Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2017
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| Cite this: | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 157-160. — англ. |
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| citation_txt | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 157-160. — англ. |
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Experimental Oncology 39, 157–160, 2017 (June) 157
Materials of a Mini-syMposiuM
“new trends in cancer research
and innovative tuMor vaccines”
held on May 11, 2017 during the vactrain suMMer school
in Kyiv, uKraine
A mini-symposium “New Trends in Cancer Re-
search and Innovative Tumor Vaccines” was held
on May, 11 of 2017 during the VACTRAIN summer
school at R.E. Kavetsky Institute of Experimental Pa-
thology, Oncology and Radiobiology (IEPOR) of the
NAMS of Ukraine, Kyiv, Ukraine.
On this day, young researchers have presented re-
sults of their current work, along with few keynote lec-
tures. The main focus of this event was to understand
the modern trends to find the new markers to diagnose
cancer disease and new targets for cancer vaccines.
Also, there was a discussion on novel cancer vaccines.
The deputy director of IEPOR Associated profes-
sor Lubov Buchynska welcomed the participants
and wished the fruitful discussion, concerning new
approaches in the development of cancer vaccines.
Professor Chanchal Mitra (Hyderabad University,
India) gave a wide overview on miRNAs that play
an important role in cancer development. Associ-
ated professor Elena Kashuba (IEPOR) presented
the new data on a role of mitochondrial ribosomal
protein S18-2 as the new oncoprotein and a putative
target for cancer vaccine. More data and the future
perspectives are described in abstracts of young
researchers from Ukraine, Belarus, Latvia, and Lithu-
ania (see below).
Concluding mini-symposium, supported by the
VACTRAIN of Horizon 2020 program, Elena Kashuba
thank VACTRAIN for the possibility of schooling the
young researchers with an aim to help them to became
the critically thinking open-minded researchers. The
importance of scientific collaboration was also enligh-
tened. Definitely, this summer school resulted in the
enhanced knowledge in medical science and helped
to plan the new fruitful scientific works with the aim
to combat cancer.
selected abstracts
coMparison of different Kind
of dendriMers for the anticancer
rnai guides delivery
Volha Dzmitruk1, Maksim Ionov2,
Dzmitry Shcharbin1, Maria Bryszewska2
1Institute of Biophysics and Cell Engineering of NAS
of Belarus, Minsk, Belarus
2University of Lodz, Lodz, Poland
E-mail: dmitruk.olga@gmail.com
One of the directions of gene therapy is curative
cancer treatment. During transformation, normal
cells start to propagate uncontrollably and lose
the ability to undergo apoptosis, resulting in tu-
mor formation. Expression of apoptosis inhibitors
(BCL-2 family proteins) can be downregulated
at a post-translational level by means of RNA inter-
ference (RNAi), i.e. by a process of highly selective
gene silencing. RNAi effectors — siRNA (small
interfering RNA) or microRNA are able to cleave
mRNA, using the cellular machinery (Burlacu,
2003). The major limit of such gene therapy appli-
cation is an absence of effective delivery of nucleic
acids (siRNA) into the target cells (Songet et al.,
2005).
Dendrimers are synthetic polymers, highlighted
for gene delivery, due to their monodispersity, pre-
determined tree-like structure, stability, low visco sity
of their solutions and a large number of charged
end groups. Cationic dendrimers can complex with
nucleic acids by self-assembly, making complexes
called “dendriplexes” (Wang et al., 2010).
Different kinds of dendrimers were tested
as delivery systems for different RNAi effectors.
It was revealed that PAMAM (carbosilane and
phosphorous-containing dendrimers) form stable
complexes with small acting RNAs by self-assem-
bling in up to 1 µm in size. In contrast to others,
phosphorous dendrimers are insensitive to heparin
impact, thus indicating not only electrostatic in-
teraction within the dendriplex (Ionov et al., 2015).
Cytotoxicity studies on HeLa (cervical cancer) and
HL-60 (promyelocytic T-leukemia) cell lines showed
the synergism in action of siRNAs specific for various
anti-apoptotic proteins, in comparison with treatment
with each single siRNA. This was observed when
siRNAs were delivered by cationic dendrimers. In our
experiments, the cytotoxicity of siRNA/dendrimer
complexes showed a dual nature, based on three
mechanisms: apoptosis, autophagy, and necrosis.
The siRNA induced apoptosis and, moreover, den-
drimers themselves may induce all above, depend-
ing on the chemical nature of dendrimer (Dzmitruk
et al., 2015).
This work was supported by 2 grants from the
Belarusian Republican Foundation for Fundamen-
tal Research No. B15RM-060 and No. B15CO-041,
a Marie Curie International Research Staff Exchange
Scheme Fellowship within the 7th European Com-
munity Framework Programme, project No. PIRSES-
GA-2012-316730 NANOGENE.
Exp Oncol 2017
39, 2, 157–160
conference report
158 Experimental Oncology 39, 157–160, 2017 (June)
siMultaneous cd150 and cd180 ligation
Mutually inhibit aKt and MapK signal
transduction pathways in the chronic
lyMphocytic leuKeMia b cells
Inna Gordiienko, Larysa Shlapatska,
Valeriia Kholodniuk, Lilia Sklyarenko,
Svitlana Sidorenko
R.E. Kavetsky Institute of Experimental Pathology,
Oncology and Radiobiology, NAS of Ukraine, Kyiv,
Ukraine
E-mail: imgordiienko@gmail.com
Combination of input signals from cell surface recep-
tors (e.g. BCR, CD140, BAFFR, TLR) in B-cell malignancies
underlies disease outcome. It was shown that cell surface
CD150 (csCD150) and CD180 (csCD180) expression
in chronic lymphocytic leukemia (CLL) correlates with
favourable prognosis.
We found that csCD150 expression has strong posi-
tive correlation with csCD180 expression in CLL cases (r =
0.75, p = 0.0001). Moreover, CD150 showed the highest
level of colocalization with CD180 (R = 0.88 ± 0.13; R[r] =
0.87 ± 0.1) at a cell membrane of CLL B cells. To determine
the biological significance of simultaneous CD150 and
CD180 expression in CLL pathobiology we performed in vi-
tro stimulation assay followed by signaling pathways analy-
sis. CD150 or CD180 crosslinking on CLL B cells alone
resulted in activation of Akt, mTORC1, ERK1/2, p38MAPK
and JNK1/2 pathways. Both CD150 and CD180 receptors
affect protein synthesis through p70S6K phosphorylation
and transmit pro-survival signals via Akt-mediated GSK3β
and FOXO1/FOXO3a transcription factors inhibition. It was
surprising that simultaneous CD150 and CD180 liga-
tion blocked Akt, mTORC1 as well as MAPK pathways
activation in CLL B cells. Moreover, in the absence
of CD150 and CD180 on cell surface, both antigens were
expressed on the protein level in the cytoplasm. Intracellu-
lar distribution of CD150 and CD180 in CLL B cells showed
different pattern. CD150 antigen was colocalized with
markers of endoplasmic reticulum, Golgi apparatus, early
endosomes, but only trace amount was detected in lyso-
somes. In contrast, CD180 was detected preferentially
in early endosomes (R = 0.60 ± 0.10; R[r] = 0.59 ± 0.10).
It could be concluded that malignant CLL B cells try to es-
cape simultaneous cell surface CD150 and CD180 expres-
sion since combined ligation of these receptors leads
to blocking of pro-survival pathways.
adaptor protein ruK/cin85 increases
tuMorigenicity of MaMMary
adenocarcinoMa cells via proMoting
eMt
Iryna Horak, Ganna Pasichnyk, Dmytro Shytikov,
Denys Geraschenko, Liudmyla Drobot
Palladin Institute of Biochemistry, NAS of Ukraine, Kyiv,
Ukraine
E-mail: iryna.horak@gmail.com
Adaptor proteins serve as molecular platforms for
multimolecular complexes assembly and thereby regulate
intracellular signaling. Our previous results demonstrated
that tumors (including breast cancer) are character-
ized by increased expression of adaptor protein Ruk/
CIN85 in comparison to normal tissues. This adaptor plays
a critical role in several cellular processes, such as ligand-
induced endocytosis of RTKs, intracellular vesicular
trafficking, adhesion, motility, and survival. In this study
we investigated the effect of Ruk/CIN85 on tumorigenicity
of mammary adenocarcinoma cells.
As a model we used mouse 4T1 mammary adeno-
carcinoma cells with stable overexpression (RukUp) and
downregulation (RukDown) of Ruk/CIN85. Motility and
invasiveness of these cells were analyzed by scratch
test and Boyden chamber assay. The effectiveness
of metastasіs formation was studied using experimental
metastasis assay. qPCR and Western blotting were carried
out to evaluate gene and protein expression.
We demonstrated that Ruk/CIN85 overexpression
leads to elevation of motility and invasiveness in vitro and in-
creased the ability to produce lung metastasis in vivo. Also,
we demonstrated increased expression of mesenchymal
markers in RukUp cells and epithelial markers in RukDown
cells that mean Ruk/CIN85 promotes EMT in 4T1 cells.
identification of novel Molecular
and genetic MarKers for early
detection of epithelial tuMors
and prognosis of the course of disease
Oksana Mankovska, Vladimir Kashuba
Institute of Molecular Biology and Genetics,
NAS of Ukraine, Kyiv, Ukraine
E-mail: mankovsska@gmail.com
The main topic of our research is the identification
of molecular and genetic markers for the early detection
of epithelial tumors and prognosis of the disease. Early de-
tection of cancer can lead to more successful treatment,
thus, significantly increasing the chances for recovery. The
investigation of non-invasive diagnostic tools, which can
be used in clinical practice, is now the main focus of on-
cology. This problem can be solved by using of molecular
nucleic acid markers that are present in biological fluids
(serum, urine, semen etc).
We have developed several useful and effective ap-
proaches for the investigation of molecular and genetic
markers. We are studying both, genetic and epigenetic
changes in cancers of different types, using a broad-
scale screening of their presence in clinical samples. The
techniques and methods we use allow us to obtain reliable
results and propose them for implementation in clinical
practice.
diagnostic value of neutrophil/
lyMphocyte ratio in hepatocellular
carcinoMa and focal nodular
huperplasia: a pilot study
Dzeina Mezale1, Ilze Strumfa1, Andrejs Vanags2,
Ilze Fridrihsone1, Polina Viktorova1
1Department of Pathology, Riga Stradins University,
Riga, Latvia
2Department of Surgery, Riga Stradins University,
Riga, Latvia
E-mail: dzeina.mezale@gmail.com
Hepatocellular carcinoma (HCC) is the sixth most
common malignancy and the third most common
Experimental Oncology 39, 157–160, 2017 (June) 159
cause of cancer death worldwide (Jain et al., 2017).
Distinguishing a well-differentiated HCC from benign
liver nodules, such as focal nodular hyperplasia
(FNH), may be difficult in some cases, particularly
in small needle core biopsies (Coston et al., 2008),
thus additional diagnostic markers are essential.
Routinely available marker of the systemic inflam-
matory response such as neutrophil — lymphocyte
ratio (NLR) has recently been reported as a predictor
of HCC; an ele vated level indicating a poor prognosis
(Xiao et al., 2014). The aim of the present study was
to examine whether NLR could be used as an ad-
ditional diagnostic marker distinguishing between
HCC and FNH.
In a retrospective pilot study, 14 patients with
morphologically confirmed HCC and 6 patients with
FNH were included. NLR was defined as the absolute
neutrophil count divided by the absolute lymphocyte
count. Complete blood count levels were used to cal-
culate NLR. Data were evaluated using IBM SPSSv23.
To assess data distribution, an independent-samples
Kolmogorov — Smirnov test was performed. Descrip-
tive statistic evaluation was performed, including
detection of median values and interquartile range
IQR = Q3–Q1. To compare groups, independent
samples median test and Fisher exact significance
test were used. The significance level was set at 0.05.
The median value of NLR in FNH group was
1.67 (N = 6; Q1 = 1.20; Q3 = 2.57; IQR = 1.37). The
median NLR of the patients with HCC was 2.49 (N = 14;
Q1 = 1.97; Q3 = 4.55; IQR = 2.58), which is higher
comparing with FNH group, but the difference was
statistically insignificant (p = 0.141).
Concluding, we showed that i) A complete
blood count with following calculation of NLR
is widely available blood test, which has a potential
to be a useful additional marker distinguishing be-
tween HCC and benign liver nodules, such as FNH;
ii) The median NLR in HCC patient group was higher
than in FNH group (1.67 and 2.49, respectively),
which corresponds to the available scientific data,
but the difference was found to be statistically insig-
nificant. However, further research in a larger group
is clearly required.
interplay between Mitochondrial
ribosoMal protein
s18-2 and retinoblastoMa
susceptibility (rb) protein
in regulation of cell steMness
and differentation
Muhammad Mushtaq1, Larysa Kovalevska2,
Elena Kashuba1, 2
1MTC, Karolinska Institutet, Stockholm, Sweden
2R.E. Kavetsky Institute of Experimental Pathology,
Oncology and Radiobiology, NAS of Ukraine, Kyiv,
Ukraine
E-mail: kreyl@yahoo.com
We have found that S18–2 is involved in regula-
tion of the RB-dependent pathway. It binds to both
hypo- and hyperphosphorylated RB. The binding be-
tween RB and S18–2 is promoted when cytoplasmic
S18–2 is targeted to the nucleus, and this disrupts
the association of E2F1 with RB, as indicated by the
increased level of free E2F1 in the nucleus (Kashuba
et al., 2008). This presumably lifts the RB-dependent
block of S phase entry. We have also found that
overexpression of the human S18–2 immortalized
primary rat embryonic fibroblasts that showed
properties of embryonic stem cells (Kashuba et al.,
2009). Elevated expression of S18–2 in stem cells
(our findings and analysis of published microarray
data) raises the question of whether this protein co-
operates with the RB protein in cell differentiation
and oncogenesis.
We aimed to seek a connection between the ex-
pression of RB and S18–2 in Rb1–/– mouse embry-
onic fibroblasts and stemness. We hypothesized that
simultaneous expression of both proteins at the high
levels might support stemness. Transfections, inocu-
lation into SCID mice, directed differentiation, qPCR,
immunostaining, immunohistochemistry, Western
blotting, zebrafish embryogenesis were used in the
present work.
We showed that S18–2 protein, together with
RB, plays a crucial role in cell de-differentiation.
We have also found that overexpression of S18–2 and
RB is needed for maintenance of cell stemness. Such
cells can differentiate into various cell lineages under
certain conditions. The presence of RB and simultane-
ous expression of S18-2 at high levels are required for
the cell stemness.
inhibition of ire1 signaling affects
the eXpression of genes encoding
glucocorticoid receptor and soMe
related proteins in u87 glioMa cells
Oksana Ratushna, Dmytro Minchenko,
Olena Riabovol, Oleksandr Minchenko
Palladin Institute of Biochemistry, NAS of Ukraine,
Kyiv, Ukraine
E-mail: oksana_ratushna@hotmail.com
Glucocorticoid receptor (nuclear receptor subfam-
ily 3, group C, member 1; NR3C1) plays an important
role in the regulation of numerous metabolic and
proliferative processes, including tumorigenesis and
metabolic diseases. Regulation of these processes
by glucocorticoids is preferentially stress-dependent,
but the involvement of the endoplasmic reticulum (ER)
stress signaling has not been explored yet.
The aim of the present investigation was to ex-
amine the effect of inhibition of ER stress signaling
mediated by IRE1 (inositol requiring enzyme 1), which
is a central mediator of the unfolded protein response,
on the expression of genes encoded NR3C1 and
some related proteins (SGK1, SGK3, NCOA1, NCOA2,
ARHGAP35, NRIP1, and NNT) and their hypoxic regu-
lation in U87 glioma cells for evaluation of their possible
significance in the control of glioma growth.
The expression of NR3C1, SGK1, SGK3, NCOA1,
NCOA2, ARHGAP35, NRIP1, and NNT genes in U87 glio-
ma cells transfected by empty vector pcDNA3.1 (control)
160 Experimental Oncology 39, 157–160, 2017 (June)
and cells without IRE1 signaling enzyme function (trans-
fected by dnIRE1) upon hypoxia was studied by qPCR.
We have shown that inhibition of IRE1 signaling en-
zyme function up-regulates the expression of NR3C1,
SGK1, NCOA1, NCOA2, ARHGAP35, and NNT genes
in U87 glioma cells in comparison with the control glioma
cells, being more significant changes for NR3C1, SGK1,
and NNT genes. At the same time, the expression
of SGK3 and NRIP1 genes is strongly down-regulated
in glioma cells upon inhibition of IRE1. We have also
shown that hypoxia increases the expression of NR3C1,
SGK1, NCOA2, ARHGAP35, and NNT genes but de-
creases SGK3, NRIP1, and NCOA1 genes expression
in control glioma cells. Moreover, the inhibition of both
enzymatic activities (kinase and endoribonuclease)
of IRE1 in U87 glioma cells enhances the effect of hy-
poxia on the expression of SGK1, SGK3, and NNT genes
but decreases the sensitivity of NR3C1 gene to hypoxic
condition. Furthermore, the expression of NCOA1 gene
is resistant to hypoxia in control glioma cells, but
NCOA2, NRIP1, and ARHGAP35 genes are resistant
to this condition in glioma cells without functional activity
of IRE1 signaling enzyme.
Results of this investigation demonstrate that in-
hibition of IRE1 signaling enzyme function affects the
expression of NR3C1, SGK1, SGK3, NCOA1, NCOA2,
ARHGAP35, NRIP1, and NNT genes in U87 glioma
cells in the gene specific manner and that all these
genes are regulated by hypoxia preferentially through
IRE1 signaling pathway of ER stress.
eXpression of cancer-associated
genes in prostate tuMors
Evgenia Rosenberg1, Ganna Gerashchenko1,
Natalia Hryshchenko1, Raisa Lytvynenko2,
Yuriy Vitruk2, Oleksandr Gryzodub2,
Eugenia Stakhovsky2, Vladimir Kashuba1
1Institute of Molecular Biology and Genetics,
NAS of Ukraine, Kyiv, Ukraine
2National Cancer Institute, Ministry of Healthcare
of Ukraine, Kyiv, Ukraine
E-mail: kalirra@mail.ru
Despite numerous studies that aimed to define the
molecular mechanisms that underlay formation and
progression of prostate tumors, markers of advanced
prostate cancer are still not elaborated. The objective
of our investigation was an analysis of relative gene
expression of a set of cancer-associated genes, previ-
ously selected based on a work with the prostate cancer
cell lines. We studied the following cancer-associated
genes in the clinical samples of prostate tumors —
TGFB1, IL1B, FOS, EFNA5, TAGLN, PLAU, and EPDR1.
To study the expression of the abovementioned
genes, a total RNA from 16 prostate adenomas, 37 ad-
enocarcinomas and 29 conventionally normal tissues
were isolated. qPCR was performed to analyze the
gene expression pattern. Immunohistochemistry was
conducted to determine the expression of the TGFβ1,
FOS, TAGLN, and PLAU proteins.
Alterations in the relative gene expression levels
were found for TGFB1, IL1B, FOS, EFNA5, TAGLN,
PLAU, and EPDR1 genes in all analyzed groups. For
TGFB1, FOS, TAGLN, and PLAU genes obtained results
were confirmed on the protein level. FOS protein was
increased during progression from adenoma to ag-
gressive prostate cancer. Meanwhile, levels of TGFβ1,
TAGLN, and PLAU proteins were decreased in ad-
vanced prostate cancer.
Concluding, the obtained results might indicate
that TGFB1, FOS, TAGLN, and PLAU are potential bio-
markers for prostate cancer. Moreover, FOS gene may
be a putative target for anti-cancer therapy. However,
these results are preliminary and it requires the further
investigations.
the role of epigenetics
in the developMent of type 2 diabetes
and effectiveness of MetforMin
therapy
Monta Ustinova, Ineta Kalniņa, Ilze Elbere,
Janis Kloviņš
Latvian Biomedical Research and Study Centre,
Riga, Latvia
E-mail: monta.ustinova@biomed.lu.lv
The incidence of type 2 diabetes mellitus (T2D) has
become an epidemic, increasing the risk of cardiovas-
cular diseases and premature death. The global T2D
prevalence is 8.3% with the highest burden in develop-
ing countries. Metformin is a first-line therapy in T2D
treatment, but pharmacodynamics of metformin is still
not clear. There is a variable efficiency observed for
metformin therapy; furthermore, nearly one-third
of T2D patients do not respond to it.
So far, there are no validated biomarkers that would
predict the effectiveness of diabetes medications
and the patient’s responses to treatment. Epigenetic
signatures are considered to be promising biomarkers
for the evaluation of the therapeutic efficiency in meta-
bolic diseases. The impact of metformin on epigenetic
regulation has not been described.
Studies on metformin’s mechanism of action and
development of new epigenetic biomarkers will allow
to predict the effectiveness and the potential toxicity
of metformin in T2D patients.
The aim of the study is to determine the epigenetic
patterns related with T2D and response to antidiabetic
drug therapy. The main objectives are the following:
i) To identify metformin-induced novel DNA meth-
ylation and RNA expression signatures in healthy
individuals and T2D patients; ii) To validate potential
epigenetic biomarkers in a large cohort of T2D patients
after long-term metformin therapy; iii) To estimate the
concordance between epigenetic patterns derived
from white blood cells with those from target tissues,
by analysis of DNA methylation and RNA expression
in mice; and iv) To develop an epigenetic-based
algorithm for prediction of efficacy and side-effects
of metformin therapy.
Copyright © Experimental Oncology, 2017
|
| id | nasplib_isofts_kiev_ua-123456789-137630 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-02T10:56:29Z |
| publishDate | 2017 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | 2018-06-17T14:21:33Z 2018-06-17T14:21:33Z 2017 Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 157-160. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/137630 en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Conference reports Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine Article published earlier |
| spellingShingle | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine Conference reports |
| title | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine |
| title_full | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine |
| title_fullStr | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine |
| title_full_unstemmed | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine |
| title_short | Materials of a mini-symposium “New trends in cancer research and innovative tumor vaccines” held on May 11, 2017 during the vactrain summer school in Kyiv, Ukraine |
| title_sort | materials of a mini-symposium “new trends in cancer research and innovative tumor vaccines” held on may 11, 2017 during the vactrain summer school in kyiv, ukraine |
| topic | Conference reports |
| topic_facet | Conference reports |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/137630 |