Clinical and Academic Research Society Inaugural
Research Presentation Evening
Friday 16th November
Lecture Theatre 2, New Hunt’s House, Guy’s Campus
Agenda
6pm Welcome
Valerie Crolley
IT Director, CARS
Professor Anne Greenough
Head of the King's College London School of Medicine & Director of Education &
Training, King’s Health Partners
6.05pm Keynote lecture
Professor Sir Robert Lechler
Vice-Principal (Health) King’s College London and Executive Director of King’s
Health Partners Academic Health Sciences Centre
6.35pm Abstract Presentations and Panel Questions
Order overleaf
8.00pm Panel Decision & Announcement
Panel members – Professor Lechler & Professor Rymer
8.05pm Drinks reception in New Hunt’s House foyer
Abstract presentations – 6.35pm – 8pm
1
Kiran Patel
In vivo screening of p38MAPK/MEK inhibitors in zebrafish
to assess their role in cardiovascular function and
development
2
Karen Bosch
Visualising neuronal plasticity using viral vectors
3
Shobit Saxena
Targeting the αvβ6 integrin in head and neck cancer
using genetically-engineered T-cells
4
Grace Grove
The impact of increased BMI on the length of stay of
women on antenatal and postnatal wards.
5
Elisabeth Howe
Is reaction time in the auditory "oddball" paradigm an
endophenotype of schizophrenia?
6
Attiya A Khan
Vitamin d, b12 and folate supplementation in diet treated
phenylketonuria need revision
7
Nada Anzak
Towards regeneration of the optic nerve: an evaluation of
the role of IGFBPL1 on RGC survival and axon growth
using an igfbpl1 knock-out mouse line.
8
Naeem Mitha
Ultrastructural localisation of nano-sized air pollutants in
human platelets by Transmission Electron Microscopy
9
Cosima Gretton
Art and the brain: a view from dementia
10
Balrik Kailey
SSTR2 is the functionally dominant somatostatin receptor
in human pancreatic β- and α-cells
11
Emma Russell-Jones
The novel use of commonly captured data to assess a
district's diabetes service that evaluates both primary and
secondary care
12
Abirami Namasivayam
Effective measurement of Proteinuria in women with
suspected Pre-Eclampsia: An Evaluation of clinical
outcome and health economic usage of women with
suspected pre-eclampsia, with and without proteinuria
Name of Author: Kiran Patel
Title of Abstract: In vivo screening of p38MAPK/MEK inhibitors in zebrafish to assess their role
in cardiovascular function and development
Text of Abstract: Introduction
There is a paucity of information regarding phenotypic responses of zebrafish embryos to MAPK
inhibitors, and if they can rescue the heart failure phenotype. A chemical screen was conducted to
address these questions.
Methods
Normal (Ekwill) zebrafish embryos were incubated with five p38MAPK and MEK1/2 inhibitors at 2hpf
and 24hpf. Phenotypic characterisation was undertaken at 24hpf, 48hpf and 72hpf as appropriate.
Dose response curves were generated and GATA4 and nppb expression was investigated using realtime qPCR. Video imaging was used to collect data on haemodynamic variables to assess the effect
of the inhibitors on cardiovascular function. Finally, an assay of nppb:luc transgenic embryos was
conducted to assess the potential of the inhibitors to rescue the heart failure phenotype.
Results
p38MAPK and MEK1/2 inhibitors produce phenotypes that are dose- and developmental stagedependent. Incubation at 2hpf vs 24hpf has important implications for the phenotype, its severity, and
the number of embryos with the phenotype. p38MAPK inhibitors produced a clustering of cardiac
phenotypes, largely comprising of pericardial oedema, absent red blood cells in the heart and valvular
regurgitation. MEK1/2 inhibitors produced a vascular and distinctive tail phenotype. These become
manifest within the first 24 hours of incubation and before 48hpf. A temporal variation in GATA4 and
nppb gene expression was noted. The tested inhibitors did not rescue to the heart failure phenotype.
Conclusions
P38MAPK and ERK signaling pathways are implicated in early cardiovascular development in the
zebrafish. Further objective quantitative data is required to assess this based on the trends presented
here.
Name of Author: Karen Bosch
Title of Abstract: Visualising neuronal plasticity using viral vectors
Text of Abstract: Following spinal cord injury (SCI) most patients undergo a degree of spontaneous
recovery believed to be due to reorganisation of neuronal circuits residing in spared neural tissue.
Many treatment strategies of experimental spinal injuries appear to promote similar plasticity. Mostly,
anatomical tracers, such as biotin dextran amines, have been used to study this phenomenon, but
these methods show axonal projections, not synaptic connections. We have developed a viral vector
expressing a genetically encoded fluorescent marker of presynaptic terminals for use as an
anatomical marker of the number and distribution of synapses in vivo. Selective expression of
synaptopHluorin (SpH) was achieved with an adeno-associated viral vector expressing SpH (AAVSpH) in peripheral and central neurons. We delivered the vector to the right sensory-motor cortex of
adult rats and achieved substantial transduction of the corticospinal tract (CST) four weeks later, seen
as diffuse labelling in the internal capsule, pyramids and spinal CST. The number of axons in the CST
in the dorsal columns at C4 averaged 2701+/-436. In addition SpH accumulated in large numbers of
bright puncta, widely taken as a measure of presynaptic specialisations. 1956+/-443 GFP-labelled
synapses were observed in each transverse section of the grey matter of the cervical enlargement.
5.3+/-1.4% of these was contralateral to the injection site. Delivery of AAV-SpH to the sciatic nerves
of naive animals led to transduction of a small number (1-5%) of primary afferent neurons, visible as
diffuse fluorescence in lumbar dorsal root ganglia. Sensory neuron synaptic puncta were visualised in
the dorsal horn of the spinal cord, providing a quatifiable method to document changes in synapse
number. This novel tool provides the opportunity to study synaptic number in a particular tract or
system and therefore measure synaptogenesis after neuronal injury and therapeutic interventions.
Name of Author: Shobhit A Saxena
Title of Abstract: Targeting the αvβ6 integrin in head and neck cancer using geneticallyengineered T-cells
Text of Abstract: Background: The 5-year survival rate for squamous cell carcinoma of the head and
neck (SCCHN) has remained unchanged for the past 30 years, at 50-60%. Two thirds of patients
present with locally advanced disease and only 40% of these are cured. αvβ6 is a cell-surface integrin
molecule expressed at low levels in normal adult epithelia. Overexpression of αvβ6 is seen in SCCHN
and many other cancers, where it is implicated in more aggressive disease. It therefore qualifies as a
novel candidate for targeted immunotherapy. This project describes the testing of T-cells genetically
engineered to target the αvβ6 integrin by expressing chimeric antigen receptors (CARs); CARs are
fusion receptors which couple HLA-independent antigen recognition to T-cell activation.
Aims: To determine the in vitro functionality of an αvβ6-targeting CAR against SCCHN tumour cell
lines.
Results: Human T-cells from healthy donors were successfully transduced to express CARs targeting
αvβ6 and a negative control CAR. CAR functionality was assessed by co-culturing T-cells with
SCCHN tumour monolayers. T-cells expressing the αvβ6-targeting CAR displayed antigen-specific
cytotoxicity and monolayer destruction on exposure to the αvβ6+ target monolayer and released large
amounts of IL-2 (2.66 ng/ml) and IFN- ɤ (178.89 ng/ml) in a strictly antigen-dependent manner. Third,
αvβ6-targeted T-cells underwent substantial enrichment following target engagement. These results
were not replicated in negative control co-cultures.
Conclusion: Our proof-of-principle data shows that αvβ6 can be potentially exploited for therapeutic
purposes; the CAR-grafted T-cells displayed potent effector functions in vitro in response to
engagement with SCCHN tumour cell lines expressing αvβ6.
Name of Author: Grace Grove
Title of Abstract: The impact of increased BMI on the length of stay of women on antenatal and
postnatal wards
Text of Abstract: Background: In 2010 WHO estimated that 63.8% of females in the UK were
overweight. Obesity has been associated with adverse effects during pregnancy, potentially leading to
increased length of stay (LOS). In an increasingly cost conscious world obesity is a pressing issue for
healthcare providers and maternity services are no exception.
Aims: To identify whether BMI had an impact on length of stay on antenatal and postnatal wards.
Methods: Data was collected at St Thomas’ Hospital, London. Information on BMI, age, gestation at
admission and LOS was collected from all the women with BMI >19 admitted to the antenatal ward
in October 2011. Information on BMI, age, birth weight of their infant and LOS was collected from a
randomly selected group of 50 women with normal BMI and 50 women of BMI >25 admitted to the
postnatal ward in October 2011.
Results: On the antenatal ward high BMI women stayed for an average of 2.41 days longer than
normal weight women. The total cost of beds for the 59 high BMI women was £115,656.00, whilst the
total cost of beds for the 56 normal BMI women was £66,992.00, a difference of £48,664.00. There
were no significant differences in LOS in the postnatal groups.
Conclusions: An increased LOS in high BMI women on the antenatal ward had substantial financial
implications for the maternity unit. Further research is necessary to assess other potential areas of
increased cost, such as more frequent obstetric appointments, scans, medication and provision of
specialist equipment.
Name of Author: Elisabetta Howe
Title of Abstract: Is reaction time in the auditory "oddball" paradigm an endophenotype of
schizophrenia?
Text of Abstract: Background: This study sought to investigate if reaction time (RT) is increased in
patients and their first-degree relatives compared to controls, and therefore, if it is a possible
endophenotype of schizophrenia. This study also sought to extend previous findings regarding the
P300 by assessing if P300 latency is associated with RT.
Method: EEG and RT recordings were taken from 68 patients with a diagnosis of schizophrenia, 62 of
their unaffected first-degree relatives, and 67 unrelated healthy controls, during the auditory “oddball”
paradigm. Mean RT was compared between groups, and correlations between P300 latency and RT
were assessed. All analyses were adjusted by age and sex.
Results: Patients’ mean RT was significantly slower than that of controls (p=0.002). Relatives
demonstrated a mean RT that was intermediate between that of patients and controls. In a combined
analysis of the whole sample adjusted by age and clinical group, RT was significantly correlated with
P300 latency (p≤0.001).
Discussion: RT may be an endophenotype of schizophrenia, as patients showed significant deviance
in RT compared to controls; and although the effect was non-significant, relatives showed a degree of
deviation in RT that may be present at a behavioural level. Genes underlying RT may also influence
P300 latency, as the two variables were highly correlated, and similar neurocognitive mechanisms are
thought to be reflected in both. If RT is confirmed to be an endophenotype, then it may benefit future
research, as it is a quick and cost-effective way of gathering data, and it can be used in conjunction
with P300 indices to further understanding of the genetic risk factors for schizophrenia.
Name of Author: Khan AU 1, Liu A 1, Walker R2, Ramachandran R2, Rahman Y2
Title of Abstract: Vitamin D, B12 and Folate Supplementation in Diet Treated Phenylketonuria
need Revision
Text of Abstract: Introduction: Phenylketonuria (PKU), an autosomal recessive metabolic disorder, is
characterised by a mutation in the gene for Phenylalanine Hydroxylase, a vital enzyme to convert
phenylalanine to tyrosine. Excess phenylalanine is neurotoxic to a developing brain. Dietary protein is
the main source of phenylalanine, and a protein-restricted diet is the cornerstone of PKU treatment.
Without adequate supplementation, dietary natural protein restriction may result in deficiency of
vitamins and essential minerals. These supplements are often the only source of vitamin D, B12 and
folate.
Aim: To evaluate the efficacy of current recommended doses of supplementation.
Method: The most recent serum vitamin D, B12 and folate in diet-restricted PKU patients (n=52)
attending a single adult Inherited Metabolic Diseases centre, were retrospectively analysed. Vitamin D
> 40 nmol/L, B12 > 400ng/L, and folate > 4ng/L were defined as sufficient.
Results: 36% of patients (n=13, 8 female, 5 male) were vitamin D deficient (median 52 nmol/L, range
12-99nmol/L). 48% (n=25, 18 female, 7 male) were vitamin B12 deficient (median 415, range [641254]). 6% (n=3, 2 female, 1 male) were deficient in folate (median 13.5ng/L, range 2.4>20.0ng/L.).
Conclusions: While most patients were folate replete, a significant number of patients are vitamin B12
or vitamin D deficient or both. These results suggest current recommended supplementation doses of
these vitamins in PKU patients may be inadequate. Poor absorption of the currently supplemented
form of the vitamins may explain the result in our cohort. Annual patient monitoring and further studies
with different form of supplementations are needed.
Name of Author: Nada Anzak
Title of Abstract: Towards regeneration of the optic nerve: an evaluation of the role of IGFBPL1
on RGC survival and axon growth using an igfbpl1 knock-out mouse line
Text of Abstract: Retinal ganglion cell (RGC) degeneration is a feature of many of the world’s leading
causes of blindness, including conditions such as glaucoma. The intrinsic regenerative capacity of
RGCs is lost during maturation of the mammalian retina. A novel protein, IGF1 binding protein-like 1
(IGFBPL1) may participate in this developmental regulation. The effect of IGFBPL1 on RGC axon
elongation was evaluated through culturing post-natal day 0 (P0) murine RGCs for 3 days and
immunostaining with β-III tubulin, both in a control medium, and in the presence of IGFBPL1. The
longest neurite, assumed to form the axon, was measured for each RGC, using Image J software.
The mean length of the longest neurite was found to be significantly greater in RGCs cultured with
IGFBPL1 than in the control medium(p<0.001). This effect was further evaluated using an igfbpl1
knock-out (KO) mouse line. RGCs were collected from P0 mice of homozygous KOs, heterozygous
KOs and wild-type (WT) genotypes. A significant reduction in neurite length was observed in both
homozygous and heterozygous igfbpl1 KOs compared to WT controls (p<0.001). The effect of
IGFBPL1 on RGC survival was also evaluated by performing a LIVE/DEAD© assay on RGCs isolated
from homozygous and heterozygous igfbpl1 KOs and WT controls. A significant reduction in RGC
survival was observed in homozygous igfbpl1 KOs (p<0.01), with a modest reduction also observed
in heterozygous KOs. We conclude that IGFBPL1 significantly promotes both RGC survival and
neurite outgrowth. It is hoped this finding may inform novel strategies towards optic nerve
regeneration.
Name of Author: Naeem Mitha
Title of Abstract: Ultrastructural localisation of nano-sized air pollutants in human platelets by
Transmission Electron Microscopy
Text of Abstract: Background: Epidemiological studies have clearly demonstrated that traffic derived
air pollution (particulate matter) is an important risk factor in lung disease, and there is increasing
evidence between the association of high levels of ambient particles and the incidence of platelet
driven cardiovascular diseases. There is evidence that combustion-derived nanoparticles, namely
diesel exhaust and carbon black particles, prevalent in the ambient UK atmosphere exert their effects
through toxicological and inflammatory processes as well as translocation into the systemic circulation
where they may interact with platelets. Therefore, this project focuses on investigating isolated human
platelet behaviour towards nanoparticle exposure in vitro using transmission electron microscopy.
Methods: The interaction between DEP and carbon black (CB) and platelets was examined by
transmission electron microscopy and the functional consequences of exposure assessed through a
cytotoxicity assay. Results: Platelets treated with 50µg/ml DEP and CB particles were shown to
interact with platelets through internalisation, although the precise ultrastructural location of these
nanoparticles as well as the uptake process remains unclear. Furthermore, this study reveals that
chronic exposure to DEP (12µg/ml) elicited a significantly higher level of platelet cytotoxicity than that
conferred by thrombin. Conclusion: The study fundamentally demonstrates that DEP and CB
pathologically interact with human platelets and provides a potential mechanism for the increased
thrombotic risk associated with exposure to ambient pollution.
Name of Author: Cosima Gretton
Title of Abstract: Art and the brain: a view from dementia
Text of Abstract: Art making encompasses a range of perceptual and cognitive functions involving
widely distributed brain systems. The dementias impact on these systems in different ways, raising
the possibility that each dementia has a unique artistic signature. Here we use a review of the visual
art of 14 artists with dementia (5 Alzheimer’s disease, 7 fronto-temporal dementia and 2 Dementia
with Lewy bodies) to further our understanding of the neurobiological constituents of art production
and higher artist function. Artists with Alzheimer’s disease had prominent changes in spatial aspects
of their art and attributes of colour and contrast. These qualities were preserved in the art of frontotemporal dementia which was characterised by perseverative themes and a shift towards realistic
representation. The art of dementia with Lewy Bodies was characterised by pronounced changes in
visual perceptual attributes and bizarre content linked to visual hallucinations. The impact of dementia
on wider aspects of art production is discussed and a novel classificatory scheme presented to help
characterise neural mechanisms of higher artistic functions in future studies.
Name of Author: Balrik Kailey
Title of Abstract: SSTR2 is the functionally dominant somatostatin receptor in human pancreatic
β- and α-cells
Text of Abstract: Introduction:
Somatostatin-14 (SST) inhibits insulin and glucagon secretion by activating G-protein coupled
somatostatin receptors (SSTRs), of which 5 isoforms exist (SSTR1-5). In mice, the effects on
pancreatic β-cells are mediated by SSTR5, whereas α-cells express SSTR2. In both cell types, SSTR
activation results in membrane hyperpolarisation and suppression of exocytosis. Here we examined
the mechanisms by which SST inhibits secretion from human β- and α-cells, and the SSTR isoforms
mediating these effects.
Methods and Results:
Quantitative PCR revealed high expression of SSTR2, with lower levels of SSTR1, SSTR3 and
SSTR5, in human islets. Immunohistochemistry showed expression of SSTR2 in both β- and α-cells.
SST application hyperpolarized human β-cells and inhibited action potential firing. The membrane
hyperpolarization was unaffected by tolbutamide but antagonized by tertiapin-Q, a blocker of Gprotein gated inwardly-rectifying K+-channels (GIRK).
The effect of SST was mimicked by an SSTR2-selective agonist while a SSTR5 agonist was
marginally effective. SST strongly (>70%) reduced depolarization-evoked exocytosis in both β- and
α-cells. A slightly weaker inhibition was observed in both cell types after SSTR2 activation. SSTR3and SSTR1-selective agonists moderately reduced the exocytotic responses in β- and α-cells,
respectively, whereas SSTR4- and SSTR5-specific agonists were ineffective.
SST also reduced voltage-gated P/Q-type Ca2+-currents in β-cells, but normalization of Ca2+ influx to
control levels by prolonged depolarizations only partially restored exocytosis.
Discussion:
We conclude that SST inhibits secretion from both human β- and α-cells by activating GIRK and
suppressing electrical activity, reducing P/Q-type Ca2+-currents and directly inhibiting exocytosis.
These effects are predominantly mediated by SSTR2 in both cell types.
Name of Author: Emma Russell-Jones
Title of Abstract: The novel use of commonly captured data to assess a district's diabetes
service that evaluates both primary and secondary care
Text of Abstract: Aims: To identify commonly captured data in the United Kingdom to look at the
performance of a district’s diabetes care that encompasses both primary and secondary care.
Methods: Data collected from primary-care quality outcomes framework (QOF) measures for
diabetes, standardised admission rates (SAR) for first appointment to OPD hospital specialists and
emergency admission rates for diabetes (Dr Foster/HES) were used to produce a new performance
index scoring system. Illustrative measures from QOF were total diabetes points, DM6 attainment of
HbA1c<7% (53mmol/mol) and its exemption-rate (number of patients excluded from analysis).
The performance index was used to study the performance of the ‘Medway’ district diabetes service,
and this was compared with other Primary Care Trusts within the Strategic Health Authority (SHA)
and Primary Care Trusts nationally
Results: Medway has the highest prevalence of Diabetes of the 8 Primary Care Trusts within the
SHA, lowest achievement of diabetes QOF points (96.1%) and lowest achievement of an HbA1c
level<7% (54.3%). Exemption reporting was 3rd highest. SAR for first diabetes OPD appointment to
the hospital was low at 281 (predicted 576) 48% of expected. The emergency-admission-rate was
high at 225 (predicted 168) 133% of expected. Thus primary-care diabetes needs to raise
performance and implement a lower threshold for OPD referral to prevent emergency-admissions.
Conclusion: It is possible to produce an assessment of diabetes care that transcends
primary/secondary-care that gives a true reflection of a district’s performance which will be useful to
plan future health service provision.
Name of Author: Abirami Namasivayam
Title of Abstract: Effective measurement of Proteinuria in women with suspected Pre-Eclampsia:
An Evaluation of clinical outcome and health economic usage of women with suspected preeclampsia, with and without proteinuria
Text of Abstract: Background: Pre-eclampsia is a leading cause of significant maternal and fetal
morbidity and mortality. Today in the UK, it is diagnosed as de novo hypertension and proteinuria.
Since diagnosis guides management of the disease, women with suspected pre-eclampsia are likely
to increase the use of health services with associated economic burden. The accuracy of proteinuria
as a diagnostic criterion and predictor of adverse complications has been widely debated. One can
question the use of proteinuria in determining management of patients with regards to accurate
diagnosis, safety and health economic costs.
Objective: To compare and evaluate the clinical outcome and health economic usage of women with
suspected pre-eclampsia, with and without proteinuria.
Method: A prospective observational study was conducted in St Thomas Hospital, Antenatal Day Unit.
Handheld notes from 100 women with suspected pre-eclampsia were collected over 10 weeks. Basic
demographics, clinical outcome and health economic usage of proteinuric women (57%) and nonproteinuric women (43%) were compared and analysed.
Results: A larger proportion of proteinuric women were either overweight, Black, or had a past
medical history. With regards clinical outcomes, proteinuric women were associated with operative
deliveries, preterm deliveries, small for gestational age babies and longer hospital stays and perinatal
deaths; they also required greater utilisation of health services compared to non-proteinuric women
and were therefore more costly.
Conclusion: Since proteinuria is currently used as a diagnostic marker, it influences the management
of women with suspected pre-eclampsia. Although its use as a diagnostic tool and indicator of
complications is debated, our results illustrate that proteinuria is associated with clinical complications
and a subsequent increase in health economic usage compared to non-proteinuric women. One could
postulate that proteinuria is a reflection of multiple organ damage and therefore identifies preeclamptic women who are more at risk of adverse outcomes. This implies that proteinuria should be
used as predictive indicator pre-eclampsia rather than a hallmark of the disease. It is apparent that
alternative markers which accurately predict pre-eclampsia and its associated complications are yet to
be established.