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CERTIFICAT DE ACREDITARE

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Determinarea capacitatii antioxidante a produselor hidrosolubile prin metoda fluorimetrica ORAC

CERCETARE

Domeniul de Cercetare:

Preocuparile actuale ale grupului de cercetatori urmaresc elaborarea si efectuarea unor modele experimentale in vivo sau in vitro (sisteme acelulare sau celulare) care sa furnizeze informatii privind proprietatile imunomodulatorii (antitumorale si/sau antiinflamatorii) ale unor extracte naturale (in special extracte vegetale) sau compusi inalt purificati izolati din extracte naturale. Cercetarile au ca scop implicarea produselor analizate in activitati biologice cum sunt: modularea pompelor de transport (Pgp, MRP) exprimate pe diverse linii celulare normale si tumorale, inhibarea activitatii unor enzime implicate in metabolismul acidului arahidonic (inhibarea activitatii de COX-2, 5-LO, precum si inhibarea expresiei mRNA pentru COX-2), modularea unor markeri celulari (PI3K-g si PPAR-g si d, PKB/Akt, NF-kB). In mod curent, testam potentiali inhibitori ai proceselor mediate de receptorii Toll-like (TLRs) si Dectin1, membrii ai familiei Pattern Recognition Receptors (PRR), cu rol esential in raspunsul imun innascut fata de bacterii, respectiv fungi.

Pornind de la ideea interdependentei intre sistemul nervos si sistemul imun, preocuparile noastre vizeaza si inflamatia neurogenica. Ne indreptam atentia in special spre receptorii TRPV1, urmarind modularea activitatii acestor receptori; efectuam studii in vitro (utilizand diverse linii celulare) si studii in vivo (Pletismometrie computerizata, soc endotoxic, modularea producerii de anticorpi, model pentru testarea capacitatii antitumorale – melanom, studii comportamentale – Capsaicin induced Eye-Wipe Test etc).

Rezultatele cercetarilor noastre permit ca produsele analizate sa fie dirijate spre noi tinte terapeutice in bolile cu componenta inflamatorie si apoi sa fie utilizate ca ingrediente bioactive in prepararea industriala de noi medicamente de uz uman sau veterinar.

O preocupare recenta vizeaza dezvoltarea/modernizarea infrastructurii existente in Laboratorul Imunomodulare din Centrul de Studii Avansate, INCDMI Cantacuzino, in scopul integrarii, validarii si acreditarii de metodologii moderne pentru testarea eficientei medicamentelor recomandate in terapia bolilor inflamatorii acute si cronice:

• Determinarea capacitatii antioxidante a produselor hidrosolubile prin metoda fluorimetrica ORAC;
• Determinarea capacitatii de inhibare a productiei de oxid nitric in macrofage activate prin masurarea cantitatii de nitriti in supernatant de cultura utilizand metoda colorimetrica Griess;
• Determinarea capacitatii antiinflamatoare a unor produse naturale sau sintetice prin chemiluminometrie in placa – metoda rapida de screening;
• Determinarea in vivo a capacitatii antiinflamatoare a unor produse naturale sau sintetice prin pletismometrie computerizata.

Activitati stiintifice:

-       Identificarea, cuantificarea si  modularea pompelor de transport ATP-dependente (Pgp, MRP) prezente pe celule tumorale, pompe responsabile de efluxul  substantelor chimioterapice; Identificarea potentialilor inhibitori ai pompelor de eflux PgP si MRP in scopul depistarii unor potentiali agenti terapeutici pentru tratarea tumorilor rezistente la chimioterapice (citometrie in flux);

- Identificarea si monitorizarea activitatilor unor compusi cu proprietati antiinflamatoare si imunosupresoare, purificati din plante medicinale, prin aplicarea unor teste de biologie celulara si moleculara, in scopul fundamentarii actiunii lor si a largirii sferei de aplicabilitate;

- Strategii moderne de abordare a studiului patogenezei bolilor autoimune/inflamatorii prin modularea retelei de citokine pro si antiinflamatoare cu ajutorul unor substante naturale/sintetice (ELISA);

- Testarea capacitatii antitumorale (melanom) a unor compusi naturali/sintetici (studii in vivo);

- Cercetari privind rolul agonistilor si antagonistilor de E-Prostanoid in diferentierea limfocitelor B si in maturarea celulelor dendritice;

- Teste de citotoxicitate/viabilitate pe linii celulare normale si tumorale (tehnica MTT)

- Identificarea compusilor activi in modularea producerii de anticorpi in vivo;

- Testarea activitatii PI3K-g determinata prin cuantificarea nivelului de fosforilare a PKB/Akt (Westernblot);

- Modularea activitatii/activarii receptorilor TRPV1 in vitro si in vivo (studii comportamentale -Capsaicin induced Eye Wipe Test; inducerea edemului plantar prin injectarea de capsaicina);

-  Identificarea capacitatii modulatorii a unor produsi asupra:

• artritei experimentale induse cu Colagen II (pletismometrie)
• artritei cronice poliarticulare induse cu Mycobacterium tuberculosis (pletismometrie)
• socului endotoxic indus de LPS

Lista publicatiilor in perioada 2005 – 2010:

-  ”The effect of the plasma needle on tumoral cell lines apoptosis” Ana Calugaru, Lidia Cremer, Aurora Herold, Andreea Lupu, G.Szegli, C.Lungu, Ana Lungu and N.Georgescu, Roum.Arch.Microbiol.Immunol., vol.64, nr.1-4, (2005).

- „COX-2 inhibitors can down-regulate in vivo antibody response against T-dependent antigens”, Andreea-Roxana Lupu, Lidia Cremer, Steliana Durbaca, Ana Calugaru, Aurora Herold, F. Kerek, G. Szegli si D.L. Radu Roum.Arch.Microbiol.Immunol., vol.65, nr.1-2, (2006).

- “The novel arthritis drug substance MCS-18 attenuates the antibody production in vivo”, F.Kerek, G.Szegli, Lidia Cremer, Andreea-Roxana Lupu, Steliana Durbaca, Ana Calugaru, Aurora Herold, D.L. Radu, Acta Microbiologica et Immunologica Hungarica, 55 (1), 15-31, (2008).

- “Local analgesic drug “MCS-18” provides high survival rates in septic-shock mice”, F.Kerek, A.Babes, Lidia Cremer, Andreea-Roxana Lupu, C.Coman, Ana Calugaru, G.Szegli, Shock, 29 (Supplement 1), 110, (2008).

- «STUDY OF APOPTOSIS INDUCED BY CYTOSTATICS AND VEGETAL EXTRACTS ON HUMAN ENDOTHELIAL CELL LINE”, Bârzu Natalia Simona, Bădulescu Maria Mihaela, Lupu Andreea Roxana, Cremer Lidia, G.Szegli, F.Kerek, Călugăru Ana,   Roum.Arch.Microbiol.Immunol.,  Vol.67, Nr.1-2, (2008).

- „Pharmacological evaluation of total grape seed polyphenolic extract as ingredient for functional food products complying with EU regulation concerning bioactive products”, B.Burghelea, G.Caraene, R.Campeanu, C.Nichita Georgeta Radulescu,  V.Vulturescu, A.Lupu, publicata in „Proceeding of the International Symposium “NEW RESEARCH IN BIOTECHNOLOGY” USAMV, Bucuresti, Romania, 2008”.

- „Pharmacological evaluation of total grape seed polyphenolic extract as ingredient for cosmetics complying with EU regulation concerning bioactive products”, B.Burghelea, G.Caraene, R.Campeanu, C.Nichita, G.Radulescu,  V.Vulturescu, A.Lupu, publicata in „Proceeding of the International Symposium “NEW RESEARCH IN BIOTECHNOLOGY” USAMV, Bucuresti, Romania, 2008”

- „Extractul polifenolic total din semintele de struguri, ingredient antioxidant in alimentele functionale”, B.Burghelea, G.Caraene, R.Campeanu, V.Vulturescu, C.Nichita, G.Radulescu, L.Cremer, R.Albulescu, R.Porumb, A.R.Lupu, G.Szegli, revista CALITA, nr. 68/ 2008.

- „Extractul polifenolic total din semintele de struguri (TGSPE), ingredient alimentar antioxidant in produse alimentare din fructe procesate”, B.Burghelea, L.Catana, M.Catana, M.Negoita, R.Campeanu, R.Porumb, A.R.Lupu, L.Cremer, revista CALITA, nr. 68/ 2008.

- „Extractul polifenolic total din semintele de struguri (TGSPE), ingredient alimentar antioxidant in produse alimentare de patiserie si brutarie”, B.Burghelea, L.Catana, M..Catana, M.Negoita, R.Campeanu, R.Porumb, A.R.Lupu, L.Cremer, revista CALITA, nr. 68/ 2008.

- “The modulation of REACTIVE oxygen SPECIES production from human polymorphonuclear cells by CURDLAN DERIVATES AS Dectin–1 agonists/antagonists”, Maria-Mihaela Bădulescu, Andreea-Roxana Lupu, Lidia Cremer, Ana Călugăru, Natalia Simona Apetrei, M. Moscovici, Georgeta Mocanu, G. Szegli, Roum.Arch.Microbiol.Immunol.,  Vol.68, Nr.2, pag.63-68, (2009).

- « THE EFFECTS OF COLD ATMOSPHERIC PLASMA JETS ON b16 AND coLO320 TUMORAL CELLS”, Andreea-Roxana Lupu, N.Georgescu, Ana Calugaru, Lidia Cremer, G.Szegli, F.Kerek, Roum.Arch.Microbiol.Immunol., Vol.68, Nr.3, (2009)

- « Recognition and MODULATION of Dectin-1 and TLR-2 receptors by Curdlan DERIVATIVES and purified natural extracts”, Ana Calugaru, Lidia Cremer, Andreea Roxana Lupu, Mihaela Maria Badulescu, Natalia Simona Apetrei, M.Moscovici, Georgeta Mocanu, Doina Mihai, F.Kerek, G.Szegli, Roum.Arch.Microbiol.Immunol., Vol.68, Nr.3, pag.22-27, (2009)

- „CURDLAN DERIVATIVES ABLE TO ENHANCE CYTOSTATIC DRUGS ACTIVITY ON TUMORAL CELLS”, Maria-Mihaela Badulescu, Natalia Simona Apetrei, Andreea-Roxana Lupu, Lidia Cremer, G.Szegli, M.Moscovici, Georgeta Mocanu, Doina Mihai, Ana Calugaru, Roum.Arch.Microbiol.Immunol., Vol.68, Nr.4, pag.11-16, (2009)

- „Tendinte terapeutice in sepsis”, Andreea-Roxana Lupu, C.Ciufecu, G.Szegli, Viata medicala, Nr.39 (1029), 2009

Lista lucrarilor comunicate in 2009-2010 :

- „Analgesic and anti-inflammatory compound MCS -18 provides high survival rates in septic-shock mice”, Andreea-Roxana Lupu,  Lidia Cremer,  F Kerek, Ana Calugaru, C Coman, A Babes, G Szegli, Sepsis 2008, Granada, Spania, 19-22 Nov.2008 si Sesiunea de Comunicari Stiintifice a INCDMI Cantacuzino, ianuarie 2009.

- „MCS-18 and d161 – TLR ANTAGONISTS OR NEGATIVE REGULATORS OF TLR INTRACELLULAR SIGNALING?”, Andreea-Roxana Lupu, Lidia Cremer, Franz Kerek, Ana Calugaru, Maria-Mihaela Badulescu, Maria-Luisa Flonta, Geza Szegli, TRIPR Conference „Innate immunity and the pathogenesis of rheumatic diseases”, 6-8 May 2009, Genoa, Italia.

- „THE MODULATION OF IL-10 AND TNF-alpha RELEASED BY ACTIVATED HUMAN PBMC IN THE PRESENCE OF NATURAL AND DERIVED BETA-GLUCANS”, Natalia Simona Apetrei, Lidia Cremer, Mihaela Maria Bădulescu, Andreea-Roxana Lupu, Ana Calugaru, Misu Moscovici, Geza Szegli, TRIPR Conference „Innate immunity and the pathogenesis of rheumatic diseases”, 6-8 May 2009, Genoa, Italia.

- „The modulation of free oxygen radicals production from human polymorphonuclear cells by Dectin–1 and TLRs agonists / antagonists”, Maria-Mihaela Bădulescu, Andreea-Roxana Lupu, Lidia Cremer, Ana Călugăru, Natalia Simona Bârzu, Misu Moscovici, Geza Szegli, TRIPR Conference „Innate immunity and the pathogenesis of rheumatic diseases”, 6-8 May 2009, Genoa, Italia.

- “Novel antirheumatic drug “MCS-18” involved in TLR signaling”, Andreea-Roxana Lupu, Lidia Cremer, Geza Szegli, Ana Calugaru, Maria-Mihaela Badulescu, Natalia-Simona Barzu, Maria-Luisa Flonta, Franz Kerek, The 2^nd European Congress of Immunology, Sept. 2009, Berlin, Germany

- “Recognition and control of Dectin-1 receptors and TLR-2 by Curdlan–like biopolymers and purified natural extracts”, Ana Calugaru, Lidia Cremer, Andreea-Roxana Lupu, Maria-Mihaela Badulescu, Natalia-Simona Barzu, Franz Kerek, Geza Szegli, The 2^nd European Congress of Immunology, Sept. 2009, Berlin, Germany

- “The role of natural compound D161 in neurogenic inflammation”, Andreea-Roxana Lupu, Lidia Cremer, F.Kerek, Ana Calugaru, Natalia-Simona Barzu, C.Coman, Maria-Luiza Flonta, G.Szegli, Pain in Europe VI, 6^th Congress of the European Federation of IASP^®, Chapters (EFIC), Sept.2009, Lisbon, Portugal

- “New natural compound MCS-18, a TLR-2 antagonist able to down-regulate inflammation-related pain” Andreea-Roxana Lupu, Lidia Cremer, F.Kerek, Ana Calugaru, Maria-Mihaela Badulescu, Maria-Luiza Flonta, G.Szegli, Sept.2009, Pain in Europe VI, 6^th Congress of the European Federation of IASP^®, Chapters (EFIC), Sept.2009, Lisbon, Portugal

-„Analgesic and anti-inflammatory compound MCS -18 provides high survival rates in septic-shock mice”, Andreea-Roxana Lupu,  Lidia Cremer,  F.Kerek, Ana Calugaru, C.Coman, A.Babes, G.Szegli, Scientific Communications Symposium of NIRDMI Cantacuzino, January 2009, Bucharest.

- “Effect of Curdlan type biopolymers and some natural extracts on Dectin-1 and TLRs”, Ana Calugaru, Lidia Cremer, Andreea-Roxana Lupu, Maria-Mihaela Badulescu, Natalia-Simona Apetrei, Franz Kerek, Geza Szegli, The 39^th National Conference of Immunology, Sept. 2009, Calimanesti

- „THE CAPACITY OF SOME β-GLUCAN BIOPOLYMERS TO MODULATE REACTIVE OXYGEN SPECIES PRODUCTION FROM HUMAN POLYMORPHONUCLEAR CELLS”, Maria-Mihaela Badulescu, Andreea-Roxana Lupu, Lidia Cremer, Ana Calugaru, Natalia Simona Barzu, Misu Moscovici, Georgeta Mocanu, Geza Szegli, The 39^th National Conference of Immunology, Sept. 2009, Calimanesti

-  „COLD ATMOSPHERIC PLASMA JETS – a new medical device acting on tumoral cells”, Andreea-Roxana  Lupu,  N.Georgescu,   Ana  Calugaru,  Lidia  Cremer, G.Szegli,  F.Kerek, The 39^th National Conference of Immunology, Sept. 2009, Calimanesti

- „MCS-Ab162 and D161 – possible therapeutic agents in neuroimmune diseases”, Andreea-Roxana Lupu, Lidia Cremer, Franz Kerek, Ana Calugaru, Coman Cristin, Vlase Ene, Maria-Mihaela, Badulescu, Natalia Simona Apetrei, Maria-Luisa Flonta, Geza Szegli, 5^th „Gheorghe Marinescu” Symposium of the National Neuroscience Society of Romania, Oct. 2009, Bucharest

- CARACTERIZAREA UNOR PIGMENTI MICROBIENI PRODUSI DE MICROORGANISME DIN CLASA ASCOMYCES – POSIBILE MATERII PRIME PENTRU OBTINEREA NANOMATERIALELOR CU APLICATII IN TERAPIE, Nicoleta Radu, Cosmin Corobea, Caloian Florentin, Andrea Lupu, Valentin Raditoiu, Sanda Doncea, Irina Dumitriu, Radu Fierascu, A 8-a Editie a Seminarului National de Nanostiinte si Nanotehnologie, 27 aprilie 2009, Bucuresti

- Inhibition of induced mouse B16 melanoma by copper microspherical particles, Ana Calugaru, Cristina Surdu Bob, Lidia Cremer, M. Badulescu, C.Coman, The 40^th National Conference of Immunology, 28 Aprilie -1 Mai  2010, Sibiu

- ENHANCEMENT OF SOME CYTOSTATIC DRUGS ACTIVITY ON TUMOR CELL LINES USING CURDLAN DERIVATIVES, Mihaela Bădulescu¹, Natalia Simona Apetrei¹, Lidia Cremer, Andreea Lupu, M. Moscovici, Georgeta Mocanu, Doina Mihai, G. Szegli, Ana Călugăru, The 40^th National Conference of Immunology, 28 Aprilie -1 Mai  2010, Sibiu

- Studies regarding the efficacy of “Natural SOD” and its fractions on PMN cells of patients with rheumatic diseases, Andreea-Roxana Lupu, Lidia Cremer, Aurora Herold, Maria Cristea, Maria-Mihaela Badulescu, Natalia-Simona Barzu, Ana Calugaru, G. Szegli, TRIPR Conference „Biological Agents and Emerging Treatments in the Management of Rheumatic Diseases”, 27-30 May 2010, Genoa, Italia.

I.N.C.D.M.I. Cantacuzino

Splaiul Independentei 103,
Sector 5, 050096 Bucuresti
Tel: 021-3069298,  021-3069251
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