plasma poster competition
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PLASMA POSTER COMPETITION. All Staff and Students are invited to use the ballot papers below to vote for your favourite plasma poster. Mitochondrial DNA?. Western Blots?. Bladders?. . Prostates?. . Urine?. . Non-invasive assessment of bladder function. Jennifer Caffarel. - PowerPoint PPT PresentationTRANSCRIPT
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
Jennifer Caffarel
1/3 of menwill have
bothersome urinary symptoms
at some point in their life
… then you might have
“Bladder Outlet Obstruction”
The prostate gland surrounds the urethra
With increasing age, it increases in size and can obstruct the urethra…
Urine flow can be quantified by a technique called “uroflowmetry”.
… and can cause a slow or “weak” urine flow.
Uroflowmetry is performed in clinics, using expensive electronic flowmeters
Attending a flow clinic can be a long and tedious
experience…
… and having to fill and empty your bladder several times in a short period of time is rather
unnatural!
The data obtained from flow clinics is variable, due to a number of factors:
• Time of day
• Volume of urine in the bladder
• Emotional state of patient
The aim is to improve the diagnosis made using uroflowmetry.
For example, we tested a very basic flowmeter, to obtain more
representative results.
vs.Basic Flowmeter
Electronic Flowmeter
This is how the basic flowmeter is used:
• The patient takes the device home
• Makes multiple measurements over several days, in his own time
• This home flowmeter provided more representative results than in-clinic flows.
• And 2 patients who could not urinate in the clinic were able to use this device.
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
Questions (and some answers) on
research into stomach cancer…
Claire Worrall
??
? ??
?
?
???
?
? ?
?
???
?
Who
It is difficult to predict.
It depends on: If you have H. pylori infection
What you eat If you have stomach ulcers
How much alcohol you drink If you smoke
Your Body Mass Index What country you live in Your genetic background
Your gender
gets stomach cancer?
So biological markers of early disease would be very useful.
Whatchanges occur in stomach cancer?
Expression levels of many proteins are altered…
We are studying one protein whose expression is lost in the majority of stomach cancers:
TFIZ1 is expressed in mucin-secreting cells and secreted into the mucous layer in the stomach.
It is virtually undetectable by RT-PCR in our panel of 7 gastric cancer cell lines.
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
70.00%
80.00%
90.00%
100.00%
Normalstomach
Stomachcancer
% of casesexpressing
TFIZ1
0 %
100 %
AGS NUGC3 MKN74 KATO III SNU1 SNU5 SNU16
% of normalTFIZ1
expression
0 %
0.005 %
AG
S
NU
GC
3
MK
N7
4
KA
TO
III
SN
U 1
SN
U 5
SN
U 1
6
Wherecan we study stomach cancer?
These behaviours in patient cancer cells help determine tumour growth and the ability to metastasize.
There is limited supply of patient samples.
However, we can grow stomach cancer cells in the lab to perform tests on.
These cells grow and divide, allowing us to measure the effects of different conditions on:How fast they grow and divideHow easily they dieHow quickly they move
Whendo we study a protein in stomach cancer?
When we have evidence linking it to stomach cancer…
TFIZ1 is bound in the stomach to TFF1, which has many links to cancer:
TFIZ1TFF1High expression
in oestrogen responsivebreast cancer
Binds to H. pylori
TFF1
Involved inulcer repair30% null mice get
stomach cancer
TFF1
Howare we studying this protein?
By introducing controllable TFIZ1 expression into stomach cancer cells:
pTet-Off
neoR tTA
tTAneoR
TRE TFIZ1pTRE-Tight TFIZ1
TFIZ1
TRE
TFIZ1Off
TFIZ1On
1. Transfect inregulator plasmid
2. Transfect inTFIZ1 plasmid
Stomach cancerCell line
Addingdoxycycline
turns off TFIZ1expression
The cells nowexpress TFIZ1
Whydo we study a protein in gastric cancer?
If we know
WHY the expression changesWHAT the protein does
In the normal stomachIn stomach cancer
WHEN the expression changes
It can
Help us decide whether it might be a good early indicator of cancer.
Help us decide whether it might be a good therapeutic target.
Acknowledgements:
Felicity May
Herbie Newell
Northern Institute for Cancer Research
Claire Worrall
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
AIM:
To develop an in vitro model system of radiogenic breat cancer, so that the molecular genetic events associated with transformation and the development of a radio-resistant phenotype can be investigated.
INTRODUCTION:
Chemotherapy and radiotherapy- induced second cancers are the leading cause of death in patients cured of Hodgkin Lymphoma (cancer of the lymphatic system). Young women treated for Hodgkin Lymphoma with radiotherapy at the chest are at a high risk of developing breast cancer
PROCEDURE:
1. This project will involve exposing transformed (MCF-7) and non-transformed (MCF-10) breast cell lines to increasing doses of ionising radiation.
2. Post treated cell lines will be compared to untreated parental cells across the whole genome to identify regions characterised by loss of heterozygosity, copy number alterations and uniparental disomy.
3. Candidate locations of interest will be further investigated in treated cell lines and in breast tissue from patients with breast cancer following radiotherapy for Hodgkin Lymphoma.
FUNDED BY: Northern Institute for Cancer Research
‘MRI scan of the breast reveals two discrete areas of abnormality, which proved to be cancer.’
http://www.breastcancer.org/pictures/diagnosis/mammogram/mri_2.jsp
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
Forkhead Proteins & Cancer
Frances Purtill
iCAMB
Forkhead Proteins & Cancer
Unregulated cell proliferation is a characteristic of all cancers
Understanding the mechanisms behind cell proliferation will help in
the treatment and prevention of cancer
Forkhead Proteins & Cancer
M
G2 G1
S
M Phase (Mitosis)Replicated DNA is equally distributed
into 2 cells
Cell proliferation is controlled by a series of events known as the Cell Cycle
1
2
The cell cycle consists of 4 phases:
G1 and G2 are ‘gap’ phases
which prepare the cell for its
next step
3 & 4
Genetic material is copied and 2 identical cells are formed
S PhaseDNA is replicated
Forkhead Proteins & CancerForkhead Transcription Factors (FKH-TF) are critical for the regulation of the cell cycle
TARGET GENE
GENE SWITCHED ONTARGET PROTEIN MADE
FKH-TF
Forkhead proteins have been associated with various human
cancers
They bind upstream of genes involved in mitosis, and activate their expression
Forkhead Proteins & CancerForkhead proteins are vital for mitosis in all eukaryotes, from yeast to man
Yeast is easy to manipulate, genetically and biochemically
Therefore it is a valuable model to study forkhead proteins, and improve our knowledge of the
cell cycle in humans
In the future, this knowledge could potentially be used to
improve existing cancer therapies
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
The Development of Small Molecule
Inhibitors of the MDM2-p53 Interaction
Junfeng Liu
p53 is a Tumour Suppressor
p53+/+
p53+/-
p53-/-
1% at 18 months
% mice with tumour
74% at 6 months
2% at 9 months
Donehower et al. Nature, 1992Donehower et al. Nature, 1992Also suggest to read Donehower et al. Nature genetics, 1993Also suggest to read Donehower et al. Nature genetics, 1993
MDM2 controls p53 through an auto- regulatory negative-feedback loop
p53
MDM2
mdm2 mRNA
mRNA of other targets
p53 MDM2
p53’s proteasomal degradation
Three mechanism to repress p53 by activation of MDM2-expression:
1. MDM2 binds p53 at transactivation domain and blocks its ability to activate transcription
2. MDM2 acts as an E3 ubiquitin ligase that promotes p53’s proteasomal degradation
3. MDM2 is involved in the nuclear export of p53
Breaking the negative-feedback loop with antagonists
Therapeutic potential Rescue of p53 Function by Disrupting the p53–MDM2 Interaction.
Blocking MDM2 Expression
Inhibiting MDM2 Ubiquitin Ligase Activity
Disruption of the p53–MDM2 binding
p53 p53 MDM2
MDM2
mdm2 mRNA
mRNA of other targets
p53’s proteasomal degradation
Possible Methods
Disruption of the p53–MDM2 Binding
Interaction Antibody microinjection
Peptide analogues corresponding to the MDM2
binding domain of p53
Small molecular weight compounds
The p53-MDM2 interaction: Unique binding site
Unique
& Good for Small Molecule Drug
Design and Development
MDM2 protein and p53 peptide structures come from Protein Data Bank.
PyMOL Software were applied for the followed cartoons.
Structure based drug design
MDM2 Binding Domain(Binding area high lighted)
Structure based drug design
8mer p53 peptide Binding with MDM2
Structure based drug design
Three key residues of p53 in the hydrophobic pocket of MDM2
Structure based drug design
Three key residues of p53 were picked out for drug design
Structure based drug design
Drug design based on the three key residues of p53
Structure based drug design
Designed small molecule scaffold
Structure based drug design
Manual docking of the compound with MDM2 binding domain
Inhibitor Screen Using ELISA
ECL detection
DECREASED LIGHT
YY
M
Incubation of MDM2with antagonist
M M
M
M
M
ECL detection
DECREASED LIGHT
YY
M
Incubation of MDM2with antagonist
M M
M
M
M
ECL detection
LIGHT
Incubation withMDM2
M
M M
MM
YYYY Y Y
BiotinylatedIP3 Peptide
Streptavidin
M M M
YHRP conjugatedsecondary antibody
YAddition of: MDM2 primary antibody,
Cellular activity Confirmation via Western Blotting Assay
Effect of MDM2 inhibitors on the cellular levels of p53, MDM2 and p21
Nutlin-3 NU8XXX
MDM2
p53
p21
Actin
DMSO 0.5 1 10 1 10 20 M
Thank you
for
your attention!
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
…and tools to diagnose Lower Urinary Tract Dysfunction
having to go in the middle of the night
inability to pass any water
1 in 3 men will suffer urinary symptoms in their life:
struggling to begin peeing
having to rush to the toilet
having to go often
pain while peeing
Poor stream
‘Terminal dribble’
Incontinence
Incomplete emptying
To diagnose the cause we need to measure:
•Bladder pressure•Urine flow rate•Volume of urine voided
… among other tests
Well, how do you take a blood pressure measurement?
How do you take a bladder pressure measurement?
Use the same principle…
…and measure the pressure to stop urine flow!
Place a small, inflatable cuff around the penis…
Pressure
Flow rate
Volume
Flow is interrupted
Pressure is measured
We can also use it to measure the bladder characteristics in healthy people to further understand the mechanics of the lower urinary tract.
Conventionally, pressure- flow measurements would require urethral catheterisation, an uncomfortable procedure with risk of infection.
Penile Cuff Machine
We can now use this non-invasive machine clinically to diagnose Bladder Outlet Obstruction in patients.
With this tool we hope to:
•Verify theoretical bladder models
•Understand better how the bladder copes with obstruction
•Apply this to diagnosis and treatment of patients
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
Heart Physiology: “The role of p50”
Silvia Gaspar PereiraPhD student with the Cell Signaling Group
Institute of Cellular Medicine
Light from the Heart Nebula - apod.nasa.gov
Heart Physiology: “The role of p50”
What does p50 do?
When the cell receives a signal, the protein p50 is released from the cytoplasm into the nucleus.
In the nucleus, p50 binds DNA and thereby induces a cellular response to the signal.
In the absence of p50, this signaling pathway is impaired.
Aim: Is p50-dependent Aim: Is p50-dependent signaling important in signaling important in heart muscle cells?heart muscle cells?
p50
p50
Heart Physiology: “The role of p50”
To answer this question, we use mice lacking p50:
In mice without p50, hearts are significantly bigger than in normal mice.
The p50-deficient hearts are more sensitive to certain injuries, e.g. lack of oxygenation in the tissue.
p50
p50
Future Work:
Does p50 help to prevent injury in the heart tissue?
Is p50 necessary during heart formation pre- or postnatally?
If p50 is directly involved in these events, it could become If p50 is directly involved in these events, it could become a potential target for heart disease treatments!a potential target for heart disease treatments!
PLASMA POSTER COMPETITION
All Staff and Students are invited to use the ballot papers below to vote for
your favourite plasma poster
Regulation of the oxidative stress response in the pathogenic fungi
Candida albicans
Miranda PattersonSupervisor – Dr Janet Quinn
http://www.chuv.ch/imul/euresfun
Candida albicans
Candida albicans is the major systemic pathogenic fungi of humansIn immunocompetent people C.albicans is the
cause of common superficial infections such as oral and vaginal thrush
But, in immunocompromised people, such as people with HIV/AIDS, some cancer patients or transplant patients, C.albicans can cause life threatening systemic infections, with ≈ a
38% mortality rate
C.albicans invasion of blood vessels in the oesophagus
C.albicans invasion of a kidney from an immunocompromised rabbit
http://pathmicro.med.sc.edu/mycology/mycology-3.htm
http://pathmicro.med.sc.edu/mycology/mycology-3.htm
Why are we interested in C.albicans oxidative stress responses?
Macrophages and neutrophils are cells of the innate immune system responsible for killing
micro-organisms
Macrophage
C.albicans
C.albicans cells are ingested by the
macrophage
After ingestion C.albicans starts growing hyphae
and escape the macrophage
MacrophageC.albicans hyphae
Neutrophils are important in killing C.albicans
Neutrophils kill by producing toxic reactive oxygen species in a respiratory burst
C.albicans lacking key proteins involved in oxidative stress responses are more sensitive to
killing by neutrophils and have reduced virulence
Full understanding of mechanisms used by C.albicans to overcome oxidative stresses
may reveal novel therapeutic targets!www.biology.uark.edu/dmcnabb/dmcnabb.htmlwww.biology.uark.edu/dmcnabb/dmcnabb.html
But C.albicans can kill macrophages!
What are we doing?
Nucleus
Cap1
Oxidative Stress
Anti-oxidant genes
Previous research shows that following oxidative stress, Cap1 translocates to the nucleus and upregulates transcription of anti – oxidant genes so that cells can survive oxidative stress.
We want to know what is regulating Cap1?
Investigating the major oxidative stress pathway in
C.albicans – the Cap1 pathway
-ive
How are we doing this?
Deleting potential key regulators of Cap1 and seeing how cells survive an oxidative
stress challenge
Control
H2O2 concentration (mM)
Wild type strainCap1 delete strainYbp1 delete strain
Wild type cells grow until high concentrations of H2O2
Cells lacking Cap1 are sensitive to H202 and can not grow even at low concentrations
Cells lacking the potential regulator of Cap1, Ybp1, are also sensitive to H202 but only at high concentrations
Have we found a regulator of Cap1 at high levels of H202 oxidative challenge?
We grow C.albicans in the presence of hydrogen peroxide (H2O2) to induce oxidative stress
Continuing research will hopefully answer this question!