Oral Presentations
Integration of Metabolism and Survival
OP-1
The ankyrin repeat and SOCS box-containing
protein Asb-9 targets creatine kinase B for
degradation
M. A. Debrincat
1
, J. G. Zhang
2
, T. A. Willson
3
, B. T. Kile
3
,
S. L. Masters
2
, L. M. Connolly
4
, R. J. Simpson
4
, H. M. Martin
2
,
N. A. Nicola
2
and D. J. Hilton
3
1
Cancer and Haematology/Molecular Medicine, The Walter
and Eliza Hall Institute of Medical Research, Melbourne,

Asb-9. Transfection of increasing concentrations of a tagged
Asb-9 construct into 293T cells increased the polyubiquitination
of CKB and resulted in a concomitant decrease in total CKB
levels within the cell. The targeting of CKB for degradation by
Asb-9 was entirely SOCS box-dependent. The interaction has
been confirmed in vivo and suggests that Asb-9 may act as a
specific ubiquitin ligase-regulating CKB abundance.
OP-2
Signal transduction of cytokinine
M. Gilmanov
1
, S. Ibragimova
1
, Sv. Sadykova
1
, Zh. Basygaraev
1
and A. Sabitov
2
1
Laboratory of Enzyme Structure and Regulation, Aithozhin’s
Institute of Molecular Biology and Biochemistry,
2
Department of
Chemistry, Al-Faraby’s Kazakn National University.
E-mail:
The cytokinine the most important phytohormone, which is con-
trolling the division of the plant, cells. We carried out the investi-
gation of cytokinine signal transduction. We were discovered the
most important participant of cytokinine signal transduction.

regulation of polypeptide synthesis in skeletal muscle. Here, the
effect of contractions on skeletal muscle eukaryotic elongation
factor 2 (eEF2) phosphorylation and eEF2 kinase activity was
investigated. In response to contractions in situ, there was a rapid
(i.e. 15s) fivefold increase in eEF2 phosphorylation at Thr56 in
the contracting gastrocnemius muscle of rats that was maintained
at this level during 30 min of contractions, with no change in the
non-stimulated contralateral muscle. Furthermore, eEF2 phos-
phorylation was higher in both soleus and extensor digitorum
longus muscles of mice when contracted ex vivo, indicating that
the mechanism behind this increase is related to local factors. No
change in in vitro eEF2 kinase activity was observed in the con-
tracted rat muscles at any time-point or when measured at pH
6.8 versus 7.2. Furthermore, the increase in eEF2 phosphoryla-
tion occurred at a time before any change in AMPK activity was
observed and was normal in contracting muscles of mice expres-
sing non-functional AMPK. However, Ca
2+
-calmodulin potently
increased the activity of skeletal muscle eEF2 kinase when meas-
ured in vitro. Taken together, these data indicate that the inhibi-
tion of protein synthesis in contracting muscle may arise from
phosphorylation of eEF2 via a Ca
2+
-calmodulin-eEF2 kinase
cascade.
Abstracts
42
Integration of Defence and Survival
OP-4

rence of malignancy after therapy is a frequently observed situation
in cancer chemotherapy. Multidrug resistance (MDR) phenomenon
is defined as the resistance of tumor cells to various cytotoxic drugs.
It is a major impediment to successful treatment of breast cancer
using chemotherapy. Cancer cells either strengthen the already pre-
sent systems necessary for the removal of toxins from cells or acquire
resistance to cytotoxic drugs. Members of the ATP-binding cassette
(ABC) transporter superfamily have an important role in MDR.
Among these, proteins coded by the ABCB1 (MDR1), ABCC1
(MRP1), and ABCG2 (BCRP) genes are the most important trans-
porters related to MDR phenotype. In this study, effects of expres-
sion levels of the MDR1, MRP1, BCRP genes on the development
of docetaxel and doxorubicin resistance using a model MCF-7
breast carcinoma cell line is evaluated. Docetaxel and doxorubicin
were applied to cell culture in dose increaments and resistant sub-
lines were developed. Cytotoxicity analysis of drugs was performed
in wild type and developed resistant sublines to test development
of resistance. Total RNA was isolated from cells, converted to
cDNA and amplified by using gene (MDR1, MRP1, and BCRP)-
specific primers by RT-PCR. Western blot analysis and immuno-
staining were performed to determine the related protein levels.
OP-5
GSK3: identification of a novel mechanism
controlling inflammation in the brain
E. Beurel
1
, S. Michalek
2
and R. Jope
1

the devastating effects of neuroinflammation.
Rhythmic Signals: the Setting of Biological Time
OP-6
Computational search of the interaction
between melanopsin and cryp tochrome-2
proteins
E. B. Unal
1
, B. Erman
2
and I. H. Kavakli
2
1
Computational Sciences and Engineering, Koc University, Istan-
bul, Turkey,
2
Chemical and Biological Engineering, Koc University,
Istanbul, Turkey. E-mail:
Circadian rhythms are the biological processes that oscillate in the
biochemical, physiological and behavioral functions of organisms
with a periodicity of approximately 24 h without any external cues.
In mammals, circadian rhythm is generated by molecular clock,
which is located at suprachiasmatic nuclei (SCN) part of brain.
Circadian rhythm is reset by external factors such as light. The
cryptochromes,which was first discovered in Arabidopsis,are the
blue-light photoreceptors. They absorb light and transmit the elec-
tromagnetic signal to blue-light dependent of signal transduction.
In mammals, the cryptochromes and melanopsin have been pro-
posed as circadian photoreceptor pigments that exist in the inner
retina to transmit signal to the SCN to tell the time of day. Both

Faculty of Medicine, Department of Physiology, University of
Erciyes, 38039 Kayseri, Spain,
2
Department of Biochemistry and
Molecular Biology, University of Balearic Islands, Mallorca,
Spain. E-mail:
Mitochondria plays a central role in energy-generating processes
within the cell thorough the electron transport chain (ETC),
the primary function which is ATP synthesis via oxidative
Abstracts
43
phosphorylation (OXPHOS) which is shown to be related to age-
ing and apoptosis when this balance is destroyed under different
circumstances. This study is performed to investigate the effects
of alterations in the physiological melatonin levels via the circa-
dian rhythm changes, on the mitochondiral ETC in brain and
eye and how these changes are correlated to the pineal gland
melatonin receptor expressions. Fifty Sprague–Dawley male rats
weighing 200–250 g were used in five groups of different circa-
dian rhythms. The control group was 12/12 h of light/dark (L/D)
cycle. Different circadian rhythms of 24/0 h L/D, 0/24 h L/D,
16/8 h L/D and 8/16 h L/D cycles were applied to the groups for
1 week, respectively, in special cages where the duration of the
light and the climate can be adjusted. The melatonin receptors,
MEL1 and MEL2 expressions were determined by real-time PCR
in the pineal gland. The mitochondria of the brain and eye tis-
sues were isolated from the homogenates and the activation of
the mitochondrial OXPHOS complexes were determined by spec-
trophotometric micro-methods described before. Plasma melato-
nin levels were also determined by ELISA kit (IBL, Turkey).

, B. Ruhin
2
, D. Hotton
3
and A Berdal
3
1
Laboratoire de Biologie Oro-Faciale et Pathologie, INSERM
U714-IFR-58, Universite
´
s Paris 7 and Paris 6 Laboratoire
d’Histologie-Embryologie, Faculte
´
de Me
´
decine Dentaire de
Monastir, Tunisia,
2
Stomatology and Maxillofacial Surgery
Department, Pitie
´
Salpe
ˆ
trie
`
re University Hospital, Paris Cedex 13,
France,
3
Laboratoire de Biologie Oro-Faciale et Pathologie
INSERM U714-IFR-58, Universite

T. I. Klokk
1
, P. Kurys
1
, C. Elbi
2
, A. K. Nagaich
2
,
A. Hendarwanto
2
, T. Slagsvold
1
, C. Y. Chang
3
, G. L. Hager
2
and F. Saatcioglu
1
1
Department of Molecular Biosciences, University of Oslo, Oslo,
Norway,
2
Laboratory of Receptor Biology and Gene Expression,
National Cancer Institute, Bethesda, USA,
3
Department of
Pharmacology and Cancer Biology, Duke University Medical
Center, Durham, MD, USA. E-mail:
Androgen receptor (AR) mediates the action of androgens, which

dwarfism
A. Arman
1
, N. Yordam
2
, A. Ozon
2
, P. Isguven
3
, A. Coker
4
and
I. Peker
1
1
The Faculty of Engineering, Marmara University, Istanbul,
Turkey,
2
The Department of Pediatrics, Division of Endocrinology,
Hacettepe University, Ankara, Turkey,
3
The Department of
Endocrinology, Goztepe Government Hospital, Istanbul,
Turkey,
4
The Department of Biology, Art and Sciences, Marmara
University, Istanbul, Turkey. E-mail:
Growth hormone (GH) mediates its growth, fat and carbohy-
drate metabolism through insulin-like growth factor-I (IGF-I).
Interaction of GH with the GH receptor (GHR) is necessary for

H. Jueppner
2
and M. Bastepe
2
1
Pediatric Endocrinology and INSERM U561, Saint Vincent de
Paul Hospital, Paris, France,
2
Endocrine Unit, Department of
Medicine, Massachusetts General Hospital and Harvard Medical
School, Boston, MA, USA. E-mail:
harvard.edu
XLas is partly identical to the a-subunit of the stimulatory G pro-
tein (Gsa) with an extended N-terminus. Using adenoviral expres-
sion and cells that endogenously lack Gsa and XLas (GnasE2–/–
cells), we investigated human XLas (hXLas). Immunofluorescence
microscopy showed membrane and punctate perinuclear staining
for hXLas. On metal affinity chromatography, hXLas co-purified
with a histidine-tagged Ga1. Furthermore, a PTH(1–15) analog,
which normally prefers binding to a Gsa-coupled form of
PTHR1, bound to membranes from GnasE2–/– cells co-expres-
sing PTHR1 and hXLas. Also, hXLas was able to mediate basal
and agonist-induced cAMP accumulation. However, while hXLas
was expressed at lower levels than Gsa in transduced cells, basal
cAMP level in cells expressing hXLas was ~twofold higher than
in cells expressing Gsa. Similarly, basal ERK1/2 phosphorylation
in GnasE2–/– cells transiently expressing hXLas was markedly
enhanced. Isoproterenol treatment also resulted in significantly
higher levels of cAMP accumulation in hXLas-expressing cells
than Gsa-expressing cells, whereas PTH-induced cAMP accumu-

tory effect of carvedilol on brain tumours and on the efficacy of
imatinib has not yet been evaluated experimentally. In the present
study, we have investigated whether carvedilol provides synergistic
or antagonistic effect on imatinib-induced cytotoxicity in mono-
layer and spheroid cultures of malignant C6 glioma cells. C6
glioma cells in monolayer and spheroid cultures were treated with
the combination of carvedilol and imatinib in concentration of
10 lM. The cell proliferation, morphology, spheroid volumes,
bromodeoxyuridine-labelling index (BrdU-LI) were evaluated.
The expression levels of caspase-3, caspase-9, hypoxia inducible
factor-1 (HIF-1), Bcl-2 and platelet-derived growth factor receptor
alpha (PDGFRa) were examined by Western blot analysis. The
statistical significance was analysed by using the Student’s t-test.
The results demonstrated that carvedilol and imatinib in combina-
tion display enhanced antitumour activity in vitro against experi-
mental rat C6 glioma in monolayer and spheroid cultures.
OP-13
Non-traditional ways of B lymphocyte
regulation: receptors for acetylcholine and
thrombin
S. Komisarenko
1
, R. Grailhe
2
, Y. Petrova
1
, J. P. Changeux
2
,
W. Bahou

proliferation, but enhanced CD40-mediated B-lymphocyte prolif-
eration. It is concluded that signaling through nAChR and PAR3
affects basic vital functions of B lymphocytes, such as prolifer-
ation and survival, and interferes with their activation pathways.
These data show the ways, by which acetylcholine, nicotine or
thrombin can regulate the humoral branch of immunity.
OP-14
Identification and characterization of a novel
transcriptionally active domain in the linker
region of the TGFb-regulated Smad3 protein
M. Siderakis, V. Prokova, S. Mavridou, P. Papakosta and
D. Kardassis
Department of Medicine, University of Crete and IMBB-FORTH,
Heraklion, Crete, Greece. E-mail:
Our structure–function analysis of human Smad3 protein, a key
mediator of TGFb signaling in mammalian cells revealed that the
middle, non-conserved, linker domain has an autonomous and
potent transactivation function. The region with the maximal
transactivation capacity was the 143–248 that consist of almost
the entire linker domain and the first 18 amino acids of the MH2
domain. The corresponding regions in Smad4 as well as in
Smad1, which is a key mediator in Bone Morphogenetic Protein
(BMP) signaling, are also transcriptionally active in mammalian
cells further supporting the important role of this domain for
Smad function. Smad3 mutants bearing an internal deletion of
the 200–230 region or single amino acid substitutions in two
highly conserved residues of this region (Q222A and P229A) had
severe defects in oligomerization and transcriptional activation of
target promoters. In contrast, mutagenesis of a non-conserved
amino acid residue in the same region (N218A) did not affect

cells,demonstrated that sigma1 receptor and beta1 integrin are
associated. Furthermore, both confocal microscopy experiments
and surface biotinylation experiments indicated that both appli-
cation of sigma receptor drugs and knock-down of the sigma1
receptor increased the beta1 integrin expression in the mem-
brane. Lipid raft fractionation experiments in MDA-MB-231
cells demonstrated that both application of the sigma receptor
drugs and the knock-down the sigma1 receptor levels, beta1 in-
tegrin protein in lipid rafts fraction of MDA-MB-231 cells were
altered. All these data suggest that sigma1 receptor is associated
with beta1 integrin and is likely modulate beta1 integrin levels.
Therefore, sigma1 receptor is likely to be a novel target for
breast cancer metastasis.
OP-16
HERG potassium channels and heart disease
S. Kuyucak
School of Physics, University of Sydney, NSW, Sydney, Australia.
E-mail:
The heartbeat is controlled by electrical signals mediated by the
flow of ions through specialized ion channels. Of the channels
that contribute to cardiac electrical activity, potassium channels
encoded by the Human ether-a-go-go-related gene (HERG) have
been of particular interest for many reasons. First, mutations in
HERG are the cause of one-third of cases of congenital long QT
syndrome, an inherited cause of sudden cardiac death. Secondly,
HERG is the molecular target for the vast majority of drugs that
cause drug-induced long QT syndrome, the commonest cause of
drug-induced arrhythmias and cardiac death. Thirdly, HERG
channels have very unusual biophysical properties, which suggest
that they may act as an endogenous antiarrhythmic agent. There-

1
Department of Pharmacology, National University of Singapore,
Yong Loo Lin School of Medicine, Singapore,
2
Department of
Biochemistry, National University of Singapore, Yong Loo Lin
School of Medicine, Singapore,
3
Department of Pathobiology,
University of Illinois at Urbana Champaign, Urbana, IL, USA.
E-mail:
We investigated the molecular mechanisms that regulate the
apoptosis of acinar cells induced by crambene (1-cyano-2-hydrox-
y-3-butene-CHB), a plant nitrile. As evidenced by annexin
V-FITC staining, crambene treatment for 3 h induced the apop-
tosis but not necrosis of pancreatic acini. Caspase 3, 8 and 9
activity in acini treated with crambene were significantly higher
than in untreated acini. Treatment with caspase 3, 8 and 9 inhibi-
tors inhibited annexin V staining, as well as caspase 3 activity,
pointing to an important role of these caspases in crambene-
induced acinar cell apoptosis. The mitochondria membrane
potential was collapsed in crambene-treated acini than in
untreated that displayed polarized mitochondria. Also, the treat-
ment of acini with crambene induced the release of cytochrome
C by mitochondria than in untreated acini. Neither TNF-a nor
Fas ligand levels were changed in pancreatic acinar cells after
crambene treatment. These results provide evidence for the induc-
tion of pancreatic acinar cell apoptosis in vitro by crambene and
suggest the involvement of mitochondrial pathway in pancreatic
acinar cell apoptosis.

well as prostate cancer progression.
OP-19
Enhancement of peptidylarginine deiminase 4
enzymatic activity assists apopt osis
C Y. Lin
1
, Y F. Liao
2
, P C. Hsu
3
, H C. Hung
2
and G Y. Liu
1
1
Institute of Immunology, Chung Shan Medical University, Tai-
chung, Taiwan,
2
Department of Life Sciences, National Chung-
Hsing University, Taichung, Taiwan,
3
Department of Medicine, Da
Chien General Hospital, Taiwan. E-mail:
PAD4 post-translationally converts peptidylarginine to citrulline.
It plays an essential role in immune cell differentiation and apop-
tosis. A haplotype of single nucleotide polymorphism (SNP) in
PAD4 is functionally relevant as a rheumatoid arthritis (RA)
gene. It could increase enzyme activity leading to raised levels of
citrullinated protein and stimulating autoantibody. Inducible
PAD4 causes haematopoietic cell death (Liu et al. 2006) apopto-

Institute of Biology, National Center for Scientific Research
‘Demokritos’, Greece,
2
Foundation for Biomedical Research of the
Academy of Athens, Athens, Greece.
E-mail: effi
Type 2 diabetes is characterized by progressive pancreatic b-cell
dysfunction and apoptosis. Recent reports provided evidence for
an autocrine role of insulin on the signalling cascade in b-cell
growth, function and survival are concerned. We examined the
effects of chronic glucose stress on insulin signalling. Exposure of
bTC-6 cells to high glucose resulted in impairment of insulin-sti-
mulated phosphorylation of IRS-2. These changes were accom-
panied by significant impairment of IRS-2-associated PI3-kinase
activation, and substantially decreased activation of Akt. We also
examined mTOR kinase, a downstream effector of Akt, which
stimulates protein synthesis in response to Akt phosphorylation.
High glucose abolished insulin-induced activation of mTOR
without affecting mTOR expression, thus protein synthesis
should also be impaired. A mechanism by which Akt promotes
cell survival includes phosphorylation of the pro-apoptotic
Abstracts
47
protein BAD, which keeps the pro-apoptotic protein BAX
engaged to Bcl-XL. Exposure of cells to high glucose also resul-
ted in suppression of insulin-stimulated phosphorylation of BAD
without affecting BAD expression. In conclusion, we demonstra-
ted that chronic exposure of pancreatic b-cells to increased glu-
cose concentration resulted in impaired activation of the IRS-2/
PI3-kinase/Akt signalling pathway in response to insulin. The

Hsp90 client proteins involved in oncogenic survival signaling are
often found to be mutated in leukemia, and the integrity of the
Hsp90 complex could therefore be important for leukemic cell
survival. We demonstrate here that the Hsp90 co-chaperone p23
is cleaved in leukemic cell lines treated with commonly used che-
motherapeutic drugs. The cleavage of p23 paralleled the activa-
tion of procaspase 3 and was suppressed by z-DEVD-FMK.
Interestingly, p23 cleavage was also observed in caspase 3-defici-
ent MCF-7 cells, and in vitro translated 35S-p23 was cleaved by
both caspase 3 and 7. Two caspase target sites were identified in
the C-terminal sequence EVD142GAD145, and only Asp to Ala
mutagenesis of both sites (D142/145A) completely blocked p23
cleavage. The Hsp90 inhibitor geldanamycin, which inhibits p23
binding to Hsp90, did not induce cell death or p23 cleavage on
its own, but enhanced anthracycline-induced caspase activation,
p23 cleavage and apoptosis. This implies that Hsp90 inhibition
amplifies caspase activation. Geldanamycin also enhanced
caspase cleavage of 35S-p23 in vitro, indicating that the associ-
ation of p23 to Hsp90 protects against cleavage. These findings
underscore the importance of the Hsp90-complex in antileukemic
treatment, and suggest that p23 may have a role in survival
signaling.
OP-22
Overview how colorectal cancer cells avoid
cell elimination; mechanisms of immune- and
chemo-resistance
B. Pajak
1
, H. Engi
1

pump in chemoresistant COLO 320 cell line. The variety of antia-
poptotic mechanisms found in colorectal cancer cells and the
knowledge how complex they are renders the anticancer therapy a
challenge but the more we know how to sensitize cancer cells to
death signals the more likely promise to eliminate them.
Embryonic Stem Cells
OP-23
Expression of the cholinergic system
components during mouse embryonic stem
cell differentiation
L. Paraoanu, G. Steinert and P. Layer
Developmental Biology, Darmstadt University of Technology,
Darmstadt, Germany. E-mail:
Expression of cholinesterases during phases of embryonic devel-
opment is a general phenomenon. However, no precise function
for cholinesterases could be described during early developmental
stages. We have examined the expression of cholinesterases and
other cholinergic components during in vitro differentiation of
CGR8 cells, an embryonic stem cell line. Undifferentiated mono-
layers of these cells can be maintained in culture, or embryoid
bodies can be generated in vitro. The embryoid bodies were
allowed to differentiate in the absence of further additives or
treated with retinoic acid, and collected at various times for fur-
ther analysis. The cholinesterase expression was analyzed by
reverse transcriptase polymerase chain reaction, histochemistry,
and activity measurement. Low levels of cholinesterase activity
and transcripts could be detected already in undifferentiated cells.
The phases of differentiation are accompanied by increased ace-
tylcholinesterase transcripts. Butyrylcholinesterase expression was
initially constant, decreasing at later differentiation stages. All

Maternity Hospital, Ankara, Turkey. E-mail: alpcan@medicine.
ankara.edu.tr
A recently isolated source for stem cells is the mesenchymal cells
of human umbilical cord stroma. Although neuronal differenti-
ation has been demonstrated in human umbilical cord stroma cells
(HUSCSs), there is a still discordance among studies about the
potential of these cells to differentiate into neurons. The aim of
this study was to differentiate HUSCSs into neurons and investi-
gate the temporal development potency of newly differentiated
neuronal cells. The spatio-temporal distribution and the quantifi-
cation of various proteins during differentiation, immunofluores-
cent single and multi-immunolabeling techniques were performed
using a series of antibodies raised against major cellular markers
for neurons, such as MAP1, MAP2, GFAP, tau, NSE, NFM, b-
III tubulin, NeuN, and thyrosine hydroxylase. MAP1, a specific
structural protein in the cytoplasm, MAP2, specific for dendrites
and NFM were all positive during the course of the entire differ-
ention period whereas tau and b-III tubulin were detected at only
certain time-points of differentiation. While NeuN stained faintly,
GFAP and thyrosine hydroxylase were both negative since no dif-
ferentiation toward astrocytes and dopaminergic neurons was
accomplished. Conclusively, it appears that throughout the differ-
entiation of HUCSCs into neurons, expression of neuronal mark-
ers present a diversity which follows a similar progress as seen in
embryologic development of neurons of the body.
Gene Therapy
OP-25
Cationic nanoparticle synthesis and their use
in gene transfer
G. Guven

Nanoparticles with such a high amount of surface charge make
these materials useful for the gene transfer especially.
OP-26
Peripheral pool of CD8+ lymphocytes contains
T cells that recognize syngeneic MHC class II
molecules
E. S. Zvezdova, T. S Grinenko, L. A Pobezinsky,
E. L. Pobezinskaya and D. B. Kazansky
Russian N.N. Blokhin Cancer Research Center.
E-mail:
A number of publications appeared, which assume that autoim-
mune cells exist in the pool of CD8+ T lymphocytes with the
potential to recognize both MHC class I and MHC class II mole-
cules. To test this, we generated autoreactive T-cell hybridomas
with the ability to recognize both allogeneic MHC class I mole-
cule and MHC class II molecules: the LN CD8+ T cells from
C57BL/6 mice were activated in the presence of allogeneic MHC
class I molecules Kbm3 and fused with BW5147 thymoma cells,
expressing CD4 coreceptor. The percentage of autoreactive hy-
bridomas in CD8+ T cells was 9.3%. One of these hybridomas
expressed two a-chains. One of these a-chains help to recognize
syngeneic MHC class II molecules, while the other allogeneic
MHC class I molecule Kbm3. To elucidate would the same situ-
ation be observed in TCRa
+/0
mice capable to express single
a-chain of TCR, we obtained hybridomas from their CD8+ T
cells. The frequency of self MHC class II reactive hybridomas
was lower in contrast with that of wild type 1.3%. We have
shown that in the peripheral pool of wild-type CD8+ T cells the

2
Faculty of Medicine, Department of
Medical Biology and Genetics, Kirikkale University, Kirikkale,
Turkey,
3
Wellcome Trust Sanger Institute, Wellcome Trust Genome
Campus, Hinxton, Cambridge, UK,
4
Faculty of Medicine,
Department of Pharmacology, Hacettepe University, Ankara,
Turkey,
5
Institute of Oncology, Department of Preventive
Oncology, Hacettepe University, Ankara, Turkey.
E-mail:
Attenuated Salmonella typhimurium (ST) strains have being
investigated as promising vectors for gene therapy. In our study,
Abstracts
49
STaroB strain was evaluated as a gene delivery vehicle for the
first time. Eukaryotic expression vectors, pTARGET and
pcDNA3, carrying murine IL-18 or CD40L genes were construc-
ted to enhance immune responses of the host for further cancer
therapy studies. mRNAs used to synthesize cDNAs to amplify
IL-18 and CD40L DNAs were obtained from lipopolysaccharide-
induced adherent peripheral blood mononuclear cells, and Phor-
bol 12-Myristate 13-Acetate and Ionomycin-induced splenocytes
respectively. A bioactive IL-18 molecule consisting of IFN-b sig-
nal and mature IL-18 sequences was constructed by overlap
extension method. The plasmids were transformed into STaroB

to occur due to the presence of a 28 bp direct repeat flanking 1.6
(pCI-neo) and 3.2 kb (pGPV-PV) intervening sequences, was
responsible for the acquisition of a kanamycin resistance pheno-
type in different E. coli strains (DH5a, Top 10F’, HB101 and
JM109). This could be explained as follows: as a result of slip-
page events, the Neo
r
gene falls under the control of a promoter-
like sequence located at the origin of replication, thus enabling
cells to strive in kanamycin. The Poisson distribution of mutants
determined by Luria-Delbru
¨
ck fluctuations tests and reconstruc-
tion experiments surprisingly, but unequivocally, indicates that
this plasmid slippage event behaves adaptively. This study high-
lights the need to carefully design plasmid vectors avoiding, for
example, repetitive sequences that are genetically unstable and
may give rise to unforeseen problems during plasmid DNA pro-
duction and subsequent utilization.
Reference:
1. Bahloul et al.Vaccine 1998; 16: 417–425.
Therapeutic Enzymes
OP-29
Engineering of nucleoside kinases to improve
the activation of nucleoside analog prodrugs
M. Konrad
1
, S. Ort
1
, C. Monnerjahn

enzyme) therapeutic treatment involving expression (or direct intra-
cellular protein transduction) of a catalytically improved human
enzyme may pave the way to the development of novel strategies in
nucleoside prodrug-dependent cancer chemotherapy.
OP-30
Effect of N2A connectin/titin on disassembly
of p94/capn3 caused by autolysis in IS2
Y. Ono
1
, F. Torii
2
, K. Ojima
3
, N. Doi
3
, K. Yoshioka
2
, D. Labeit
4
,
S. Labeit
4
, K. Suzuki
5
, K. Abe
2
, T. Maeda
6
and H. Sorimachi
1

regulated by its binding protein connectin/titin. In vitro analysis
of p94 autolysis revealed that autolytic cleavage in each p94-spe-
cific insertion sequence, NS, IS1, and IS2, causes different impact
on molecular integrity of p94 as a protease; autolysis in IS2, but
not in IS1, causes disassembly of autolyzed fragments. Using
proteinase trapping system to semiquantitatively assay p94 auto-
lysis, where p94 was expressed in yeast as a hybrid protein
between DNA binding and activation domains of the yeast tran-
scriptional activator Gal4, N2A connectin was shown to suppress
p94 autolytic disassembly. Proximity of IS2 autolytic and connec-
tin-binding sites in p94 suggested that N2A connectin interferes
with IS2 autolysis. It was also shown that N2A connectin with
mdm mutation that is causative of dystrophy in mice and abol-
ishes connectin–p94 interaction was incapable of suppressing p94
autolytic disassembly. These data indicate that p94–connectin
interaction plays an important role in the control of p94 func-
tions by regulating autolytic decay of p94.
OP-31
Prevention of thermal aggregation of yeast
alcohol dehydrogenase by arginine and
a-cyclodextrin
A. Barzegar and A. A. Moosavi-Movahedi
Institute of Biochemistry and Biophysics, University of Tehran,
Tehran, Iran. E-mail:
Yeast alcohol dehydrogenase (YADH) is a tetrameric enzyme
and is a widely studied as a metaloenzyme for its well-known
biotechnological importance. Arginine (Arg) and a-cyclodextrin
(a-CD) are the universal reagents that are effective in assisting re-
folding of recombinant proteins from inclusion bodies. The
results showed that YADH (0.5 mg/ml) initiate to aggregation

Medical Center, Tainan, Taiwan,
2
Division of Hepatogastroentero-
logy, Department of Internal Medicine, Chi Mei Medical Center,
Tainan, Taiwan,
3
Department of Pathology, Chi Mei Medical
Center, Tainan, Taiwan,
4
Core Facilities for Proteomics Research
and Institute of Biological Chemistry, Academia Sinica, Taipei,
Taiwan. E-mail:
The link of proto-oncogenic protein Wnt-1 with NF-lkappaB
activity has been reported in PC12 cells, a rat pheochromocytom-
a cell line of neural crest lineage, showing that the Wnt-1-medi-
ated survival of these cells is dependent on NF-lkappaB
activation, and that stable expression of Wnt-1 increases NF-
lkappaB activity. In Wnt-1-mediated breast tumorigenesis, Wnt-1
is reported to be a fundamental signaling event in metastatic pro-
gression of human cancers. We have shown that enhanced NF-
lkappaB-associated Wnt-1 protein expression is detected in the
majority of hepatoma samples from both hepatitis B and C
patients. Insight into how NF-lkappaB regulates Wnt-1 protein
production may help design highly effective therapeutic agents in
treating Wnt-1-producing cancers and prevent their metastatic
progression. RNA interference (RNAi) was used in this study to
assess the role of p50 and p65 of NF-lkappaB in the regulation
of Wnt-1 protein production by Het-1A cells, a human esopha-
geal cancer cell line with Wnt-1 protein production. Cultured
Het-1A cells were transfected with sequence-specific double-stran-

50 nM Pac. Cytotoxicity was determined with MTT assay after
cells were treated with the drug for 24–72 h. 20 nM Pac treat-
ment for 24 h inhibited growth of MCF-7 by 30% and MDA-
MB-231 by 44%. Cytotoxic effect was dose-dependent as 50 nM
Pac caused growth inhibition by 52% and 64% for 24 h in both
cell line. DNA fragmentation experiments showed that 20 and
50 nM Pac induced apoptosis. To understand the role of Bcl2 in
apoptotic response Bcl2 mRNA levels was checked by qPCR.
Pac at both concentrations decreased Bcl2 expression by 1.8- and
2.1-fold in ER+ and 1.2- and 1.7-fold in ER– cells compared to
control.To further clarify the role of Bcl2 we have prepared Bcl2
siRNA cells and the experiments are in progress.
Oxidative Stress
OP-34
ANKRD1 gene shows opposite atrial versus
ventricular expression kinetics at heart failure
M. Torrado
1
, B. Nespereira
1
, Y. Bouzamayor
1
, A. Centeno
2
,
E. Lo
´
pez
2
and A. Mikhailov

atrium (twofold change in mRNA/protein levels). Owing to these
opposite expression kinetics, the normal atrio-ventricular pattern
of ANKRD1 distribution was abolished in failing hearts. Conclu-
sions: (a) ANKRD1 expression is distinctly regulated in cardiac
chambers in the postnatal heart; (b) the inverse gene expression
response in atria when compared to ventricles at advanced heart
failure may be considered a molecular sigh of severity of cardiac
disease.
OP-35
Peroxynitrite induces red blood cell membrane
reorganization
M. N. Starodubtseva
1
, T. G. Kuznetsova
1
, J. C. Ellory
2
,
T. A. Kuznetsova
3
and S. O. Abetkovskaya
3
1
Gomel State Medical University, Gomel, Belarus,
2
Department of
Physiology, Anatomy and Genetics, University of Oxford, Oxford,
UK,
3
A. V. Lykov Heat and Mass Transfer Institute of NASB,

ti
Department of Medical Chemistry, Semmelweis University,
Budapest, Hungary. E-mail:
Molecular chaperones or stress proteins guard the cellular con-
formational homeostasis of proteins, protecting against a num-
ber of stresses. The strongest inducer of stress proteins is the
accumulation of misfolded proteins. Proper function of chaper-
ones as well as a robust mounting of the stress response is
required for longevity. Indeed, a hallmark of ageing is the
increase of aggregated and oxidized proteins. However, little is
known about the causal relationship between protein misfold-
ing, oxidative stress and chaperone induction. In our present
studies using different cell lines we examined the relationship
between protein misfolding, aggregation and chaperone induc-
tion. As expected, heat shock, the archetype of stress as well
as geldanamycin induced the uniform elevation of the major
chaperones, Hsp70 and Hsp90. In contrast, the oxidative stres-
sor hydrogen peroxide induced only Hsp90. Investigating the
hydrophobic surface exposure to detect protein misfolding in
live cells, the fluorescence of ANS, an apolar probe was
applied. In contrast to heat shock, hydrogen peroxide was
unable to induce the ANS signal of purified model proteins,
cytosolic extracts and live cells. Moreover, did not result in
significant protein aggregation. These results suggest that short-
term oxidative stress does not induce bulk protein denaturation
and stress response, raising the question what is the depend-
ence of protein oxidation and misfolding during ageing and in
degenerative diseases.
Abstracts
52

2. Zhang W, Carneiro MJVM, Turner IJ, Allen S, Roberts CJ,
Soultanas P. J Mol Biol 2005; 351: 66–75.
OP-38
Coordination of DNA base excision repair
studied by photoreactive DNA probes and
functional assay
O. I. Lavrik
Institute of Chemical Biology and Fundamental Medicine,
Novosibirsk, Russia. E-mail:
Photoaffinity-labeling technique has been developed to study
coordination of base excision repair (BER) proteins. Photoreac-
tive DNA intermediates of BER pathways were created in sys-
tems reconstituted of purified proteins and in cellular/nuclear
extracts to identify proteins interacting with damaged DNA. The
main target proteins interacting with branch point BER interme-
diate were identified as Poly(ADP-ribose)polymerase1 (PARP1),
flap endonuclease1 (FEN1), DNA polymerase beta (Pol beta)
and apurinic/apyrimidinic endonuclease1 (APE1). The results
indicate that APE1 and PARP1 interact preferentially with
nicked BER intermediate carrying 3’-photoreactive dNMP resi-
due and the 5’-sugarphosphate moiety, whereas intermediate with
5’-phosphate is less favourable interaction partner. Thus, PARP1
and APE1 can discriminate intermediates of SP and LP BER
pathways to regulate BER. DNA repair synthesis catalysed by
Pol beta is modulated by the interplay between Pol beta, APE1,
PARP1 and XRCC1. APE1 can perform stimulation of strand-
displacement synthesis catalysed with Pol beta and proofreading
function while PARP1 inhibits these reactions. PARP1 inhibits
activity of FEN1 preventing formation of intact DNA structure.
The inhibition by PARP1 of these activities was not completely

2
MolCat Bt, Buda-
pest, Hungary,
3
Proteomics Laboratory, Biol.Res.Ctr., Szeged,
Hungary,
4
Immunology Department, Eotvos University, Budapest,
Hungary. E-mail:
Uracil in DNA may arise by cytosine deamination or thymine
replacement and is removed by members of the uracil-DNA gly-
cosylase (UDG) superfamily. The surprising lack of the major
UDG-superfamily member UNG from the fruitfly genome may
allow the fly to tolerate thymine-replacing incorporation of uracil
in its DNA. Such incorporation is usually prevented by the
enzyme dUTPase that removes dUTP from the DNA polymeriza-
tion pathway. However, fruitfly larval tissues do not contain any
detectable dUTPase (the enzyme being confined to the imaginal
disks, progenitors of adult fly organs; Be
´
ke
´
si et al. J Biol Chem
2004; 279: 22362). Here, we asked if the putative presence of ura-
cil-DNA in larval tissues, potentially accumulated due to lack of
both UNG and dUTPase, might trigger any physiological
response. Such response would require macromolecular (e.g. pro-
tein) factors specifically recognizing uracil-DNA; we therefore set
out to identify such proteins from Drosophila extracts. We show
that the most intensive hit of uracil-DNA pull-down experiments

nucleophile was concluded to be of primary importance for the
reaction, while the removal of metal ion B was less dramatic.
Thus, we propose a unified catalytic scheme involving only one
obligatory metal ion at the active site that serves to stabilize the
attacking nucleophile, which is coming from the bulk phase and
not assisted by a general base or substrate. A second, variable
group is required to facilitate the nucleophilic attack via interac-
tions with the pentavalent transition state.
DNA Repair in Health, Disease and Aging
OP-41
Base excision repair pathway at telomeric
DNA
M. Muftuoglu, P. Opresko and V. Bohr
Laboratory of Molecular Gerontology, National Institute on Aging,
National Institutes of Health, Baltimore, MD, USA.
Telomeres consist of protein–DNA complexes that protect the
ends of linear chromosomes from inappropriate chromosome
fusions by DNA repair pathways. Telomere-repeat binding fac-
tors 1 and 2 (TRF1 and TRF2) bind specifically to duplex telo-
meric DNA and regulate telomere length. When bound to
telomeres, TRF2 allows the cell to recognize the telomeres as
chromosome ends rather than double strand breaks in DNA.
Defects in TRF2 induce telomere dysfunction that causes telo-
mere end fusions, replicative senescence, apoptosis, and genomic
instability. Oxidative damage causes loss of telomeric DNA, and
we found previously that oxidative DNA base damage disrupts
TRF1 and TRF2 binding to telomeric DNA, emphasizing the
importance of DNA repair at telomeres. We are investigating the
function of base excision repair (BER) in preserving telomeres,
and specifically roles for TRF2 in cooperation with BER proteins

FANCA gene by SSCP/HD analysis. Ironically while sequencing
of the variant bands led to the identification of many homozygous,
non-pathogenic mutations including missense/silent mutations,
base substitutions, and deletions/insertions in introns, only a few
pathologic mutations could be detected. Despite much effort, the
molecular defects in the most of Turkish group A patients go
undetected. Given that all of the exons could be amplified by PCR
in almost all of the patients, it is unlikely that large deletions could
be cause of the disease either. This study proved that mutation
analysis of FANCA gene is quite difficult and inefficient, which
makes pre-natal diagnosis and detection of carriers in Turkish
population almost impossible. Acknowledgment: This study was
supported by Hacettepe University Research Fund (02G116).
Diabetes, Obesity and Metabolic Syndrome
OP-43
The role of TGFb1 in glycosaminoglycan
metabolism in diabetes mellitus
N. Yu. Yevdokimova
Molecular Immunology, Institute of Biochemistry, Kyiv, Ukraine.
E-mail:
The dysregulation of glycosaminoglycan (GAG) metabolism is a
typical feature of renal diabetic complications. TSP-1-dependent
TGFb1 activation is known to mediate numerous ECM disor-
ders in diabetic kidney. However, its role in the metabolism of
GAGs is studied insufficiently. We observed that high glucose
enriched the mesangial ECM with hyaluronic acid (HA) of
high-molecular weight (>2000 kDa). We detected the upregula-
tion of hyaluronan synthase 2 (HAS2) mRNA without altera-
tions of HAS3 expression. High glucose also stimulated the
production and TSP-1-dependent activation of TGFb1. The

Pediatric Molecular Genetics Department, Ankara University
School of Medicine, Ankara, Turkey,
2
Pediatric Endocrinology
Department, Ankara University School of Medicine, Ankara,
Turkey,
3
The Laboratory for Translational Research, Harvard
University School of Medicine, Boston, MA, USA.
E-mail:
Plasminogen Activator Inhibitor I (PAI-1), a member of the ser-
ine proteinase inhibitor family is not only inhibits fibrinolysis but
also has complex interactions with cellular matrix, further inhib-
its proteolysis and important in cardiovascular diseases. Obesity
is a metabolic disorder, associated with increased PAI-1 concen-
tration in the circulation. This increase is also related to insulin
resistance, dyslipidemia and cardiovascular diseases. In our study
we investigated the frequency of –675 4G/5G PAI-1 polymorph-
ism located at –675 in promoter of the gene in Turkish pediatric
obese patients and the effect of this polymorphism on PAI-1 gene
expression and on the adipocyte differentiation. The frequency of
PAI-1 4G/5G genotype was determined as previously described.
We constructed a dual-glo luciferase effect reporter assay to
investigate of this deletion on the PAI-1 promoter activity in pre-
adipocytes, adipocytes, and HUVEC cells. We also investigated
the effects of troglitazone and ciglitazone treatments on PAI-1
promoter activity. 4G/4G genotype was significantly higher than
the 5G/ 5G genotype in Turkish pediatric obese patients. In the
present study, we demonstrated that 4G/4G genotype increases
the PAI-1 promoter activity. PAI-1 increases the differentiation

insulin-induced Akt and ERK1/2 phosphorylation. These results
show that modifications in Grb14 expression level can rapidly
modulate insulin signalling and action, suggesting that Grb14 can
be considered as a new target for the development of insulin-sen-
sitizing agents.
Lipid-related Disorders and Atherosclerosis
OP-46
Associations of TSH levels with bloo d lip ids,
metabolic syndrome, coronary risk factors,
coronary heart disease in Turkish adult
G. Hergenc
1
, A. Onat
2
, S. Albayrak
3
, A. Karabulut
4
,
S. Turkmen
5
, I. Sari
4
and G. Can
6
1
Yildiz Tech University, Department of Biology, Istanbul,
Turkey,
2
Turkish Society Cardiology, Istanbul, Turkey,

sion analysis revealed TChol as the sole independent covariate of
TSH levels in men and both sexes combined. To conclude, TSH
was found not to contribute to CHD, hypercholesterolaemia, MS
and DM risk in logistic regression analyses. Prospective follow
ups may give a better understanding concerning the thyroid sta-
tus and CHD risk in Turkish adults.
OP-47
The carboxy-terminal region of apoA-I is
important for the biogenesis of HDL in vivo
A. Chroni
1
, A. Duka
2
, G. Koukos
2
and V. Zannis
2
1
Institute of Biology, National Center for Scientific Research
Demokritos, Athens, Greece,
2
Boston University School of
Medicine, Boston, MA, USA. E-mail:
Apolipoprotein A-I (apoA-I) promotes ABCA1-mediated lipid
efflux that results in the initial lipidation of apoA-I. This leads to
the formation of discoidal HDL and, after cholesterol esterifica-
tion by the enzyme LCAT, of spherical a-HDL particles. Follow-
ing adenovirus-mediated gene transfer in apoA-I-deficient mice
the plasma HDL levels were greatly reduced in mice expressing
the C-terminus deletion mutants apoA-I[del(185–243)] and apoA-

1
Institute for Medical Biochemistry, University School of
Pharmacy & Clinical Center of Serbia, Belgrade, Serbia &
Montenegro,
2
Institute of Neurology , Clinical Center of Serbia,
Belgrade, Serbia & Montenegro,
3
Laboratory for Radiobiology and
Molecular Genetics, VINC
ˇ
A Institute of Nuclear Sciences,
Belgrade, Serbia & Montenegro. E-mail:
The aim of this study was to determine whether the DNA poly-
morphism in apolipoprotein (apo)B, E and angiotensin-converting
enzyme (ACE) is associated with occurrence of ischemic stroke
(IS) in young adults and to find out the relationship between IS,
lipids, apolipoproteins, blood pressure (BP) levels and those poly-
morphisms. The possible association of apoB (XbaI, EcoRI,
MspI, I/D, 4311 A/G), apoE (HhaI) and ACE (I/D) polymorph-
isms were analyzed in 65 IS survivors younger than 65 and 591
age-matched healthy subjects. The occurrence of stroke was pro-
ven by computed tomography or magnetic resonance of the brain.
The results showed that patients with IS presented statistically sig-
nificant higher frequencies of X (XbaI), I (I/D), M1 (MspI), A
(4311 A/G), E4 allele, and lower E2 allele than control subjects
(P < 0.01). These results are discussed in the sense of finding
risk/protective haplotype for IS, lighting the role of circulating
lipids, BP levels in the pathogenesis of IS subtypes in different
genotypes in this study. ApoB, triglycerides and MAP were the

31.2% at 10 years (SE: 0.05, 0.06, and 0.07, respectively). All-
cause mortality rate increased with an elevated fibrinogen level.
Eighty percentage of patients with a fibrinogen level >3.4 g/l
had a survival time of <3 years (P = 0.002). This relation was
also demonstrated within patients with critical ischemia. The
plasma fibrinogen level was thus identified as an independent risk
factor for mortality in PAD patients after adjusting for con-
founding factors.
Oncogenes and Tumor Suppressors
OP-50
An approach for deciphering molec ular
patterns involved in genistein effects in
human prostate
R. Laslo
1
, H. Klocker
2
, I. Rowland
3
, R. L. Hancok
4
,
R. S. Pardini
4
and A. I. Baba
5
1
Nutrition and Toxicology, University of Agricultural Sciences and
Veterinary Medicine of Cluj, Cluj-Napoca, Romania,
2

esting features that are presented here. We discuss the results of
genes of interest in order to temp establishing of preventive gene
Abstracts
56
set. They concern the coverage quality, in terms of consistency
and reproducible, of the protein-encoded sequences of genes of
interest, an EST analysis, the validation of splice types, and
finally the link between gene expression analysis/alternative spli-
cing and prostate cancer prevention.
OP-51
Ornithine decarboxylase interferes with tumor
necrosis factor alpha-mediated matrix
metalloproteinase- 9 productions on
macrophage-like differentiation
Y. Liao
1
, H. Hung
1
and G. Liu
2
1
Department of Life Sciences, National Chung-Hsing University,
Taichung, Taiwan,
2
Institute of Immunology, Chung-Shan Medical
University, Taichung, Taiwan. E-mail:
Proteolytic activity of matrix metalloproteinase-9 (MMP-9) is
necessary for a variety of macrophage functions such as extrava-
sation, migration, and tissue remodeling. Ornithine decarboxylase
(ODC) is a rate-limiting enzyme of polyamine biosynthesis. Poly-

dence that the receptor-type PTP epsilon (RPTPe) supports
transformation by linking Neu with downstream events. Cells
from mammary epithelial tumors induced in vivo by Neu in mice
genetically lacking RPTPe are less transformed and proliferate
slowly. At the molecular level RPTPe activates Src, a known col-
laborator of Neu in mammary tumorigenesis, as well as the Src-
related kinases Yes and Fyn. Accordingly, absence of RPTPe
reduces Src activity and alters Src phosphorylation in tumor cells,
RPTPe dephosphorylates and activates Src in heterologous sys-
tems, and Src binds a substrate-trapping mutant of RPTPe. The
role of Neu is central, since Neu phosphorylates RPTPe specific-
ally at Y695 at its C-terminus, thereby directing RPTPe to acti-
vate Src. Finally, the altered morphology of Neu-induced
mammary tumor cells lacking RPTPe is corrected by exogenous
Src or RPTPe. We conclude that a Neu-RPTPe-Src pathway
exists in mammary tumor cells, and that phosphorylation of
RPTPe by Neu directs the phosphatase to activate Src in a man-
ner required for complete transformation. Inhibition of RPTPe
may be useful to augment direct pharmacologic inhibition of Src
in fighting mammary tumors.
OP-53
Both C and N-terminal regions of p53 are
required for its optimal Mdm2-mediated
degradation
R. Hjerpe
1
, F. Aillet
1
, T. Hay
2

by other cellular and viral E3s.
OP-54
In vivo dynamics of MN1TEL suggest that its
immobility may be crucial in leukemi as caused
by this fusion protein
M. Ter Haar, M. Meester-Smoor, M. Janssen, A. Houtsmuller
and E. Zwarthoff
Department of Pathology, Erasmus MC, Rotterdam,
the Netherlands. E-mail:
The MN1TEL fusion gene is created by a leukemia-associated
translocation and combines the MN1 gene from chromosome 22
with the TEL (ETV6) gene from chromosome 12. TEL is a mem-
ber of the ETS family of transcription factors and contains a
DNA-binding domain (DBD). TEL represses transcription from
ETS-responsive elements. MN1 contains transcription-activating
domains. MN1 can stimulate transcription from several promot-
ers by interaction with coactivators such as p300 and members of
the p160 family and can enhance expression driven by the reti-
noic acid receptor. In the MN1TEL protein the transactivating
domains from MN1 are combined with the TEL DBD.
MN1TEL stimulates transcription but is unable to synergize with
the retinoic acid receptor and is not stimulated by p300 or p160
coactivators. To get an insight in the physical properties of the
proteins, we decided to examine their behaviour in vivo. For this
Abstracts
57
we generated inducible cell lines expressing GFP-tagged proteins.
Protein mobility was examined using the fluorescence recovery
after photobleaching (FRAP) technique. It was found that MN1
is completely mobile and that a small, but significant fraction of

stably transfected with EWS-FLI1 enhanced the anchorage-inde-
pendent phenotype of EWS-FLI1 alone. Reduction of RHA pro-
tein levels by siRNA in ESFT cell lines decreased their growth
rate. Our results provide strong evidence for EWS-FLI1 and
RHA interaction and its oncogenic consequences. This finding
may lead to development of better therapeutic agents that may
target EWS-FLI1 and RHA interaction.
OP-56
Role of p38 MAP kinases in oncogene-induc ed
malignant transformation
A. Nebreda, I. Dolado, A. Cuadrado, V. Lafarga and A. Swat
CNIO, Melchor Ferna
´
ndez Almagro, Madrid, Spain.
E-mail:
We are investigating the role of p38 MAPKs in the processes of
cell proliferation, apoptosis and malignant transformation
induced by oncogenes. Oncogenic H-RasV12 induces a more dra-
matic transformed phenotype in p38a-deficient mouse fibroblasts
than in their wild type counterparts. The inhibitory effect of p38
MAPKs on H-RasV12-driven transformation was confirmed in
NIH3T3 fibroblasts overexpressing the p38 MAPK activator
MKK6. Interestingly, p38 MAPKs do not inhibit transformation
induced by all oncogenes but seem to rather specifically modulate
signalling pathways activated by oncogenic Ras. We have also
observed that, in fibroblasts expressing H-Ras V12, there is an
inverse correlation between p38a activity levels and the ability of
the cells to lose contact inhibition and grow in multiple layers.
Consistent with these results, p38a–/– fibroblasts grow to a
higher saturation density than wild-type fibroblasts. Moreover,

Penn State University, Hershey, PA, USA
We previously developed a cell line from a heart metastases of
4T1 murine breast carcinoma. This cell line named as 4THMpc
behaved more aggressively than the parental cell line (4T1) and
formed multiple macroscopic liver metastases. Expressions of
over 12 000 genes were determined in liver metastases, 4T1 and
4THMpc primary tumors. Comparison of gene array results
demonstrated that expression of adherence junctions proteins
were markedly decreased in liver metastases compared to primary
tumors formed by 4T1 and 4THMpc. Among these proteins
gamma catenin is examined at protein level. Immunoblots from
tumor lysates demonstrated the presence of gamma catenin in
both 4T1 and 4THMpc primary tumors. 4T1 primary tumors dis-
played more gamma catenin than 4THMpc. Immunohistochemi-
cally, patches of cells were stained in both groups. Interestingly,
cytoplasmic staining was observed in both groups but membrane
staining was detectable only in 4T1 tumor cells. Instead, nuclear
and perinuclear staining was observed in 4THMpc tumor.
Gamma catenin expression was undetectable in liver metastases.
Similarly, hepatocytes were mostly unstained, except around
metastases and the central vein. These findings demonstrate that
abnormal expression of gamma-catenin correlates with the more
aggressive tumor and an indicator of liver metastatic breast can-
cer. We here also demonstrated that liver metastases of breast
carcinoma do not express gamma catenin.
OP-58
Molecular and cellular characterization of
GPCR mas-induced tumour formation
W. Z. Lin, S. Y. Tsang, S. S. T. Lee and W. T. Cheung
Department of Biochemistry, The Chinese University of Hong

hepatocellular carcinoma cells
E. Kavak
1
,K.C¸ avus¸ og
˘
lu
1
,N.O
¨
ztu
¨
rk
2
, A. D. Duru
1
, T. Saygılı
1
,
T. Aslan
1
, E. K. Bayrakc¸ eken
1
, C. Bilgir
1
,M.C.Hız
1
, B. Ergel
1
,
D. O

tumors in nude mice. We used Serial Analysis of Gene Expres-
sion (SAGE) to compare gene expression between low and
high TCF activity cells. We have presently identified 429
differentially expressed tags, 326 show decreased expression pro-
file in high activity cells, whereas the remaining 103 tags show
increase. We compared these tags with the SAGE data of several
different types of tumors including liver and brain tumors. We
identified several molecules whose expression profile is similar in
several different tumors and in TCF4 over-activation. Some of
these molecules have not previously been related to cancer and
may be important candidates as tumor markers.
Acknowledgments: Bog
˘
azic¸ i University Scientific Research Pro-
jects, 04B105; Scientific and Technological Research Council of
Turkey, TBAG-2417; State Planning Organization of Turkey,
DPT 01K120290.
Intracellular Trafficking in Health and Disease
OP-60
The role of nucleoside diphosphate kinase in
ADP transport across the outer mitochondrial
membrane
V. V. Voinova and T. Yu. Lipskaya
Department of Biochemistry, Faculty of Biology, Lomonosov
Moscow State University, Mosow, Russia.
E-mail:
In aerobic cells cytoplasmic ADP is transformed into ATP
mainly in mitochondria, but the outer mitochondrial membrane
has a limited permeability for ADP. In the present study, the role
of nucleoside diphosphate kinase (NDPK) in transport of ADP

from the endoplasmic reticulum (ER) and degraded by the
proteasomes in the cytosol, a process called ER-Associated
Degradation (ERAD). Aberrant ERAD contributes to patho-
genesis of cystic fibrosis, a-1-antitrypsin deficiency, neurodegen-
erative diseases like Huntington disease and diabetes. ERAD
requires modification of misfolded proteins by chains of ubiqu-
itin. p97/VCP ATPase, which is recruited by its receptor in the
ER, then recognizes ubiquitinated misfolded proteins via its
ubiquitin-binding cofactor Ufd1-Npl4 heterodimer. gp78, ori-
ginally identified as the tumor autocrine motility factor recep-
tor promoting tumor metastasis, is also an ubiquitin ligase
(E3) involved in ERAD. We have identified a novel VCP-inter-
acting domain (VID) in gp78 and SVIP, another VCP-interact-
ing protein with unknown function. VID is transferable and
interferes with the binding of Ufd1 to VCP. Our data suggest
that VCP forms two mutually exclusive complexes, VCP-gp78
and VCP-Ufd1-Npl4. Functionally, both complexes are compet-
ent for CD3d (an ERAD substrate) binding and degradation.
Both E3 and VCP-recruitment activities of gp78 are required
for CD3d degradation. This study reveals a novel Ufd1-inde-
pendent ERAD mechanism that operates in parallel with the
previously described VCP-Ufd1-Npl4-mediated pathway.
Abstracts
59
OP-62
Ibogaine affects brain energy metabolism
R. Pas
ˇ
kulin
1

electrophoresis. Individual proteins were identified by MALDI-
TOF mass spectrometry. Levels of 12 proteins were observed to
increase, four of which were identified as metabolic enzymes
involved in glycolysis and the TCA cycle. The results suggest that
the vitalizing effect of ibogaine could be mediated by change in
energy availability. Since energy dissipating detoxification and
reversion of tolerance to opioids requires underlying functional
and structural changes in the cell, higher metabolic turnover
would be favourable. Understanding the pharmacodynamics of
antiaddiction drugs highlights the subcellular aspects of addiction
diseases, in addition to neurological and psychological perspec-
tives.
OP-63
Galectin-3: a lectin essential for apical protein
sorting
D. Delacour
1
, A. Koch
1
, C. I. Cramm-Behrens
1
, A. Le Bivic
2
,
H. Y. Naim
3
and R. Jacob
1
1
Department of Cytobiology, Philipps University Marburg,

a conclusion our data suggest that galectin-3 is the first lectin
sorting receptor identified in raft-independent apical trafficking.
OP-64
Role of ADP-ribosylation factor1 (ARF1) in
vesicular trafficking in plants
A. R. Memon and S. Usluer
TUBITAK, Institute for Genetic Engineering and Biotechnology,
Gebze, Kocaeli, Turkey. E-mail:
ADP-ribosylation factors (ARFs) are small monomeric GTP-
binding proteins, implicated in vesicle trafficking and secretion
pathway in yeast, mammalian and plant cells. ARFs have been
implicated in the formation of COPI-coated vesicles in eukaryotic
cells and ARF-GEFs are a target of Brefeldin A (BFA). We have
investigated the interaction between ARF and BFA and the
effect of a dominant inhibitory ARF [TN] mutant on Golgi-ER
transport in tobacco epidermal and BY2 cells. Our data show
that N-ST-GFP and GmMan1-RFP markers were transported to
Golgi in tobacco leaves and BY2 cells when expressed with wild-
type ARF1 or ARf1 [QL]. Most of the N-ST-GFP and
GmMan1-RFP markers were redistributed in ER when leaves or
BY2 cells were infiltrated with BFA or ARF-TN mutant. The
BFA effect was suppressed when the tobacco leaves expressed
with either ARF1 wild type or ARF1 [QL]. ARF1 wild type and
ARF1 [QL] also rescued the ARF1 [TN] phenotype in both
tobacco leaves and BY2 cells. Coexpression of either N-HDEL-
GFP or N-Sec-YFP with ARF [TN] mutant in tobacco leaves
resulted the appearance of an Endo H-resistant population of
GFP and YFP. This effect was overcome when leaves were coex-
pressed with ARF wild type or ARF [QL] mutant and was
enhanced by low concentrations of BFA. In contrast, Rab1b

lin and DOG caused differential activation of ChEs and ERK
proteins. Forskolin treatment increased AChE expression and
caused R28 cells to develop neurites causing cells to be choline-
receptive. DOG treatment increased both BChE expression and
activity along with neurite extension. Our studies display associ-
ation of ChEs with different signaling pathways and cell cycle
events.
Abstracts
60
Neurodegenerative Disorders
OP-66
The effect of beta-boswellic acid on
microtubule polymerization
O. Karima, G. H. Riazi, A. Moosavi Movahedi, F. Mokhtari
and B. Fakurian
Biochemistry Department, Institute of Biochemistry and
Biophysics, University of Tehran, Tehran, Iran.
E-mail:
Tau protein has been shown to be essential in Alzheimer’s disease
(AD). This important protein maintains the structural integrity
of microtubule. Hyperphosphorylation of tau protein observed in
AD and several mutations of the tau genes have been considered
to result in reduced binding ability of tau to microtubule and
resulted in a lack of microtubule dynamic instability. Therefore,
disruption of its structure leads to impairment of axonal trans-
port and finally loss of synaptic function in neural cells and cell
death. Therefore the key factor, which induces the loss of mem-
ory, could be in microtubule destabilization. The gum resin of
the Boswellia species (Frankincense) has long been in use for the
prevention of the amnesia and enhancing the power of memory

1
1
Institute of Reproductive and Developmental Biology, Clinical Sci-
ences Division, Imperial College London, Hammersmith Hospital
Campus, London, UK,
2
Department of Obstetrics and Gynaecology,
Perinatal Centre, Sahlgrenska Academy, Go
¨
teborg, Sweden.
E-mail:
c-Jun N-terminal kinase 3 (JNK3) is a member of the stress-acti-
vated group of mitogen-activated kinases (MAPK). Although
JNK3 is a potent mediator of apoptosis and neurodegenaration,
little is known about the role of JNK3 and its mechanism of
action in neonatal brain injury. The aim of the present study was
to compare the vulnerability of neonatal JNK3-KO and wild-
type mice to cerebral hypoxic-ischaemic injury (HII) using unilat-
eral acrotid occlusion combined with transient hypoxia. The
degree of neural tissue loss in JNK3-KO mice was substantially
reduced compared to wild-type mice (JNK3-KO 27.8 ± 2.8%
versus wild type 48.3 ± 2.0%, P < 0.0001) following HII. Mole-
cular analysis of JNK3 signalling revealed that HII increased
JNK phosphorylation and activation, and that JNK3 was the
major activated JNK isoform. Significantly, in JNK3-KO animals
there was no difference in the activation of either MKK4 or
MKK7. Downstream of JNK3, HII lead to increased phosphory-
lation of the transcription factors c-Jun and ATF-2. Interestingly,
although this phosphorylation was attenuated in JNK3-KO mice,
the effect on ATF-2 was significantly greater. In JNK3-KO mice

motor-cortex (MC), nucleus caudatus (NC) and SN from con-
trols (C) and patients with PD. Materials and Method: The ini-
tial samples consisted of 18 subjects of PD and C. Brains were
matched with age and postmortem time. Brain tissue was homo-
genized in a phosphate buffer with pH 7.3 and separated with
two-step centrifugation at 15 000 rpm for 30\,min and
15 000 rpm for 10 min at 4 °C. Antioxidant capacity in the sup-
ernatants was measured using oxygen radical absorbance assay
(ORAC) on a Perkin-Elmer spectrometer LS 55 with a fluores-
cent filters (Ex: 485 nm; Em: 520 nm). Hydroxyl and peroxyl rad-
ical generators were used. Fluorescence was used as a target of
free radical attack. Results: No changes in the antioxidant capa-
city of postmortem MC and NC of parkinsonian’s brain in com-
parison with C were found. In the SN of parkinsonians brain,
antioxidant capacity seems to be lower in comparison with C
(P < 0.05). Conclusion: Our results showed that in the SN of
parkinsonians brain the balance between production of free radi-
cals and the neutralization by a complex antioxidant system is
disturbed.
Abstracts
61
Bioinformatics and Proteomics
OP-69
Atomic hydration potenti als using Monte
Carlo reference state advance protein
solvation modeling
S. V. Rahmanov
Bioinformatics Department, GosNIIGenetika, Moscow, Russia.
E-mail:
We present a new method for constructing knowledge-based pot-

and B. Wittmann Liebold
2
1
Faculty of Engineering, Chemical Engineering Department,
Marmara University, Istanbul, Turkey,
2
Wita GmbH, Teltow,
Germany. E-mail:
The optimum living condition for a micro-organism is in its nor-
mal physiological environment. When any change occurs, this
change inflicts a stress on the organism. The type and the severity
of the stress factor determine the survival possibility and the con-
ditions for the micro-organism. Since the proteins are directly
responsible for maintenance of correct cellular function and con-
sequently the viability of the organism, the identification of pro-
teins and protein expression pattern under such given
physiological conditions by proteomic analysis is important for
functional studies of cellular processes. In our study, moderately
halophilic micro-organisms isolated from Anatolian saltern areas
are studied with traditional proteome tools, to determine the
characteristic proteins of the halophilic micro-organisms and to
have information of the microbial life in saltern parts of Anato-
lia. The isolates and reference strains are grown at different salt
concentrations (5% and 20% NaCl) and different temperatures
(10 and 40 °C). After growing and cell disrupture, the entire pro-
teins of the organisms are subfractionated as membrane proteins
and cytosolic proteins. An acidic character of most of the whole
cell proteins is detected both in isolates and the reference strains
(H. salina DSMZ5928 and H. halophila DSMZ4770). The 2-DE
maps of the new halophiles are compared with the reference

process involves sequence and literature curation. 1 In order to
achieve non-redundancy and to improve sequence reliability, all
protein sequences encoded by the same gene are merged into a
single entry. Sequence comparison allows us to find and show the
most reliable sequence. 2 Literature curation involves a critical
review of the current knowledge for each protein. Information is
extracted from scientific publications and continuously updated
thanks to external scientific expertise. A special emphasis is laid
on the annotation of biological events, which generate protein
diversity but are not predicted at the genomic level. Alternative
products, post-translational modification (PTMs) and polymor-
phisms are extensively annotated. 3 UniProtKB/Swiss-Prot is
highly cross-referenced to about 100 database.
OP-72
Fluorescent immunoprecipitation assay – a
proteomic analysis of cell surface proteins
A. Filatov
1
, G. Krotov
1
, M. Krutikova
1
and V. Zgoda
2
1
Institute of Immunology, Moscow, Russia,
2
V.N. Orechovich
Institute of Biomedical Chemistry RAMS, Moscow, Russia.
E-mail:

1
, Z. Kasap
1
and E. Kavak
2
1
Biological Sciences and Bioengineering Program, Sabanci Univer-
sity, Istanbul, Turkey,
2
Molecular Biology and Genetics, Bogazici
University, Istanbul, Turkey. E-mail:
G-protein-coupled receptors (GPCR) are a superfamily of cell
membrane proteins that play a vital role in transmitting extracel-
lular signals into the cytosol. These signals (ligands) bind GPCRs
to control the activation of various mechanisms inside the cell.
Due to their wide range of regulatory effects, GPCRs are one of
the main targets of pharmaceutical research today. However, the
structural information on these molecules are quite scarce, and
ligand specificity for most of them are unknown. In this study,
we describe the utilization of support vector machines to predict
the ligand specificity of a given class of GPCRs, based on total
and partial primary sequence information. We used three-letter
word composition of GPCRs using reduced amino acid alpha-
beth representation and Support Vector Machines for classifica-
tion. Using these compositions we could predict whether a
protein is a GPCR with 97% accuracy. We were also able to pre-
dict the ligands of GPCRs belonging to classes A, B and C with
an accuracy of 93%, 100% and 100%, respectively. Moreover,
we could also determine structural segments that were important
for ligand binding in GPCRs.

ence intervals) for these data sets and estimate that the complete
yeast interactome will have approximately 24 000–30 000 interac-
tions. We then apply this formalism to different data sets from
P. falciparum, D. melanogaster, C. elegans and H. sapiens to see
if predicted interactome size reflects the complexity of these
organisms. We obtain consistent interactome size estimates of
approximately 19 000, 70 000, 230 000 and 680 000 interactions,
respectively. Thus, the human protein interaction network
appears to be approximately an order of magnitude larger or
more complex than the fruitfly’s interactome. We conclude with
a discussion of the implications of these results for comparative
and functional analyses in systems biology.
1. Stumpf et al. PNAS 2005; 102: 4221–4224.
OP-75
Probable evolutionary pathways towards the
tubulo-interstitial nephritis antigen protein
family
C. Paliakasis, V. Kaltezioti, V. Sideraki, C. Zervas, A. Charonis
and S. Kossida
Foundation for Biomedical Research of the Academy of Athens,
Athens, Greece. E-mail:
Tubulo-interstitial Nephritis Antigen (TIN-ag) is involved in auto-
immune renal disease. It is a multidomain protein located at the
basement membrane of renal proximal tubule epithelium and
intestinal epithelium. Another family named ‘TIN-ag-like’ proteins
shares extensive similarity despite a different expression pattern.
More distant homologues are present in invertebrates. All mem-
bers of these two families exhibit similarity to relevant domains of
well-characterized protein families. The C-terminal half is largely
similar to the Cathepsin family of proteases; in the N-terminal

Cornell University, Ithaca, NY, USA.
E-mail:
Because of exponentially growing number of known protein
sequences, only about 30% of structure of proteins which are
potentially significant in biomedical sciences can be determined,
which calls for reliable algorithm for protein-structure prediction.
Local interactions play a very significant role in determining pro-
tein structure, because they determine the geometry of secondary
structure elements (helices and sheets), as well as that of turns
and loop regions. Therefore, accurate representation of local
interaction terms in the empirical force fields for physics-based
protein-structure prediction is of utmost importance. In this com-
munication, we present the determination of local interaction
terms for our physics-based united-residue UNRES force field [1]
for protein-structure prediction. We computed the corresponding
potentials of mean force from the energy surfaces of terminally
blocked amino acid residues calculated by the ab initio MP2/6-
31G** approach and the semiempirical AM1 method. Analytical
formulas derived based on generalized cumulant expansions were
subsequently fitted to the potentials of mean force and the result-
ing expressions were implemented in the UNRES force field. The
ability of the UNRES force field with new terms to predict pro-
tein structure by global energy minimization was tested on a
number of benchmark proteins with size from 50 to 100 residues.
1. Liwo A, Czaplewski C, Pillardy J, Scheraga HA. J Chem Phys
2001; 115: 2323–2347.
Abstracts
63
Clinical Proteomics
OP-77

identified by MALDI-TOF-TOF mass spectrometry. These pro-
teins were: albumin, a-enolase, calgranulin B, galectin 7, myoglo-
bin, myosin light chain 1 and light chain 2, and tropomyosin
b-chain. Also, differences in allele frequencies of a Wnt gene fam-
ily member between aggressive and non-aggressive cases and in
alternative splicing isoform expression between normal and meta-
static lymph nodes were observed. Our results showed different
gene and protein expression pattern and polymorphism frequen-
cies between aggressive and non-aggressive tumors, which may be
used as a predictor of outcome, and rational targets in HNSCC
(FAPESP/Ludwig Institute).
DNA and Protein Microarrays
OP-78
Redox-labelled nucleotides for molecular
biology and bioelectronics
W. Wlassoff
1
, D. Di Giusto
2
and J. Gooding
3
1
Biomedical Research Centre, Sheffield Hallam University,
Sheffield, UK,
2
School of Biotechnology and Biomolecular Sciences,
The University of New South Wales, Sydney, Australia,
3
School of
Chemical Sciences, The University of New South Wales, Sydney,

Chemistry, Goettingen, Germany. E-mail:
Polycomb group (PcG) genes are essential genes in higher euk-
aryotes for the maintenance of the spatially distinct repression of
developmentally important regulators. Their absence, as well as
overexpression, causes transformations in the axial organization
of the body. Little is known about their stability or the exact
mechanism of repression in vivo. Using fluorescence recovery
after photobleaching (FRAP), we determined the translational
diffusion coefficients of two GFP fusion proteins of the PcG pro-
teins as well as the binding equilibrium and dissociation con-
stants and residence times for complexes of the proteins in vivo at
different developmental stages in whole living Drosophila organ-
isms and tissues. The small nuclei of Drosophila cells as well as
the movement of chromatin within the nuclear volume presented
technical difficulties. Recently a method, RICS, was developed
that utilizes the temporal information encoded in CLSM data
and time correlation analysis of the fluorescence signals (FCS) to
determine diffusion constants. We use RICS to analzye the diffu-
sion and binding equilibria of another GFP-labeled PcG protein,
ESC, which is part of the histone methylation complex that
directs PcG protein binding to chromatin. We compare the pros
and cons of FRAP and RICS as well as the results for measure-
ments of Polycomb-GFP in both systems in living embryos and
larval tissues.
1. Ficz et al. Development 2005; 132.
2. Digman et al. Biophys J 2005; 89.
Abstracts
64
OP-80
Periodic twisted structure of amyloid fibrils

which were virtually homogenous along the fiber, the r and
FDLD images revealed a periodicity of about 1.5 lm. The
strong, periodic variations in the dichroism signals might origin-
ate from a twisted macrostructure of the amyloid fibrils. In addi-
tion, r and FDLD images yielded values for the order parameters
associated with the emission and absorption dipoles, respectively.
By also using data of P imaging, we determined the preferential
orientation of the transition dipoles and the static and dynamic
fluctuations of the dye molecules with respect to the long axis of
the fibrils.
OP-81
Capillary electrophoretic analysis of the
superoxide production by individual
mitochondria isolated from cybrid cells
D. Li
2
, M. Navratil
2
, B. G. Poe
2
, C. T. Moraes
1
and
E. A. Arriaga
2
1
Department of Neurology, University of Miami School of
Medicine, Miami, FL, USA,
2
Department of Chemistry, University

H. I. Canter
1
, P. Korkusuz
2
, I. Vargel
1
, F. Oner
3
, B. Cil
4
and
Y. Erk
1
1
Faculty of Medicine, Department of Plastic and Reconstructive
Surgery, Hacettepe University, Ankara, Turkey,
2
Faculty of
Medicine, Department of Histology and Embryology, Hacettepe
University, Ankara, Turkey,
3
Faculty of Pharmacy, Department of
Pharmaceutical Technology, Hacettepe University, Ankara,
Turkey,
4
Faculty of Medicine, Department of Radiology, Hacettepe
University, Ankara, Turkey. E-mail:
Bone grafts, used to provide structural integrity of cranial vault
remodeling, could not always integrate with the remaining bone
structures. They may sequestrate forming fibrous tissue around

M. Legorreta-Herrera
1
, A. Ramos-Avila
1
,
J. L. Ventura Gallegos
2
and A. Zentella-Dehesa
2
1
Divisio
´
n de Posgrado, FES Zaragoza, Universidad Nacional
Auto
´
noma de Me
´
xico, Me
´
xico,
2
Instituto de Investigaciones
Biome
´
dicas, Universidad Nacional Auto
´
noma de Me
´
xico, Mexico.
E-mail:

2+
pool in Leishmania donovani was iden-
tified by permeabilizing plasma membrane with digitonin. In Fura-
2 loaded cells Ca
2+
was released synergistically when mitochond-
rial function was blocked by antimycin and oligomycin. Vanadate
did not have any effect if applied before incorporation of these mit-
ochondrial poisons. However, the same inhibitor, which inhibits
Ca
2+
-ATPase activity of endoplasmic reticulum, was able to
release Ca
2+
at a slow rate when added after antimycin and oligo-
mycin. Alkalization of cytoplasmic pH allowed further release of
Ca
2+
essentially from the acidocalcisome. Purified glycosomes
could mediate Ca
2+
uptake mechanism in presence of vanadate
whereas bafilomycin, a specific and potent inhibitor of vacuolar
proton pump did not have any effect. Glycosomal Ca
2+
-ATPase
activity was optimum at pH 7.5. The apparent K
m
for calcium in
presence of vanadate was 12 nM. Taken together, it may be sug-

Antisense (AS) oligodeoxynucleotides (ODNs) have shown prom-
ise as chemotherapeutic agents. Exogenous delivery of phos-
phorothioate AS ODNs against different regions of P. falciparum
topoisomerase II gene to P. falciparum K1 strain between 0.01
and 0.5 lM significantly inhibited parasite growth compared with
sense sequence control suggesting sequence-specific inhibition by
fluorescence-activated cell sorter assay or by microscopic assay.
This inhibition was shown to occur during maturation stages, to
improve stability and to increase intracellular penetration, ODNs
were complexed with the biodegradable polymer chitosan to form
solid nanoparticles with an initial diameter of ~55 nm. AS nano-
particles showed stronger inhibition of parasite growth compared
with free AS ODNs. These results should prove useful in future
designs of novel antimalarial agents and use of nanoparticles to
delivery ODNs.
Immune Intervention
OP-86
Induction of NK lymphocytes against cancer
cell lines by administration of Morinda
citrifolia
T. I. Toliopoulos
1
, D. B. Bougiouklis
1
, D. T. Daskalou
1
,
K. S. Karkabounas
1
, G. S. Gerou