lung and liver cancer were significantly under-reported.

These results may allow researchers to selectively target the press in disseminating their research. The low level of cancer research reporting, particularly for Irish research, provides an opportunity to journalists, researchers and research bodies to increase the promotion of cancer research in the press.

Reference:

1.  Lewison, G; Tootell, S; Roe, P; Sullivan, R. How do the media report cancer research? A study of the UK's BBC website. Br. J Cancer. 2008; 99: 569–576

  • Presenting Author: Ms Sharon Ryan
  • Supervisor:  Dr Sinead Walsh
  • Co-Supervisor:  Dr Marion Rowland

6. The relationship between gestational weight gain and gestational diabetes mellitus

Fennessy AM, O’Higgins A1, Rowan A, Daly N, Mullaney L, Turner MJ1

1UCD Centre for Human Reproduction, Coombe Women and Infants University Hospital, Cork St, Dublin 8

Maternal obesity (Body Mass Index (BMI) ≥ 30 kg/m²) is increasing worldwide. The development of gestational diabetes mellitus (GDM) is strongly associated with obesity which increases the risk for both the woman and her baby. Recent guidelines concerned about rising rates of maternal obesity have therefore made recommendations for gestational weight gain (GWG) in obese women stricter. We analysed the relationship between GWG and the development of GDM. 

White women were recruited in their first trimester of pregnancy after sonographic confirmation of a singleton pregnancy. BMI was calculated accurately at recruitment. Screening for GDM was performed according to national guidelines at 28 weeks gestation. A repeat measurement of weight was performed at this time. Clinical details were collected from medical records and the Hospital’s computerised databases.

Of the 494 women recruited, GWG data at 28 weeks was available in 321 women (65.0%). Of the 321 women, 41% were primigravidas and 15.6% were obese; 39.9% (197/494) qualified for GDM screening and 7.3% (36/494) were diagnosed with GDM. GDM was diagnosed in 26.0% of the obese population, compared to 1.2% of those with a normal BMI (p<0.0002). Obese women who developed GDM had a lower weight gain at 28 weeks gestation than those who did not develop GDM (p < 0.05) (Table 1).

Obesity is the most important risk factor for the development of GDM. Focus on reducing weight gain during pregnancy before GDM screening may not lead to reductions in GDM incidence and could be harmful to the baby.

Table 1: Mean Gestational Weight Gain (GWG) (kg) at 28 weeks gestation by BMI category

BMI Category (kg/m²)

Total                              (n=321)

GWG without OGTT (n=196)

GWG with normal OGTT (n=103)

GWG with abnormal OGTT  (n=22)

BMI < 18.5

7.9 ±3.5

 (n=9)

7.3 ±3.2

(n=8)

13.0

(n=1)

n/a

(n=0)

BMI 18.5-25

8.1 ±3.0

(n=171)

8.0 ±2.9

(n=130)

8.3 ±3.1

(n=39)

4.6 ±5.2

(n=2)

BMI ≥ 25

7.6 ±3.4

(n=91)

8.3 ±3.4

(n=58)

6.8 ±3.1

(n=26)

5.0 ±1.6

(n=7)

BMI ≥ 30

4.3 ± 3.3

(n = 50)

n/a

(n=0)

4.8 ± 3.3

(n = 37)

2.7 ± 3.0

(n = 13)

Total

7.3 ± 3.4

(n=321)

8.1 ± 3.1

(n=196)

6.7 ± 3.5

(n=103)

3.6 ± 2.9

(n=22)

 

  • Presenting Author: Ms Anne Fennessy
  • Supervisor: Prof Michael J Turner
  • Co-Supervisor: Dr Amy O’Higgins

 

7. The effect of bariatric surgery on insulin dynamics and glucose homeostasis

McCarthy A2, Chen M3, Jackson S1,2, Neff KJ1,2, Le Roux CW1,2, Andrews RC3,

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2Experimental Pathology, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4.
3School of Clinical Sciences, University of Bristol. 

Obesity is an important cause of insulin resistance and impaired pancreatic β cell function, resulting in type 2 diabetes mellitus (T2DM). Bariatric procedures such as laparoscopic Roux-en-Y gastric bypass (RYGB) and laparoscopic adjustable gastric banding (LAGB) can result in remission of T2DM. This may be due to effects on glucagon-like-peptide 1 (GLP-1), and consequential effects on insulin dynamics.

Objective:- To quantify the effect of RYGB and LAGB on remission of type 2 diabetes, GLP-1 and insulin dynamics

Patients were recruited from a bariatric clinic and serum and plasma samples were taken before and 18 months after RYGB and LAGB.  Oral glucose tolerance tests (OGTT), intravenous glucose tolerance tests with arginine stimulation (IVGTT), and hyperinsulinaemic euglycaemic clamps were completed at each time point.

Samples were tested for insulin and c-peptide using immunoassays and glucose using a chemiluminesence assay (ARCHITECT, Abbott Laboratories). GLP 1 was measured using an ELISA (Millipore). Area under the curve (AUC) for each variable was compared. Differences were analysed between groups with Student’s t-test. Chi-square test and Fishers Exact test were used to analyse changes in diabetes remission.

RYGB resulted in remission (defined as a fasting glucose <5.6mmol/L) of diabetes in 74% of recipients compared to 50% after LAGB (p<0.001). GLP-1 AUC and first phase insulin response were greater after RYGB (both p<0.001). Insulin resistance as measured by HOMA-IR was reduced to a greater extent after RYGB (p=0.02). 

RYGB improves glycaemia and insulin secretion more compared to LAGB. The enhanced GLP-1 secretion seen after RYGB may partly explain our findings.

  • Presenting Author: Ms Andrea McCarthy
  • Supervisor: Prof Carel Le Roux
  • Co-Supervisor: Dr Karl Neff

8. Diabetic kidney disease after metabolic surgery

Lucey H, Elliot J, Higgins DF1, Neff KJ1,2, le Roux CW1,2

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2Diabetes Complications Research Centre, UCD Conway Institute of Biomedical and Biomolecular Research, University College Dublin, UCD, Belfield, Dublin 4.

Roux-en-Y gastric bypass (RYGB) is the most effective treatment to control type 2 diabetes(T2DM).(1) Diabetic kidney disease (DKD) is a common complication of T2DM which increases mortality.(2) Our study aims to determine whether RYGB can ameliorate DKD in a rat model of T2DM.

18 week old Zucker Diabetic Fatty (ZDF) rats were randomly assigned to either RYGB (N=15) or one of four control groups; sham operated (SAL, N=6), sham operated glucose matched (SGM, N=8), sham operated body weight matched (SBW, N=8), un-operated non-diabetic controls (NSX, N=5). Rats were studied 12 weeks post-operatively. Kidneys were examined using Sirius Red and Periodic Acid Schiff staining for fibrosis and glomerulosclerosis respectively, and quantitative PCR for markers of fibrosis. Results were compared using Kruskal Wallis testing for groups, and appropriate post hoc tests between groups.

Sirius Red staining showed no differences. Glomerulosclerosis was more pronounced in SAL compared to RYGB groups (p>.05). Vimentin expression was higher in SBW(p=0.03) and SGM rats (p=0.04) than RYGB rats. Collagen1A2 (COLL1A2) expression was up-regulated in SGM as compared to RYGB rats (p=0.004).

RYGB appears to attenuate early renal damage, such as glomerulosclerosis. RYGB induces positive changes in the expression of genes that are consistent with early disease onset, as compared to control groups. Lack of histological differences between groups suggests that despite profound hyperglycaemia, control rats were terminated before significant fibrosis developed.  The ZDF model may be appropriate to determine mechanisms by which RYGB prevents renal damage, but rats will need more than 30 weeks to develop DKD.

References:

1. Mingrone, G. et al. (2012) ‘Bariatric surgery versus conventional medical therapy for type 2 diabetes’. New England Journal of Medicine 366(17):1577-85.

2. Afkarian, M. et al. (2013) ‘Kidney Disease and Increased Mortality Risk in Type 2 Diabetes’. Journal of the American Society of Nephrology 24:302-308. 2002;347(7):284-7. 

 

  • Presenting Author: Ms Hannah Lucey
  • Supervisor: Prof Carel Le Roux
  • Co-Supervisor: Dr Karl Neff

9. Biomarkers of monocyte activation (scd163 and scd14) in human immunodeficiency virus (HIV)-positive subjects

Gurwith MMP1, O’Halloran JA1, Mallon P1

1HIV Molecular Research Group, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4

HIV infection is associated with increased risk of coronary artery disease (CAD). Elevated biomarkers of monocyte activation, such as soluble clusters of differentiation 163(sCD163) and sCD14 have been associated with atherosclerosis in HIV-negative subjects. We aim to examine changes in biomarkers post-initiation of antiretroviral therapy (ART) and to compare with HIV-negative controls.

Plasma samples from HIVpos subjects at 0,4 and 12 weeks post-ART initiation were analysed using enzyme linked immunosorbent assay for sCD163 and sCD14 and compared to HIVneg controls. Data was analysed using Wilcoxon signed-rank sum tests or Mann-Whitney-U test where appropriate. Correlations were determined using Spearman’s rank correlation coefficient.

Baseline characteristics are described in Table 1. Values represent median (IQR) unless otherwise stated. Baseline sCD163 levels of HIVpos subjects were significantly higher than HIVneg subjects (table 1) P=0.001. sCD163 significantly decreased by week 12 post-ART (-22.3% median change(-38.3%,-4.8%),P=0.001). sCD163 was not significantly different by  week 12 to HIVneg. Baseline sCD14 levels were not significantly different from HIVneg (table 1) P=0.138. sCD163 did not significantly decrease by week 12 post-ART (-3.0% median change(-16.2%,8.2%),P=0.584). HIVpos baseline sCD163 correlated with Log10HIVRNA (r=0.638,P=0.001) and lower baseline CD4+T cell count (r= -0.429,P=0.032).

Soluble CD163 levels are elevated in HIV-pos subjects compared with HIVneg controls and decrease with ART initiation. Higher sCD163 correlates with more advanced HIV disease as measured by CD4+ T cell and LogHIV RNA. These results suggest evidence of ongoing monocyte activation in HIVpos subjects in the absence of ART, which would contribute to increased CAD seen in this population.

  • Presenting Author: Ms Meredith Gurwith
  • Supervisor: Dr Paddy Mallon
  • Co-Supervisor: Dr Jane O’Halloran

 

 

 

 

 

10. The effects of HIV protease inhibitors on insulin signalling in hepatocytes.

Lee S, Maughan R, Mallon P1

1HIV Molecular Research Group, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

HIV protease inhibitors (PIs) are a class of antiretroviral drugs regularly used in the treatment of HIV infection. Therapy with PIs has been directly associated with an increased risk of insulin resistance (1). The effects of PIs on insulin signalling in hepatocytes are not well characterised. My aim thus was to explore the effects of two commonly prescribed PIs, Lopinavir (LPV) and Ritonavir (RTV) on insulin signalling in hepatocytes.

HepG2 (human hepatocarcinoma) cells were grown to confluence and were exposed to different concentrations of LPV and RTV in independent triplicates for 14 days. After 14 days, cells were harvested in fasting state, RNA was extracted and cDNA libraries were prepared. The expression of peroxisome proliferator activated receptor gamma (PPARG), glucose transport type 4 (GLUT4) and the insulin receptor (INSR) were measured by Real Time PCR using the Lightcycler 480 (Roche Applied Science, Mannheim, Germany). Expression levels were normalised to beta actin mRNA. 

In the study, LPV was seen to inhibit GLUT4 in a dose dependent manner. PPARG was not inhibited by LPV, however a small dose of RTV induced a non-significant decrease in PPARG. A significant inhibition of INSR occurred at the highest concentration of LPV.

Our results show an inhibition of GLUT4 expression in hepatocytes which may contribute to the insulin resistance associated with PIs. Decreased levels of INSR were noted at high concentrations of LPV leading to a possible imbalance of metabolism. Future experiments will measure the expression of other genes downstream of insulin signalling in hepatocytes.

References:

1. Heath KV, Hogg RS, Chan KJ, Harris M, Montessori V, O'Shaughnessy MV, et al. Lipodystrophy-associated morphological, cholesterol and triglyceride abnormalities in a population-based HIV/AIDS treatment database. AIDS (London, England). 2001;15(2):231-9.

2. Murata H, Hruz PW, Mueckler M. Indinavir inhibits the glucose transporter isoform Glut4 at physiologic concentrations. AIDS. 2002 Apr 12;16(6):859-63. PubMed PMID: 11919487. Epub 2002/03/29. eng.

  • Presenting Author: Mr Samuel Lee
  • Supervisor: Dr Robert Maughan
  • Co-Supervisor: Dr Paddy Mallon

11. Children in Slovenia - understanding of x-ray

Harding J1, Davis M1,2, Starc T3

1UCD School of Medicine and Medical Science University College Dublin, UCD Belfield, Dublin 4.

2Diagnostic Imaging, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

3Univerza v Ljubljani.

The aim of this study was to see what perception children aged between 6 and 10 years had of X-ray. It is thought that a better knowledge of a child’s understanding of X-ray can lead to a better experience for them in the X-ray department.

The method used was the draw and write technique as described by Bradding and Horstman (1999). This technique was chosen as it is seen as child friendly and it facilitates the expression of views by the child without the adult being continuously involved. The children in the study were asked to draw what they thought of when they heard the word “X-ray”. No other information was given to them, and they all had a free choice of the same colours to use. The pictures were drawn in summer camps with consent given by the camp leader, after full ethical approval was granted. The children were given the right to withdraw at any time. A pilot study was conducted on seven children. 

73 children took part in the research. Of these, 60 (82%) drew their experience in an x-ray room, 31 (42%) drew negative aspects such as crying or fear and 42 (57%) drew their experience with no healthcare professional. 6 (8%) drew the healthcare professional as a female nurse.

The findings show that there are similarities (only 20% (n=15) of the children drawing a dedicated radiographer) and differences (only 3% (n=2) of the children drawing syringes) to previous research that has been conducted. 

  • Presenting Author: Mr James Harding
  • Supervisor: Dr Michaela Davis
  • Co-Supervisor: Ms Tina Starc

12. A study to investigate the ability of diagnostic radiographers in a dedicated paediatric hospital to recognise typical non-accidental injury fractures which may present in children.

Ahmed S1, Davis M2, Grehan J2

1UCD School of Medicine and Medical Science, University College Dublin, UCD Belfield, Dublin 4.

1,2 Diagnostic Imaging, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

The aim of this research was to measure radiographers’ ability to recognise typical non-accidental injury (NAI) fractures that occur in paediatric patients on plain film radiographs.

Participants were chosen by convenience sampling. The participants for this study were 17 radiographers who worked in a dedicated paediatric centre and 1 paediatric radiologist. They were shown 18 plain film radiographic images which were selected from the university radiographic database (8 images had typical NAI fractures/10 images had common accidental injury fractures). Participants viewed the images on a laptop calibrated to DICOM greyscale standard display function. Viewing conditions were standardised and optimised for each participant using a calibrated photometer. Each participant viewed the 18 images and rated their confidence of the injury on a scale from 1 to 5 denoting whether they thought the injury was accidental (1, 2), non-accidental (4, 5) or unsure (3).

Sensitivity and specificity values were calculated for each participant. The level of agreement between the radiographers and the radiologist for recognising NAI was measured using kappa measure of agreement. Results are illustrated in table 1.

Mean linear weighted kappa score.

0.43= moderate agreement beyond chance

Mean sensitivity of the radiographers as a group in recognising NAI.

75%

Sensitivity of the radiologist in recognising NAI.

87.5%

Specificity of radiographers and radiologist in recognising accidental injuries

90%

 

It is possible to conclude that radiographers can recognise typical NAI fractures better than chance and can differentiate between accidental and non-accidental injury fractures.

  • Presenting Author: Ms Shafq Ahmed
  • Supervisor: Dr Michaela Davis
  • Co-Supervisor: Ms Jennifer Grehan

 

13. Decision making and variation in radiation exposure factor selection by radiographers: an eye-tracking study

Darcy S, Rainford, L1, Toomey R1

1 Diagnostic Imaging, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

The goal of radiographic imaging is to produce a diagnostically useful image while minimising patient radiation dose. This study aimed to review variations in exposure factor selection by radiographers for a selection of virtual patients with varying body mass index characteristics. Patient dose is impacted upon by a number of exposure parameters including: kVp, mAs, source to image receptor distance (SID) and anti-scatter grids.

Eleven radiographers were asked to assign exposure parameters (kVp; mAs; SID; grid) for forty computer-generated patient images. Patient images represented five BMI categories – underweight; healthy weight; overweight; obese; super-obese. There were four examination categories: antero-posterior (AP) shoulder; AP lumbar; lateral lumbar; AP ward chest. As participants assigned exposures their visual patterns were recorded by a Tobii ® TX300 eye-tracker.

Significant (p<0.05) correlation was found between radiographer age/experience and assignment of mAs for the AP shoulder, and lumbar examinations. Greater age/experience correlated with higher mAs for the AP shoulder exam, but with lower values for the lumbar examinations. Strong linear correlation between times to first fixations on relevant anatomical areas and kVp/mAs values existed for the AP portable chest examination.

Exposure selection differences related to age/experience highlight inconsistencies in the practice of exposure parameter setting. The reasons for these inconsistencies requires further investigation, and how to address deficiencies practice requires consideration to optimise safe patient care. Due to the small sample size used, further research into the relationship between visual factors and individual examinations is suggested, following the findings regarding the AP portable chest examination.

  • Presenting Author: Ms Sarah Darcy
  • Supervisor: Dr Rachel Toomey
  • Co-Supervisor: Dr Louise Rainford

14. The development of diagnostic accuracy and search pattern behavior in the interpretation of chest radiographs

Kelly BS1, Toomey RJ1, Kavanagh EC2, Rainford LA1

1Diagnostic Imaging, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Department of Radiology, Mater Misericordiae University Hospital, Eccles St, Dublin 7.

To investigate the development of radiological image interpretation skill through medical training by measuring both diagnostic accuracy and eye movements during visual search.

Five Consultant Radiologists, deemed the reference expert group, four Radiology Registrars, five Senior House Officers (SHOs) and six Interns formed four clinician groups. Participants were shown 30 chest radiographs, 14 of which had a pneumothorax and were asked to give their level of confidence as to whether a pneumothorax was present. Receiver Operating Characteristic (ROC) Analysis was carried out on diagnostic decisions. Eye movements were recorded by a Tobii TX300 eye tracker. Four Eye-tracking metrics were analysed. Variables were compared to identify any differences between groups.

All data were compared using the Friedman non-parametric method. The average area under the ROC Curve for the groups increased with experience (p=0.009). Statistically significant difference in diagnostic accuracy was found between Consultants and Registrars (p=0.046). All four eye-tracking metrics reduced with experience, this was statistically significant for Registrars compared with SHOs. The total reading time reduced with experience; significantly for Registrars compared to SHOs (p=0.046) and between SHOs and Interns (0.025).

Reader performance increased with experience. The level of experience at which there was a statistically significant difference was higher for diagnostic accuracy than for eye-tracking metrics. This data would suggest that specific training is needed to improve radiology expertise and that the development of an “expert” search pattern predates the development of “expert” levels of diagnostic accuracy.

  • Presenting Author: Mr Brendan Kelly
  • Supervisor: Dr Louise Rainford
  • Co-Supervisor: Dr Rachel Toomey

15. Acute infections in current and past injecting drug users presenting to an inner –city emergency department

Connolly S1,2, O’Connor G1,2, Mallon  P2,3

1Emergency Department, Mater Misericordiae University Hospital, Eccles Street, Dublin;

2Department of Infectious Diseases, Mater Misericordiae University Hospital, Eccles Street, Dublin 7. 

3HIV Molecular Research Group, UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Injecting drug use (IDU) brings its own host of acute infection risks.

The PRESIDNT (Prospective Epidemiological Study in Injecting Drugs Users in North Dublin) is a prospective cohort study that records Emergency Department attendances to the Mater Misericordiae University Hospital by those with any history of IDU. The cohort’s records were interrogated to describe acute infections in both active and past injecting drug users. Details of attendees, including basic demographics, acute diagnoses, drug use and co-morbid medical conditions were recorded. The nature of acute infective diagnoses and the associated microbiology was also described.

Over six months 1377 presentations were identified prospectively with histories of past or current IDU.  Of the 687 patients studied, 63.2% were male while the mean age was 35 years and was normally distributed. The overall employment rate was 6.7%.  65.8% were ‘housed’; the remainder either residing in hostels or temporary shelter (13.2%); ‘sleeping rough’ (10.0%); or being in other lodgings, including prison (10.9%). Acute infections comprised 15.1% of presentations. Skin and soft tissue infections and respiratory tract infections proved commonest, involving 65.4% and 33.7 of infectious cases, respectively. Staphylococcus aureus and coliforms were the most common virulent bacterial isolates in invasive infections, and were cultured in 17.7% and 16.1% of such infections respectively. Significant contribution from virulent Gram-negative organisms was noted, accounting for 49% of invasive infections.

The high numbers of Gram-negative infections demonstrated contrast with established literature published in the area, and may have implications for prescribing empirical antimicrobial therapy for infections in this population (1, 2).

References:

1. Sumanen PH, Talan DA, Strong ,. McTeague, Bennion R, Thompson JE, Väisänen ML, Moran G, Winer M, Finegold SM. Bacteriology of Skin and Soft-Tissue Infections: Comparison of Infections in Intravenous Drug Users and Individuals with No History of Intravenous Drug Use. Clin Inf Dis. 1995:20(6):279-282.

2. Scheidegger C, Zimmerli W. Infectious complications in drug addicts: seven- year review of 269 hospitalized narcotics abusers in Switzerland. Rev Infect Dis. 1989:11(3):486-93.

  • Presenting Author: Mr Stephen Connolly
  • Supervisor: Dr Paddy Mallon
  • Co-Supervisor: Dr Gerard O’Connor

16. Hypoplastic left heart syndrome: an overview and techniques

Lantz, N1, Brown N2, Connolly, B2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada M5G1X8

The current surgical techniques for the treatment of hypoplastic left heart syndrome are reviewed and compared with new techniques. The usefulness of additional techniques for improving outcomes in surgical cases is reviewed.

A literature review was conducted regarding these topics, using research from the governing hospital and others.

The Norwood procedure is currently the most widely used surgical intervention. A hybrid procedure has been developed to replace the Norwood but presently, is only used in high-risk cases, and therefore its efficacy over the Norwood cannot be determined. Treatment with phenoxybenzamine at the beginning of surgical intervention has been shown to improve outcomes by decreasing systemic vascular resistance. Maintenance of oxygen saturation over 50%, using superior vena cava oximetry, is associated with a lower mortality rate. In a retrospective study, completed at The Hospital for Sick Children, hyperglycemia was defined as a blood glucose concentration over 15mmole/litre, and was significantly associated with negative outcomes.

Currently at The Hospital for Sick Children, multiple techniques are used in surgical cases to maintain an unadjusted mortality rate of 10%. These include intensive phenoxybenzamine treatment and anastomosis of the descending aorta, aortic arch and main pulmonary artery (1).

Reference:

1.  A Azakie, S Merklinger, B McCrindle, G Van Arsdell, K-J Lee et al. Involving strategies and improving outcomes of the modified Norwood procedure: A 10-year single-institution experience. Ann Thorac Surg 2001;72:1349-1353.

  • Presenting Author: Ms Natasha Lantz
  • Supervisor: Dr Bairbre Connolly
  • Co-Supervisor: Ms Nicole Brown

17. A case of retinoblastoma: image guided therapy’s role in one patient’s care

A case of an 18 month old female presenting with a two week history of leukocoria and subsequent diagnosis of unilateral retinoblastoma was examined. In particular the Image Guided Therapy (IGT) department’s involvement in her care was reviewed, and how interventional radiologists at the hospital may be involved in such cases in the future.

Post-enucleation the patient was found to have swelling on the ipsilateral side of the face. IGT was involved in performing a soft tissue biopsy guided by sonography which aided in diagnosis of a tumour extension. IGT then inserted a Port-A-Cath to aid the administration of chemotherapy.

Following this, a femorally inserted central venous line was inserted with the aid of ultrasound and fluoroscopy. This line was placed to facilitate the harvesting of cells for a bone marrow transplant. Unfortunately prior to the transplant the patient was found to have focal metaphyseal lesions in some long bones. IGT became involved in her care again to perform an MRI guided bone biopsy as the lesions could not be clearly identified on CT scans. A core needle biopsy from the proximal tibia was performed using real time MR guidance, which determined that the lesion was a retinoblastoma metastasis..

Primary treatment for retinoblastoma is currently enucleation with some focal therapies used for small intraocular tumours. The interventional radiologists at SickKids are intending to assist in the treatment of retinoblastoma via intra-arterial chemotherapy. A chemotherapeutic agent will be delivered directly into the ophthalmic artery guided by fluoroscopy which will increase the concentration of the drug delivered to the tumour and reduce systemic side effects.

  • Presenting Author: Ms Ruth Ellard
  • Supervisor: Dr Bairbre Connolly
  • Co-Supervisor:  Dr Simal Bindra

18. Paediatric interventional radiology observership: osteoid osteoma diagnosis and treatment

Sertic A1, Connolly2 B, Brown, N2

1UCD School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin 4.

2Image Guided Therapy, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada

Osteoid osteoma is a benign skeletal neoplasm, typically smaller than 2 cm in diameter.  It is characterised by its osteoid rich nidus, surrounded by a fibrovascular rim of stroma and perilesional sclerosis.  The nidus may contain variable amounts of calcification, depending on time of diagnosis. 

The tumour has an unknown aetiology, although the nidus is associated with high levels of prostaglandin synthesis and is most prevalent in the second decade of life.  Patients often describe a deep and continuous pain which worsens at night and is relieved by non-steroidal anti-inflammatory drugs.  The lesion is found in the cortex of long bones in approximately 80% of cases. 

Osteoid osteoma may be diagnosed with the use of various imaging modalities; however due to its sensitivity and specificity, computed tomography (CT) is the gold standard. Further scans (such as scintigraphy) or a biopsy may be necessary in order to differentiate osteoid osteoma from other bone tumours or osteomyelitis.

Performed by interventional radiologists at the Hospital for Sick Children, CT guided thermal ablation (either radiofrequency or laser) is a minimally invasive technique that has replaced the historical method of en-bloc surgical resection in the treatment of osteoid osteoma.  Thermal ablation probes are able to target the nidus of the lesion directly, causing the least possible damage to the surrounding healthy bone. These techniques have primary clinical success rates of 89-95%, and reduce the risk of fracture and the length post-procedural hospitalisation, and should be the first choice in treating intolerable osteoid osteoma[1]. 

Reference:

Motamedi D, Learch TJ, Ishimitsu DN, Motamedi K, Katz MD, Brien EW, et al.  Thermal Ablation of Osteoid Osteoma: Overview and Step by Step Guide.  Radiographics. 2009; 29: 2127-2141.

  • Presenting Author: Mr Andrew Sertic
  • Supervisor: Dr Bairbre Connolly
  • Co-Supervisor: Ms Nicole Brown

20. A pilot survey of current management of atrial fibrillation in patients ≥ 70 years of age in general practice, as part of a national atrial fibrillation screening study

Cahill D1, Swan D1, Carberry C1, Tobin H1, Gaughan B1, Keane D1, Kelleher C1, Cullen W2, Fitzmaurice D3, Barnes J1, Bury G1.

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Graduate Entry Medical School, University of Limerick, Limerick, Ireland.

3Primary Care Clinical Sciences, School of Health and Population Sciences, College of Medical and Dental Sciences, University of Birmingham, UK.

Around 10,000 people suffer a stroke in Ireland annually and 30,000 live with consequent disability. Atrial Fibrillation (AF) creates a five-fold risk of stroke for those with AF.Prevalence of AF doubles each decade after 50 years of age. Anti-coagulation of AF patients can reduce stroke risk by up to 60%1 yet AF is frequently unrecognised and undertreated[1] [i]. A common algorithm used to calculate stroke risk for AF patients is the CHA2DS2- VASC score. Guidelines advise anticoagulation where score >2. Management of AF includes prevention of thromboembolic events, rhythm/rate control and therapy of concomitant cardiac disease. Main risk factors for AF include advancing age, hypertension, valvular heart disease, heart failure, obesity, diabetes, and sleep apnoea.

We have recently completed a national screening study in general practice to establish the prevalence rate of AF in a cross-sectional sample of 1003 patients aged ≥70 years, not known to have AF, using 3 lead ECG technology.  This identified 149 AF patients through chart review, ECG screening and GP reporting of whom 116 were followed up in this phase to establish relevant events and AF therapy. These patients are highly representative of patients with AF aged ≥ 70 years in Irish general practice.

116/149 (78%) patients agreed to allow their data to be included in this phase of the study. The pilot study involved doing chart reviews of 26 patients by research staff. Data was extracted using a structured instrument, collated centrally and analysed using SSPS v20.

50% of the 26 patients received anticoagulant therapy to prevent thromboembolic events (46% warfarin, 8% New Oral Anticoagulant agent (OAC)). Seven of the 26 patients (27%) suffered a stroke, of whom four were not taking anticoagulant due to contraindicating co-morbidity. GPs considered various factors before prescribing anticoagulant therapy including: cardiac co-morbidities, social status, general health/mobility/predisposition to falls and cost. Hypertension was the most prevalent risk factor for AF in this cohort (81%). 

When the full study is completed, we will get a clearer picture of anticoagulation rates in a likely representative sample of over 70s in Ireland. Further studies should explore barriers to anticoagulation and attitudes to new OACs amongst GPs.

Acknowledgement:

This project has ethical approval from the ICGP, and is funded by the Health Research Board.

References:

1. Ir Med J. 2004 Jan;97(1):10-2.Community-based study of atrial fibrillation and stroke prevention. White SFeely JO'Neill D

2. Ir Med J. 2003 Nov-Dec;96(10):296-9.Implementation of anticoagulation guidelines in general practice: a practice report. Dubhghaill CTMoonen LO'Loughlin AO'Keeffe SEgan DMurphy AW

  • Presenting Author: Ms Deirdre Cahill
  • Supervisor: Dr Crea Carberry
  • Co-Supervisor: Dr Davina Swan

21. Impact on growth of a nutrition policy to improve early oral and parenteral nutrition for preterm infants: retrospective audit

Low CS1, Ho JJ2, Nallusamy R3

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Department of Paediatrics, Penang Medical College, Penang 10450, Malaysia

3Department of Paediatrics, Penang Hospital, Penang, Malaysia. 

The objective of this study was to compare growth and feeding of preterm infants before and after the introduction of a new aggressive feeding policy using early Parenteral Nutrition and standardised counselling aimed at providing early expressed mother’s milk. This is a retrospective study between 2009 and 2012 in Penang Hospital, a tertiary referral hospital.

80 preterm babies (34 weeks and below) discharged from NICU were included (40 pre- and 40 post-intervention). The primary outcome was growth and secondary outcomes were time to full feeding, breastfeeding on discharge, and adverse events. Normally distributed data were expressed as means and standard deviation and analysed using independent T-test. Non-normal data were expressed as medians and interquartile range and analysed using Mann-Whitney-U. Categorical data were analysed using Chi-Square.

Weight at 15 and 28 days was not available for some babies discharged earlier. Otherwise complete data was available for all babies. Baseline data were similar in the two groups. There was no overall significant weight difference at 7, 14, and 28 days. There was a post-intervention trend to earlier full oral feeding (13(11) versus 11(6) days p=0.058). More post-intervention babies were breastfed at discharge (8 versus 21, p=0.005). Nosocomial infection (11 versus 4 p=0.045), and blood transfusion (31 versus 19 p=0.01) were significantly lower post-intervention. There was a trend for reduced necrotising enterocolitis (5 versus 0, p=0.055 (Fishers exact test))

Our finding of no improvement in growth has been previously described.(1) However we found other important benefits associated with the new nutrition policy.  

 

Reference:

1. Ibrahim HM, Jeroudi MA, Baier R, Dhanireddy R, Krouskop RW. Aggressive early total parental nutrition in low-birth-weight infants. Journal of perinatology. 2004;24(8):482.

  • Presenting Author: Mr Chuen-Siang Low
  • Supervisor: Prof Jacqueline Ho
  • Co-Supervisor: Dr Revathy Nallusamy

22. Investigation of potential anti-inflammatory and antifibrotic effects of glp-1 receptor agonists

Gorman DM1,2, Nolan E1,2, Gaffney A1,2,  Godson C1,2

1Diabetes Complications Research Centre, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4.

2UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Glucagon-like peptide-1 receptor agonists (GLP-1-RA) are a class of incretin drugs used to treat type 2 diabetes (T2D). Recent evidence suggests GLP-1-RA exert renoprotective and systemic anti-inflammatory effects independent of glycemic control.[1][2] The aim of this study was to investigate whether the renoprotective effects of GLP-1-RA result from anti-inflammatory and antifibrotic effects on infiltrating and resident kidney cells.

THP-1 human monocytes were treated with GLP-1-RA (exenatide or liraglutide) at various concentrations (0.1nM-1μM) for 30 minutes, before exposure to LPS (1ng/ml) for 24 hours. TNF-α and IL-6 levels in cell supernatants were determined by ELISA. Human proximal tubular epithelial (HK-2) and human mesangial cells were similarly treated with exenatide or liraglutide at various concentrations (0.1nM-1μM), before stimulation with TGF- β1 (10ng/ml). Protein was extracted and assayed by western blotting for expression of fibronectin, CTGF, and JAG1.

Exenatide and liraglutide attenuate the production of TNF-α and IL-6 by LPS-stimulated monocytes. The effect is seen at all concentrations, with the greatest reduction seen at 1μM for exenatide and 0.1nM for liraglutide (n=1). Exenatide and liraglutide attenuate the expression of fibronectin and CTGF in TGF- β1-stimulated HK-2 and mesangial cells, while exenatide also reduces the expression of JAG1 in HK-2 cells (n=1).

In conclusion, our results, albeit preliminary, support our hypothesis that GLP-1-RA may exert anti-inflammatory and/or fibrosuppressant effects. Investigation is ongoing to consolidate these preliminary data and to investigate the potential mechanism of action and relative efficacies, which may suggest additional therapeutic benefit of GLP-1-RA in the context of diabetic kidney disease.

References:

1. Hogan A.E. et al. Glucagon-like peptide-1 (GLP-1) and the regulation of human invarient natural killer T-cells: lessons from obesity, diabetes and psoriasis. Diabetologia. 2011 Nov;54(11):2745-54.

2. Kodera R. et al. Glucagon-like peptide-1 receptor agonist ameliorates renal injury through its anti-inflammatory action without lowering blood glucose level in a rat model of type 1 diabetes. Diabetologia. 2011 Apr;54(4):965-78

  • Presenting Author: Ms Dora Gorman
  • Supervisor: Prof Catherine Godson

23. Igf-1-r and egfr signalling crosstalk in triple negative breast cancer cell survival, migration and invasion

C McDermott 1, S McClendon 2, T Alcaide,2, T Liu 2, T McGlothen 2, L Taliaferro-Smith2, and R O’Regan 2,3

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2Department of Haematology and Medical Oncology, Winship Cancer Institute

3Georgia Cancer Centre for Excellence at Grady Memorial Hospital, Atlanta, GA, 30303.

Triple negative breast cancers (TNBCs) are a heterogeneous group of breast cancers characterised by the lack of three receptors; oestrogen receptor, progesterone receptor and HER2/neu. The diagnosis of TNBC is associated with a poor prognosis and a highly metastatic and aggressive disease; furthermore there are no targeted therapies available for TNBCs.  The aim of the project was to investigate how TNBC cells are impacted in terms of protein expression, survival, migration and invasion, and ultimately metastasis when IGF-1-R and EGFR signaling pathways are inhibited.

Cell lines:HCC1806, HCC70, MDA-MB-231, Hs578T, and BT549 TNBC cell lines were cultured in DMEM media supplemented with 10% FBS. Western Blot: 30μg total protein lysates were resolved on 8-12% polyacrylamide gels, transferred to PVDF membranes, and probed with antibodies specific for IGF-1-R, pEGFR, EGFR, pERK, pAKT, pS6, and β-actin. Matrigel Invasion Assays: Cells treated with DMSO or inhibitors for 24h were plated in the upper chambers of Transwell units coated with Matrigel. Fibronectin and complete media were used as a chemoattractant in the lower chambers. After 24 hours, cells that invaded the membranes were fixed in methanol, stained with crystal violet, and counted.

The combination of an IGF-1-R inhibitor (PPP) and an EGFR inhibitor (lapatanib/erlotinib) were found to be more effective than either drug in isolation in terms of reducing invasion, migration and cell survival.

These results suggest that IGF-1-R/EGFR signaling crosstalk stimulates cell survival, migration and invasion and may ultimately promote metastasis. Further investigation of the mechanisms of this potentially pro-metastatic interaction is merited.   

Acknowledgement:

This research was supported by the Glenn Family Breast Funds, Winship Cancer Institute of Emory University

  • Presenting Author: Mr Christopher McDermott
  • Supervisor: Dr LaTonia Taliaferro-Smith
  • Co-Supervisor: Dr Tongrui Liu

24. Identification of a gene causing epileptic encephalopathy in a single irish family

Nealon J1,   Conroy  J1,  McCreary  D1, McGettigan  P2, Crushell E3,  Lynch  SA4, Ennis S1

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4. 

2UCD School of Agriculture, Food Science and Veterinary Medicine, University College Dublin, Belfield, Dublin 4.

3Children's University Hospital Temple Street, Temple street North, Dublin.

4The National Centre of Medical Genetics Ireland, Our Lady’s Hospital for Sick Children, Crumlin, Dublin.

Epileptic encephalopathies (EE’s) are rare but devastating forms of epilepsy, which present as frequent and severe seizures with cognitive and behavioural disturbances. The cohort in this study was a single family consisting of two sisters with idiopathic EE in addition to an unaffected mother, father and brother. The objective was to identify the potential genetic cause for this familial form of EE using next-generation sequencing. 

Exome sequencing was performed on DNA samples of each family member at Atlas biolabs (http://www.atlas-biolabs.com/home). Following bioinformatics analysis and rare/novel variant identification, the resulting data was analysed based on three patterns of inheritance. These were recessive, compound heterozygous and de-novo models. The list of rare variants was compared to a list of epilepsy-linked genes. The resulting candidates were screened in control populations (Irish exome control database, Seattle exome database, and 1000 genomes project). In silico investigations to predict mutation effects were undertaken using SIFT, Polyphen and MutationTaster. Literature reviews and further analysis were performed using OMIM, Pubmed, UCSC and Orphanet. High priority candidate variants were confirmed by Sanger sequencing.

Eight variants were identified in five candidate genes (1 recessive, 3 compound heterozygous  and 1 EE gene). Additional variants of unknown effect were also present. Currently these cannot be ruled out as EE candidates. Furthermore, it is possible that the disease causing mutation is not present in the exome and therefore cannot be identified using this technique.

Although potential candidate variations have been reported, further work is required to confirm these as disease causing mutations. 

Acknowledgements:

We sincerely thank the families for the use of clinical information and genetic samples. We would like to thank Dr Ellen Crushell and Dr Sally Ann Lynch for their collaboration in this project. This project was funded by The Children’s Fund for Health - Children’s University Hospital, Temple Street and the Pathological Society of Great Britain and Ireland Undergraduate Bursary.

  • Presenting Author: Mr John Nealon
  • Supervisor: Dr Sean Ennis
  • Co-Supervisor: Dr Judith Conroy

25. Suicide in the children of Ireland from 2003-2008

McLoughlin AB1, Kelleher C2, Malone KM1

1Department of Psychiatry, Psychotherapy and Mental Health Research, St. Vincent's University Hospital and School of Medicine and Medical Science, University College Dublin, Elm Park, Dublin 4.

2UCD School of Public Health, Physiotherapy and Population Science, University College Dublin, UCD Belfield, Dublin 4. 

Worldwide, suicide is a leading cause of death among children.  Despite the prevalence of childhood suicide in Ireland, research in this area remains scant, with an almost complete focus on the interpretation of quantitative data.

To voice the stories of the suicides of children as told by their caregivers, siblings and friends. The aim of this research project is to provide an understanding and meaning beyond simple vital statistics for suicide in children.

We used a Psycho-biographical Autopsy template and a grounded theory approach to analyse qualitative interviews with a volunteer sample of suicide-bereaved parents, siblings, and friends of 14 children who had died by suicide. Also included were personal documents (diaries/letters/notes) belonging to the deceased to identify age-related themes concerning adolescent risks for suicide, as well as factors independent of age, such as severe mental illness. A qualitative case study of a cluster within this sample is also undertaken using an ecological approach.

Themes to emerge include risk factors of peer/adult/statutory violation, displacement, lack of connectedness, mirroring/clusters, humiliation, mental distress, substance abuse, and family history of mental illness and suicidality. Within the Cluster Study, themes to emerge include connectivity and contagion.

This report offers a holistic reflection of child suicide in Ireland that draws together sociological, psychological and relational domains of the suicidal process that may critically inform policy development in Child Health.

  • Presenting Author: Ms Aoibheann McLoughlin
  • Supervisor: Prof Kevin Malone
  • Co-Supervisor: Prof Cecily Kelleher

 

 

26. Elucidating the molecular mechanism underlying bmp-7 protection against renal injury

Tennant S1, Godson C1,2, Higgins D1,2.

1Diabetes Complications Research Centre, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4.

2UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Bone morphogenetic proteins, BMPs, are glycosylated ECM-associated molecules that belong to the transforming growth factor beta (TGFb) superfamily. BMP growth factors play a crucial role in normal developmental processes such as bone growth. Dysregulated BMP signalling is a key factor in many pathological processes and loss of BMP-7 signalling is associated with the pathophysiology of renal disease. Exogenous BMP-7 administration offers protection against renal fibrosis in vivo however the precise molecular mechanisms underlying this protection are unclear (1, 2).

The aim of this study was to examine BMP-7 modulation of TGFβ-responsive signalling pathways in human renal tubular epithelial HK-2 cells. HK-2 cells were stimulated with TGFb1 (5ng/ml, 30-120 minutes) or hypoxia (1% O2, 30-120 minutes) in the presence or absence of  BMP-7 (5-100ng/ml, 30 minutes). TGFb1-activation of pAkt, pGSK3b, pERK  and pSMAD2 and effects of BMP-7 on this activation were analysed by Western blot.

TGFb1 or hypoxia induced phosphorylation and hence activation of pAkt and its downstream target pGSK3b along with pERK and pSMAD2 after 60 minutes of stimulation. Pre-treating cells with BMP-7 for 30 minutes prior to TGFb1 or hypoxia inhibited activation of pAkt and pGSK3b but had no effect on pERK and pSMAD2 in renal tubular epithelial cells.

This study has described a novel signalling mechanism underlying the protective and anti-fibrotic effects of BMP-7 in renal epithelial cells. As similar results were recently observed in human renal collecting duct cells, BMP-7 inhibition of pAkt and pGSK-3b likely represents a general mechanism of BMP-7 protection.

Acknowledgment:

Funding from the UK Biochemical Society

References:

1. Hruska, KA, et al., Osteogenic protein-1prevents renal fibrogenesis associated with

ureteral obstruction. Am J Physiol Renal Physiol, 2000. 279:F130-F143

2. Zeisberg, M, et al., BMP-7 counteracts TGFbeta1-induced epithelial-to- mesenchymal transition and reverses chronic renal injury. Nature Medicine, 2003. 9(7):964-968

  • Presenting Author: Ms Sadhbh Tennant
  • Supervisor: Dr Deborah Higgins
  • Co-Supervisor: Prof Catherine Godson

27. Role of hypoxia inducible transcription factors hif-1 and hif-2 in macrophage differentiation towards pro-inflammatory m1 or pro-resolution m2 phenotypes

Uwadiae H1, Godson C1,2, Higgins D1,2

1Diabetes Complications Research Centre, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4.

2UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

Hypoxia occurs when oxygen supply is insufficient for cellular demand. Hypoxia inducible transcription factors (HIF-1α and HIF-2α) play a primary role in mediating cellular adaptation to hypoxia and inflammation through regulation of a number of genes. The HIF-α subunit binds HIF-β forming functional HIF-1 or HIF-2 transcription factors. Recent studies have shown evidence of differing roles between HIF-1 and HIF-2 in hypoxic and inflammatory process.

The aim of this study was to determine whether macrophage differentiation towards either pro-inflammatory M1 or pro-resolution M2 is influenced by HIF-1 and HIF-2, which may impact progression or regression of inflammation in renal injury. A number of strategies were employed to optimise transfection-efficiency of THP-1 monocytes and THP-1 derived macrophages with either HIF-1a or HIF-2a. Transfection-efficiency was monitored using a reporter luciferase assay for HIF-transcriptional activity. Effect of over-expression of HIF-1a and HIF-2a was determined by monitoring changes in expression of M1 marker MCP-1 and the M2 markers TGFb and IL-10 using ELISA and quantitative real time PCR analysis.

Transfection-efficiency was highest in THP-1 monocytes transfected prior to differentiation into macrophages using Fugene HD (Invitrogen) and HiPerfect (Qiagen) transfection reagents. Over-expression of HIF-1a enhanced MCP-1 mRNA expression whereas over-expression of HIF-2a increased TGF-b mRNA expression. While MCP-1 protein levels were not significantly changed with HIF-1a or HIF-2a over-expression, HIF-2a significantly increased expression of the M2 marker IL-10 in THP-1-derived macrophages.

The data support a role for the promotion of an M1 macrophage phenotype via HIF-1 mediated processes whereas HIF-2a promotes an M2 macrophage phenotype.

References:

1. Schofield, C.J. and P.J. Ratcliffe, Oxygen sensing by HIF hydroxylases. Nat Rev Mol Cell Biol, 2004. 5(5):343-54.

2. Takeda, N., et al., Differential activation and antagonistic function of HIF-{alpha} isoforms in macrophages are essential for NO homeostasis. Genes Dev, 2010. 24(5): 491-501.

Acknowledgements:

Biochemical Society Summer Vacation Studentship

Irish Health Research Board Translational Medicine Fellowship

  • Presenting Author: Mr Harmony Uwadiae
  • Supervisor: Dr Deborah Higgins
  • Co-Supervisor: Prof Catherine Godson

28. Evaluation of prostate cancer risk calculators: a prospective multi- institutional irish study

Foley RW­1, Lundon DJ1,3, O’ Brien F4, Galvin D1,2,3, Watson RW1

1UCD School of Medicine and Medical Science, UCD Conway Institute of Biomedical and Biomolecular Research, University College Dublin, Belfield, Dublin 4.

2Department of Urology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7.

3Department of Urology, St. Vincents University Hospital, Elm Park,Dublin 4.

4Department of Urology, Cork University Hospital, Cork.

The gold standard of diagnosing prostate cancer is a biopsy, determining who should go for this invasive and expensive test is difficult. Our objective was to assess the predictive accuracy of two widely used Prostate Cancer Risk Calculators, the Prostate Cancer Prevention Trial Prostate Cancer risk calculator (PCPT-RC), and the European Randomized Study of Screening for Prostate Cancer risk calculator (ERSPC-RC); to inform the need for a biopsy in an Irish cohort in order to identify the most accurate tool which will improve patients care and outcomes.

We prospectively collected the data of 1721 men who underwent TRUS biopsies in three Irish institutions. Receiver operating characteristic (ROC) curves for each calculator were generated and decision curve analysis was utilised to assess the net benefit of each calculator.

The area under the ROC curve for the ERSPC-RC and PCPT-RC was 0.643 and 0.609 respectively, this shows a significantly greater efficacy of the ERSPC-RC (p<.01). The PCPT-RC and the ERSPC-RC both demonstrate statistically significant prediction of both prostate cancer and high grade disease diagnoses in an Irish Cohort. Decision Curve Analysis demonstrates superior performance of the ERSPC-RC in this Irish cohort, and demonstrates a net benefit to the patient with a calculated risk between 45-75% for prostate cancer and 35-75% for high grade disease.

ERSPC-RC can most accurately identify those who will most benefit from a biopsy. In a time of healthcare cutbacks the introduction of the ERSPC-RC into clinical practice could help streamline access to prostate biopsies for the diagnosis of prostate cancer.

  • Presenting Author: Mr Robert Foley
  • Supervisor: Dr Dara Lundon
  • Co-Supervisor: Prof R William G Watson

29. Family-professional partnership: evaluating the effectiveness of joint parent-professional conference

Teng JA1, Lawlor A2, Rowland M1, Crotty F2

1UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

222q11 Ireland,  Carmichael House, Brunswick Street, Dublin 7. 

Family-Professional partnership is essential in providing support to individuals with Velo-Cardio-Facial Syndrome (VCFS) or 22q11.2 Deletion Syndrome who have complex medical, psychological and social needs. VCFS conferences continue this partnership with conference sessions opened to parents and professionals. A study was carried out during the 20th International Scientific Meeting of VCFS Education Foundation in Dublin, with the aim of establishing if expectations of conference attendees were met and determining the level of healthcare services available internationally.

Conference attendees were invited to complete an anonymous survey.

39 professionals and 66 families completed the survey from 12 different countries. The survey highlighted substantial changes in parental needs with education and psychosocial development as the current key issue for parents. Neither clinical information nor new research findings were the main reasons for parents and professionals attending the conference.

Dedicated VCFS clinics are available in 8 countries. There is no dedicated VCFS clinic in Ireland with families attending an average of 4 specialty clinics per year (range 1-8). In ranking services to be developed, families ranked a dedicated multidisciplinary clinic first followed by doctors with expertise in the area second.

In planning future conferences organisers should consider a more broad-based programme with more parental input. Pre-conference surveys could help tailor conference structure according to attendee requirements. It is planned to replicate this survey prior to the European conference in 2015. A well-planned joint conference contributes in strengthening parent-professional collaboration while maintaining priority emphasis on individuals with VCFS/22q11.2 Deletion Syndrome.

References:

1. ES Jeppson, J Thomas. Essential Allies:  Families as Advisors. US: Institute for Family-Centred Care; 1995.

2. CJ Dunst, I Dempsey. Family-Professional Partnerships and Parenting Competence, Confidence, and Enjoyment. International Journal of Disability, Development and Education. 2007; 54(3):305-318.

  • Presenting Author: Ms Jo-Ann Teng
  • Supervisor: Dr Fiona Crotty
  • Co-Supervisor: Ms Anne Lawlor

30. Oncotype dx testing reduces the number of patients receiving chemotherapy for invasive breast carcinoma

Dakin A1, Cotter M2,  Walshe J3, Quinn C2

1 UCD School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4.

2 Department of Histopathology, St. Vincent’s University Hospital,Elm Park, Dublin 4.

3 Department of Oncology, St. Vincent’s University Hospital,Elm Park,  Dublin 4.

Oncotype DX, a multi-gene assay, quantifies risk of disease recurrence in women with early-stage estrogen receptor (ER) positive breast cancer. Women are stratified as low, intermediate or high risk according to a Recurrence Score (RS). This result is used to predict benefit from adjuvant chemotherapy.

Our aims were to determine if Oncotype DX testing results in fewer women receiving adjuvant chemotherapy and to evaluate the concordance between RS, histopathological characteristics, Nottingham prognostic index (NPI) and Adjuvant! Online. 

220 women with ER positive, node negative, invasive breast carcinoma had Oncotype Dx testing between November 2011 & June 2013.  Tumour type was ductal (n=170), lobular (n=34) and other (n=16). Each woman was a candidate for chemotherapy based on tumour grade (grade 1: 24, grade 2: 134, grade 3: 62), size (range 10-50mm) and lymphovascular invasion (LVI) (n=79). Bivariate statistical analysis was carried out using SPSS Statistics.

Results of RS analysis were as follows – low: 138, intermediate: 61, high: 21.  Four patients were excluded. 160/216 patients (74%) avoided adjuvant chemotherapy and received hormonal therapy alone based on their Oncotype RS.  This included 50% of patients with grade 3 tumours, 61% with tumour size >20mm and 79% with LVI, characteristics previously considered to be strong indicators for chemotherapy. Tumour grade was highly predictive of RS (p < 0.001).  There was significant concordance between RS, NPI and Adjuvant!Online. RS was not significantly correlated with tumour type, size or LVI. 

Acknowledgement:

The authors would like to thank the Pathological Society of Great Britain and Ireland for funding this work.

References:

1. Paik S, Shak S, Tang G, et al: A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N Engl J Med 351: 2817-2826, 2004.

2. Paik S, Tang G, Shak S, et al: Gene Expression and Benefit of Chemotherapy in Women with Node-Negative, Estrogen Receptor-Positive Breast Cancer. J Clin Oncol 24: 3726-34, 2006

  • Presenting Author: Ms Alex Dakin
  • Supervisor: Dr Cecily Quinn
  • Co-Supervisor: Dr Maura Cotter