first multidisciplinary conference of the italian ... · 17) test of genetic code evolution...

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FIRST MULTIDISCIPLINARY CONFERENCE OF

THE ITALIAN RESEARCHERS

IN CZECH REPUBLIC

Italian Institute of Culture

Sporkova 14, Mala Strana, Prague

18-20 June, 2019

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SCIENTIFIC PROGRAM AND ABSTRACTS

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PROGRAM

18/6/2019

17.00 – 19.00 Opening ceremony with the participation of the Italian Ambassador

H.E. Francesco Saverio Nisio, the Representative of the Italian Institute of Culture in

Prague Dr. Mauro Ruggiero, the President of the Czech Immunological Society and

organizer of the Conference Dr. Luca Vannucci, and authorities from the Czech

Universities, Czech Academy of Sciences, Ministry of Education, Youth and Sports.

Concert of Johanna Hanikova, pianist

19/6/2019

Section 1: Bio-medicine, physiology and biotechnologies

Chair: Luca Vannucci, Giancarlo Forte

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Part 1 - 9.00-10.40

Luca Vannucci. Morphology and immunity in transforming tissues: looking for the

tumor niche. Laboratory of immunotherapy, Institute of Microbiology CAS, v.v.i.,

Prague 20‘

Giancarlo Forte. International Clinical Research Center: a science excellence center

and an incubator for international talents. Center for Translational Medicine (CTM)

International Clinical Research Center (ICRC), St. Anne’s University Hospital, Brno,

Czech Republic 20’

Guido Caluori. Basic to clinical cardiological research in FNUSA-ICRC

electrophysiological center Interventional Cardiac Electrophysiology, FNUSA-ICRC,

Brno and Nanobiotechnology, CEITEC-MU, Brno, Czech Republic 20’

Alice Abbondanza. Understanding the role of nicotinic cholinergic receptors in

striatal-based behaviour Department of Neurochemistry, Institute of Physiology, Czech

Academy of Sciences, Prague, Czech Republic 20’

Anna Malandra. Effect of Bordetella pertussis virulence factors increasing cAMP

levels on mucin production in human airways. Institute of Microbiology of the CAS,

v.v.i., Prague, Czech Republic 20’

Question time: 10.40-10.55

Coffee break 10.55-11.10

Part 2 - 11.10-11.50

Vincenzo Tarallo. Evolved Methionine Sulfoxide Reductase A for Biotechnological

Applications. Department of Organic Chemistry, Faculty of Science, Charles University

20’

Paolo Tenti. Lysyl Oxidases and formation of the early Tumoral Niche. Laboratory of

immunotherapy, Institute of Microbiology of the CAS, v.v.i., Czech Republic and

Faculty of Science, Charles University, Prague, Czech Republic 20’

Question time: 11.50 – 11.55

Part 3 – 11.55- 12.50

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Section 2: Physics, chemistry and engineering

Chair: Benjamin Irving

Mini-session: Computational Materials Science

Benjamin Irving, Antonio Cammarata, Paolo Nicolini (55 min)

Advanced Materials Group, Department of Control Engineering, Faculty of Electronic

Engineering, Czech Technical University in Prague, Prague, Czech Republic

a) Overview of the mini-session (B. Irving) 10 min

b) Nanoscale friction properties of ordered and disordered molybdenum disulfide (P.

Nicolini) 15 min

c) Tailoring friction and energy dissipation at nanoscale (A. Cammarata) 15 min

d) Nanoscale properties of TMDs: an in-silico study (B. Irving) 15 min

Lunch 12.50 – 14.00

Part 4 - 14.00-15.20

Chair: Alessandra Picchiotti, Gabriele Maria Grittani

Francesco Piana. Polymers and composite materials for flexible electronics. Ústav

makromolekulární chemie AV ČR, v. v. i. - Oddělení polymerů pro elektroniku a

fotoniku Prague, Czech Republic 20’

Lorenzo Giuffrida. Brilliant and energetic alpha-particle source based on proton-

boron nuclear fusion. Institute of Physics ASCR, v.v.i (FZU), ELI-Beamlines project,

Prague, Czech Republic 20’

Daniele Margarone. Laser-driven Ion Acceleration and Societal Applications at the

Extreme Light Infrastructure (ELI). Institute of Physics ASCR, v.v.i (FZU), ELI-

Beamlines project, Prague, Czech Republic 20’

Gabriele Maria Grittani. Radiotherapy with high energy electrons generated by high

power lasers: a potential new technology to treat cancer. Institute of Physics ASCR,

v.v.i (FZU), ELI-Beamlines project, Prague, Czech Republic 20’



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Part 5 - 15.40-16.40

POSTER SESSION

1) Role of APP axonal transport in Alzheimer’s disease and Chronic Traumatic

Encephalopaty. Valentina Lacovich, PhD, FNUSA-ICRC, Victorio Martin Pozo

Devoto, PhD, FNUSA-ICRC, Maria Novaková, PhD, FNUSA-ICRC, Kateřina Texlová,

Bc, FNUSA-ICRC, Monica Feole, MS, FNUSA-ICRC, Gorazd Bernard Stokin, MD,

PhD, FNUSA-ICRC

2) In vitro model of cardiac fibrotic process to unveil the mechanisms of cardiac

cells crosstalk. Pamela Mozetic, Ana Rubina Perestrelo, Jorge Oliver-De La Cruz and

Giancarlo Forte. International Clinical Research Center (FNUSA-ICRC), St. Anne's

University Hospital, Brno, Czech Republic; Competence Center for Mechanobiology in

Regenerative Medicine, Brno, Czech Republic.

3) hnRNPC: a linker between ECM mechanics and mRNA homeostasis in cardiac

diseases. Fabiana Martino1,2

, Ana Rubina Perestrelo1, Stefania Pagliari

1, Jan Vrbský

1,

Vladimír Vinarský 1,3

, Jorge Oliver De La Cruz1,3

, Francesca Cavalieri3, Giancarlo

Forte1,3,5;

1Center for Translational Medicine (CTM), International Clinical Research

Center (ICRC), St. Anne´s University Hospital, Brno, Czech Republic0; 2Department of

Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; 3Competence Center for Mechanobiology in Regenerative Medicine, INTERREG

ATCZ133, CZ-62500 Brno, Czech Republic; 4Department of Chemical and

Biomolecular Engineering, University of Melbourne, Australia; 5Department of

Biomaterials Science, Institute of Dentistry, University of Turku, Finland.

4) Identifying regulators of axonal transport. Monica Feole; Victorio M. Pozo Devoto;

Mária Čarná; Valentina Lacovich; and Gorazd B. Stokin. Translational Neuroscience

and Aging Program (TAP), Center for Translational Medicine (CTM),

International Clinical Research Center (ICRC), St. Anne's University Hospital, Brno, Czech

Republic

5) The role of PV+ GABAergic interneurons in the medial prefrontal cortex on a

rat model of psychosis-related cognitive inflexibility as revealed by optogenetic

stimulations. E. Patrono1, K. Hruzova1, J. Svoboda1, A. Stuchlík1. 1Institute of

Physiology - Academy of Sciences Czech Republic, Neurophysiology of Memory,

Praha, Czech Republic

6) Molecular principles of Cajal body formation. Davide Alessandro Basello1,2

,

Michaela Efenberková1, Radek Macháň

2, Nicola Maghelli

3, David Stanek

1.

1Institute of

Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic, 2Charles

University in Prague, Faculty of Science, Prague, Czech Republic ,3Max Planck

Institute of Molecular Cell Biology and Genetics, Dresden, Germany

7) NFAT as a novel pathway in mesenchymal stem cell response to inflammation.

Tidu F. (1, 2), Jose S. S. (1, 2), De Zuani, M. (1), Bendíčková K.(1), Pompeiano A. (1),

Bělašková S. (1), Frič J. (1); (1) St. Anne’s University Hospital Brno International

Clinical Research Center, Brno, Czech Republic; (2) Department of Biology, Faculty of

Medicine, Masaryk University, Czech Republic

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8) Understanding nanoparticles-cells' interactions through mechanobiology. Marco

Cassani, Jorge Oliver-De La cruz, Soraia Fernandes, Giancarlo Forte. Center for

Translational Medicine (CTM) International Clinical Research Center (FNUSA-ICRC)

St. Anne's University Hospital

9) Femtosecond Stimulated Raman Spectroscopy. Alessandra Picchiotti, ELI-

beamlines/ELIBIO - Institute of Physics ASCR, v.v.i (FZU), ELI-Beamlines project,

Prague, Czech Republic

10) Pulse Radiolysis. Martin Precek, ELI-beamlines/ELIBIO - Institute of Physics

ASCR, v.v.i (FZU), ELI-Beamlines project, Prague, Czech Republic

12) Ultrafast dynamics of plasmon and magnetic resonances in nanoparticles.

Carlo Maria Lazzarini, Tadzio Levato, Jamie M. Fitzgerald, José A. Sánchez-Gil,

Vincenzo Giannini. Institute of Physics of the ASCR, ELI Beamlines Project, Prague,

Czech Republic

13) In vitro and in vivo evaluation of ferric-based and gadolinium-based

nanoparticles in theranostic perspective. Lenka Rajsiglova. Laboratory of

immunotherapy, Institute of Microbiology of the CAS, v.v.i., Faculty of Science,

Charles University, Prague, Czech Republic

14) Immunity and remodelling of human colon mucosa in colorectal cancer. Pavol

Lukac. Laboratory of immunotherapy, Institute of Microbiology of the CAS, v.v.i. ;

Faculty of Science, Charles University, Prague, Czech Republic

15) Mechanism underlying axonal swelling formation. Victorio M Pozo Devoto,

Valentina Lacovich, Maria Carna, Monica Feole, Katerina Texlova & Gorazd B Stokin.

Translational Neuroscience and Aging Program (TAP), Center for Translational

Medicine (CTM), International Clinical Research Center (ICRC), St. Anne's University

Hospital, Brno, Czech Republic

16) Mechanical thrombectomy results variability per occluded artery in ischaemic

stroke Luca Mengozzi. 3rd

Medical Faculty, Charles University, Prague Czech Republic

17) Test of genetic code evolution hypotheses: Reverse evolution of specific target proteins

by mRNA-display technique. Valerio Guido Giacobelli1, Kosuke Fujishima

2, Vyacheslav

Tretyachenko1, Klara Hlouchova

1;

1Department of Cell Biology, Faculty of Science, Charles

University, Prague, Czech Republic, 2Earth-Life Science Institute, Tokyo Institute of

Technology, Tokyo 1528550, Japan

18) Glutamate receptors as therapeutic targets for glioblastoma multiforme

(GBN). Daniela Arduini. Delegazione Nazionale Biotecnologi ONB, Rome, Italy

19) mGlu3 metabotropic glutamate receptors as candidate targets for

neuroprotective drugs in the MTPT mouse model of Parkinsonism. Marika

Alborghetti. Delegazione Nazionale Biotecnologi ONB, Rome, Italy

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20/6/2019

Part 1 - 9.00-10.10

Section 2: Physics, chemistry and engineering (continuation)

Chair: Daniele Margarone, Antonio Cammarata

Roberto Versaci. Radiation transport Monte Carlo simulations at ELI Beamlines.

Institute of Physics ASCR, v.v.i (FZU), ELI-Beamlines project, Prague, Czech Republic

20’

Alessandra Picchiotti, Jakob Andreasson. Bio and material applications at ELI-

beamlines. Institute of Physics ASCR, v.v.i (FZU), ELI-Beamlines project, Prague,

Czech Republic 20’

Federico Brivio. Computational approach to materials modelling. Charles University

20’



Section 3: Humanities, economics, social sciences, archaeology

Chair: Luca Vannucci

10.20-11.15

Dino Numerato. Corruption and Public Secrecy: An ethnography of football match-

fixing. Faculty of Social Sciences, Charles University, Prague 20’

Massimiliano Nuzzolo. Archaeological works in Egypt: the latest results of the Italian

expedition at the solar temples. Czech Institute of Egyptology, Charles University

Prague 20’


11.15-11.30 Conclusions and farewell ceremony with Authorities

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18.06.2019 19.06.2019 20.06.2019

09:00 Session Luca Vannucci Session Roberto Versaci

One Five Alessandra Picchiotti

Giancarlo Forte Federico Brivio

Guido Caluori

Alice Abbondanza Q/A

10:10 Anna Malandra COFFEA BREAK

10:20 Q/A Session Dino Numerato

Six Massimiliano Nuzzolo

10:55 COFFEA BREAK

11:10 Session Vincenzo Tarallo Q/A

11:15 Two Paolo Tenti Closing

Q/A

11:55 Session Benjamin Irving

Three Paolo Nicolini

Antonio Cammarata

Benjamin Irving

Q/A PROGRAM AT A GLANCE

12:50 LUNCH

14:00 Section Francesco Piana

Four Lorenzo Giuffrida

Daniele Margarone

Gabriele M. Grittani

Q/A

15:30 COFFEA BREAK

POSTER SESSION

17:00 Opening

Authorities/Vannucci

Piano Concert

Johanna Hanikova

(Scarlatti, Chopin, Liszt)

Refreshments

20:00

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ABSTRACTS OF THE PRESENTATIONS AND POSTERS

Section 1: Bio-medicine, physiology and biotechnologies

Morphology and immunity in transforming tissues: looking for the tumor niche Luca Vannucci

1, Pavol Lukac

1,3, Paolo Tenti

1,3, Fabian Caja

1,3, Dmitry Stakheev

1,3, O. Chernyavskiy

2, D.

Vondrasek2, Lenka Rajsiglova

1,3, Tomas Hudcovic

1, Hana Kozakova

1, Jiri Dvorak

1, Petr Sima

1, Pavol

Makovicky4, Miroslav Levy

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1. Institute of Microbiology of the CAS, v.v.i. Prague/Novy Hradek, 2. Institute of

Physiology of the CAS,

v.v.i. Prague, and 3. Faculty of Science, Charles University, Prague, Czech Republic; 4. Department of Biology, Pedagogical Faculty, Selye Janos University, Komarno, Slovak Republic; 5. Thomayer’s Teaching Hospital, Prague, Czech Republic E-mail: [email protected]

The cell transformation leading to uncontrolled replication and initial tumor clone formation determines a physical event inducing stress and alteration of the structural homeostasis of the tissue. The elicited expression of stress molecules and release of alarmins start the early immune reaction and tissue changes that can drive either tumor clone ablation or its establishment and progression. In this play the collagen scaffold appears particularly sensible to the local immune changes. In our animal models in which we simulate various conditions of immune activation involving the colon mucosa, this relationship and its high dynamism appear evident. Thanking the 2-photon microscopy approach in second harmonic generation, the evidenced changes in the collagen scaffold appear linked to the changes of pro-inflammatory signals in the tissue microenvironment. This was seen in DSS-induced colitis in the rat and in the mouse, in the AOM-induced carcinogenesis in the rat and in the bacterial colonization of germ-free mice bowel in which was evident the different and integrated role of IL-6, IL-1, IL-10 and TGF-β depending the type of inflammatory process involving the microenvironment, since early period. The comparison with human samples from colon cancer surgical specimens - including normal mucosa, near tumor mucosa and cancer tissue, showed the morphological changes were paralleling the immunological features in the mucosa. Here, increase of gene expression for collagen I, LOX2L, IL-1-β, IL-6, IL-13 were detected especially in the near tumor mucosa, taken as a tissue under tumor evolution. The condition of progression to tumor can be hypothesized by a downregulated or deficient expression of TGF-β versus IL-10 in the initial phases of tumor microenvironment establishment. Acknowledgements: grants RVO 61388971 (CZ), UniCredit Bank s.r.o. (CZ), Generali CEE Holding B.V. (CZ), CAMIC CZ, Michael Cukier Fund (CA), Roberto Di Cursi Fund (CZ), Italinox a.s. and Eurinox a.s. (CZ), Iginio Longo, Roberto and Olga Magini Fund, Manghi Czech Republic (CZ), Paul’s Bohemia (CZ) and ARPA Foundation (IT).

International Clinical Research Center (FNUSA-ICRC): a science excellence center and an incubator for

international talents.

Giancarlo Forte, PhD.

Center for Translational Medicine (CTM), International Clinical Research Center (ICRC), St. Anne’s

University Hospital, Brno, Czech Republic

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The International Clinical Research Center (ICRC) is the european project of excellence of St. Anne’s University Hospital (FNUSA) in Brno, Czech Republic. Based within the historical faculty hospital of Masaryk University which served as primary recovery for the soldiers wounded during the Austerlitz battle in 1815, FNUSA-ICRC was founded in 2011 thanks to a massive investment of structural funds by the European Union. After the first 5 years, which served to establish state-of-the-art research facilities endowed both with clinical and translational reseach premises, FNUSA-ICRC managed to set the conditions to attract a number of foreign researchers. With its newly established, top notch research facilities, and its unique and convenient connection with FNUSA clinical departments, ICRC qualifies as the only clinical research center of that kind in Czech Republic. FNUSA-ICRC currently has 35% foreign representatives among the 400 employees. This number includes quite a few italian students and researchers. Thanks to the succesful fundraising campain of the last years and the support of Czech and european govenrments, the Institute gained sustainability and regularly produces science at the highest international standards. So it quickly became a reference center for the whole Country and a hub for clinical studies in Europe. The presentation will highlight the contribution of italian students and researchers to the establishement and continuous growth of FNUSA-ICRC into a reference center in czech medical science.

Basic to clinical cardiological research in FNUSA-ICRC electrophysiological center

Guido Caluori1,2

, Tomasz Jadczyk1,3

, Eva Odehnalova1, Martin Pesl

1,4,5, Jakub Krenek

1, Zdenek Starek

1,4

1. Interventional Cardiac Electrophysiology, FNUSA-ICRC, Pekarska 53, 65691, Brno; 2.

Nanobiotechnology, CEITEC-MU, Kamenice 5 62500, Brno; 3. Department of Cardiology and Structural

Heart Diseases, Medical University of Silesia, Katowice, Poland; 4. 1st Department of Cardiovascular

Diseases, St. Anne's University Hospital and Masaryk University, Brno, Czech Republic; 5. Department of

Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.

Cardiovascular diseases hold the primate in western world mortality. Cardiac arrhythmias are a broad

class of life threatening and life impairing conditions that are increasing their prevalence due to

population aging and lifestyle changes. For instance, atrial fibrillation has a prevalence of 4% and it is the

most common arrhythmia. Due not only to the intrinsic cardiovascular risks, but also to the collateral

systemic damages and economic burden of chronic hospitalization, population education and research

forces towards this class of cardiac diseases has become a milestone for national healthcare systems.

In our center, we regularly perform state of the art (manual and magnetically navigated) catheter

radiofrequency ablation of drug-resistant cardiac arrhythmia. We enroll up to 300 patients for

endovascular ablation procedures every year, spanning from supraventricular tachycardias to rare cases

of hypertrophic cardiomyopathy. We further contribute to the surgical approach to persistent atrial

fibrillation, a procedure known as hybrid ablation. Patients can be enrolled in clinical trials according to

the expert consensus of the ethical commission of the first cardioangiology clinic in FNUSA, and

following study designs provided by our Biostatistics core facility in ICRC.

In the fields of basic and translational science, we design and perform experimenter-initiated and

contracted research of novel devices and advanced ablation therapies on large animal models (swine,

sheep). We perform a wide range of feasibility, safety and efficacy trials. The animal lab in the

Veterinary and Pharmaceutical Sciences University in Brno allows us to perform ablation with a unique

magnetic navigation system for animals, and a 1.5T MRI for chronic imaging of live animals. Through our

national and international collaborating institutions, we are also involved in testing novel imaging and

ablation strategies, such as high-intensity MRI irreversible electroporation of arrhythmic foci.

Understanding the role of nicotinic cholinergic receptors in striatal-based behaviour

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Abbondanza A1, Höfflin J

2, Janickova H

1

1. Department of Neurochemistry, Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic; 2. Rheinische Friedrich-Wilhelms-Universität Bonn

Striatum is the basal ganglia nucleus responsible for motor control, reward related processes, learning and motivation. The vast majority of its cell population is represented by medium spiny neurons (MSNs), which form the striatal output. The fine regulation of MSNs activity is given by cholinergic transmission, exerted by cholinergic (CINs) and GABAergic interneurons (GABAINs). While CINs modulation trough nicotinic and muscarinic receptors has been extensively studied, the role of nicotinic receptors on GABAergic interneurons (GABAINs) is still largely unknown.

To determine the function of nicotinic acetylcholine receptors (nAChRs) expressed by GABAINs, we selectively deleted β2 subunit in the dorsal striatum by injecting β2-floxed mice with Cre-expressing AAV viral vector. After confirming that the deletion only occurs in the injected area, we tested the mice in a battery of behavioral tasks focused on striatal-based behavior such as open field test, cued Morris water maze, cross maze task, nest building and social preference test, followed by histological and biochemical measurements.

First data obtained with one cohort of mice consisting of both males and females showed the effect of deletion might differ between sexes. While male mutants showed impairment of goal-directed behavior, motivation and sociability, females did not differ from the non-deleted controls.

We confirmed the successful deletion of 2 nicotinic subunit in transduced cells by immunofluorescence and RT-qPCR. We assessed that the deletion is limited to the striatum and the viral vector is not retrogradely transported to other brain regions. We conclude that nAChRs expressed by GABAINs in the striatum have a functional role in the control of striatal-based behavior.

This work was supported by the Grant Agency of the Czech Republic grant 19-07983Y. J.H. was supported by DAAD RISE program during her internship.

Effect of Bordetella pertussis virulence factors increasing cAMP levels on mucin production in human

airways

Anna Malandra, Shakir Hasan, Peter Sebo and Radim Osicka

Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic

Bordetella pertussis is a human respiratory pathogen and causative agent of whooping cough, a highly contagious disease which is increasing in incidence despite high vaccination coverage worldwide. The toxicity associated with the whole-cell pertussis vaccines has led to the development of a new generation of acellular vaccines containing one or more virulence factors. However, the acellular vaccines have shown reduced effectivity contributing to the new rise of the disease in developed countries where, despite the widely applied vaccination programs, Bordetella pertussis continues to kill; about 300,000 children every year worldwide are still killed by the bacterium. Anti-vaccine pseudo-ideologies and the misdiagnosis of the disease also contributed to the new rise of it. In this scenario, current studies are focused on determining the composition of an efficient acellular vaccine or a non-toxic new whole-cell vaccine. Our contribution in the field is currently focused on studying the interaction of Bordetella pertussis bacterium with the airways epithelium with particular focus on the effect of Adenylate cyclase and Pertussis toxin virulence factors on mucous secretion and production in order to verify the hypothesis that Bordetella pertussis exploits the mucosal layer as a binding substrate to inject its virulence factors into the epithelial cells cytosol and, probably achieve translocation of the whole bacterium inside the cells enhancing its colonisation ability and its survival in the host. So far, we have demonstrated that both adenylate cyclase and pertussis toxin are able to upregulate mucous production and secretion when human airways epithelial layers are intoxicated with the purified toxins for 24 h in vitro. Furthermore, we showed that the upregulation of mucous production is achieved through the cAMP/PKA/CREB signalling pathway, demonstrating that both adenylate cyclase and pertussis toxin are able to translocate inside the cells after being secreted by the bacterium and both

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are able to increase cytosolic concentration of cAMP in the host’s airways epithelial cells. Future studies will be focused on investigating the effect of the bacterial infection of the human airways epithelial layer on mucous secretion using the wild type bacterium and mutants alternatively lacking different virulence factors in vitro and in vivo. Evolved Methionine Sulfoxide Reductase A for Biotechnological applications Tarallo Vincenzo Department of Organic Chemistry, Faculty of Science, Charles University Chiral sulfoxides are important biologically active compounds, several of which constitute the active component of marketed pharmaceuticals. They are also privileged chiral ligands, catalysts and auxiliaries in asymmetric synthesis. Ever since the pivotal work of Kagan, a large number of chemicocatalytic methods have been developed for asymmetric synthesis of chiral sulfoxides. Despite the wealth of these methods, truly general protocols that provide sulfoxides with high enantiomeric purity (≥99% ee) remain scarce. Recently, we have developed of a highly efficient deracemization method for the preparation of chiral sulfoxides. This chemoenzymatic process utilizes the enantioselective enzyme methionine sulfoxide reductase A (MsrA). In spite of the excellent enantiooselectivity of the enzyme, there is a limitation in terms of substrate scope. Therefore, we aimed to develop a new high-throughput assay for laboratory evolution of MsrA. A novel fluorescent probe has been synthesized and utilized for the assay development. The newly established assay enables to screen large libraries of mutants of the enzyme (~107 variants). This high-throughput format allowed us to find an active mutant with significantly broader substrate scope while maintaining its enantioselectivity. Thanks to our new screening, MsrA`s mutants were selected in order to use them for racemic resolution of molecules of biotechnological interest. Lysyl Oxidases and formation of the early Tumoral Niche 1, 2

P. Tenti, 1, 2

L. Rajsiglova,1, 2

P. Lukac and 1

L. E. Vannucci 1 Laboratory of Immunotherapy, Institute of Microbiology of the CAS, v.v.i., Prague 4, Czech Republic 2 Faculty of Science, Charles University, Prague, Czech Republic The Lysyl Oxidases (LOXs) are a family of enzymes deputed to cross-link collagen and elastin, shaping the structure and strength of the extracellular matrix (ECM). Additional functions have also been recently described, suggesting a multifaceted role of LOXs within a complex network of signals regulating many cell functions including survival/ proliferation/ differentiation. Among these signaling pathways, TGF-β and PI3K/Akt/mTOR, in particular, cross-talk extensively with each other and with LOXs also initiating complex feedback loops. According our preliminary data, the tissue microenvironment remodelling starts early, already at the beginning of the carcinogenesis process, as a result of the altered balance of pro-inflammatory and regulatory signals and altered colon mucosa homeostasis. In particular, the thickening of the collagen scaffold and the increase in the tissue stiffness is largely dependent on LOXs activity. Tissue stiffening is a well-known mechanism leading to epithelial-to-mesenchymal transition and metastatic tumoral progression. While LOXs association with advanced and metastatic cancer is well established, there is enough experimental evidence to also support a significant role of LOXs in promoting the transformation of normal epithelial cells. Working on a mice experimental model of colitis-induced colorectal carcinoma, we are trying to better define the role of LOXs on the early establishment of the tumour microenvironment and the formation of the early tumoral niche, also trying to elucidate the network with crucial signaling pathways possibly involved, e.g. PI3K/Akt/mTOR, IL-13, non-canonical TGF-ß pathway. Section 2: Physics, chemistry and engineering

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Mini-session on Computational Materials Science

by Advanced Materials Group, Department of Control Engineering, Faculty of Electronic Engineering,

Czech Technical University in Prague, Prague, Czech Republic

A. Overview of the mini-session B. Irving The Advanced Materials Group (AMG) at the Czech Technical University in Prague (Department of Control Engineering, Faculty of Electrical Engineering) is a young group which started in January 2009 pursuing purely experimental research. Since its inception, the group has actively contributed to the fabrication of novel materials with superior tribological properties. Owing to its high quality research output, AMG rapidly established itself as one of the most successful groups in the faculty. In 2013, computational and theoretical activities started in the group, leading to important achievements in the field of computational materials science in general, with a particular focus on computational tribology. For a more comprehensive overview of AMG research activities and publications, please refer to the group webpage. Within the AMG, a fundamental area of research is the mitigation of friction across all size scales from the nano to the macro. Of the many ways to achieve this goal, solid lubricants have come to the fore, especially transition metal dichalcogenides (TMD). These are a family of layered compounds kept

together with weak van der Waals interactions (Figure 1). TMD-based materials are largely used as catalysts (e.g. hydrodesulfurization and CO hydrogenation reactions), as materials for microelectronics applications (e.g. single-layer transistors, sensitive photodetectors, piezotronics and superconducting applications) and, most importantly for our purposes, as solid lubricants and photovoltaic materials. Concerning tribology, TMDs show a very low friction coefficient, approaching the so-called ‘superlubrication’ regime, i.e., an apparent loss of frictional force, which decreases below detection limits in macroscopic measurements. This has been proven experimentally and also predicted by theoretical investigations.

B. Nanoscale friction properties of ordered and disordered molybdenum disulfide

P. Nicolini

In this contribution I will present the results of a study on the nanometric sliding of molybdenum

disulfide against itself, both from an experimental and computational point of view. The differences

between ordered material (single crystal) and disordered material (sputtered coating) in both humid

and dry environments have been investigated. Tribological experiments were performed using lateral

force microscopy. Atomic force microscopy tips modified by sputter deposition of molybdenum disulfide

were used for the first time. This feature opened up the possibility for a close comparison with classical

molecular dynamics simulations. In both cases, the coefficient of friction for the ordered system in inert

Figure 1. Layered structure of a prototypical TMD

https://dce.fel.cvut.cz/en/advanced-materials-group

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conditions was found to be smaller than for the disordered system. This result demonstrates the impact

of morphology at the nanoscale and highlights the importance of molecular dynamics as a diagnostic

and predictive tool in nano-friction. Furthermore, experiments show that the effects of the environment

on nanoscale friction are reduced with respect to the macroscale case. These findings can expedite the

process of fabricating molybdenum disulfide-based coatings with superior tribological properties, with

the ultimate aim of reducing the energy dissipation due to friction.

C. Tailoring friction and energy dissipation at nanoscale

A. Cammarata

One of the main difficulties in understanding and predicting frictional response is the intrinsic

complexity of highly non-equilibrium processes in any tribological contact, which include breaking and

formation of multiple interatomic bonds between surfaces in relative motion.

To understand the physical nature of the microscopic mechanism of friction and design new tribological

materials, we conduct a systematic quantum mechanical investigation at the atomic scale on

prototypical TMDs. We combine structural and dynamic information from group theoretical analysis and

harmonic phonon calculations with the characterization of the electronic features using advanced

methods such us orbital polarization, bond covalency and cophonicity analyses. We formulated a

theoretical framework which is able to drastically reduce the computational resources needed to

characterize and predict frictional properties at the nanoscale.

Thanks to the general formulation of our framework, the present outcomes can be promptly used to

finely tune physical properties for the design of new materials with diverse applications beyond

tribology such as energy harvesting/storage, optoelectronics, and catalysis among others.

D. Nanoscale properties of TMDs: an in-silico study

B. Irving

Extensive series of calculations have been performed within the framework of density functional theory

in order to investigate the dependence of important nanomechanical and electronic properties of TMDs

on their chemical composition and physical assembly. In particular, our calculations underscore the

intrinsic relationship between incommensurate layers and superlubricity, and the influence of charge

density distribution on nano-frictional properties. Our latest calculations are focused on the design of

TMD-based van der Waals heterostructures, which are candidate materials for the next generation of

thin-film solar cell assemblies.

Acknowledgements: This work was supported by the Czech Science Foundation (Grants No. 1724164Y

and No. 16-11516Y); by the Ministry of Education, Youth and Sports from the Large Infrastructures for

Research, Experimental Development and Innovations project “IT4Innovations National Supercomputing

Center – LM2015070”; by the project “Centre for Advanced Photovoltaics” funded by the Ministry of

Education, Youth and Sport (CZ.02.1.01/0.0/0.0/15_003/0000464); and partly by COST Action MP1303.

---------------

Polymers and composite materials for flexible electronics

Francesco Piana, Jiří Pfleger

Institute of Macromolecular chemistry, Czech Academy of Sciences – Department of Polymers for

Electronics and Photonics.

14

Flexible versatile electronic devices based on organic and hybrid materials are developed and

investigated in our laboratories for a variety of applications like biosensors, solar plants, smart textiles,

electronic labels and more. In this regard, polymers are the fundamental construction tools, which can

be easily modified to obtain electrically conductive, semi-conductive or insulating properties. They can

be used as inert supports or active components, they can be solubilised in various solvents and

deposited in thin layers to form nano-structures but also applied in solvent-free processes.

Combining polymers with small molecules, ceramic nanoparticles and thin metal layers, we prepare

basic components of electrical circuits like capacitors, solar cells, transistors and memristors. On one

side, we point to achieve novel products suitable for industry, on the other side, we study in deep the

electrical and optical phenomena occurring in the materials. As example, by techniques like dielectric or

transient optical absorption spectroscopy we investigate photoexcitation, energy conversion, or

intramolecular relaxation processes. These results lead us to improve the devices we prepare but also

help to clarify theoretical aspects and to broaden our knowledge about the electronic behaviour of

molecules, and the light-matter interaction.

All these efforts are pointed to the technological challenge of developing devices which are not only

high performing, but also ecologically sustainable for mass production. This means to make use of easily

available raw materials, low energy production processes, long lifetime of the devices and easy

recycling. Acknowledgment: Technology Agency of the Czech Republic, project No. TE01020022.

Brilliant and energetic alpha-particle source based on proton-boron nuclear fusion.

Lorenzo Giuffrida1, Fabio Belloni

2, G. A. P. Cirrone

1,3, Giada Petringa

3, Valentina Scuderi

1,3, Giuliana

Milluzzo3, Andriy Velyhan

1, Marcin Rosinski

4, Antonino Picciotto

5, Milan Kucharik

6, Jan Dostal

7, Josef

Krasa1, Valeria Istokskaia

1, Roberto Catalano

3, Pierluigi Bellutti

5, Georg Korn

1, Daniele Margarone

1

1Institute of Physics ASCR, v.v.i (FZU), ELI-Beamlines project, Prague, Czech Republic;

2European Commission, Directorate-General for Research and Innovation, Directorate Energy, Brussels,

Belgium; 3Laboratori Nazionali del Sud, INFN, Catania, Italy;

4Institute of Plasma Physics and Laser

Microfusion, Warsaw, Poland; 5Micro-Nano Facility, Center for Materials and Microsystems, Fondazione

Bruno Kessler, Trento, Italy; 6Czech Technical University in Prague, FNSPE, Prague, Czech Republic;

7Institute of Plasma Physics of the ASCR, PALS Laboratory, Prague, Czech Republic

The nuclear reaction known as proton-boron fusion has been triggered with a record yield using a sub-nanosecond laser system focused onto a thick boron nitride target at modest laser intensity (~10

16

W/cm2) at the PALS laser facility in Prague (Czech Rep.). The estimated yield leads to values around 10

11

alpha particles per laser pulse, thus orders of magnitude higher than any other result reported in literature. We used a kinetic model to explain our experimental achievements in terms of total amount of α particles generated in the laser-matter interaction. In such a model, protons generated inside the plasma, moving forward into the bulk of the target, can interact with

11B atoms, thus triggering fusion

reactions. An overview of results obtained with different laser parameters, experimental setups and target compositions present in literature is reported for a comprehensive understanding of the mechanisms involved in this neutron-less fusion reaction occurring in a laser-generated plasma.

be used for treat cancer in a more efficient way than the traditional proton therapy. Acknowledgments: This work has been supported by the Ministry of Education, Youth and Sports of the Czech Republic (project No. LQ1606) and by the projects “advanced research using high intensity laser produced photons and particles“ (CZ.02.1.01/0.0/0.0/16_019/0000789).

Laser-driven Ion Acceleration and Societal Applications at the Extreme Light Infrastructure (ELI) D. Margarone

http://aip.scitation.org/action/doSearch?field1=Affiliation&text1=INFN&field2=AllField&text2=&Ppub=&Ppub=&AfterYear=&BeforeYear=&access=

http://aip.scitation.org/action/doSearch?field1=Affiliation&text1=Center%20for%20Materials%20and%20Microsystems&field2=AllField&text2=&Ppub=&Ppub=&AfterYear=&BeforeYear=&access=

15

ELI-Beamlines, Institute of Physics (Czech Academy of Science), Prague, Czech Republic Acceleration of high energy ion beams by ultrahigh intensity laser-matter interaction is an emerging field of interest both for fundamental and applied science. Laser-driven ion acceleration occurs inside a high temperature, high density plasma, which transfers unique characteristics to the accelerated particle beam, e.g. ultrahigh dose rate (~10

9 Gy/s) and short bunch duration (~ps). Furthermore, typical

accelerating fields sustainable in laser-beamlines compared to conventional accelerator facilities. Recently the ELIMAIA (ELI Multidisciplinary Applications of laser-Ion Acceleration) beamline has been installed at ELI-Beamlines in the Czech Republic. The main goal of ELIMAIA is to offer short ion bunches accelerated by lasers with high repetition rate to users from different fields (physics, biology, material science, medicine, chemistry, archaeology) and, at the same time, to demonstrate that this source can be delivered through innovative and compact approaches. In fact, ELIMAIA will provide stable, fully characterized and tunable particle beams accelerated by PW-class lasers and will offer them to a broad community of users for multidisciplinary applied research, as well as fundamental science investigations. An international scientific network, called ELIMED (ELI MEDical applications), particularly interested in future applications of laser-driven ions for hadrontherapy, has already been established. In such a perspective ELIMAIA will enable to use laser-driven proton/ion beams for medical research thanks to the reliability and accuracy of its particle beam transport and dose monitoring devices. An overview of the ELI-Beamlines international facility in the Czech Republic, along with the ELIMAIA

user beamline and ELIMED objectives will be given.

This work has been supported by the Ministry of Education, Youth and Sports of the Czech Republic

(project No. LQ1606) and by the projects “advanced research using high intensity laser produced photons

and particles” (CZ.02.1.01/0.0/0.0/16_019/0000789).

Radiotherapy with high energy electrons generated by high power lasers: a potential new technology

to treat cancer

G. Grittani, T. Levato, C. Lazzarini & G. Korn

Institute of Physics of the Czech Academy of Sciences, ELI-Beamlines Project, Dolni Brezany, Czech

Republic

The use of 150-250 MeV electrons (very high energy electrons, VHEE) for radiotherapy has been

proposed for the first time two decades ago by DesRosiers et al. Such beams are of interest for

radiotherapy since they present two major dosimetric advantages when compared to photons or

protons. These are a sharper penumbra (resulting in a better transverse sparing of healthy tissue) and a

flatter longitudinal dose profile (which makes the treatment more robust against tumor imaging

reconstruction errors). The design of a VHEE radiotherapy device based on conventional rf technology is

being investigated nowadays, and the major technical issue is the size and complexity of such device. A

possible alternative is represented by laser plasma accelerator (LPA) technology, which has been

demonstrated for the first time in 2004. LPA allow for compact dimensions due to their extremely high

accelerating gradients (>100 MeV/mm). The use of LPA for VHEE radiotherapy has been patented in

2006 by the group of Prof. Malka at LOA. Our research group at ELI-Beamlines has patented in 2016 a

radiotherapy device based on LPA which allows for real time monitoring of the tumor by means of X-ray

or MRI. This imaging capability makes this device unique, since with conventional technology such

monitoring cannot be easily achieved.

16

Radiation transport Monte Carlo simulations at ELI Beamlines

Roberto Versaci


Republic

Monte Carlo simulations for radiation transport are a common tool in high energy physics, radiation

protection, and even medical physics. Recent developments in laser research, namely the capacity to

produce a large amount of ionizing radiation, are opening a new field of application. The ELI Beamlines

laser facility is home of a relatively large Monte Carlo group which has close links with other scientific

groups in Europe. The ELI Monte Carlo group's activities span over a large area: from simulations for the

evaluation of the radiation damage to people and machines, to the identification of mitigation options,

from the benchmark between experimental data and simulations to better understand the experimental

results, from the development of new detectors to the supervision of students. We will shortly describe

the use of Monte Carlo simulations at the ELI Beamlines facility and try to highlight possible

interconnections with other research areas.

Bio and material applications at ELI-beamlines

Alessandra Picchiotti and Jakob Andreasson


Republic

Laser-driven secondary sources at ELI-Beamlines is used for applications in molecular, biomedical, and

materials (MBM) sciences. Applications include coherent diffractive imaging, atomic, molecular, and

optical (AMO) sciences, soft X-ray materials science, hard X-ray scattering, diffraction, spectroscopy and

imaging, advanced optical spectroscopic techniques, and pulse radiolysis.

The research group for applications in molecular, biomedical, and materials sciences works to develop

experimental capabilities using the secondary sources that are driven by the uniquely powerful ELI-

Beamlines lasers. The MBM group mainly develops beam lines and end stations for time-resolved

photon science applications in the THz-to-hard X-ray range. This group also aims to combine different

laser-driven sources (of photons and particles) for unique applications in research and development.

The MBM group is currently in an active phase of growth and procurement. This phase is expected to

continue throughout 2019, and during this time the group will also welcome national and international

collaborators for user-based commissioning experiments. In ELI-Beamlines’ operational phase of 2019,

the MBM group will mainly support user operations but will also develop its own research interests.

In particular, we developed or are developing the following capabilities for time-resolved experiments

covering the femtosecond-to-millisecond time scales:

A multi-purpose end station for atomic, molecular, and optical (AMO) sciences and coherent diffractive

imaging (CDI)

A soft X-ray materials science station based on a time-resolved IR-to-VUV magneto-optical ellipsometer

A modular station for hard X-ray sciences covering applications in diffraction, spectroscopy and imaging

Advanced optical spectroscopy capabilities in the THz-to-UV range, including a setup for stimulated

Raman scattering and a wide array of pump beams coupled with all experimental stations for pump

probe experiments.

Computational approach to materials modeling

Federico Brivio

Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Czech

Republic

17

The advancement of computer power reshaped the approach to different fields of science. The

revolution affected new discoveries in both quantitative and qualitative way. As computer calculation

power increased so did our ability to process data and pin-point unnoticed trends. Contemporaneously

models could be refined and better theories became part of the scientist palette. We focus our

attention to different methodologies applied in material science. Nowadays we are spectators of a

transition from the calculation of materials property towards design of new materials with specific

requested properties. With means based on ab-initio density functional theory (DFT) we investigate the

properties of different class of materials. Our scope includes collaboration with experimental groups to

investigate catalytic materials and to support the design of new materials for different applications and

improvements of performances.

Section 3: Humanities, economics, social sciences, archaeology

Corruption and Public Secrecy: An ethnography of football match-fixing

Dino Numerato

Faculty of Social Sciences, Charles University, Prague

The topic of corruption has recently moved from the periphery to the centre of social scientific

attention. Notwithstanding the increased interest, research into corruption has been empirically limited

and under-theorized. This study addresses that gap by providing an ethnographic account of football

match-fixing in the Czech Republic. By qualitatively analysing both primary and secondary data, this

study examines matchfixing and corruption through the lens of the concept of public secrecy. Three

different, narrowly intertwined forms of match-fixing are identified: direct corruption, mediated

corruption and meta-corruption. By conceptualizing match-fixing as a public secrecy, the study explores

how the publicly secret nature of match-fixing is normalized and how the match-fixing complex is

reinforced by a compromising complicity of social actors who are both victims and principals. Although

this study focuses on a sportrelated example, it has both theoretical and empirical implications for a

sociological understanding of corruption outside the sphere of sport.

Previously published: Numerato D (2016). Corruption and Public Secrecy: An ethnography of football

match-fixing. Current Sociology, 64(5): 699-717.

Funding: The data collection was funded by the European Commission FP6 Marie Curie Excellence Grant

MEXT-25008. This article was further drafted under a FP7 Marie Curie Intra-European Fellowship, FP7-

PEOPLE-2012-IEF, Grant number 331097.

Archaeological works in Egypt: the latest results of the Italian expedition at the solar temples

Massimiliano Nuzzolo

Czech Institute of Egyptology, Charles University Prague

In 2010 an Italian archaeological mission from the University of Naples L’Orientale started field-works in

the area of Abu Ghurab, in Egypt. This is the area of the so-called solar temples, the first cult places ever

built in ancient Egypt (and one of the very first in the history of mankind) for the cult of the sun (2550-

2450 BCE). Since January 2017 this investigation has become a joint-research project (Rise and

Development of the Solar Cult and Architecture in Third Millennium BC Egypt) with the Czech Institute

of Egyptology – Charles University in Prague, and with the auspices of the Czech Science Foundation

18

(GACR grant no. 17-10799S). One of the final goals is to search for the missing sun temples. In fact, out

of the six temples known from historical sources only two have been discovered so far. The present

paper thus wishes to resume the main results of these three years of work, as well as to pinpoint the

major research objectives and perspectives for the coming years.

Role of APP axonal transport in Alzheimer’s disease and Chronic Traumatic Encephalopaty

Valentina Lacovich, Victorio Pozo Devoto, Maria Novakova, Kateřina Texlová, Monica Feole, Gorazd

Bernard Stokin

Translational Neuroscience and Aging Program (TAP), Center for Translational Medicine (CTM), St.

Anne's University Hospital Brno, International Clinical Research Center (FNUSA-ICRC), Brno

Although amyloid precursor protein (APP) and microtubule-associated protein tau have been shown to

play a major role in the pathogenesis of Alzheimer’s disease (AD) and more recently in the chronic

traumatic encephalopathy (CTE), the mechanisms underlying their role in the pathogenesis of these

neurodegenerative disorders remain unclear. Accumulating evidence suggests that perturbed axonal

transport plays an early and possibly causative role in the pathogenesis of AD and CTE. This project tests

further the role of perturbed axonal transport in neurodegeneration with particular emphasis on

dynamic changes in APP and tau, since knowledge about these changes will also contribute to a better

understanding of frequently equally dynamic behavioral and cognitive changes in AD and CTE.

We have developed a novel cell culture paradigm to assess in vivo dynamic changes in axonal APP and

tau following injury. This paradigm consists in specifically designed microfluidic chamber coupled to an

electronically controlled syringe pump, which induces sheared stress in the axons. Moreover,

microfluidic chamber is populated by human stem cell derived mature neurons. This set up altogether

allows for real time APP imaging in response to axonal injury. Results obtained with this cell culture

paradigm are coupled to a well-established mouse traumatic brain injury model to allow for

confirmation of cell culture findings in an in vivo setting.

Neurons differentiated from H9 derived human neural stem cells were characterized by RT-PCR, FACS,

immunochemistry and electrophysiology. We found that sheared stress induced axonal injury results in

selective and immediate perturbation of the axonal transport of APP and these finding were further

confirmed in a mouse model of traumatic brain injury. Considering our recent results identified subtle

perturbations in tau isoforms as sufficient to trigger perturbations in the axonal transport of APP we

next addressed whether axonal perturbations of APP also result in changes in tau. We found that

perturbations of APP also result in changes in tau.

Our finding disclose an intimate bidirectional link between APP and tau in AD and CTE, which sheds new

light to the mechanisms involved in the development of neurodegeneration.

In vitro model of cardiac fibrotic process to unveil the mechanisms of cardiac cells crosstalk. Pamela Mozetic, Ana Rubina Perestrelo, Jorge Oliver-De La Cruz and Giancarlo Forte. International Clinical Research Center (FNUSA-ICRC), St. Anne's University Hospital, Brno, Czech Republic; Competence Center for Mechanobiology in Regenerative Medicine, Brno, Czech Republic. Myocardial remodeling following myocardial infarction is emerging as a key cause of chronic infarct mortality. Hypoxia is one of the causes of ischemic damage, and cardiac fibroblasts (CFs) have a key role in adverse myocardial remodeling. Ischemia-associated CFs have a critical influence on myocardial remodeling progression and fibrosis. Injurious stimuli induce cellular and molecular signaling pathways dysregulation which, in turn, lead to the activation of CFs. Activated fibroblasts (myofibroblasts) produce ECM proteins and matrix metalloproteinases, as well as autocrine and paracrine factors (i.e., inflammatory cytokines), and mediate tissue remodeling processes. The aim of this work is to establish an in vitro hypoxia-reperfusion model to study the crosstalk among cardiac fibroblasts, cardiomyocytes and immune cells, and to unveil the mechanisms that modulate the response of cardiomyocytes.

19

hnRNPC: a linker between ECM mechanics and mRNA homeostasis in cardiac diseases

Authors: Fabiana Martino1,2


1, Jan Vrbský

1, Vladimír Vinarský

1,3, Jorge Oliver De La Cruz

1,3, Francesca Cavalieri

3, Giancarlo Forte

1,3,5

1Center for Translational Medicine (CTM), International Clinical Research Center (ICRC), St. Anne´s

University Hospital, Brno, Czech Republic; 2Department of Biology, Faculty of Medicine, Masaryk

University, Brno, Czech Republic; 3Competence Center for Mechanobiology in Regenerative Medicine,

INTERREG ATCZ133, CZ-62500 Brno, Czech Republic; 4Department of Chemical and Biomolecular

Engineering, University of Melbourne, Australia; 5Department of Biomaterials Science, Institute of

Dentistry, University of Turku, Finland.

-----------

Identifying regulators of axonal transport

Monica Feole, Victorio M. Pozo Devoto, Mária Čarná, Valentina Lacovich and Gorazd B. Stokin

Translational Neuroscience and Aging Program (TAP), Center for Translational Medicine (CTM),

International Clinical Research Center (ICRC), Fakultní nemocnice u sv. Anny v Brně / St. Anne's

University Hospital Brno

The intracellular transport is a regulated process, involving several proteins controlling the trafficking in

all the cells, particularly in neurons. The onset of several neurodegenerative diseases, such as

Alzheimer’s, Parkinson’s or Amyotrophic Lateral Sclerosis, exhibits significant axonal pathology where

transport impairment plays a crucial role. However, the mechanism underlying the disruption of

neuronal transport is still unclear. In the present work we aim to characterize transport dynamics of

specific proteins and the molecular machinery that is regulating this complex process. Human Neural

Stem Cells derived neurons were terminally differentiated in multichannel μ-Slide VI (Ibidi) for 30 DIV.

GFP-coupled cargos involved in many functions (development, pre- and post-synaptic activity, disease

signaling) were transfected in neurons, in order to describe possible differences in movement. Time-

lapse movies of the various cargos were acquired, and then analyzed with Imaris, obtaining a description

of motion parameters: mean velocity, directionality, segmental velocity, average run length, pauses, and

reversions. In addition, immunocytochemistry was performed after imaging, in order to describe the

specific localization (axon vs dendrite) of our cargos. Our preliminary results showed different behaviors

during transport, mainly in mean velocities and directionality proportions: for Amyloid Precursor Protein

an high velocity both in anterograde and retrograde movement was observed, with a small fraction of

stationary particles; a similar pattern was also visible for Synaptophysin, an important protein involved

in synapses maturation. Relying on the description of movement dynamics in neurons, we decided to

screen for specific protein-protein interactions of our cargos. The combination of GFP-trap and

Immunoprecipitation (IP) of the endogenous proteins, both followed by Mass Spectrometry analysis, will

shed light on possible regulators involved in neuronal transport. Preliminary results were obtained using

GFP-trap approach: protein lysate from transfected neurons, was used to perform GFP-IP. The isolated

fractions were then analyzed by mass-spec to highlight the population of specific interactors for our

cargos. This study describes an in vitro model for neuronal transport studies, which highlights the

different behavior of several proteins involved in crucial physiological pathway for the neuronal

homeostasis. Ongoing experiment will elucidate more about the regulation of these process and will

open a way to understand how the transport can be part of the pathophysiology of neurodegenerative

diseases.

20

The role of PV+ GABAergic interneurons in the medial prefrontal cortex on a rat model of psychosis-

related cognitive inflexibility as revealed by optogenetic stimulations.

E. Patrono, K. Hruzova, J. Svoboda, A. Stuchlík

Institute of Physiology - Academy of Sciences Czech Republic, Neurophysiology of Memory, Praha, Czech

Republic.

Cognitive flexibility, i.e., the ability to adjust a response based on changed conditions, is one of the

prominent executive functions that is disrupted in schizophrenia-like psychosis. Further, it is known that

parvalbumin-positive (PV+) interneurons in the medial prefrontal cortex (mPFC) have a critical role in

executive functions, as well as that poor PV+ interneuron function in mPFC is linked to schizophrenia.

One aim of the study was to create a new rat model of cognitive inflexibility induced by acute systemic

injections of MK-801, an NMDA receptor antagonist, and by using a modified version of Attentional Set-

Shifting Task (ASST). Another aim of this study was to investigate the therapeutical role of PV+

interneurons in mPFC in the acute model of cognitive inflexibility, using in vivo optogenetics PV+

stimulation in channelrhodopsin-2 (ChR2) transfected rats.

In the ASST, animals learned to dig and retrieve food reward from a cup, after a "two dimension-

operant-association" - relevant (odor) and irrelevant (digging medium). Reversal learning (RL-odor

switch) and extra-dimensional shift (EDS-relevancy switch) sessions measured the cognitive flexibility in

the rats. AAV-containing-ChR2 transfections and optic fiber implant were stereotactically applied onto

rat mPFC, unilaterally, 2 weeks before assessing ASST.

Results showed that acute MK-801 injections during RL and EDS induced cognitive inflexibility compared

to the saline. Moreover, PV+ optogenetic ChR2-stimulation in mPFC recovers the ability to switch odor

and/or medium relevancy.

We observed that enhancing the inhibitory activity of PV+ interneurons in mPFC is a new experimental

therapeutic opportunity to rescue cognitive flexibility in schizophrenia-like psychosis. This work was

supported by GACR grant 17-04047S and AZV grant 17-30833A. Institutional support for IPHYS was

provided by RVO: 67985823. Additional support came from MEYS (LM2015062) Czech-BioImaging and

H2020 INFRADEV-01-2017 project ID-EPTRI (European Paediatric Translational Research Infrastructure -

EPTRI Grant Agreement 777554).

Molecular principles of Cajal body formation

Davide Alessandro Basello1,2, Michaela Efenberková1, Radek Macháň2, Nicola Maghelli3, David Stanek1

1. Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic; 2. Charles

University in Prague, Faculty of Science, Prague, Czech Republic; 3. Max Planck Institute of Molecular

Cell Biology and Genetics, Dresden, Germany

The cell nucleus is a highly heterogeneous environment crowded with numerous macro-molecules. Part

of the nuclear complexity rises from the presence of a number of different bodies, non-membrane

bound structures, which accumulate various proteins and, often RNAs. The molecular principles behind

bodies assembly and maintenance are recently a matter of an intensive debate. One of the "classical"

examples of a nuclear body is the Cajal body (CB). CBs are involved in biogenesis, quality control and

recycling of spliceosomal snRNPs. Coilin, the essential scaffolding protein of CBs, self-oligomerize and

interacts with numerous proteins including snRNPs, and these interactions are important for CB

formation. However, the basic information regarding its structure and function are lacking. To uncover

molecular principles of CB formation we determined coilin dynamics in the nucleoplasm and CBs and

analyzed the snRNP influence on coilin self-oligomerization and CB formation. To eliminate the effect of

endogenous coilin, we generated a coilin KO cell line. Then we prepared several different mutated

versions of coilin that prevent interaction with snRNPs and express them in coilin KO cells. Our results

21

show that coilin self-interaction does not depend on snRNP binding. However, the mutants that do not

associate with snRNPs fail to reconstitute CBs in coilin KO cells. We also apply different fluorescence

microscopy techniques (FRAP, single-point and imaging FCS and 3D-SIM SPT) to determine coilin

dynamics in the nucleus and the CB. The data confirm our biochemical results and show that abolishing

coilin interaction with snRNPs does not inhibit coilin self-association. Interestingly coilin dynamics inside

the CB does not differ from the nucleoplasmic coilin, suggesting that the CB nucleation and maintenance

are not based on different coilin mobility inside and outside CBs.

NFAT as a novel pathway in mesenchymal stem cell response to Inflammation.

Tidu F. (1, 2), Jose S. S. (1, 2), De Zuani, M. (1), Bendíčková K.(1), Pompeiano A. (1), Bělašková S. (1), Frič

J. (1)

(1) St. Anne’s University Hospital Brno International Clinical Research Center, Brno, Czech Republic;

(2) Department of Biology, Faculty of Medicine, Masaryk University, Czech Republic

Mesenchymal stem cells (MSCs) are multipotent cells present in the stromal fraction of many tissues. It

is now clear that they play an important role in immune response control during inflammation, due to

their ability to migrate to inflamed site and secrete immunoregulatory factors.

MSCs-expressed pattern recognition receptors (PRRs) allow direct sensing of infection or damage in

tissue microenvironment. So far most of the studies pointed at TLR NF-κB axis as the main responsible

for controlling MSCs inflammatory profile. However a clear insight on PRRs activation patterns and

cross-talks between NF-κB and other inflammatory pathways has not been sufficiently described, thus

hampering

MSCs therapeutic applications. Here we show that zymosan recognition by MSCs leads to the activation

of calcineurin-NFAT axis, which shapes MSCs response to inflammation.

The present work aims to shed light on NFAT role in MSCs biology, in order to better describe the

mechanisms underlying MSCs immunomodulation, which is of utmost importance in a translational

perspective.

Understanding nanoparticles-cells' interactions through mechanobiology

Marco Cassani, Jorge Oliver-De La cruz, Soraia Fernandes, Giancarlo Forte

Center for Translational Medicine (CTM) International Clinical Research Center (FNUSA-ICRC) St. Anne's

University Hospital

The new era of medicine requires patient-customized nanomaterials in which different cure

strategies,such as targeting and drug delivery, can be combined into a unique platform. As material

science progresses, a more complex understanding and deeper knowledge of the mechanisms involved

in the cells’ response to nanoparticles exposure are needed. Once the nanoparticles target the tumor

site, they are expected to induce significant changes in the extracellular microenvironment.

Mechanobiology, which studies the mechano-transduction by which the cells convert the outer

mechanical stimuli into inner biochemicals responses, is now emerging as the leading discipline for

understanding cancer development. The possibility to implement mechanobiology into nanomedicine

can spread new light on how healthy and unhealthy cells respond to nanoparticle-based treatments

helping to drive the nanomaterial’s activity to reach the desired specificity and efficiency in cancer

treatment.

Femtosecond Stimulated Raman Spectroscopy

Alessandra Picchiotti

22

ELI-beamlines/ELIBIO - ELI Beamlines project, Institute of Physics (FZU) of the Czech Academy of

Sciences, Dolní Břežany, Czech Republic

The use of femtosecond stimulated Raman probes make it possible to measure the vibrational spectra

of liquid, gaseous, and transparent solid state samples with femtosecond time resolution. This setup is

provided in the facility ELI-Beamlines in Dolni Brezany both as an independent beam line and as an

optional add-on to the other experiments.

Recording time-resolved structures of complex molecules (such as protein complexes) by femtosecond

X-ray pulses is among the important goals of ELI-Beamline’s project. However, these techniques are still

in the process of being developed and suffer from a high level of damage that is inflicted on a sample. In

contrast, time-resolved vibrational spectra can map molecular conformations in a highly non-invasive

way. The possibility of running these experiments simultaneously with X-ray experiments would allow

for valuable supplementary information to be obtained to cross-validate the conclusions drawn from the

X-ray experiments. A crucial advantage of the stimulated Raman probe is the great flexibility it offers in

terms of beam delivery, because it is based on pulses in the optical domain.

Pulse Radiolysis

Martin Precek



Radiation chemistry is a scientific field that is involved with studying the chemical effects of ionizing

radiation of matter to advance knowledge both in fundamental atomic and molecular science and in

applied radiation methods. In the latter case, research in radiation chemistry has strong connections to

areas that study and employ radiation effects in living systems, with important medical applications in

radiotherapy and radiation dosimetry, and also with areas dealing with the development of applied

radiation processing methods, such as radiation sterilization, radiation sanitation, radiation

polymerization, and food irradiation.

The ELI-Beamlines facility of the Institute of Physics of the Czech Academy of Sciences will be capable of

using laser interactions in matter to directly generate multiple kinds of pulsed ionizing radiation (soft

and hard X-rays, electrons, or protons and possibly even heavier ions). Capabilities are being developed

at ELI Beamlines for building experiments enabling ultrafast pulse radiolysis at user end-stations. As

opposed to approaches using conventional accelerators (LINACs, cyclotrons, etc.) the laser-driven

secondary radiation sources make it possible to achieve a temporal resolution of tens of femtoseconds

due to the short length of the laser-generated radiation pulse and the near-perfect synchronization of

the temporal overlap of the pulse of radiation and of a probing optical pulse in the studied sample

(usually liquid). This will then allow to perform measurements far below the famous "picosecond

barrier" in radiolysis, experimentally revealing poorly charted time scales where primary radical

products are generated from the interactions of ionizing radiation with matter.

Ultrafast dynamics of plasmon and magnetic resonances in nanoparticles

Carlo Maria Lazzarini, Tadzio Levato, Jamie M. Fitzgerald, José A. Sánchez-Gil, Vincenzo Giannini

Here I present a study of nanoparticle response to ultrafast laser excitation. I work in the electron

acceleration group at ELI-Beamlines, where we study and develop new particle acceleration techniques

and diagnostics methods for laser-plasma and laser-matter interaction.

The nanoplasmonics world is built around the fundamental concept of the collective resonant response

of the conduction electrons of a nanostructure, known as plasmon resonances, induced by an incident

electric field. Recent developments in nanofabrication techniques, high-sensitivity single-particle optical

characterization techniques, and fast numerical modeling tools for simulating complex nanostructures

have led to an increasing interest of the scientific community.

23

However, usually it is considered single only the monochromatic, continuous-wave regime. Nonetheless,

in the last decades, due to the development of ultrashort (fs timescale) laser pulses, many attempts

have been made to combine ultrashort laser pulses with nanostructures, generating the fast expanding

field of ultrafast nanoplasmonics. In which the the final aim is the control of the strong-field

enhancement localized in space on the nanometer scale and in time on the fs and sub-fs scale.

I present an analytical description of the ultrafast localized surface plasmon and magnetic resonance

dynamics in a single nanoparticle (Ag or Si), driven by an ultrashort (fs time scale) Gaussian pulse. Three

possible scenarios have been found depending on the incident pulse duration compared to the

resonance lifetime.

Nanoparticle resonance dynamics may lead to a wealth of new phenomena and applications in

nanophotonics such as multipole order resonance interference, pulse-induced delay or temporal

shaping on the fs scale, high harmonic generation, attosecond near-field pulse sources, and electron

acceleration from metasurface or 3D engineered nanostructures.

Mechanisms underlying axonal swelling formation Victorio M Pozo Devoto, Valentina Lacovich, Maria Carna, Monica Feole, Katerina Texlova & Gorazd B Stokin International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic Axonal swellings (AxS) are focal enlargements of axons found post-mortem in a range of biological and pathological settings including traumatic brain injury, Alzheimer's disease and multiple sclerosis. Despite their description in a range of settings, the mechanisms involved in the formation of AxS remain poorly understood. Here we developed a novel in vitro experimental paradigm to test for mechanisms underlying AxS formation. Human neuronal progenitor cells were seeded into microfluidic chambers and terminally differentiated to neurons. In these chambers axons grow into microchannels, which are crossed by a perpendicular channel to which we connected a syringe pump. Syringe pump generates force, which subjects axons to bending stress. After full characterization of the channel fluid dynamics, we tested how axons respond to the stress. Detailed analysis of the kinetics by time-lapse imaging showed a significant increase in the number and size of axonal enlargements during and after stress as visualised by transducing neuronal culture with a membrane targeted Cherry. We have first studied these axonal enlargements by scanning electron microscopy to reveal and distinguish physiological versus pathological enlargements following axonal injury. We next performed super-resolution microscopy to demonstrate loss of Spectrin BII periodicity of the sub-axolemmal cytoskeleton. We then investigated whether enlargements form as a result of membrane leakage in response to shear stress. Considering many reports found increased Ca2+ following axonal injury, we then asked whether Ca2+ concentrations increase in axonal enlargements. We found a significant increase in Ca2+ within the enlargements as visualised by the Fluo 4AM Ca2+ sensor. Furthermore, to understand the role of Ca2+ in enlarged axons following injury further we next depleted Ca2+ from media and blocked different axonal membrane and ER/mitochondrial Ca2+ channels. In summary, we created a unique cell culture paradigm to study the response of axons to physical injury and provide novel insight into mechanisms responsible for the formation of axonal swellings. Mechanical thrombectomy results variability per occluded artery in ischaemic stroke. L. Mengozzi, J. Vavrova, B. Koznar, F. Rohac, J. Sulzenko, J. Kroupa, I. Stetkarova, T. Peisker, P. Vasko, P. Widimsky University Hospital Kralovske Vinohrady and Third Faculty of Medicine, Charles University, Prague, Czech Republic.

24

INTRODUCTION. Although mechanical thrombectomy is a rapidly developing technique worldwide in ischaemic stroke, the success rate according to the affected vessel is still not fully established. METHODS. A total of 190 patients presenting with acute ischaemic stroke undergoing mechanical thrombectomy were divided into 5 subgroups: (a) isolated occlusion of the middle cerebral artery (MCA), (b) isolated occlusion of the proximal internal carotid artery (ICAprox), (c)isolated occlusion of the distal ICA (T-occlusion), (d) tandem occlusion of ICAprox and MCA, (e) basilar or vertebral artery occlusion. Immediate reperfusion success rate and 3-months clinical outcomes were analysed according to the occluded artery involved. RESULTS. Isolated occlusions of the MCA represented alone one of the largest subgroups with 42% of the patients, whereas 46% presented with ICA occlusions, of which 19% ICA distal occlusions (T-occlusion), 14% ICA proximal occlusions and 13% ICA proximal/ MCA tandem occlusions; only 12% of cases manifested ischaemia of the vertebral/basilar artery (VB). The TICI grading system was used to assess the success rate of the thrombectomy and grade 2b/3 was achieved in 84% of patients with obstructions of the MCA and 92% of cases with ICA proximal occlusions. Altogether, the success rate at TICI 2b/3 for MCA and ICA occlusions reached 96%. Although not the least common, ICA distal occlusions had a postthrombectomy success rate of 56% and mortality at 3 months of 42%, not quite unlike vertebral/basilar occlusions (VB) with a 48% reperfusion success and a 52% mortality in the first trimester, the latter being the rarest encountered type. Overall, 16% and 19% of patients died within 3 months, previously presenting with respectively MCA and ICA proximal occlusions, of which 53% and 46% scored between 0-2 with the modified Rankin scale (mRs). Only approximately ¼ of patients treated for VB and ICA distal occlusions manifested no significant disability despite some symptoms at 3 months (mRs 0-2). Test of genetic code evolution hypotheses: Reverse evolution of specific target proteins by mRNA-display technique Valerio Guido Giacobelli

1, Kosuke Fujishima

2, Vyacheslav Tretyachenko

1, Klara Hlouchova

1

1. Department of Cell Biology, Faculty of Science, Charles University, Hlavova 2030, 128 00, Prague 2, Czech Republic; 2. Earth-Life Science Institute, Tokyo Institute of Technology, Tokyo 1528550, Japan Although extant proteins consist of 20 different amino acids, it has been proposed that primordial proteins consisted of a smaller set of “early” amino acids and that additional “modern” amino acids have gradually been recruited into the genetic code. This naturally leads to the questions: can structured and functional proteins be constructed using the “early” amino-acid alphabet? Can extant proteins be reverse-evolved while preserving their structure/function? To test this, protein databases have been inspected to select model extant protein candidates with different structural folds. Our preliminary search contains proteins with both catalytic and binding/interaction functions. The selected protein targets were “reverse-evolved” in vitro into variants where the “modern” amino acid were randomized by “early” ones. The libraries of randomized genes were incorporated into a genotype-phenotype linkage to be compatible with an appropriate library display (mRNA display) and selection method. The selection of successful candidates was based on conservation of structure and/or function and the most “successful” variants will be characterized. This research will inform us of the essentiality of “modern” amino acids for building protein structure/function and thus will provide a direct test of the hypotheses about early proteins. In addition, proteins constructed from a limited amino acid alphabet are of importance in protein engineering and synthetic biology. Finally, this area touches upon the very basic link of protein sequence-structure-function that lies at the core of many biotechnological and biomedicine problems and has express implications for construction of artificial biochemistries.

ABSTRACTS OF POSTERS

25

Role of APP axonal transport in Alzheimer’s disease and Chronic Traumatic Encephalopaty

Valentina Lacovich, Victorio Pozo Devoto, Maria Novakova, Kateřina Texlová, Monica Feole, Gorazd

Bernard Stokin

Translational Neuroscience and Aging Program (TAP), Center for Translational Medicine (CTM), St.

Anne's University Hospital Brno, International Clinical Research Center (FNUSA-ICRC), Brno

Although amyloid precursor protein (APP) and microtubule-associated protein tau have been shown to

play a major role in the pathogenesis of Alzheimer’s disease (AD) and more recently in the chronic

traumatic encephalopathy (CTE), the mechanisms underlying their role in the pathogenesis of these

neurodegenerative disorders remain unclear. Accumulating evidence suggests that perturbed axonal

transport plays an early and possibly causative role in the pathogenesis of AD and CTE. This project tests

further the role of perturbed axonal transport in neurodegeneration with particular emphasis on

dynamic changes in APP and tau, since knowledge about these changes will also contribute to a better

understanding of frequently equally dynamic behavioral and cognitive changes in AD and CTE.

We have developed a novel cell culture paradigm to assess in vivo dynamic changes in axonal APP and

tau following injury. This paradigm consists in specifically designed microfluidic chamber coupled to an

electronically controlled syringe pump, which induces sheared stress in the axons. Moreover,

microfluidic chamber is populated by human stem cell derived mature neurons. This set up altogether

allows for real time APP imaging in response to axonal injury. Results obtained with this cell culture

paradigm are coupled to a well-established mouse traumatic brain injury model to allow for

confirmation of cell culture findings in an in vivo setting.

Neurons differentiated from H9 derived human neural stem cells were characterized by RT-PCR, FACS,

immunochemistry and electrophysiology. We found that sheared stress induced axonal injury results in

selective and immediate perturbation of the axonal transport of APP and these finding were further

confirmed in a mouse model of traumatic brain injury. Considering our recent results identified subtle

perturbations in tau isoforms as sufficient to trigger perturbations in the axonal transport of APP we

next addressed whether axonal perturbations of APP also result in changes in tau. We found that

perturbations of APP also result in changes in tau.

Our finding disclose an intimate bidirectional link between APP and tau in AD and CTE, which sheds new

light to the mechanisms involved in the development of neurodegeneration.

In vitro model of cardiac fibrotic process to unveil the mechanisms of cardiac cells crosstalk. Pamela Mozetic, Ana Rubina Perestrelo, Jorge Oliver-De La Cruz and Giancarlo Forte. International Clinical Research Center (FNUSA-ICRC), St. Anne's University Hospital, Brno, Czech Republic; Competence Center for Mechanobiology in Regenerative Medicine, Brno, Czech Republic. Myocardial remodeling following myocardial infarction is emerging as a key cause of chronic infarct mortality. Hypoxia is one of the causes of ischemic damage, and cardiac fibroblasts (CFs) have a key role in adverse myocardial remodeling. Ischemia-associated CFs have a critical influence on myocardial remodeling progression and fibrosis. Injurious stimuli induce cellular and molecular signaling pathways dysregulation which, in turn, lead to the activation of CFs. Activated fibroblasts (myofibroblasts) produce ECM proteins and matrix metalloproteinases, as well as autocrine and paracrine factors (i.e., inflammatory cytokines), and mediate tissue remodeling processes. The aim of this work is to establish an in vitro hypoxia-reperfusion model to study the crosstalk among cardiac fibroblasts, cardiomyocytes and immune cells, and to unveil the mechanisms that modulate the response of cardiomyocytes.

hnRNPC: a linker between ECM mechanics and mRNA homeostasis in cardiac diseases

26

Authors: Fabiana Martino1,2


1, Jan Vrbský

1, Vladimír Vinarský

1,3, Jorge Oliver De La Cruz

1,3, Francesca Cavalieri

3, Giancarlo Forte

1,3,5

1Center for Translational Medicine (CTM), International Clinical Research Center (ICRC), St. Anne´s

University Hospital, Brno, Czech Republic; 2Department of Biology, Faculty of Medicine, Masaryk

University, Brno, Czech Republic; 3Competence Center for Mechanobiology in Regenerative Medicine,

INTERREG ATCZ133, CZ-62500 Brno, Czech Republic; 4Department of Chemical and Biomolecular

Engineering, University of Melbourne, Australia; 5Department of Biomaterials Science, Institute of

Dentistry, University of Turku, Finland.

--------------------------------------

Identifying regulators of axonal transport

Monica Feole, Victorio M. Pozo Devoto, Mária Čarná, Valentina Lacovich and Gorazd B. Stokin

Translational Neuroscience and Aging Program (TAP), Center for Translational Medicine (CTM),

International Clinical Research Center (ICRC), Fakultní nemocnice u sv. Anny v Brně / St. Anne's

University Hospital Brno

The intracellular transport is a regulated process, involving several proteins controlling the trafficking in

all the cells, particularly in neurons. The onset of several neurodegenerative diseases, such as

Alzheimer’s, Parkinson’s or Amyotrophic Lateral Sclerosis, exhibits significant axonal pathology where

transport impairment plays a crucial role. However, the mechanism underlying the disruption of

neuronal transport is still unclear. In the present work we aim to characterize transport dynamics of

specific proteins and the molecular machinery that is regulating this complex process. Human Neural

Stem Cells derived neurons were terminally differentiated in multichannel μ-Slide VI (Ibidi) for 30 DIV.

GFP-coupled cargos involved in many functions (development, pre- and post-synaptic activity, disease

signaling) were transfected in neurons, in order to describe possible differences in movement. Time-

lapse movies of the various cargos were acquired, and then analyzed with Imaris, obtaining a description

of motion parameters: mean velocity, directionality, segmental velocity, average run length, pauses, and

reversions. In addition, immunocytochemistry was performed after imaging, in order to describe the

specific localization (axon vs dendrite) of our cargos. Our preliminary results showed different behaviors

during transport, mainly in mean velocities and directionality proportions: for Amyloid Precursor Protein

an high velocity both in anterograde and retrograde movement was observed, with a small fraction of

stationary particles; a similar pattern was also visible for Synaptophysin, an important protein involved

in synapses maturation. Relying on the description of movement dynamics in neurons, we decided to

screen for specific protein-protein interactions of our cargos. The combination of GFP-trap and

Immunoprecipitation (IP) of the endogenous proteins, both followed by Mass Spectrometry analysis, will

shed light on possible regulators involved in neuronal transport. Preliminary results were obtained using

GFP-trap approach: protein lysate from transfected neurons, was used to perform GFP-IP. The isolated

fractions were then analyzed by mass-spec to highlight the population of specific interactors for our

cargos. This study describes an in vitro model for neuronal transport studies, which highlights the

different behavior of several proteins involved in crucial physiological pathway for the neuronal

homeostasis. Ongoing experiment will elucidate more about the regulation of these process and will

open a way to understand how the transport can be part of the pathophysiology of neurodegenerative

diseases.

The role of PV+ GABAergic interneurons in the medial prefrontal cortex on a rat model of psychosis-

related cognitive inflexibility as revealed by optogenetic stimulations.

27

E. Patrono, K. Hruzova, J. Svoboda, A. Stuchlík

Institute of Physiology - Academy of Sciences Czech Republic, Neurophysiology of Memory, Praha, Czech

Republic.

Cognitive flexibility, i.e., the ability to adjust a response based on changed conditions, is one of the

prominent executive functions that is disrupted in schizophrenia-like psychosis. Further, it is known that

parvalbumin-positive (PV+) interneurons in the medial prefrontal cortex (mPFC) have a critical role in

executive functions, as well as that poor PV+ interneuron function in mPFC is linked to schizophrenia.

One aim of the study was to create a new rat model of cognitive inflexibility induced by acute systemic

injections of MK-801, an NMDA receptor antagonist, and by using a modified version of Attentional Set-

Shifting Task (ASST). Another aim of this study was to investigate the therapeutical role of PV+

interneurons in mPFC in the acute model of cognitive inflexibility, using in vivo optogenetics PV+

stimulation in channelrhodopsin-2 (ChR2) transfected rats.

In the ASST, animals learned to dig and retrieve food reward from a cup, after a "two dimension-

operant-association" - relevant (odor) and irrelevant (digging medium). Reversal learning (RL-odor

switch) and extra-dimensional shift (EDS-relevancy switch) sessions measured the cognitive flexibility in

the rats. AAV-containing-ChR2 transfections and optic fiber implant were stereotactically applied onto

rat mPFC, unilaterally, 2 weeks before assessing ASST.

Results showed that acute MK-801 injections during RL and EDS induced cognitive inflexibility compared

to the saline. Moreover, PV+ optogenetic ChR2-stimulation in mPFC recovers the ability to switch odor

and/or medium relevancy.

We observed that enhancing the inhibitory activity of PV+ interneurons in mPFC is a new experimental

therapeutic opportunity to rescue cognitive flexibility in schizophrenia-like psychosis. This work was

supported by GACR grant 17-04047S and AZV grant 17-30833A. Institutional support for IPHYS was

provided by RVO: 67985823. Additional support came from MEYS (LM2015062) Czech-BioImaging and

H2020 INFRADEV-01-2017 project ID-EPTRI (European Paediatric Translational Research Infrastructure -

EPTRI Grant Agreement 777554).

Molecular principles of Cajal body formation

Davide Alessandro Basello1,2, Michaela Efenberková1, Radek Macháň2, Nicola Maghelli3, David Stanek1

1. Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic; 2. Charles

University in Prague, Faculty of Science, Prague, Czech Republic; 3. Max Planck Institute of Molecular

Cell Biology and Genetics, Dresden, Germany

The cell nucleus is a highly heterogeneous environment crowded with numerous macro-molecules. Part

of the nuclear complexity rises from the presence of a number of different bodies, non-membrane

bound structures, which accumulate various proteins and, often RNAs. The molecular principles behind

bodies assembly and maintenance are recently a matter of an intensive debate. One of the "classical"

examples of a nuclear body is the Cajal body (CB). CBs are involved in biogenesis, quality control and

recycling of spliceosomal snRNPs. Coilin, the essential scaffolding protein of CBs, self-oligomerize and

interacts with numerous proteins including snRNPs, and these interactions are important for CB

formation. However, the basic information regarding its structure and function are lacking. To uncover

molecular principles of CB formation we determined coilin dynamics in the nucleoplasm and CBs and

analyzed the snRNP influence on coilin self-oligomerization and CB formation. To eliminate the effect of

endogenous coilin, we generated a coilin KO cell line. Then we prepared several different mutated

versions of coilin that prevent interaction with snRNPs and express them in coilin KO cells. Our results

show that coilin self-interaction does not depend on snRNP binding. However, the mutants that do not

associate with snRNPs fail to reconstitute CBs in coilin KO cells. We also apply different fluorescence

microscopy techniques (FRAP, single-point and imaging FCS and 3D-SIM SPT) to determine coilin

28

dynamics in the nucleus and the CB. The data confirm our biochemical results and show that abolishing

coilin interaction with snRNPs does not inhibit coilin self-association. Interestingly coilin dynamics inside

the CB does not differ from the nucleoplasmic coilin, suggesting that the CB nucleation and maintenance

are not based on different coilin mobility inside and outside CBs.

NFAT as a novel pathway in mesenchymal stem cell response to Inflammation.

Tidu F. (1, 2), Jose S. S. (1, 2), De Zuani, M. (1), Bendíčková K.(1), Pompeiano A. (1), Bělašková S. (1), Frič

J. (1)

(1) St. Anne’s University Hospital Brno International Clinical Research Center, Brno, Czech Republic;

(2) Department of Biology, Faculty of Medicine, Masaryk University, Czech Republic

Mesenchymal stem cells (MSCs) are multipotent cells present in the stromal fraction of many tissues. It

is now clear that they play an important role in immune response control during inflammation, due to

their ability to migrate to inflamed site and secrete immunoregulatory factors.

MSCs-expressed pattern recognition receptors (PRRs) allow direct sensing of infection or damage in

tissue microenvironment. So far most of the studies pointed at TLR NF-κB axis as the main responsible

for controlling MSCs inflammatory profile. However a clear insight on PRRs activation patterns and

cross-talks between NF-κB and other inflammatory pathways has not been sufficiently described, thus

hampering

MSCs therapeutic applications. Here we show that zymosan recognition by MSCs leads to the activation

of calcineurin-NFAT axis, which shapes MSCs response to inflammation.

The present work aims to shed light on NFAT role in MSCs biology, in order to better describe the

mechanisms underlying MSCs immunomodulation, which is of utmost importance in a translational

perspective.

Understanding nanoparticles-cells' interactions through mechanobiology

Marco Cassani, Jorge Oliver-De La cruz, Soraia Fernandes, Giancarlo Forte

Center for Translational Medicine (CTM) International Clinical Research Center (FNUSA-ICRC) St. Anne's

University Hospital

The new era of medicine requires patient-customized nanomaterials in which different cure strategies,

such as targeting and drug delivery, can be combined into a unique platform. As material science

progresses, a more complex understanding and deeper knowledge of the mechanisms involved in the

cells’ response to nanoparticles exposure are needed. Once the nanoparticles target the tumor site, they

are expected to induce significant changes in the extracellular microenvironment. Mechanobiology,

which studies the mechanotransduction by which the cells convert the outer mechanical stimuli into

inner biochemicals responses, is now emerging as the leading discipline for understanding cancer

development. The possibility to implement mechanobiology into nanomedicine can spread new light on

how healthy and unhealthy cells respond to nanoparticle-based treatments helping to drive the

nanomaterial’s activity to reach the desired specificity and efficiency in cancer treatment.

Femtosecond Stimulated Raman Spectroscopy

Alessandra Picchiotti



29

The use of femtosecond stimulated Raman probes make it possible to measure the vibrational spectra

of liquid, gaseous, and transparent solid state samples with femtosecond time resolution. This setup is

provided in the facility ELI-Beamlines in Dolni Brezany both as an independent beam line and as an

optional add-on to the other experiments.

Recording time-resolved structures of complex molecules (such as protein complexes) by femtosecond

X-ray pulses is among the important goals of ELI-Beamline’s project. However, these techniques are still

in the process of being developed and suffer from a high level of damage that is inflicted on a sample. In

contrast, time-resolved vibrational spectra can map molecular conformations in a highly non-invasive

way. The possibility of running these experiments simultaneously with X-ray experiments would allow

for valuable supplementary information to be obtained to cross-validate the conclusions drawn from the

X-ray experiments. A crucial advantage of the stimulated Raman probe is the great flexibility it offers in

terms of beam delivery, because it is based on pulses in the optical domain.

Pulse Radiolysis

Martin Precek



Radiation chemistry is a scientific field that is involved with studying the chemical effects of ionizing

radiation of matter to advance knowledge both in fundamental atomic and molecular science and in

applied radiation methods. In the latter case, research in radiation chemistry has strong connections to

areas that study and employ radiation effects in living systems, with important medical applications in

radiotherapy and radiation dosimetry, and also with areas dealing with the development of applied

radiation processing methods, such as radiation sterilization, radiation sanitation, radiation

polymerization, and food irradiation.

The ELI-Beamlines facility of the Institute of Physics of the Czech Academy of Sciences will be capable of

using laser interactions in matter to directly generate multiple kinds of pulsed ionizing radiation (soft

and hard X-rays, electrons, or protons and possibly even heavier ions). Capabilities are being developed

at ELI Beamlines for building experiments enabling ultrafast pulse radiolysis at user end-stations. As

opposed to approaches using conventional accelerators (LINACs, cyclotrons, etc.) the laser-driven

secondary radiation sources make it possible to achieve a temporal resolution of tens of femtoseconds

due to the short length of the laser-generated radiation pulse and the near-perfect synchronization of

the temporal overlap of the pulse of radiation and of a probing optical pulse in the studied sample

(usually liquid). This will then allow to perform measurements far below the famous "picosecond

barrier" in radiolysis, experimentally revealing poorly charted time scales where primary radical

products are generated from the interactions of ionizing radiation with matter.

Ultrafast dynamics of plasmon and magnetic resonances in nanoparticles

Carlo Maria Lazzarini, Tadzio Levato, Jamie M. Fitzgerald, José A. Sánchez-Gil, Vincenzo Giannini

Here I present a study of nanoparticle response to ultrafast laser excitation. I work in the electron

acceleration group at ELI-Beamlines, where we study and develop new particle acceleration techniques

and diagnostics methods for laser-plasma and laser-matter interaction.

The nanoplasmonics world is built around the fundamental concept of the collective resonant response

of the conduction electrons of a nanostructure, known as plasmon resonances, induced by an incident

electric field. Recent developments in nanofabrication techniques, high-sensitivity single-particle optical

characterization techniques, and fast numerical modeling tools for simulating complex nanostructures

have led to an increasing interest of the scientific community.

However, usually it is considered single only the monochromatic, continuous-wave regime. Nonetheless,

in the last decades, due to the development of ultrashort (fs timescale) laser pulses, many attempts

30

have been made to combine ultrashort laser pulses with nanostructures, generating the fast expanding

field of ultrafast nanoplasmonics. In which the the final aim is the control of the strong-field

enhancement localized in space on the nanometer scale and in time on the fs and sub-fs scale.

I present an analytical description of the ultrafast localized surface plasmon and magnetic resonance

dynamics in a single nanoparticle (Ag or Si), driven by an ultrashort (fs time scale) Gaussian pulse. Three

possible scenarios have been found depending on the incident pulse duration compared to the

resonance lifetime.

Nanoparticle resonance dynamics may lead to a wealth of new phenomena and applications in

nanophotonics such as multipole order resonance interference, pulse-induced delay or temporal

shaping on the fs scale, high harmonic generation, attosecond near-field pulse sources, and electron

acceleration from metasurface or 3D engineered nanostructures.

Mechanisms underlying axonal swelling formation Victorio M Pozo Devoto, Valentina Lacovich, Maria Carna, Monica Feole, Katerina Texlova & Gorazd B Stokin International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic Axonal swellings (AxS) are focal enlargements of axons found post-mortem in a range of biological and pathological settings including traumatic brain injury, Alzheimer's disease and multiple sclerosis. Despite their description in a range of settings, the mechanisms involved in the formation of AxS remain poorly understood. Here we developed a novel in vitro experimental paradigm to test for mechanisms underlying AxS formation. Human neuronal progenitor cells were seeded into microfluidic chambers and terminally differentiated to neurons. In these chambers axons grow into microchannels, which are crossed by a perpendicular channel to which we connected a syringe pump. Syringe pump generates force, which subjects axons to bending stress. After full characterization of the channel fluid dynamics, we tested how axons respond to the stress. Detailed analysis of the kinetics by time-lapse imaging showed a significant increase in the number and size of axonal enlargements during and after stress as visualised by transducing neuronal culture with a membrane targeted Cherry. We have first studied these axonal enlargements by scanning electron microscopy to reveal and distinguish physiological versus pathological enlargements following axonal injury. We next performed super-resolution microscopy to demonstrate loss of Spectrin BII periodicity of the sub-axolemmal cytoskeleton. We then investigated whether enlargements form as a result of membrane leakage in response to shear stress. Considering many reports found increased Ca2+ following axonal injury, we then asked whether Ca2+ concentrations increase in axonal enlargements. We found a significant increase in Ca2+ within the enlargements as visualised by the Fluo 4AM Ca2+ sensor. Furthermore, to understand the role of Ca2+ in enlarged axons following injury further we next depleted Ca2+ from media and blocked different axonal membrane and ER/mitochondrial Ca2+ channels. In summary, we created a unique cell culture paradigm to study the response of axons to physical injury and provide novel insight into mechanisms responsible for the formation of axonal swellings. Mechanical thrombectomy results variability per occluded artery in ischaemic stroke. L. Mengozzi, J. Vavrova, B. Koznar, F. Rohac, J. Sulzenko, J. Kroupa, I. Stetkarova, T. Peisker, P. Vasko, P. Widimsky University Hospital Kralovske Vinohrady and Third Faculty of Medicine, Charles University, Prague, Czech Republic. INTRODUCTION. Although mechanical thrombectomy is a rapidly developing technique worldwide in ischaemic stroke, the success rate according to the affected vessel is still not fully established.

31

METHODS. A total of 190 patients presenting with acute ischaemic stroke undergoing mechanical thrombectomy were divided into 5 subgroups: (a) isolated occlusion of the middle cerebral artery (MCA), (b) isolated occlusion of the proximal internal carotid artery (ICAprox), (c)isolated occlusion of the distal ICA (T-occlusion), (d) tandem occlusion of ICAprox and MCA, (e) basilar or vertebral artery occlusion. Immediate reperfusion success rate and 3-months clinical outcomes were analysed according to the occluded artery involved. RESULTS. Isolated occlusions of the MCA represented alone one of the largest subgroups with 42% of the patients, whereas 46% presented with ICA occlusions, of which 19% ICA distal occlusions (T-occlusion), 14% ICA proximal occlusions and 13% ICA proximal/ MCA tandem occlusions; only 12% of cases manifested ischaemia of the vertebral/basilar artery (VB). The TICI grading system was used to assess the success rate of the thrombectomy and grade 2b/3 was achieved in 84% of patients with obstructions of the MCA and 92% of cases with ICA proximal occlusions. Altogether, the success rate at TICI 2b/3 for MCA and ICA occlusions reached 96%. Although not the least common, ICA distal occlusions had a postthrombectomy success rate of 56% and mortality at 3 months of 42%, not quite unlike vertebral/basilar occlusions (VB) with a 48% reperfusion success and a 52% mortality in the first trimester, the latter being the rarest encountered type. Overall, 16% and 19% of patients died within 3 months, previously presenting with respectively MCA and ICA proximal occlusions, of which 53% and 46% scored between 0-2 with the modified Rankin scale (mRs). Only approximately ¼ of patients treated for VB and ICA distal occlusions manifested no significant disability despite some symptoms at 3 months (mRs 0-2). Test of genetic code evolution hypotheses: Reverse evolution of specific target proteins by mRNA-display technique Valerio Guido Giacobelli

1, Kosuke Fujishima

2, Vyacheslav Tretyachenko

1, Klara Hlouchova

1

1. Department of Cell Biology, Faculty of Science, Charles University, Hlavova 2030, 128 00, Prague 2, Czech Republic; 2. Earth-Life Science Institute, Tokyo Institute of Technology, Tokyo 1528550, Japan Although extant proteins consist of 20 different amino acids, it has been proposed that primordial proteins consisted of a smaller set of “early” amino acids and that additional “modern” amino acids have gradually been recruited into the genetic code. This naturally leads to the questions: can structured and functional proteins be constructed using the “early” amino-acid alphabet? Can extant proteins be reverse-evolved while preserving their structure/function? To test this, protein databases have been inspected to select model extant protein candidates with different structural folds. Our preliminary search contains proteins with both catalytic and binding/interaction functions. The selected protein targets were “reverse-evolved” in vitro into variants where the “modern” amino acid were randomized by “early” ones. The libraries of randomized genes were incorporated into a genotype-phenotype linkage to be compatible with an appropriate library display (mRNA display) and selection method. The selection of successful candidates was based on conservation of structure and/or function and the most “successful” variants will be characterized. This research will inform us of the essentiality of “modern” amino acids for building protein structure/function and thus will provide a direct test of the hypotheses about early proteins. In addition, proteins constructed from a limited amino acid alphabet are of importance in protein engineering and synthetic biology. Finally, this area touches upon the very basic link of protein sequence-structure-function that lies at the core of many biotechnological and biomedicine problems and has express implications for construction of artificial biochemistries.

HOSTS’ POSTERS:

mGlu3 METABOTROPIC GLUTAMATE RECEPTORS AS CANDIDATE TARGETS FOR NEUROPROTECTIVE

DRUGS IN THE MPTP MOUSE MODEL OF PARKINSONISM

32

M. Alborghetti*1, L. Di Menna*2, A. Traficante2, F. Pontieri1, F. Nicoletti2,3, V. Bruno2,3, G.

Battaglia2,3.

1Department of Neuroscience, Mental Health and Sensory Ogans (DESMOS), 2I.R.C.C.S. Neuromed,

Pozzilli, and 3Department of Physiology and Pharmacology, University Sapienza of Rome, Italy. *Equally

contributed to the work.

Introduction. mGlu3 metabotropic glutamate receptors are candidate targets for neuroprotective drugs

owing to their ability to stimulate the production of TGF-β and GDNF. We have found that systemic

treatment with a mixed mGlu2/3 receptor agonist, LY379268, in mice enhanced GDNF levels in the

striatum, and this effect was abrogated in mice lacking mGlu3 receptor. In addition, treatment with

LY379268 protected nigro-striatal neurons against neurodegeneration caused by an acute dose of MPTP

(Battaglia et al., Plos One 2009). The recent availability of subtype-selective agonists of mGu3 receptors

gave us the impetus to further examine the role of the receptor in mechanisms of neurodegeneration.

Aim:To establish whether the genetic deletion of mGlu3 receptors amplifies nigro-striatal damage and

whether selective pharmacological activation of mGlu3 receptors is neuroprotective in a chronic MPTP

mouse model that recapitulates the progressive degeneration occurring in Parkinson’s disease.

Methods: We are using mGlu3 or mGlu2 receptor knockout mice and their wild-type counterparts

treated with 20 mg/kg of MPTP each other day for 15 or 30 days. In addition, we are planning

experiments with a novel, selective, and brain permeant mGlu3 receptor agonist using the same mouse

model. Neurodegeneration is assessed by stereological cell counting in the substantia nigra, and HPLC

measurements of dopamine and its metabolites in the striatum. Results: So far, we have found that

chronic treatment with MPTP led to a substantial drop in dopamine levels in the striatum. This effect

was amplified in mGlu3 receptor knockout mice, whereas it was attenuated in mGlu2 receptor knockout

mice. Studies with the mGlu3 receptor agonist are ongoing. Conclusion: The use of the chronic MPTP

model in mice with genetic deletion supports the hypothesis that mGlu3 receptors extert a

neuroprotective activity attenuating the progressive degeneration of nigro-striatal dopaminergic

neurons. Whether selective pharmacological activation of Glu3 receptors causes neuroprotection and by

which mechanism (production of GDNF or reduction of neuroinflammation) is currently under

investigation.

MAGNESIUM IN INFLAMMATORY BOWEL DISEASES: AN OVERLOOKED ACTOR

Daniela Arduini

National delegate to biotechnologies of the National Chamber of the Biologists (Italy)

Background: Magnesium (Mg) is essential for human health and is absorbed mainly in the intestine. In

view of the likely occurrence of an Mg deficit in inflammatory bowel disease (IBD) and the documented

role of Mg in modulating inflammation, the present study addresses whether Mg availability can affect

the onset and progression of intestinal inflammation. Methods: To study the correlation between Mg

status and disease activity, we measured magnesemia by atomic absorption spectroscopy in a cohort of

IBD patients. The effects of dietary Mg modulation were assessed in a murine model of dextran sodium

sulfate (DSS)-induced colitis by monitoring magnesemia, weight, fecal occult blood, diarrhea, colon

length, and histology. Expression of the transient receptor potential melastatin (TRPM) 6 channel was

assessed by real-time reverse transcription polymerase chain reaction and immunohistochemistry in

murine colon tissues. The effect of Mg on epithelial barrier formation/repair was evaluated in human

colon cell lines. Results: Inflammatory bowel disease patients presented with a substantial Mg deficit,

and serum Mg levels were inversely correlated with disease activity. In mice, an Mg-deficient diet

caused hypomagnesemia and aggravated DSS-induced colitis. Colitis severely compromised intestinal

Mg2+ absorption due to mucosal damage and reduction in TRPM6 expression, but Mg supplementation

resulted in better restoration of mucosal integrity and channel expression. Conclusions: Our results

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highlight the importance of evaluating and correcting magnesemia in IBD patients. The murine model

suggests that Mg supplementation may represent a safe and cost-effective strategy to reduce

inflammation and restore normal mucosal function.

In vitro and in vivo evaluation of ferric-based and gadolinium-based nanoparticles in theranostic

perspective

Lenka Rajsiglova1,5, Pavol Lukac1,5, Paolo Tenti1,5, Michal Babic2, Daniel Jirak3, Peter Makovicky4,

Radislav Sedlacek4 and Luca Vannucci1

1 Institute of Microbiology, and 2 Institute of Macromolecular Chemistry of the CAS, v.v.i.; 3 Institute of

Clinical and Experimental Medicine - IKEM, Prague, Czech Republic; 4 Czech Centre for Phenogenomics,

Institute of Molecular Genetics of the CAS, v.v.i., Vestec, Czech Republic, 5 Faculty of Science, Charles

University, Prague, Czech Republic

With the progress of nanomedicine, a variety of nanoparticles (NPs) have been widely studied and

applied in biomedicine. Depending on their unique properties, NPs have become a hotspot in

theranostic approaches. Despite the growing interest, their possible immunotoxicity and long term

toxicity still need better elucidation. In our study, tumour and non-tumour cell lines (B16 melanoma and

3T3 fibroblasts) were challenged with 3 types of NPs. γ-Fe2O3-based NPs (polymer coated/naked), γ-

Fe2O3-based NPs with nickel (polymer coated/naked) and gadolinium silica-coated NPs, were tested for

effects on cell viability in vitro, via MTT assay. Their effect on immune cells in vivo was evaluated by

FACS on murine separated splenocytes. Our results indicate that no type of NPs altered the B16 cells

viability. Instead, almost all NPs affected the viability of 3T3 cells. After in vivo administration in mice,

the ferric NPs affected the activation (CD69 expression) of B lymphocytes and NK/NKT cells and also

slightly decreased the percentage of NK and NKT cells in the spleen. Ferric NPs, tested in magnetic

resonance (NMR), produced signal similar to standard contrast. Gadolinium-based NPs appeared to

localize to little vessels, even in the tumour, by evaluation with LA-ICP-MS (Laser Ablation Inductively

Coupled Plasma Mass Spectrometry). In conclusion, in vitro and in vivo data suggest that NPs can

interfere with fibroblasts viability but do not significantly affect the immune cells after systemic

administration, and generate signal in NMR and LA-ICP-MS. In the future, we will concentrate on

proliferation of the cells and their production of free radicals after NPs challenge in vitro and possible

theranostic potential of ferric and Gd NPs in vivo.

IMMUNITY AND REMODELING OF HUMAN COLON MUCOSA IN COLORECTAL CANCER

Pavol Lukac1,3, Paolo Tenti1,3, Fabian Caja1,3, Dmitry Stakheev1,3, O. Chernyavskiy2, D. Vondracek2,

Lenka Rajsiglova1,3, Pavol Makovicky4, Miroslav Levy5 , Peter Makovicky6, Radislav Sedlacek6 and Luca

Vannucci1

1Institute of Microbiology of the CAS, v.v.i. Prague, 2Institute of Physiology of the CAS, v.v.i. Prague, and

3Faculty of Science, Charles University, Prague, Czech Republic, and 4Department of Biology,

Pedagogical faculty, Selye Janos University, Komarno, Slovak Republic, 5Thomayer’s teaching Hospital,

Prague, Czech Republic, 6Czech Centre for Phenogenomics, Institute of Molecular Genetics of the CAS,

v.v.i., Vestec, Czech Republic

The tumor stroma is an important modulator of cancer cell behaviour. Looking by 2-photon microscopy

(second-harmonic generation imaging - SHG) to the colorectal cancer specimens from patients, we

found differences in the stroma organization of mucosa far from the tumor (apparently normal), near

the tumor border (transitional) mucosa and tumor. PCR analysis showed the progressive increase in

expression of COL1A1, IL-1 beta, IL-13 and LOXL2, all involved in the tissue remodelling. IL-6 appeared

increased especially at the transition from mucosa to tumor in correlation with higher inflammatory cell

34

infliltrate. IL-6 participation was also immunohistochemically evident. To model in vitro the tumor

development, 3D cultures of colorectal tumor cells as spheroids are undergoing to better explore the

appearance of PDL-1 immune check-point ligand for PD-1. This is in the view to elucidate PDL-1

expression and immune escape mechanisms in early stages of tumor development. Acknowledgements:

grants RVO 61388971 (CZ), Generali/Ceska Pojistovna Foundation (CZ), UniCredit Bank s.r.o., Prague

(CZ), AERO s.r.o. (CZ), CAMIC CZ, Eurinox s.r.o (CZ) and ARPA Foundation (IT).

first multidisciplinary conference of the italian ... · 17) test of genetic code evolution...

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