annual report 2016 - biodyn.ro · implications for ecology, food industry, bio-medicine and...

The International Centre of Biodynamics

1

ICB Annual Report 2016

Annual Report 2016

International Centre of Biodynamics Bucharest - Romania

www.biodyn.ro


2


Content Content ................................................................................................................................................................... 2

About us ................................................................................................................................................................. 3

Our Mission ............................................................................................................................................................ 4

Important Dates in the Institute’s History ................................................................................................... 5

Director’s Message ............................................................................................................................................... 6

The Managing and Advisory Boards .......................................................................................................... 10

… and us .................................................................................................................................................................. 11

Cell Cultures Laboratory .................................................................................................................................. 12

Electrochemistry Laboratory .......................................................................................................................... 14

Applied Electrochemistry Laboratory.......................................................................................................... 15

Microfabrication & Prototyping Laboratory ............................................................................................. 18

Bioanalysis Laboratory ..................................................................................................................................... 19

(including a mobile laboratory) .................................................................................................................... 19

Electrical Measurements Laboratory ..........................................................................................................20

Impedimetric and Plasmonic Bio Sensors ................................................................................................... 21

Optical Microscopy Laboratory .................................................................................................................... 23

Modeling and Data Analysis Laboratory .................................................................................................. 25

Pilot Laboratory on monitoring fish behavior for quality assessment of aquatic environments ................................................................................................................................................................................. 26

Atomic Force Microscopy Laboratory ......................................................................................................... 28

International projects ....................................................................................................................................... 30

National projects ............................................................................................................................................... 32

Visibility actions .................................................................................................................................................. 34

Emergent Applications fostered by ICB ..................................................................................................... 35

Relevant publications ...................................................................................................................................... 38


3


About us

According to the Agreement between UNESCO and Romanian Government, the

International Centre of Biodynamics-ICB has been established through the decision No.

1378/2000 of the Romanian Government, under the aegis of UNESCO, as a non-profit

organisation of general interest and public utility to pursue graduate training, research and

development initiatives in the field of Biodynamics.

ICB focuses on noninvasive, real time

assays and (non) linear data analysis for

appraisal of biosystems or/and of their

dynamics.

In a multidisciplinary approach, it

integrates precise measurements into

detailed models (of the dynamics) of living

systems and biointerfaces, taking into

account the complexity of the interaction

between biological structures and the

measuring systems.

The potential applications of these

methodologies have far reaching

implications for Ecology, Food industry,

Bio-Medicine and Pharmaceutical

industry. Some examples are: fast

detection of analytes, e.g. contaminants

or/and toxic compounds in liquid media,

appraisal the efficacy and cytotoxicity of

chemical compounds, biomass monitoring

etc.


4


Our Mission

The International Centre of Biodynamics (ICB) is a flexible, highly dynamic structure active

on the national and international research areas. It initiates and coordinates research related

to noninvasive, sensitive and cost effective methods to analyze and control biosystems. The

applicative domains span BioMedicine, Food Industry and Ecology. ICB aims to maintain a

balance between fundamental and applied research.

ICB conducts and is involved in a number of collaborative national and international

research projects having the following headlines:

Characterization techniques for cellular systems, with emphasis on noninvasive multi-

parameter real-time monitoring

Detection of contaminants and adulterates in food products and water

(microorganisms, heavy metals, antibiotics & toxins)

Monitoring cell cycle progression & Biomass assessment

Biomedical applications - pre-clinical, in vitro assays to assess the interaction

mechanisms between selected drugs and cellular structures as well as related cytotoxic effects

Monitoring & Nonlinear Analysis of the external stimuli (e.g. drugs, toxins, pathogens,

EM radiation, pollutants) effects on: (1) evolution of cellular systems and (2) fish behavior

ICB aims to provide a competitive research environment supporting the (re)

integration of valuable researchers and formation of new ones.


5


Important Dates in the Institute’s History

November 1996: The proposal to create the International Centre of

Biodynamics, ICB, was discussed at the UNESCO Centre of Membrane Science and Technology,

UNSW, Sydney, Australia

April 1998: The first International Symposium on Biodynamics

October 1999: The Agreement between Romania and UNESCO to establish the ICB

December 2000: Romanian Government issued the Decision No. 1378/2000 to establish ICB

January 2001: The ICB commences its independent activities; starts AFRAMILK, the first

European Research Project (under FP5) in ICB’s portfolio

March 2001: Romanian Parliament passes the law 110/2001 to ratify the Agreement between

Romania and UNESCO to establish the ICB

December 2001: In Agreement with UNESCO the first Management Board of the ICB is

appointed by Romanian Ministers of Education and Research

April 2004: Appointment of the first International Advisory Board of ICB

April 2005: Starts ROBIOS, the first European Project (under FP6) coordinated by ICB

April 2006: ICB activities start in the new lodging

May 2006: The International Conference on Biosensing and Biodynamics: From Basics to

Applications co-financed by UNESCO

April-May 2008: The IInd European Young Investigator Awardees Symposium EURYIAS 2008-

Self-organization and Selection in Evolution of Matter, Molecules and Life

July 2012: Starts EXTRACELL – the first ERC- like project “Monitoring the extracellular space

with catalytic self-propelled nanomotors”, hosted by ICB

July 2012: Starts BIOSCOPE – the first Complex project “Electro-Plasmonics for the analysis of

the dynamics of cellular processes and biomolecular interactions”, coordinated by ICB

April 2016: the first international Patent awarded to ICB members, U.S. Patent 9,315,855/2016,

Systems and Methods for Detection and Quantitation of Analytes Using an Oscillating Stimulus


6


Director’s Message This Annual Report highlights the major ICB accomplishments during 2016 with emphasis on both the progress of ICB research and the capabilities of ICB staff and laboratories for continuous development. I acknowledge with pride the landmarks of 2016:

Two international projects started in 2016:

FLAG ERA Graphene Flagship - GRAPHTIVITY: Graphene-based optoelectrochemical sensor for the simultaneous monitoring of the electrical and chemical activity of single cells, Project Director: Szilveszter Gaspar

NATO-SPS 985042: Cell Biosensors for Detection of Chemical and Biological Threats. Project Director: Eugen Gheorghiu Complemented by Manunet II project SENS4WINE which has been proposed and recommended for funding in 2016: New automated system based on biosensors for winemaking monitoring and assessment of allergen risk along the wine production chain

The projects supporting ICB activity: o BIOSCOPE, Electro-Plasmonics for the analysis of the dynamics of cellular processes

and biomolecular interactions, Contract No. 11/2012, ID: PN II-ID-PCCE-2011-2-0075, coordinated by ICB; Project Director: Eugen Gheorghiu;

o ESPRim - Single-molecule detection of DNA hybridization, based on electrochemical surface plasmon resonance microscopy and magnetic tweezers, Contract No. 29/2015, ID: PN-II-RU-TE-2014-4-2363, Project Director: C. Polonschii

o ANTIOXWIN-Exploiting the antioxidant capacity of black grapes for producing wines with high authentic quality – Contract No. 101/2012, ID: PN-II-PT-PCCA-2011-3.1-1809; Project Director: A. Vasilescu;

And 5 applicative projects proposed and approved in 2016:

FIND PATHOGEN - Advancement of a Portable System for Fast and Sensitive Detection of Pathogenic Cells, Contract No. 2PED/2017, ID: PN-III-P2-2.1-PED-2016-1041, Project Director: E. Gheorghiu

GRAPHTOOL - Graphene-based, miniaturized, electrochemical tool for the

investigation of tumor cell pH regulation (GrapHtool), Contract No. 110PED/2017, ID: PN-III-P2-2.1-PED-2016-1106,Project Director: S. Gaspar


7


SENSECELL - Sensitive Platform for Dynamic Assessment of

cellular effects of target endo- and exogenous compunds: case study Amyloidß, Contract No. 161PED/2017, ID: PN-III-P2-2.1-PED-2016-1137, Project Director: M. Gheorghiu

EPSIS - Electro-plasmonic system with improved sensitivity for

analyte detection, Contract No. 111PED/2017, ID: PN-III-P2-2.1- PED-2016-1279, Project Director: C. Polonschii

COLDSENSOR - Novel aldehyde dehydrogenase from Antarctic bacterium as highly

efficient catalyst for low temperature biosensing and biotechnologies, Contract No. 1PED/2017 PN-III-P2-2.1-PED-2016-0116, Coordinator: The Institute of Biology of the Romanian Academy, Partner institution: International Center of Biodynamics Partner responsible: A. Vasilescu

New scientific developments

International Patents: U.S. Patent 9,315,855/2016, Systems and Methods for Detection and Quantitation of Analytes Using an Oscillating Stimulus, Authors: E. Gheorghiu, S David, C. Polonschii, D.Bratu Papers:

Published: 1. Vasilescu A, Marty J-L, Electrochemical aptasensors for the assessment of food quality

and safety, Trends in Analytical Chemistry, 2016, 79, 60-70 2. Andrei V, Sharpe E, Vasilescu A, Andreescu S, A single use electrochemical sensor

based on biomimetic nanoceria for the detection of wine antioxidants, Talanta, 2016, 156-157, 112-118.

3. Chekin F., Singh S.K., Vasilescu A., Dhavale V.M., Kurungot S., Boukherroub R. , Szunerits S., Reduced Graphene Oxide Modified Electrodes For Sensitive Sensing of Gliadin in Food Samples, ACS Sensors, 2016, 1 (12), pp 1462–1470

4. Vasilescu A., Nunes G. , Hayat A., Latif U., Marty J-L., Electrochemical Affinity Biosensors Based on Disposable Screen-Printed Electrodes for Detection of Food Allergens, Sensors, 2016, 16, 1863.

5. Vasilescu A., Wang Q., Li M., Boukherroub R., Szunerits S., Aptamer-Based Electrochemical Sensing of Lysozyme, Chemosensors, 2016, 4, 10; doi:10.3390/chemosensors4020010

6. Wang Q, Palaniappan S, Schechter A, Teblum E, Yemini R, Nessim G.D, Vasilescu A, Li M, Boukherroub R, Szunerits S, Vertically Aligned Nitrogen-Doped Carbon Nanotube Carpet Electrodes: Highly Sensitive Interfaces for the Analysis of Serum from Patients with Inflammatory Bowel Disease, ACS Applied Materials & Interfaces , 2016, 8(15), 9600–9609.

7. Vasilescu A., Purcarea C., Popa E., Zamfir M., Mihai I., Litescu S., David S., Gaspar S., Gheorghiu M., Marty J-L., Versatile SPR aptasensor for detection of lysozyme dimer in oligomeric and aggregated mixtures, Biosensors & Bioelectronics, 2016, 83, 353-360.

Accepted: 8. Vasilescu A, Gáspár S, Gheorghiu M, David S, Dinca V, Peteu S, Wang Q, Li M,

Boukherroub R, Szunerits S, Surface Plasmon Resonance based sensing of lysozyme in


8


serum on Micrococcus lysodeikticus-modified graphene oxide surfaces, accepted, Biosensors and Bioelectronics

Submitted: 9. Vasilescu A, Gheorghiu M, Peteu S, Nanomaterial-based

electrochemical sensors and optical probes for detection and imaging of peroxynitrite: a review, Microchimica Acta

Book chapters: 1. S. David, C. Polonschii, M. Gheorghiu, D. Bratu, E. Gheorghiu, Biosensing Based on

Magneto-Optical Surface Plasmon Resonance, in MiMB series, Biosensors and Biodetection: Methods and Protocols, IInd Ed., A. Rasooly & B. Prickril Eds. submitted

2. Vasilescu A, Marty J-L, Aptasensors, an analytical solution for mycotoxin detection, Comprehensive Analytical Chemistry, Elsevier, submitted

3. Vasilescu A, Vezeanu A, David S, Gaspar S, Development of an aptasensors for lysozyme based on graphene oxide through the layer-by-layer method, in “Nanomaterials, nanoparticles, Nanodevices”, Series in Micro and Nanoengineering, 2016, vol 24, p. 145-160, Edited by Maria Zaharescu, Horia Chiriac, Dan Dascalu, Publishing House of the Romanian Academy. ISBN: 978-973-27-2643-3.

4. Vasilescu A, Dinca V, Filipescu M, Rusen L, Hosu I.S, Boukherroub R, Szunerits S, Dinescu M,. Peteu S.F, Recent Approaches to Enhance the Selectivity of Peroxynitrite Detection, in “Peroxynitrite Detection in Biological Media : Challenges and Advances”, 2016, editors: Royal Society of Chemistry, DOI: 10.1039/9781782622352-00166

5. Bucur B, Hosu I.S, Vasilescu A, Biosensors for Characterisation of Bioactive Compounds from Wines in “Bioactive compounds: natural sources, physicochemical characterization, applications”, Editor: Constantin Apetrei, Bentham Science Publishers, in Frontiers in Bioactive compounds, 2016, ISSN: 2468-6395.

6. Szunerits S, Wang Q, Vasilescu A, Li M, Boukherroub R, Graphene/gold nanoparticles for electrochemical sensing, in “Nanocarbons for Electroanalysis”, accepted for publication, editors: Sabine Szunerits, Rabah Boukherroub, Alice Downard, Jun-Jie Zhu, Wiley.

These accomplishments could not have been possible without the constant support, motivation and efforts of ICB staff and collaborators as well as of ICB Management and International Advisory Boards that fostered the continuous increase of Centre’s RTD capabilities and visibility during 2016.

Thank you!

Eugen Gheorghiu

http://pubs.rsc.org/en/content/ebook/978-1-78262-085-3


9


Organizational Chart

ICB Functional Structure

Cell Cultures Laboratory Electrochemistry Laboratory Applied Electrochemistry Laboratory Microfabrication & Prototyping Laboratory Bioanalysis Laboratory (including a mobile laboratory) Electrical Measurements Laboratory Impedimetric and Plasmonic Biosensors Optical Microscopy Laboratory Modeling and Data Analysis Laboratory Pilot Laboratory on Monitoring Fish Behavior Atomic Force Microscopy Laboratory


10


The Managing and Advisory Boards The International Advisory Board, the Managing Board and the

Director coordinate the activity of the International Centre of Biodynamics.

According to ICB statute, the current Managing Board was appointed in 2013 by the International Advisory Board.

Managing Board members (in alphabetical order):

Mircea Dumitru - Rector, University of Bucharest, Corresponding Member of Romanian Academy

Alexandru Morega - University Politehnica of Bucharest, Corresponding Member of Romanian Academy

Romeo Resiga - Polytechnic University of Timisoara

Octavian Popescu – Babes-Bolyai University of Cluj-Napoca, Member of Romanian Academy

Nicolae Zamfir- Director General, Institute for Nuclear Physics and Engineering “Horia Hulubei”, Member of Romanian Academy

The members and observers of the International Advisory Board (in alphabetical order):

Prof. Dr. Koji ASAMI Japan

Prof. Dr. Anton ANTON Representative of Romanian University Research Council

Prof. Dr. Jean-Michel KAUFFMANN Universite Libre de Bruxelles, Belgium

Prof. Dr. Maciej NALECZ Director, Division of Basic and Engineering Sciences UNESCO –Paris, France Director, UNESCO – CEPES

Minister of Education and Research

Representative of the European Commission, to be nominated


11


… and us

Eugen Gheorghiu -2001* ICB Director Research Professor PhD, Physicist, theoretical physics EIS&SPR assays Modeling & Data analysis

Dumitru Bratu-2001* Engineer Electronics Development of Electrochemical devices

Szilveszter Gáspár-2006* Head of Electrochemistry Laboratory PhD, Chemist Electrochemical biosensors

Mihaela Gheorghiu -2001* Head of (Bio) Sensors, (Bio) Surface EIS&SPR biosensing Analyses and Cell culturing Department Research Professor, Physicist Electro - Optical Analysis of Biointerfaces

Mihai Sorin David-2003* PhD, Biophysics/Biodynamics EIS&SPR biosensing Bioimpedance Measurements, Biosensor Development and Nonlinear Analysis

Cristina Polonschii-2006* PhD, Biophysics/Biodynamics EIS&SPR biosensing Development of analytical methods to interrogate bio interfaces

Alina Vasilescu-2011* PhD, Analytical Chemistry EIS&SPR biosensing Practical applications of Biosensors

Luciana Stanica – 2012* PhD student Biologist Electrophysiological analysis

Mihnea Rosu Hamzescu-2012* PhD student EIS&SPR biosensing

Florica Moranescu – 2011* Cell culture technician

Cezar Giubalca-2001* Head of the Administrative Office Economist

Valeria Nane-2001* Chief Accountant Economist

* Date of employment at ICB


12 ICB Annual Report 2016

Cell Cultures Laboratory

Team: Mihaela Gheorghiu Luciana Stanica Florica Moranescu

Facilities

- Cell cultures facilities that enable cultivation of different epithelial tissues: cell collection

which contains renal epithelial cells (A6, MDCK-I, MDCK-II, MDCK parental, LLC-PK1),

intestinal epithelial cells (CaCo-2), epithelial cells from a cervical carcinoma (HeLa), cells

derived from an adrenal medulla (PC12); Neuroblast from human neural tissue SH SY-5Y,

optogenetically engineered HEK 293 cells containing bacterial channel rhodopsin; HT29

cancer cells expressing Carbonic anhydrase IX under hypoxic conditions; CO2 incubators

(from Sanyo Biomedical Division–U.K), cryogenic storage facilities and deep freezer, Two

microelectrodes Voltage Clamp (TEVC) system, Osmometer (from Gonotec Germany),

Transepithelial, impedance and noise system (TINS), inverted microscope; microinjection

setup and support peripherals (pumps, pipette puller, Faraday cages, dedicated

measurement chambers and antivibration table); hypoxic chamber, computer controlled

LED illuminators.

- other lab utilities (purified air, microbiological hood class II, ultra pure water)

Current Research

In the Cell Cultures Laboratory we have developed a battery of methods to study the effects of

external factors (pharmaceutical compounds or bacteria) assessing several useful parameters.

The methods consider:

A. Analysis of transport and cellular permeability;

B. Toxicity studies at cellular level (changes in transport properties and cellular viability);

C. Changes in cell surface attachment and cell- to cell communication upon interaction

with stressors;

D. Cell behavior under hypoxic conditions;

This expertise has been advanced with optogenetically engineered HEK 293, hypoxic

conditions and multisensory approaches towards continuous opto-electrical evaluation and

integration of cells in biosensing platforms.

Accomplishments

Impact of Controlled Illumination on stable Cell Lines Containing Light Activated Channels (Channel Rhodopsin)



A novel biosensing method based on an exogenous driven rhythm induced using optogenetic tools and assessment of the dynamics of cell response and recovery as reporter processes of bioactive analytes’ presence

Cell behavior under hypoxic conditions and carbonic anhydrase inhibition.

2 articles submitted:

L. Stanica*, M. Rosu-Hamzescu*, M. Gheorghiu, M. Stan, L. Antonescu, C. Polonschii, E. Gheorghiu Bioanalytical electro-optical platform for evaluation of bioeffects; Case study – cancer hypoxia

L. Stanica*, M. Gheorghiu*, M. Stan, C. Polonschii, S. David, D. Bratu, A. Dinischiotu, C. T. Supuran, E. Gheorghiu Quantitative assessment of specific carbonic anhydrase inhibitors effect on hypoxic cells using electrical impedance assays submitted Biosens Bioelectron 3 project applications submitted (1 PED projects, one PCE project, one PCEexploratory)

2 oral and 1 poster presentations at international conferences

Oral A new twist for optogenetics: light driven dynamics for cell based sensing M. Gheorghiu, L. Stanica, C. Polonschii, D. Bratu, E. Gheorghiu IC-ANMBES, Brasov, June29-July 01, 2016

Oral L. Stanica, M. Gheorghiu, E. Gheorghiu Electro-optical evaluation of cellular dynamics of ChR2 expressing HEK293 cells induced by light stimulation The EMBO Meeting 2016, 10–13 September 2016 Mannheim, Germany

Poster L. Stanica, M. Gheorghiu, E. Gheorghiu Characterization of light responsive, genetically modified cells as cellular sensors, 10th FENS Forum of Neuroscience, July 2-6 2016, Copenhagen, Denmark



Electrochemistry Laboratory

Team: Szilveszter Gáspár

Facilities

The resources of the laboratory include the following equipment: VSP modular

potentiostat/ galvanostat (from Bio-Logic S.A., France), CellTest multi-channel

potentiostat/ galvanostat (from Solartron Analytical, UK), Nanoband Explorer II anodic

stripping voltammetry-based heavy metal analyzer (from TraceDetect, USA), 797 VA

Computrance PC controlled system for voltammetry (from Metrohm AG, Switzerland),

Scanning Electrochemical Microscopy (SECM, from Sensolytics, Germany), trinocular stereo

microscope (from World Precision Instruments, USA) equipped with a Coolpix 995 digital

camera (from Nikon, Japan), EG-40 micropipette beveller (from Narishige, Japan), and 3D

micropositioner (HS6 from World Precision Instruments, USA).

Current Research and associated projects

The Electrochemistry Laboratory was mainly focused on Graphtivity a project developing

electrochemical sensors which i.) allow monitoring small molecules (e.g. hydrogen peroxide,

dopamine, glucose, etc.) at cellular level, ii.) are also compatible with optical microscopy

methods (e.g. Surface Plasmon Resonance microscopy), and iii.) build on the advantageous

electrical and optical properties of graphene. The laboratory has also made contributions to

projects on protein aggregation and on electro-plasmonics.The following papers were

published during 2016 with the involvement of the Electrochemistry

Laboratory:

(1) Vasilescu, A.; Purcarea, C.; Popa, E.; Zamfir, M.; Mihai, I.; Litescu, S.; David, S.;

Gaspar, S.; Gheorghiu, M.; Jean-Louis Marty. Versatile SPR Aptasensor for Detection of

Lysozyme Dimer in Oligomeric and Aggregated Mixtures. Biosensors and Bioelectronics

2016, 83, 353–360.



Applied Electrochemistry Laboratory

Team: Alina Vasilescu

Facilities

Main resources are represented by an 8-channel potentiostat from Dropsens, Spain and a

portable SPR system (SPREETA, Nomadics). The group shares with the Electrochemistry

group the VSP modular potentiostat/ galvanostat (from Bio-Logic S.A., France).


For the Applied Electrochemistry group, 2016 marked the end of the Partnership Project

ANTIOXWIN. In 2016, the focus of the group was on publishing the research results and

strengthening collaborative ties with other groups from Romania (Institute of Biology of the

Romanian Academy, National Institute for R&D in Biological Sciences, Bucharest, Faculty of

Chemistry-University of Bucharest, National Institute for Laser, Plasma and Radiation

Physics (INFLPR), Magurele), France (Prof. Jean-Louis Marty’s group at BAE, University of

Perpignan and Prof. Sabine Szunerits group at Institute of Electronics, Microelectronics and

Nanotechnology-IEMN, University of Lille 1, France) and the US (dr. Serban Peteu at

Michigan State University and Prof. Silvana Andreescu at Clarkson University). This was

reflected in papers published or accepted, done in collaboration with the above groups. An

important achievement was the CSII title obtained by Alina Vasilescu.

Accomplishments:

CSII title accorded to Alina Vasilescu 7 papers published, 1 accepted and 1 submitted for publication;

5 book chapters –three published, one accepted, one submitted for publication; 2 oral and 1 poster presentations at national/international conferences in Romania 1 national research project finished in 2016 (ANTIOXWIN, PN-II-PT-PCCA-2011-3.1-

1809). 1 research project accepted for funding (Horizon 2020, Manunet II SENS4WINE); 4 project applications submitted (2 PED projects, one PCE project, one Bilateral

Romania-France Brancusi) Workshop “Modern approaches for producing high quality wines rich in antioxidants”

organized in the frame of ANTIOXWIN project on July 05, 2016 at the International Centre of Biodynamics.



Articles: Published:

3 1. Vasilescu A, Marty J-L, Electrochemical aptasensors for the assessment of food quality and safety, Trends in Analytical Chemistry, 2016, 79, 60-70

4 2. Andrei V, Sharpe E, Vasilescu A, Andreescu S, A single use electrochemical sensor based on biomimetic nanoceria

for the detection of wine antioxidants, Talanta, 2016, 156-157, 112-118. 3. Chekin F., Singh S.K., Vasilescu A., Dhavale V.M., Kurungot S., Boukherroub R., Szunerits S., Reduced Graphene Oxide Modified Electrodes For Sensitive Sensing of Gliadin in Food Samples, ACS Sensors, 2016, 1 (12), pp 1462–1470 4. Vasilescu A., Nunes G., Hayat A., Latif U., Marty J-L., Electrochemical Affinity Biosensors Based on Disposable Screen-Printed Electrodes for Detection of Food Allergens, Sensors, 2016, 16, 1863. 5. Vasilescu A., Wang Q., Li M., Boukherroub R., Szunerits S., Aptamer-Based Electrochemical Sensing of Lysozyme, Chemosensors, 2016, 4, 10; doi:10.3390/chemosensors4020010 6. Wang Q, Palaniappan S, Schechter A, Teblum E, Yemini R, Nessim G.D, Vasilescu A, Li M, Boukherroub R, Szunerits S, Vertically Aligned Nitrogen-Doped Carbon Nanotube Carpet Electrodes: Highly Sensitive Interfaces for the Analysis of Serum from Patients with Inflammatory Bowel Disease, ACS Applied Materials & Interfaces , 2016, 8(15), 9600–9609. 7. Vasilescu A., Purcarea C., Popa E., Zamfir M., Mihai I., Litescu S., David S., Gaspar S., Gheorghiu M., Marty J-L., Versatile SPR aptasensor for detection of lysozyme dimer in oligomeric and aggregated mixtures, Biosensors & Bioelectronics, 2016, 83, 353-360.

Accepted: 8. Vasilescu A, Gáspár S, Gheorghiu M, David S, Dinca V, Peteu S, Wang Q, Li M, Boukherroub R, Szunerits S, Surface Plasmon Resonance based sensing of lysozyme in serum on Micrococcus lysodeikticus-modified graphene oxide surfaces, accepted, Biosensors and Bioelectronics

Submitted: 9. Vasilescu A, Gheorghiu M, Peteu S, Nanomaterial-based electrochemical sensors and optical probes for detection and imaging of peroxynitrite: a review, Microchimica Acta, Book chapters:

10. Vasilescu A, Vezeanu A, David S, Gaspar S, Development of an aptasensors for lysozyme based on graphene oxide through the layer-by-layer method, in “Nanomaterials, nanoparticles, Nanodevices”, Series in Micro and Nanoengineering, 2016, vol 24, p. 145-160, Edited by Maria Zaharescu, Horia Chiriac, Dan Dascalu, Publishing House of the Romanian Academy. ISBN: 978-973-27-2643-3. 11. Vasilescu A, Dinca V, Filipescu M, Rusen L, Hosu I.S, Boukherroub R, Szunerits S, Dinescu M,. Peteu S.F, Recent Approaches to Enhance the Selectivity of Peroxynitrite Detection, in “Peroxynitrite Detection in Biological Media : Challenges and Advances”, 2016, editors: Royal Society of Chemistry, DOI: 10.1039/9781782622352-00166 12. Bucur B, Hosu I.S, Vasilescu A, Biosensors for Characterisation of Bioactive Compounds from Wines in “Bioactive compounds: natural sources, physicochemical characterization, applications”, Editor: Constantin Apetrei, Bentham Science Publishers, in Frontiers in Bioactive compounds, 2016, ISSN: 2468-6395.





13.Szunerits S, Wang Q, Vasilescu A, Li M, Boukherroub R, Graphene/gold nanoparticles for electrochemical sensing, in “Nanocarbons for Electroanalysis”, accepted for publication, editors: Sabine Szunerits, Rabah Boukherroub, Alice Downard, Jun-Jie Zhu, Wiley. 14. Vasilescu A, Marty J-L, Aptasensors, an analytical solution for mycotoxin detection, Comprehensive Analytical Chemistry, Elsevier,

submitted Conferences: Oral presentations:

1. Vasilescu A, “Assessing the antioxidant potential of wines with electrochemical” sensors, “Modern approached for producing high quality wines rich in antioxidants” workshop, International Centre of Biodynamics, July 5, 2016

2. Vasilescu A, Andrei V, Vezeanu A, Pircalabu L, Tudor G, Ficiu L, Porumb R, Tudorache A, Branduse E, “Application of electrochemical methods for optimizing the production and characterization of red wines from Valea Calugareasca vineyard”, Annual Session of Scientific Communications, ICDVV Valea Calugareasca, June 16, 2016

Posters: 1.Danet AF, Parcalabu L, Popa CV, Porumb R, Brinduse E, Vasilescu A, Antioxidant potential of aronia-enriched wines assessed by chemiluminometric,spectrophotome tric and electrochemical methods, IC-ANMBES, Brasov, June29-July 01, 2016 .



Microfabrication & Prototyping Laboratory

Team: Mihai Sorin David Dumitru Bratu

Facilites

CNC Shape Cutting Machine-2000 series 8 direction mill (from Sherline Products, USA),

P6700 Spin coater (from Specialty Coating Systems, USA), lathe and rectifier lathe. Physical

Vacuum Deposition PVD 75 (Kurt J Lesker, USA). Modular design configured to suit a

variety of thin film deposition applications, typically for research and development or small

batch production.

Physical Vapor Deposition-PVD 75

PVD 75 thermal evaporator from KJ Lesker is currently used

for manufacture of different configuration of electrodes and

for different applications requiring precise deposition of

several layers of materials. For example: SPR – we

manufacture SPR chips by depositing 2 nm of

Chromium/Titanium and 50 nm of Gold; Magnetic SPR – we

prepare metallic “sandwiches” comprising of Chromium,

Cobalt and Gold; Electrode passivation – we deposit thin

layer of insulating materials (e.g. SiO).

Additional facilities: Spin coating machine within glove box, laminar flow hood for

specific surface functionalization, ultrasonic processors and UV ozone cleaning system. The

spin coater is used for controlled deposition of thin polymer layers on solid substrates for

specific applications including electrode preparation for impedance measurements,

investigation of cell adhesion to various materials, controlled local electrical insulation for

electrode design, customized PCB boards, optical coupling for SPR measurements.

Current Applications

Developing user defined parts and accessories from holders and test fixtures electro-

optically compatible measurement cells (static and flow through) and tailored plasmonic

surfaces, with sizes down to tens of microns.



Bioanalysis Laboratory (including a mobile laboratory)

Team: Mihaela Gheorghiu Szilveszter Gáspár Mihai Sorin David Cristina Polonschii

Facilities

Anodic Stripping Voltammeter Nanoband Explorer II (from Tracedetect U.S.A) Dual system

EIS/SPR (ICB patent), Glomax Luminometer 20/20 (from Promega U.S.A), pH-meter and

conductometer (from WTW Germany), portable UV-VIS Spectrophotometer Nova 60A

(from Merck, Germany), Mobile autolaboratory equipped with working areas, separate

access and electrical connections.

Current Status

The Mobile and Fixed Bioanalysis Laboratory was accredited in 2010 by RENAR as a third

party testing laboratory, in compliance to the ISO/IEC 17025:2005 clauses and to the

provisions of European directives and national legislation on water quality.

Following the reduction of the commercial activities within the accredited laboratories

during 2011, the activity of this laboratory was suspended in 2012, at our request.



Electrical Measurements Laboratory

Team: Eugen Gheorghiu Mihai Sorin David Cristina Polonschii Mihnea Rosu Hamzescu Dumitru Bratu

Facilities

Impedance analyzer Agilent 4294A (from Agilent Technologies, USA), Impedance / gain

phase Analyzer Solartron; 1470 E Cell Test System(both from Solatron Analytical, U.K),

Digital Oscilloscope TDS 3052 (from Tektronix SA-France), Signal generators, Phasemeter,

Spectral Analyzer HP 3585; 8 channel low frequency impedance spectrometer (designed

and produced in ICB);

Dedicated working lab and current utilities (purified air, ultra pure water, Faraday and

thermostated chambers, 4 point electrode configurations and wide range of surface

electrodes – circular and/or interdigitated, compatible with complementary optical assays).

Current Research and related accomplished projects

DYNANO (FP7), BioScope

- Investigation of lipid film formation in view of lipid sensors development - mimics for

actual biological membranes.

- Investigation of the dynamics of electrical parameters of cellular platforms in relation to

engineered interfaces (with attractive/repulsive chemical and morphological features),

- Investigation of the dynamics of electrical parameters of cellular platforms in relation to

chemical and biological stressors (heavy metals, pathogen cells).

- Monitoring cell cycle progression on non/synchronized cell suspensions – an on line system for

electro-optical evaluation of yeast cell suspensions has been developed.

- Evaluation of functionalization protocols (deposition of thin polymeric layers, biorecognition

compounds or ligand – e.g. thiol layers).

- Development of a multi channel dual SPR impedance set-up.



Impedimetric and Plasmonic Bio Sensors

Team: Mihaela Gheorghiu Mihai Sorin David Cristina Polonschii Dumitru Bratu Luciana Stanica

Facilities

-Biacore 3000 system (from Biacore AG Sweden)

-Dual spectrometer EIS/SPR (ICB patent),

-3 channel Spreeta Modules, Contact Angle Meter CAM 100(from KSV, Finland).

-Dedicated working lab and current utilities (purified air / N2, chemical hood and ultra

pure water, thermostated chambers)

-Advanced set-up for monitoring the dynamics of aquatic species and the quality of growth

environment.

-Advanced set-up for high speed impedance assay with synchronization with light pulses.

-Custom design generators for multifrequency assays.

Additional facilities: Evolution 600 UV-VIS Spectrophotometer (Thermo Scientific USA) with

variable angle specular module, AFM module Nano Wizzard II(JPK, Germany) Multimode

readers Promega / Turner.


The design of novel (bio) sensing interfaces for selectively recognition of the analyte of

interest, from low molecular compounds to cells, and for providing a concentration-

dependent phenomenon that is easy to translate into useful analytical signal) towards a

new generation of analytical tools. These analytical tools will extend our ability of detecting

compounds of interest:

in very low concentrations,

in complex samples without prior separation,

in small sample volumes (microliters) or exotic environments (such as cellular sub

compartments), and

in a timely manner traditional analytical approaches are very often lacking such

abilities.



The integration of complementary analytical tools (SPR, electrochemistry) with micro-

technologies (fluidics, electronics) for the development of portable, sensitive solutions for

real time detection of target analytes.

Accomplishments

Book chapter S. David, C. Polonschii, M. Gheorghiu, D. Bratu, E. Gheorghiu,

Biosensing Based on Magneto-Optical Surface Plasmon Resonance, in MiMB series,

Biosensors and Biodetection: Methods and Protocols, IInd Ed., A. Rasooly & B. Prickril

Eds., Springer submitted

Supported by:

2 national research projects:

o BIOSCOPE, Electro-Plasmonics for the analysis of the dynamics of cellular processes and biomolecular interactions, Contract No. 11/2012, ID: PN II-ID-

PCCE-2011-2-0075, coordinated by ICB; Project Director: Eugen Gheorghiu; o Sensitive quantitation of target microorganisms using dual electro plasmonic

analysis and magnetic actuation, Contract No. 82-30.04.2013 , ID: PN-II-RU-PD-2012-3-0467, Project Director: Mihai Sorin David

1 international research project:

o Tumoranalyzer- Response of in vitro hypoxic tumor models to potentially therapeutic compounds as revealed by an advanced analytical platform, Romanian‐Swiss Research Programme IZERZ0_142236, 2012, Promoter from Switzerland, École Polytechnique Fédérale de Lausanne: Hubert Girault and from ICB: Eugen Gheorghiu;

Participation in the Cost Action Proposal “European Perspectives Towards Bioelectronics”



Optical Microscopy Laboratory

Team: Mihaela Gheorghiu Mihai Sorin David Luciana Stanica

Facilities

Zeiss Total Internal Reflection Microscope (TIRFM), Zeiss AxioObserver Z1, fully motorized

with xyz nm resolution, equipped with EMCCD ANDOR IXon DU-885K camera, with

facilities for epifluorescence, laser (3 line Ar) and white light TIRF, and various contrast

methods (Ph, DIC) and for cell cultivation (OkoLab controlled environmental cage);

Extended image processing facilities (AxioVision, Andor IQ and Andor IQ Tracker);

dedicated chambers for combined (optical and electric) assays.; Additional filter sets 02, 09,

14. NIKON Eclipse 400 microscope with epiflourescence system and CCD camera for data

acquisition.

Additional facilities:

- Spectrophotometer Evolution 600 (Thermo Scientific, USA), with variable angle specular

reflectance accessory and thermostated carrousel.

- GloMax 20/20 Multimode reader (Promega), with 2 injectors and UV and Blue

fluorescence modules.

- Compatible AFM module fully integrable with advanced optical assessment.


- Dual electro-optical measurements based on dedicated flow cells with optimized

electrode configurations.

- Time lapse assessment of the dynamics of cells (morphology, attachment and metabolism)

and cell organelles (pH, ions, morphology) in response to external stimuli and engineered

bio-interfaces cues; membrane processes in normal and pathologic conditions; manipulation

and characterization of natural and synthetic lipid environments (including cholesterol).

ATP & Cytotoxicity assays; Intracellular Ca2+;

The push - pull microfluidic probe developed by the Swiss Partners within

TUMORANALYZER Project has been integrated within the AxioObserver Z1 set-up and



experiments enabled 1. Precise control of probe positioning with optical

feedback; 2. Fluidic and electric cell stimulation 3. Experimental

validation of modelling studies.

Accomplishments

2 manuscripts on cell actuation and imaging to be submitted

Participation in the 14th EMBO Practical Course in Advanced Optical Microscopy, 6-16 April

2016, Plymouth England (L. Stanica, fellowship)



Modeling and Data Analysis Laboratory

Team: Eugen Gheorghiu Cristina Polonschii Sorin David

Current research and associated projects

We are currently developing fast, efficient methods to asses target analytes using periodic

actuation.

Dielectric modeling and nonlinear time series analysis

Strong emphasis is placed on both experimental and theoretical aspects regarding:

Development of new Plasmonic-EIS instrumentation;

Development of microscopic models of dielectric behavior with emphasis on:

(1) Membrane Potential (discussion of GHK approach) and the role effect of ionic channels

(2) non-spherical cells (focusing on yeasts, red blood cells, and gap junction connected cells).

A quantitative approach providing shape evolution of budding yeasts during the cell cycle,

consistent with experimental findings, is available.

Time series analysis of dielectric data yielding quantitative measures of the system

dynamics (revealing changes in the tissue structure and function and the invariants of the

cell cycle

3 project applications submitted (2 PED projects, one PC Exploratory)

An international patent application was granted: U.S. Patent 9,315,855/2016, Systems and Methods for Detection and Quantitation of Analytes Using an Oscillating Stimulus, Authors: E. Gheorghiu, S David, C. Polonschii, D Bratu Participation to international conferences

Oral – E. Gheorghiu, Advances in Magneto-Optical Surface Plasmon Resonance concepts for Affinity based biosensing in liquid media, IC-ANMBES, Brasov, June29-July 01, 2016 Oral Gheorghiu E, Polonschii C, David S, Gheorghiu M, Rosu-Hamzescu M, Bratu D, “Plasmonic based EIS: towards a sensitive analytical method gathering the virtues of both SPR and EIS”, The IVth International Conference on Analytical and Nanoanalytical Methods for Biomedical and Environmental Sciences “IC-ANMBES 2016”, Brasov, June 2016 Book chapters S. David, C. Polonschii, M. Gheorghiu, D. Bratu, E. Gheorghiu, Biosensing Based on Magneto-Optical Surface Plasmon Resonance, in MiMB series, Biosensors and Biodetection: Methods and Protocols, IInd Ed., A. Rasooly & B. Prickril Eds., Springer submitted



Pilot Laboratory on monitoring fish behavior for quality assessment of aquatic environments

Team: Eugen Gheorghiu Cristina Polonschii Dumitru Bratu

Facilities

- A platform consisting in a 3D array of

ultrasound transducers and of supporting

analytic modules to monitor fish positions and

assess fish dynamics (ICB patent);

- Dedicated water tank with adequate

infrastructure for controlling water

temperature, composition, including dissolved

Oxygen (DO);

- A module for controlled dispensing of

nutrients;

- Facilities for complementary video

observations of fish behavior;

- A set of sensors for continuous monitoring of water parameters: pH, DO, conductivity and

temperature.

Current Research

Extending the capabilities of the platform to remotely monitor and analyze the

behavior/dynamics of various fish species (including sturgeons) both in tanks and cages

immersed in aquatic media (lakes, rivers). The monitoring system fosters detection of

behavioral changes of aquatic species triggered by harmful compounds that might incidentally

occur in water (even in very small concentrations, well below the lethal threshold of those

species).

Accomplishments

A patent application was granted:

Patent Ro 128065/2015: “Platform and method to monitor the quality of an aquatic

environment based on analysis of the behavior of a fish population”, Authors: E. Gheorghiu,

C. Polonschii, D. Bratu



Oral Presentation Gheorghiu E, Polonschii C, “A multitiered approach for monitoring water quality”, International Conference on Sustainable Solutions for Energy and Environment, “EENVIRO 2016”, Bucharest, October 2016



Atomic Force Microscopy Laboratory

Team: Szilveszter Gáspár Mihaela Gheorghiu Sorin David

Facilities

The major piece of equipment of the laboratory is a NanoWizard® II Atomic Force

Microscope JPK AG (Berlin, Germany) combined with optical microscope Zeiss AxioObserver

D1 fully equipped with phase contrast, fluorescence and dark field for complementary

optical observations.


Our AFM instrument has the necessary features to work with biological samples (i.e. soft

samples in liquid), and thus is often used to observe small cellular structures (e.g. tight

junctions, microvilli, etc.) or surface absorbed / immobilized proteins and bacteria (see

Figure 1A). In addition to the study of biological samples, it is also used to investigate the

quality and thickness of thin metal films deposited by PVD or metal sputtering and the

structure of graphene-modified surfaces (see Figure 1B).



Figure 1A: AFM image of a Micrococcus

lysodeiktikus-modified surface.

Figure 1B: AFM image of a graphene oxide-

modified surface;

Acomplishments: 1 published paper Vasilescu A., Purcarea C., Popa E., Zamfir M., Mihai I., Litescu S., David S., Gaspar S., Gheorghiu M., Marty J-L., Versatile SPR aptasensor for detection of lysozyme dimer in oligomeric and aggregated mixtures, Biosensors & Bioelectronics, 2016, 83, 353-360.

1 Accepted: Vasilescu A, Gáspár S, Gheorghiu M, David S, Dinca V, Peteu S, Wang Q, Li M, Boukherroub R, Szunerits S, Surface Plasmon Resonance based sensing of lysozyme in serum on Micrococcus lysodeikticus-modified graphene oxide surfaces, accepted, Biosensors and Bioelectronics



International projects ONGOING

Cell biosensors for detection of chemical and biological threats, Contract: NATO SPS 985042, Project director: Eugen Gheorghiu, 2016-2019

GRAPHTIVITY - Graphene based optoelectrochemical sensor for the simultaneous monitoring of the electrical and chemical activity of single cells, Contract: Nr. 40 din 2016, ID: FLAG-ERA-Grafitivity, Project director: S. Gaspar, 2016-2018

ACCOMPLISHED

CAPTALLERGENES- Aptamer based sensors for allergen detection, Cooperation Romania-France, PN-II-CT-RO-FR-2012-1-0012, Project Director: Alina Vasilescu;

Tumoranalyzer-Response of in vitro hypoxic tumor models to potentially therapeutic compounds as revealed by an advanced analytical platform, Romanian‐Swiss Research Programme IZERZ0_142236, 2012, Promoter from Switzerland, École Polytechnique Fédérale de Lausanne: Hubert Girault and from ICB: Eugen Gheorghiu;

“DYNANO” Dynamic Interactive Nanosystems, FP7-PEOPLE-2011-ITN N°289033 7th FP (2011-2015);

“PROARGUS” Protein Aggregation: a quantitative assessment, PIRG08-GA-2019-277126, Call FP7-PEOPLE-2010-RG (2011-2015) – MC grant researcher- Alina Vasilescu;

“Non-invasive electro-optical methods to assess the effect of nano/micro substrattopography on living cell dynamics”, Academic Cooperation between Romania and Russian Federation, Project Director: Eugen Gheorghiu

“NANOMAGMA” - NANOstructured active MAGnetoplasmonic MAterials”, Funded under 7th FWP (Seventh Framework Programme), Contract no.: 214107 /2008, 2008-2011;

„CHARPAN” – „CHARged PArticle Nanotech” 2005-2009;

„ROBIOS”- Strengthening ROmanian Research and Training Capacities in BIOSensing and Related Areas”, 2005-2008;

„Effective Biosensing Platform for Rapid Detection of Environmental Pollutants”, NATO Reintegration Grant, Grantee: PhD Szilveszter Gaspar, Supervisor PhD. Eugen Gheorghiu, 2006-2008;

Role of membrane organization and dynamics on cell behavior and response to external stimuli; Romania -Flanders Bilateral Agreement, 2005-2006;

Novel Impedimetric Affinity Biosensors (IAB) for toxicological applications exploiting E.Coli membrane protein LamB - Romania-Flanders bilateral agreement, 2003-2005

Development of an Impedance Assay for Immunocapture - Brancusi Romania -France bilateral agreement, 2003-2005

Towards the development of a rapid portable immunoassay device for the detection of Microorganisms and Toxins in Food industry & Ecology - COBASE Grant Program 2001

http://www.biodyn.ro/cgi-bin/biodyn/biodyn.cgi?act=Main-Projects&idm=4&idsm=9&locale=en








Impedance Spectroscopy of biological membranes: modeling the epithelial tissues and the dielectric dispersion of the plasma membrane; Romania-Flanders bilateral agreement 2001-2003;

Biomonitoring quantitative evaluation of biological systems, Romanian-German Bilateral Agreement reference number RUM 003-98;

AFRAMILK -Antifraud impedimetric and ultrasonic control system to detect fraud (adulteration) of milk GRD1-2000-25801, within the 5th Framework Program of the European Commission 2001-2004 ;

Fluid Rheology – Application to biology and medicine, financed by the World Bank

Coordinator: Bucharest Politehnic University (UPB), Faculty of Energetics;











National projects ONGOING

Financing period Project title and managing/partnership status

2015-2017

2012-2016

2012-2016

ESPRim - Single-molecule detection of DNA hybridization, based on electrochemical surface plasmon resonance microscopy and magnetic tweezers, Contract No. 29/2015, ID: PN-II-RU-TE-2014-4-2363, Project Director: C. Polonschii BIOSCOPE - Electro-Plasmonics for the analysis of the dynamics of cellular processes and biomolecular interactions, Contract No. 11/2012, ID: PN II-ID-PCCE-2011-2-0075, coordinated by ICB; Project Director: E. Gheorghiu ANTIOXWIN-Exploiting the antioxidant capacity of black grapes for producing wines with high authentic quality – Contract No. 101/2012, ID: PN-II-PT-PCCA-2011-3.1-1809; Project Director: A. Vasilescu;

ACCOMPLISHED*

Financing period 2013-2015

Project title and managing/partnership status Sensitive quantitation of target microorganisms using dual electro plasmonic analysis and magnetic actuation, Contract No. 82-30.04.2013 , ID: PN-II-RU-PD-2012-3-0467, Project Director: S. David

2011 – 2014

2011-2014

2011-2013

2012-2014

Tracing proteins through food processing with biosensors (PN II-RU-TE-3-0302), Project suppporting development of Young Teams, Principal Coordinator ICB - director A. Vasilescu

SENSMOTION Sensing using the electrochemically-triggered motion of catalytic nanomotors- Contract No. 16/05.10.2011, ID: PNII-RU-TE-2011-3-0237, Project Director: Szilveszter Gaspar

A new strategy for effective assessment of DNA hybridization, based on periodical magnetic actuation, Post Doctoral Grant- director C. Polonschii

EXTRACELL- Monitoring the extracellular space with catalytic self-propelled nanomotors, ERC- like project Contract No. 9/02.07.2012, Project Director: Szilveszter Gaspar



2010-2011 Improvement of Management and Marketing Abilities within the International Centre of Biodynamics (MANMAR) – Structural Funds, Economic Competitveness Improvement, Axis 2.2.4.

2007-2010 Dielectric modeling of biological cells and heterostructures with fast and efficient algorithms for boundary integral method; Principal Project Coordinator ICB

2007-2010

DEMENTJUNCTION - Expression and function of the tight junction proteins – a study in experimental models and dementia diagnosed patients Partnership with “Victor Babes” National R&D Institute

2007-2010 BIOSADN – Development of biosensors based on nucleic acids for the evaluation and monitoring of some toxic agents with applications in bioterrorism; Partnership with Faculty of Chemistry, University of Bucharest

2007-2010

PROPETHAD – Advanced Research towards medical applications of nuclear technologies; Partnership with “Horia Hulubei” National Institute of Physics and Nuclear Engineering

* Full list is given on the institution website.



Visibility actions

Oral presentations of ICB representatives

1. Gheorghiu E, David S, Polonschii C, Bratu D, Gheorghiu M, “Biosensing Based on

Magneto-Optical Surface Plasmon Resonance”, The IVth International Conference on Analytical and Nanoanalytical Methods for Biomedical and Environmental Sciences “IC-ANMBES 2016”, Brasov, June 2016

2. Gheorghiu M, Stanica L, Polonschii C, Bratu D, Popescu O, Gheorghiu E “A new twist for optogenetics: light driven dynamics for cell based sensing”, The IVth International Conference on Analytical and Nanoanalytical Methods for Biomedical and Environmental Sciences “IC-ANMBES 2016”, Brasov, June 2016

3. Gheorghiu E, Polonschii C, “A multitiered approach for monitoring water quality”, International Conference on Sustainable Solutions for Energy and Environment, “EENVIRO 2016”, Bucharest, October 2016

4. Gheorghiu E, Polonschii C, David S, Gheorghiu M, Rosu-Hamzescu M, Bratu D, “Plasmonic based EIS: towards a sensitive analytical method gathering the virtues of both SPR and EIS”, The IVth International Conference on Analytical and Nanoanalytical Methods for Biomedical and Environmental Sciences “IC-ANMBES 2016”, Brasov, June 2016

5. Stănică L, Gheorghiu M, Gheorghiu E - Electro-optical evaluation of cellular dynamics of ChR2 expressing HEK293 cells induced by light stimulation, The 7th EMBO meeting, Mannheim, Germany, 10-13 Sept. 2016

Posters

1. Stănică L, Gheorghiu M, Gheorghiu E, Poster- Cellular sensing platform using non-excitable optogenetically modified cells, “14th EMBO Practical Course in Advanced Optical Microscopy” 06April 2016, Plymouth, England

2. Stănică L, Gheorghiu M, Gheorghiu E, Poster „New twist for optogenetics - characterization of light responsive, genetically modified cells as cellular sensors”10th FENS Forum of Neuroscience 02-7July 2016 Copenhagen, Denmark .



Emergent Applications fostered by ICB 1. Sensors and biosensors

Developing of rapid, sensitive, cost effective detection platforms to determine low

concentration of target compounds In this context, surface plasmon resonance (SPR) based

biosensors represent attractive solutions in environmental monitoring and for quality-

control, due to their sensitivity, miniaturization amenability and wide range of detectable

compounds.

Since 2006, the SPR technology is operational within ICB through several instrument

including: Biacore 3000 (Biacore AB, Sweden), SPREETA based devices (complemented by

EIS and Magneto-Optic SPR) and a recently developed bench system allowing for flexible

illumination at various wavelengths.

The analysis methods already implemented allow:

- multianalytes detection (mixture of compounds e.g. with toxicogen potential)

SPR-based techniques enable real time, label-free assessment of the interaction between

an analyte (e.g. chemicals, DNA, RNA, proteins) and a covalently immobilized specific

ligand, simultaneously with a high sensitivity in determination of concentrations and kinetic

dates, and also the possible integration with other analysis and fluidic techniques (FIA) or

with approaches like Lab-On-a Chip.

In this respect, ICB has developed cellular platforms for assessment of food quality and

cytotoxicity of water samples. Including specific electrochemical instrumentation

(impedance spectrometer, potentiostat) and a detection unit based on electrochemical

sensors, electrical components and fluidic elements to maintain the temperature at 37°C,

and applying a previously validated measurement method, multiple cell parameters can

be potentially determined.

The ICB proprietary technology based on periodic actuation supports major sensing

advantages of ICB platform versus the current state of the art

1.) Portability in a lab on a chip format;

2.) High sensitivity & specificity

3) Reduced analysis time;



The analytical platform is versatile, so it can provide relevant

information not only for water monitoring activities, but also in

chemical or pharmaceutical industry, efficiently responding to the

Romanian and European socio-economic needs regarding the

environmental quality control and / or biosensing applications in other specified domains.

2. Measurement and control systems for optimization of aquaculture activities

An issue of real interest facing both our country and other European countries is

represented by the strong demand for automated systems that will lead to optimization of

aquaculture activity through increasing productivity and reducing costs. Systems already on

the market do not optimize feeding process as a function of fish behavior, which is one of

the causes that generate many losses both in terms of quantity and quality of food (about

30% of the costs of aquaculture operations).

Taking into consideration these trends and the promising results obtained by ICB in research

and experimental studies undertaken in this direction, some of the most important

applications developed in this field are as follows:

- support system of automation and optimization of feeding

- pilot system for non-invasive , multichannel assessment of behavior (dynamics) of aquatic

species

- integrated, remote monitoring system for water (aquatic environment) quality control

able to activate a specific system based on biosensors whenever the behavior of aquatic

species deviates from a normal pattern;

- (non) linear - analysis method which provides a set of quantitative measures,

characteristic of the behavior of species with high economic value (e.g., sturgeons, carps) in

relation to actual environmental conditions (including feeding conditions).

3. Implementation of light driven dynamic processes for cell based sensing

Cell-based biosensors traditionally exploit living cells capability to alter physiologic processes

or cellular rhythms emphasized by cell cycle or exhibited by excitable cells in response to

extracellular stimuli. They are often slow responding, nonspecific and irreproducible. We

highlight for the first time the analytic potential of fast, noninvasive electrical assessment of

the discrete changes of characteristic cell response to an externally induced stimulus as

modulated by bioactive (e.g. pharmacologic or toxic) compounds. The sensing concept

highlights in 30 minutes, signature evolutions of impedance data characteristic for cellular

status and the nature of extracellular stimulus.



4. Quantitative assessment of specific carbonic anhydrase inhibitors

effect on hypoxic cells using electrical impedance assays

Carbonic anhydrase IX (CAIX) is an important orchestrator of hypoxic

tumour environment, associated with tumour progression, high

incidence of metastasis and poor response to therapy. Due to its tumour

specificity and involvement in associated pathological processes: tumorigenicity,

angiogenesis, inhibiting CA IX enzymatic activity has become a valid therapeutic option.

Dynamic cell-based biosensing platforms can complement cell-free and end-point analyses

and support the process of design and selection of potent and selective inhibitors. The

effectiveness of recently emerged CA IX inhibitors (sulfonamides and sulfocoumarins) and

their antitumour potential is assessed using electrical impedance spectroscopy (EIS)

biosensing platform. The analysis allows discriminating between the inhibitory capacities of

the compounds and their inhibition mechanisms.



Relevant publications

S. David, C. Polonschii, M. Gheorghiu, D. Bratu, E. Gheorghiu, Biosensing Based on Magneto-Optical Surface Plasmon Resonance, in MiMB series, Biosensors and Biodetection: Methods and Protocols, IInd Ed., A. Rasooly & B. Prickril Eds., Springer

Vasilescu A., Purcarea C., Popa E., Zamfir M., Mihai I., Litescu S., David S., Gaspar S.,

Gheorghiu M., Marty J-L., Versatile SPR aptasensor for detection of lysozyme dimer in oligomeric and aggregated mixtures, Biosensors & Bioelectronics, 2016, 83, 353-360.

Vasilescu A, Marty J-L, Electrochemical aptasensors for the assessment of food quality and safety, Trends in Analytical Chemistry, 2016, 79, 60-70

Andrei V, Sharpe E, Vasilescu A, Andreescu S, A single use electrochemical sensor based on biomimetic nanoceria for the detection of wine antioxidants, Talanta, 2016, 156-157, 112-118.

Mihai I, Vezeanu A, Polonschii C, Albu C, Radu G-L, Vasilescu A, Label-free detection of lysozyme in wines using an aptamer based biosensor and SPR detection, Sensors Actuators B Chemical (2015), 206, 198-204

A. Bondarenko, F. Cortes-Salazar, M. Gheorghiu, S. Gaspar, D. Momotenko, L. Stanica, A. Lesch, E. Gheorghiu, H. H. Girault, Electrochemical push-pull probe: from scanning electrochemical microscopy to multimodal altering of cell microenvironment, Anal. Chem., (2015) vol.87, p.4479–4486

S. David, C. Polonschii, C. Luculescu, M. Gheorghiu, S. Gaspar, E. Gheorghiu, Magneto-Plasmonic Biosensor with Enhanced Analytical Response and Stability, Biosens. Bioelectron. (2015), 63, 525–532

Polonschii C., David S., Gáspár S., Gheorghiu M., Rosu-Hamzescu M., Gheorghiu E., “Complementarity of EIS and SPR to Reveal Specific and Nonspecific Binding When Interrogating a Model Bioaffinity Sensor; Perspective Offered by Plasmonic Based EIS”, Anal. Chem.(2014), 86 (17), 8553–8562

Gheorghiu M., David S., Polonschii C., Olaru A., Gaspar S.,Bajenaru O., Popescu O. B, Gheorghiu E.,” Label free sensing platform for amyloid fibrils effect on living cells", Biosensors and Bioelectronics (2014) 52, 89–97

Pavel, I.-A.; Bunea, A.-I.; David, S.; Gáspár, S. Nanorods with Biocatalytically Induced Self-Electrophoresis. ChemCatChem 2014, 6, 866–872.

Bunea, A.-I.; Pavel, I.-A.; David, S.; Gáspár, S. Sensing Based on the Motion of Enzyme-Modified Nanorods. Biosensors and Bioelectronics 2014, 67, 42-48. .

Gaspar, S. Enzymatically Induced Motion at Nano- and Microscale. Nanoscale 2014, 6, 7757–7763.

Mihai, I.; Vezeanu, A.; Polonschii, C.; David, S.; Gáspár, S.; Bucur, B.; Blaszykowski, C.; Sheikh, S.; Thompson, M.; Vasilescu, A. Low-Fouling SPR Detection of Lysozyme and Its Aggregates. Anal. Methods 2014, 6, 7646–7654.

Andrei, V.; Bunea, A.-I.; Tudorache, A.; Gáspár, S.; Vasilescu, A. Simple DPPHC-Based Electrochemical Assay for the Evaluation of the Antioxidant Capacity: A Thorough Comparison with Spectrophotometric Assays and Evaluation with Real-World Samples. Electroanalysis. 2014, 26(12), 2677–2685.



Vasilescu A, Vezeanu A, Badea M, Electrochemical Impedance Spectroscopy investigations focussed on food allergens, Sensing in Electroanalysis, Vol. 8 (K. Kalcher, R. Metelka, I. Švancara, K. Vytras; Eds.), pp. 59-83.2013/2014 University Press Centre, Pardubice, Czech Republic. ISBN 978-80-7395-782-7 (print); ISBN 978-80-7395-783-4 (pdf)

Andrei V., Ngounou Wetie A.G, Mihai I., Darie C.C, Vasilescu A., Detection of Biomedically Relevant Stilbenes from Wines by Mass Spectrometry, Advances in Experimental Medicine and Biology, 2014; 806, 361-382. DOI:10.1007/978-3-319-06068-2_17

Vasilescu A, Vezeanu A., Liu Y., Hosu I.S., Worden M.R., Peteu S.F, Meat Freshness: Peroxynitrite's Oxidative Role, Its Natural Scavengers, and New Measuring Tools, in Instrumental Methods for the Analysis and Identification of Bioactive Molecules, ACS Symposium Series, vol .1185, 2014, 303-332

Vasilescu A, Mihai I, Vezeanu A, Andrei V, Nedelcu G, Popa E, Purcarea C, Subramanian P, Coffinier Y, Boukherroub R, Szunerits S, A new nanomaterial for the study of protein interactions in New applications of Nanomaterials, Series in Micro and Nanoengineering vol 22, editors: Alina Catrinel Ion, Dan Dascalu, Gabriela Carja, Magdalena Lidia Ciurea, publisher: Editura Academiei Romane, 2014, 19-32, ISBN: 978-973-27-2434-7.

Gheorghiu M., Enciu A.M., Popescu O. B, Gheorghiu E., Functional and molecular characterization of Aβ42 effect on an in vitro epithelial barrier model, Journal of Alzheimer's Disease (2014) 38, 787–798

David S., Polonschii C., Gheorghiu M., Bratu D., Dobre A., Gheorghiu E., Assessment of pathogenic bacteria using periodic actuation, Front Cover, Lab on a Chip (2013), 13, 3192–3198

Bunea A.I., Pavel I.A., David S., Gáspár S., Modification with hemeproteins increases the diffusive movement of nanorods in dilute hydrogen peroxide solutions, Chem. Comm. (2013) 49, 8803-05

Subramanian P, Alesniewski A, Kaminska I, Vlandas A, Vasilescu A., Niedziolka-Jonsson J., Pichonat E, Happy H, Boukherroub R, Szunerits S., Lysozyme detection on aptamer functionalized graphene-coated SPR interfaces, Biosens. Bioelectron. (2013) 50, 239-24,3

Gheorghiu M., David S., Polonschii C., Olaru A., Gáspár S., Bajenaru O., Popescu O. B, Gheorghiu E., Label free sensing platform for amyloid fibrils effect on living cells, Biosens. Bioelectr. (2014) 52, 89–97

Olaru A., Gheorghiu M., Polonschii C., David S. and Gheorghiu E., ”Quality assessment of SPR sensors chips; case study on L1 chips” Biosens. Bioelectr. (2013) 45C, 77-81

M. Gheorghiu, S. David, C. Polonschii, D. Bratu, E. Gheorghiu “Dynamic assessment of Amyloid oligomers – cell membrane interaction by advanced impedance spectroscopy” J. Phys.: Conf. Ser. 434 (2013) 012090

M. Gheorghiu, D. Bratu, A. Olaru, C. Polonschii, E. Gheorghiu “Revealing membrane potential by advanced impedance spectroscopy: theoretical and experimental aspects” J. Phys.: Conf. Ser. 434 (2013) 012087

Gáspár S., Marty J. L., Gheorghiu E., Cytochrome c-Based Amperometric Sensors for Superoxide Detection: Where Their Signal Comes From?, Electroanalysis (2013) 25, No. 2, 448 – 452;

Polonschii C., Bratu D., Gheorghiu E., Appraisal of fish behaviour based on time series of fish positions issued by a 3D array of ultrasound transducers, Aquacultural Eng (2013), 55, 37– 45



Vasilescu A., Gáspár S., Mihai I., Tache A. and Litescu S.C., Development of a label-free aptasensor for monitoring the self-association of lysozyme, Analyst, (2013), 138 (12), 3530 – 3537

Wang. Q, Vasilescu A, Subramanian P, Vezeanu A., Andrei V., Coffinier Y, Li M, Boukherroub R., Szunerits S., Simultaneous electrochemical detection of tryptophan and tyrosine using boron-doped diamond and diamond nanowire electrodes, Electrochem

Comm, 35 (2013) 84–87 Vasilescu A., Gáspár S., Schuhmann W, Recent progress in the electrochemical detection

of disease-related diagnostic biomarkers in " Detection challenges in clinical diagnostics", editors Vadgama P& Peteu S, publisher: RSC, 2013

A. Vasilescu., E. Sharpe, S. Andreescu “Nanoparticle-Based Technologies for the Detection of Food Antioxidants”, Current Analytical Chemistry, (2012), 8 , 495-5058

S. Gáspár, S. David, C. Polonschii, I. Marcu, M. Gheorghiu, E. Gheorghiu “Simultaneous impedimetric and amperometric interrogation of renal cells exposed to a calculus-forming salt”, Anal Chim Acta. (2012) 713: 115-20

M. Gheorghiu, C. Polonschii, S. David, A. Olaru, E. Gheorghiu “SPR Bioanalytical platform to appraise the interaction between antimicrobial peptides and lipid membranes”, In Optical Nano- and Microsystems for Bioanalytics, Series Chemo- and Biosensors 10 (Series Editor Gerald Urban), Springer (2012) pp 183-210

S. Andreescu, M. Gheorghiu, R. E. Ozel, K. Wallace Methodologies for Toxicity Monitoring and Nanotechnology Risk Assessment, chapter 7 in ACS books series "Biotechnology and

Nanotechnology Risk Assessment: Minding and Managing the Potential Threats around Us" Editor(s) Steven Ripp and Theodore B. Henry, Vol. 1079 Publication Date (Web) October 18, (2011)

S. Gáspár, "Detection of Superoxide and Hydrogen Peroxide from Living Cells Using Electrochemical Sensors" in "Oxidative Stress: Diagnostics, Prevention, and Therapy", vol. 1083, American Chemical Society, 2011, pp. 289–309

S. Peteu, S. Szuneritz, A. Vasilescu, W. Knoll "Nanoscale Architectures for Smart Bio-Interfaces: Advances and Challenges" in "Nanofabrication", editor: Yoshitake Masuda, INTECH, ISBN 978-953-307-912-7, 2011

E. Gheorghiu “Relating membrane potential to impedance spectroscopy”, Journal of Electrical Bioimpedance, (2011) 2, 93–97

T. Sandu, D. Vrinceanu, E. Gheorghiu “Surface Plasmon Resonances of Clustered Nanoparticles”, Plasmonics, (2011) 6, 2, 407–412

S. Gaspar, C. Niculite, D. Cucu, I. Marcu "Effect of calcium oxalate on renal cells as revealed by real-time measurement of extracellular oxidative burst", Biosensors and Bioelectronics (2010) 25, 1729-1734

T. Sandu, D. Vrinceanu, E. Gheorghiu “Linear dielectric response of clustered living cells” PHYS. REV. E (2010) 81, 021913 1-11

C. Polonschii, S. David, S. Tombelli, M. Mascini, M. Gheorghiu "A novel low-cost and easy to develop functionalization platform. Case study: Aptamer-based detection of thrombin by surface plasmon resonance" Talanta 80 (2010) 2157–2164

A. Olaru, M. Gheorghiu, S. David, T. Wohland , E. Gheorghiu "Assessment of the multiphase interaction between a membrane disrupting peptide and a lipid membrane", J. Phys Chem B (2009), 113, 14369

M. Gheorghiu, A. Olaru, A. Tar, C. Polonschii, E. Gheorghiu "Sensing based on assessment of non-monotonous effect determined by target analyte: Case study on pore-forming compounds", Biosens. Bioelectron. 24 (2009) 3517

E. Gheorghiu, M. Gheorghiu, S. David, C. Polonschii "Biodynsensing: sensing through dynamics of hybrid affinity / cellular platforms; towards appraisal of Environmental and



Biological Risks of Nanobiotechnology" in NATO Science for Peace and Security Series B: Physics and Biophysics , Magarshak, Yuri; Kozyrev, Sergey; Vaseashta, Ashok K. (Eds.) 2009, ISBN: 978-90-481-2522-7

L. Muresan, M. Nistor, S. Gaspar, I.C. Popescu, E. Csoregi “Multianalyte monitoring using enzyme microstructures and

Scanning Electrochemical Microscopy”, Bioelectrochemistry 2009, 76, 81-86 C. Balan, D. Broboana, E. Gheorghiu, L. Vekas “Rheological characterization of complex

fluids in electro-magnetic fields” Journal of Non-Newtonian Fluid Mechanics 154 (2008) 22–30

E. Gheorghiu “Appraisal of cellular systems using impedance spectroscopy – theoretical and experimental aspects”, IFMBE Proceedings, Springer, vol 17, 2007, part 4, 81-85

M. Gheorghiu, S. David, C. Polonschii, D. Bratu, E. Gheorghiu “Biosensing and controlled interaction with cellular systems via structured interfaces”, Eur. Cel. Mater. Vol14 , S3, 2007, p 63

M. Gheorghiu, S. David and E. Gheorghiu “Sensing the cell- substrate interaction towards development of “smart” surfaces, IFMBE Proceedings, Springer, vol 17, 2007, part 4, 86-89

C. Balut , M. Vandeven, S. Despa, I. Lambrichts, M. Ameloot, P. Steels, I. Smets “Measurement of cytosolic and mitochondrial pH in living cells during reversible metabolic inhibition”, Kidney Int. 2007, 17978815

S. David, M. Gheorghiu, C. Polonschii and E. Gheorghiu “Dual SPR-Impedance Measurement System for detection of bioaffinity interactions”, IFMBE Proceedings, Springer, vol 17, 2007, part 4, 106-109

C. Polonschii, D. Bratu and E. Gheorghiu “Multi Frequency, Multi Channel, Differential Impedance Analyzer for Rapid Assays”, IFMBE Proceedings, Springer, vol 17, 2007, part 4, 229-231

F. Jans , C. Balut, M. Ameloot , P. Wouters, P. Steels “Investigation of the Ba-Sensitive NH(4)(+) Transport Pathways in the Apical Cell Membrane of Primary Cultured Rabbit MTAL Cells”, Nephron Physiol. 2007 Jun 15;106 (3):p45-p53 17570948

K. Zor, S. Gaspar, M. Hashimoto, H. Suzuki, E. Csoregi “High temporal resolution monitoring of fermentations using an on-line amperometric flow-through microdetector”, Electroanalysis, 2007, 19(1), 43-48.

M. Hedstrom, C.E. Grey, S. Gaspar, B. Mattiasson “Miniaturized on-line digestion system for the sequential identification and characterization of protein analytes” J. Chromatography A, 2007, 1146(1), 17-22.

M. Schuvailo, S. Gaspar, A.P. Soldatkin, E. Csoregi “Ultramicrobiosensor for the selective detection of glutamate”, Electroanalysis, 2007, 19(1), 71-78

M. Gheorghiu, S. David, C. Polonschii, E. Gheorghiu “Sensing at nanoscale via structured interfaces” Eur Biophys J. (2007) 36 S157

C.M. Balut, P. Steels, M. Radu, M. Ameloot, W. Van Driessche, D. Jans "Membrane Cholesterol Extraction decreases Na+ Reabsorption in A6 Renal Epithelia", AJP - Cell, 290 (1), C87-94, 2006

annual report 2016 - biodyn.ro · implications for ecology, food industry, bio-medicine and...

Documents