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Ippazio Antonio RUBERTO

Personale tecnico amministrativo
Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze sede ex-Sc. Biomediche
Referente informatico
Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze sede ex-Sc. Biomediche

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Pubblicazioni

2015 - NDP-α-MSH attenuates heart and liver responses to myocardial reperfusion via the vagus nerve and JAK/ERK/STAT signaling [Articolo su rivista]
Ottani, Alessandra; Giuliani, Daniela; Neri, Laura; Calevro, Anita; Canalini, Fabrizio; Vandini, Eleonora; Cainazzo, Maria Michela; Ruberto, Ippazio Antonio; Barbieri, Alberto; Rossi, Rosario; Guarini, Salvatore
abstract

Melanocortin peptides afford cardioprotection during myocardial ischemia/reperfusion via janus kinases (JAK), extracellular signal-regulated kinases (ERK) and signal transducers/activators of transcription (STAT) pathways. Here we investigated whether melanocortin-induced modulation of the JAK/ERK/STAT signaling occurs via the cholinergic anti-inflammatory pathway, focusing our study on cardiac and hepatic responses to prolonged myocardial ischemia/reperfusion. Ischemia was produced in rats by ligature of the left anterior descending coronary artery for 30min; effects of ischemia/reperfusion were evaluated using Western blot of heart and liver proteins. Intravenous treatment, during coronary artery occlusion, with the melanocortin analog (Nle(4), D-Phe(7))α-melanocyte-stimulating hormone (NDP-α-MSH) induced a left ventricle up-regulation of the cardioprotective transcription factors pJAK2, pERK1/2 and pTyr-STAT3 (JAK-dependent), and a reduction in the levels of the inflammatory mediators tumor necrosis factor-α (TNF-α) and pJNK (a transcription factor also involved in apoptosis), as assessed at the end of the 2-h reperfusion period. Further, these beneficial effects of NDP-α-MSH were associated with heart over-expression of the pro-survival proteins heme oxygenase-1 (HO-1) and Bcl-XL, and decrease of ventricular arrhythmias and infarct size. In the liver NDP-α-MSH induced a decrease in the pJAK2 and pTyr-STAT3 levels, and strongly reduced pERK1/2 expression. In the liver of ischemic rats NDP-α-MSH also blunted pJNK activity and TNF-α expression, and up-regulated Bcl-XL. Bilateral cervical vagotomy prevented all effects of NDP-α-MSH, both in the heart and liver. These results indicate that melanocortins inhibit heart and liver damage triggered by prolonged myocardial ischemia/reperfusion likely, as main mechanism, via the vagus nerve-mediated modulation of the JAK/STAT/ERK signaling pathways.

2008 - Mepacrine antagonises tumour cell growth induced by natural polyamines. [Articolo su rivista]
Rossi, Tiziana; Coppi, A; Bruni, E; Ruberto, Ippazio Antonio; Giudice, Stefania; Baggio, Giosuè Gabriele
abstract

BACKGROUND: Mepacrine is an antiproliferative agent, characterised by an aliphatic chain similar to that of natural polyamines whose activation is closely associated with cell proliferation and may lead to malignant transformation and neurodegenerative diseases. This study aims to investigate a possible antagonism between mepacrine and polyamines in tumour proliferation. MATERIALS AND METHODS: MCF-7 and Vero cells were cultured in Eagle's minimum essential medium and then subjected to graded concentrations of putrescine, spermine and spermidine alone and in combination with mepacrine. Methyl thiazole tetrazolium test and Western-blotting were performed. RESULTS: Putrescine and spermidine at 0.5 mg/l significantly stimulated cell growth, whereas mepacrine treatment confirmed the enhanced p21 expression previously reported by a recent study and growth inhibition. When used in combination, mepacrine antagonized MCF-7 growth induced by polyamines. CONCLUSION: Our results suggest that mepacrine may represent a choice in the treatment of tumours induced by the modified concentration of polyamines.

2007 - Effects of Anti-malarial Drugs on MCF-7 and Vero Cell replication. [Articolo su rivista]
Rossi, Tiziana; Coppi, Andrea; Bruni, Elisa; Ruberto, Ippazio Antonio; Santachiara, Saverio; Baggio, Giosuè Gabriele
abstract

Previous in vivo studies performed in our laboratories demonstrated that anti-malarial drugs may or enhance or slow down Ehrlichs ascites tumour progression in infected mice. In the light of these observations, an in vitro study was undertaken to assess the response of human tumour cells to various anti-malarial drugs and consequently the safety of the anti-malarial therapy. Materials and Methods: MCF-7 cells and Vero cells (control line) were cultured in Eagles minimum Essential medium (EMEM) and then subjected to graded concentrations of different anti-malarial drugs. Trypan-blue exclusion, MTT and Western blotting tests were performed. Results: The findings showed that pyrimethamine (12.5 mg/L), chloroquine (12.5 mg/L) and primaquine (1.56 mg/L) stimulated MCF-7 cell growth. The proliferative effect was inhibited by doxorubicin only in cultures treated with chloroquine and primaquine. These results might indicate that some anti-malarial drugs have a worrying tumour-promoting effect which should not be underestimated when undertaking anti-malarial prophylactic measures.

2006 - Antibacterial agent discovery using thymidylate synthase biolibrary screening [Articolo su rivista]
Costi, Maria Paola; A., Gelain; D., Barlocco; S., Ghelli; Soragni, Fabrizia; F., Reniero; Rossi, Tiziana; Ruberto, Ippazio Antonio; C., Guillou; A., Cavazzuti; Casolari, Chiara; Ferrari, Stefania
abstract

Thymidylate synthase (TS, ThyA) catalyzes the reductive methylation of 2'-deoxyuridine 5'-monophosphate to 2'-deoxythymidine 5'-monophosphate, an essential precursor for DNA synthesis. A specific inhibition of this enzyme induces bacterial cell death. As a second round lead optimization design, new 1,2-naphthalein derivatives have been synthesized and tested against a TS-based biolibrary, including human thymidylate synthase (hTS). Docking studies have been performed to rationalize the experimentally observed affinity profiles of 1,2-naphthalein compounds toward Lactobacillus casei TS and hTS. The best TS inhibitors have been tested against a number of clinical isolates of Gram-positive-resistant bacterial strains. Compound 3,3-bis(3,5-dibromo-4-hydroxyphenyl)-1H, 3H-naphtho[1,2-c]furan-1-one(5) showed significant antibacterial activity, no in vitro toxicity, and dose-response effects against Staphylococcus epidermidis (MIC = 0.5-2.5 mu g/mL) clinical isolate strains, which are resistant to at least 17 of the best known antibacterial agents, including vancomycin. So far this compound can be regarded as a leading antibacterial agent.

2006 - Valutazione tossicologica di due nuovi inibitori della DHFR del P. falciparum e del P. vivax [Relazione in Atti di Convegno]
Rastelli, Giulio; Baggio, Giosuè Gabriele; Ulivi, P.; Ruberto, Ippazio Antonio; Rossi, Tiziana
abstract

Secondo una segnalazione del 2004 dell'UNiCEF "La malaria uccide più di un milionedi persone all'anno". Gli antifolato sono una classe di antimalarici cheagiscono sulla via biosintetica del folato dei vari Plasmodi inibendo gli enzimi in essacoinvolti. Sono farmaci estremamente sfruttati e, per questo motivo spesso inefficaci acausa dell'insorgenza di ceppi di Plasmodium divenuti resistenti. Oltre a ciò, nostri studirecenti in vitro hanno evidenziato la capacità di alcuni antimalarici di interferire con lareplicazione di cellule sane e cellule tumorali. Per lapresenza di gravi effetti collaterali e di ceppi resistenti, diviene sempre piiì urgente lanecessità di disporre di nuovi farmaci antimalarici. Nel presente studio sono state utilizzatedue molecole di nuova sintesi non analoghe del folato, ma in grado di inibire la DHFR(diidrofolato reduttasi) del Plasmodium falciparum e del Plasmodium vivax con una IC50compresa nel range micromolare e sub-micromolare. I composti, siglati come 1B e 6G sonorispettivamente una molecola N-idrossiamidina simile e un derivato della tiourea e sonostati saggiati in modo comparativo con l'antifolato pirimetamina nei confronti di celluleVERO e cellule MCF-7 (tumore mammario). Le cellule sono state coltivate sterilmente inpiastre con terreno EMEM arricchito. Al raggiungimento della semiconfluenza, sono stateaggiunte alle colture concentrazioni crescenti di 1B, 6G e pirimetamina (1,56-3,125-6,25-12,5-25-50 mg/l). Dopo 48 ore di incubazione sono stati eseguiti test: MTT e WesternBlotting. I risultati preliminari indicano che il composto 1B non interferisce in alcun modosulla crescita delle cellule sane o tumorali, mentre il derivato tioureico (6G) esercita unsignificativo stimolo sulla crescita delle cellule VERO (+ 60%), ma non sulle MCF-7. Ilfarmaco di riferimento pirimetamina stimola in modo dose dipendente la replicazione deileMCF-7. L'espressione delle proteine p53 e p21 immodificata nelle colture di MCF-7trattate con le due nuove molecole ne conferma l'assenza di tossicità. Questi risultatipreliminari, associati alla conferma che le due molecole sono attive specialmente verso iceppi di Plasmodium multiresistenti sono incoraggianti per il proseguimento dello studio dinuovi strumenti terapeutici caratterizzati anche da scarsa citotossicità.

2005 - Effects of glycyrrhizin on UVB-irradiated melanoma cells [Articolo su rivista]
Rossi, Tiziana; Benassi, Luisa; Magnoni, Cristina; Ruberto, Ippazio Antonio; A., Coppi; Baggio, Giosuè Gabriele
abstract

It is known that liquorice root is rich in compounds which exert several pharmacological actions. In the present study, we evaluated the effect of glycyrrhizin (the main constituent of liquorice root) and of its metabolite aglycone, 18 beta-glycyrrhetinic acid, on UVB-irradiated human melanoma cells: SKMEL-2 from metastatic tissue and SKMEL-28 from primary malignant melanoma. Tests performed (Trypan blue exclusion test, MTT and Western blot) showed that glycyrrhizin is not toxic for both types of cells. In SKMEL-28 cells, Bcl-2 expression was low after UVB irradiation, but it was increased when treated with glycyrrhizin. On the contrary, in the SKMEL-2 cell culture, Bcl-2 expression was not modified by the substances under study. The results show that g cyrrhizin treatment might offer protection from the damage induced in humans by UVB radiation, while it seems to be ineffective on metastatic cells. Further studies must be performed to understand the mechanism of the protective effect.

2005 - FLAVONOIDS AND ANTIMYCOTICS: A POSSIBILE SYNERGISTIC COMBINATION FOR THE TREATMENT OF CANDIDIASIS AND CRYPTOCOCCOSIS. [Relazione in Atti di Convegno]
Coppi, A.; Baggio, Giosuè Gabriele; Ruberto, Ippazio Antonio; Santachiara, S.; Casolari, Chiara; Rossi, Tiziana
abstract

The treatment of candidiasis and cryptococcosis is based on the use of antimycotic drugs such as amphotericin B, 5-fluorocytosine, miconazole and fluconazole. Isoliquiritigenin is a member of the flavonoids and data in scientific literature assert that these compounds can modulate the biosynthesis of endogenous sterols, which are known to be very important for the survival of mycete. In this study we investigated the antifungal effect of isoliquiritigenin on strains of Candidae spp and C.neoformans cultured on Sabouraud medium. Amphotericin B, 5-fluorocytosine, miconazole and fluconazole were chosen as reference drugs. Sensitivity tests were performed in according with the NCCLS method for agar dilution and MIC values were calculated for all the substances. No antifungal activity was evident in cultures treated with isoliquiritigenin (MIC values >100mg/L) while, very interesting results came from the combination: isoliquiritigenin/ antifungal drugs used at inhibitory and sub-inhibitory concentrations on yeast cultures (isoliquiritigenin 50-10mg/L, amphotericin B 10-0.1mg/L, 5-fluorocytosine 2.5-0.5mg/L, miconazole 0.25-0.05, fluconazole10-0.1mg/L). The FIC index used to estimate the interactions outcomes was also calculated. The association of isoliquiritigenin with the antimycotic often resulted in a synergic effect with addition (0.55<FIC index<1). In few cases we observed indifference (1<FIC index<2). Antagonism never appeared. Finally, we performed the MTT test on Vero cells to evaluate the toxic effect of the dosages used in the combinations. The results from the MTT test performed on Vero coltures added with the drugs used alone or in association showed the low toxicity of the combinations.

2005 - Interazione in vitro tra isoliquiritigenina e farmaci antifungini su ceppi di Candida spp e C. neoformans [Relazione in Atti di Convegno]
Casolari, Chiara; Rossi, Tiziana; A., Coppi; Ruberto, Ippazio Antonio; Fabio, Giuliana; Baggio, Giosuè Gabriele
abstract

Il trattamento delle rnicosi sistemiche quali candidosi e criptococcosi si basa sull'impiego di antibiotici comeI'amfotericina B o di agenti di sintesi come gli azoli e la flucitosina. L'effetto tossico e l'emergenza diresistenze possono vanificare il successo terapeutico e determinare ricadute negative per il paziente. Daqueste osservazioni scaturisce l'importanza di individuare nuove sostanze ad azione antirnicotica caratterizzateda una buona selettività d'azione oltre che da bassa tossicità. L'isoliquiritigenina, flavonoide ricavabiledalla radice di Glycyrrhiza glabra, presenta la capacità di inibire gli enzirni deputati alla sintesi deglisteroli endogeni. Considerando che la membrana cellulare dei rniceti è ricca di ergosterolo, è stata studiata in vitro l'attività di isoliquiritigenina , singolarmente e in associazione con arnfotericina B e miconazolo,su 16 ceppi di Candida spp e C. neofonnans d'isolamento clinico. I test di sensibilità sono stati eseguiti colmetodo delle diluizioni in agar secondo le indicazioni NCCLS, calcolando le MIC per amfotericina B, miconazoloe isoliquiritigenina. Non è stata riscontrata alcuna azione antirnicotica da parte del flavonoide(MIC>lOOmg/L), mentre sono emersi dati interessanti dalle associazioni isoliquiritigenina /amfotericina Be isoliquiritigenina/ rniconazolo utilizzati a dosi inibenti e sub-inibenti (arnfotericina B e miconazolo: 0.1-10 mg/L, isoliquiritigenina: 10-50 mg/L). L'efficacia delle associazioni e stata valutata calcolando l'indiceFIC. Ne è risultato che isoliquiritigenina potenzia l'effetto deil'antimicotico denotando prevalentemente sinergismocon additività (0.55< FIC index < 1), in particolare su C.neoformans. Questi risultati preliminari,pur evidenziando una mancata attività antimicotica del flavonoide se utilizzato singolamente, suggerisconol'ipotetico vantaggio di una terapia combinata con isoliquiritigenina e antifungini a più basse concentrazioni,sfruttando l'effetto sinergico delle molecole per indurre minori effetti collaterali. Sarà interessantevalutare l'effetto di tali associazioni anche su ceppi che presentano caratteri di farmaco-resistenza.

2004 - Farmaci antimalarici e progressione tumorale: Studio “in vitro” su cellule di adenocarcinoma mammario MCF-7 [Articolo su rivista]
Rossi, Tiziana; A., Coppi; Ruberto, Ippazio Antonio; Baggio, Giosuè Gabriele
abstract

Il talco P un jhsilicato estratto hiie rocce ed usato sia per la cura personale, sia per scopi industriali. Ogni giornoun numero ekvato di hvoratmi inala polvere di talco esponendosi ad un concreto rischio per la propria salute, specie se nella polvm sono presenti consistenti trarce di minerali inquinanti come ad esempio làmianto. La presente ricerca riporta cenni di mineralogiadel talco, d e hs ua u t i l ~ i o nicnd ustriale ed una raccolta di i n f 0 m i o n i m lle più comunipatologie &vanti dd una esposizioneoccasionale, acaccidentale o lavorativa. I h f t a t i raccolti evùhziano che la tossicità del talco inalato t indipmdcnte dal tempodi esposizione e dal sesso, mentre t strettamente correlata a h presenza di contaminanti. La pneumoconiosi P la patologh più fiquentmateriscontm~m, entm il cancro polmonare pub derivare solo dalla presenza di fibre di amianto. La maggior parte deiiemalattie t causata da esposizione occupazionale, tuttavia anche lirbuso di talco cosmetico pub siciare in patologe polmonari.Nonostante le severe regolamentazioni in materia di sicurezza sul lavoro, si riscontmno ancora casi di talcosi che potrebbero diminuiresolo con la conoscenza del rischio e I'applicazione delle vigenti normative.

2004 - In vitro interaction of Saquinavir and Azidovudine with some antimycotic drugs on human pathogen yeast [Relazione in Atti di Convegno]
Casolari, Chiara; Rossi, Tiziana; Baggio, Giosuè Gabriele; Fabio, Giuliana; Ruberto, Ippazio Antonio; C., Farina; Castelli, Mario
abstract

Candidiasis and eryptococcosis are the most common fwigal diseases among patientssuffixhg from HIV infection. Amphotericin B, 5-fiuorocytosine and imidazole derivatives are thedrugs prescribed in such cases. The aim of the present work is to assess whether thecontemporaneous treatment of HIV-positive patients with combined therapies including reversetnmscriptase and proteinase inhiiitors could cause an interaction able to modify the Uierapeutic&ect of antirnycotic agents. Therefore an in vifro study to evaluate the antifimgal effect of antiretroviraiand antimycotic h g s association on yeasts growth was conducted. Methods: The strainsused for this study comprisai C.albicanr ( ~ 1 6a)n d C.neofonnam (n=16).Aii isolates were fkomclinicai specimens from HIV-seropositive subjects. Susceptibility tests of yeasts to amphotericin B,5-fluroqtosine, miwnazole and fluconazole, singuiarly and in association with saquinavir andazidovudine, were performeci according to the NCCLS method for agar dilution. The MIC dueswere obtained by applying the method of increasing scalar concentrations. To esthate theassociation inhibitory effect on yeasts growth, saquinavir and azidovudine were testkd respectively ata concentration of 18.75 pM.5 and 28 CLM/L, while the antimycotic agents were tested at subinhibitoryconcentrations. The FIC index used to estimate the interactions outcomes was alsocalculated. Results: The i n t d o n of saquinavir with al1 the antimycotic drugs caused on yeastsgrowth a synergy effect with potentiation (FIC indexc0.55) , addition (0.55<FIC index<l) andindifference effect (l<FIC index~2), w hereas azidovudine - antirnycotics association alwaysshowed an indifference effect. Antagonism was never obtained. Conclusion: The lack of antagonismindicates that the anti-retrovisal and antimycotic drugs association can be administrated withoutreducing the therapeutic antiibgai effects. The synergy shows that combined therapy could becaxried out using lower doses of antimycotic drugs, with a consequent reduction of toxic side effects.

2004 - Inhibition of the putrescine-mediated cell growth by the antimalarial mepacrine. [Relazione in Atti di Convegno]
Coppi, A.; Rossi, Tiziana; Zandomeneghi, G.; Ruberto, Ippazio Antonio; Baggio, Giosuè Gabriele
abstract

A problem strictly related to the spreading of malaria, is immunocompromission whichmakes patients more vulnerable to bacterial and viral infections and to other illness suchas tumour. We recently demonstrated that some antimalarials can modify the growth ofnormal and tumoural cells (1). Mepacrine, was found to be a potent inhibitor of VERO andMCF-7 cell growth. Like chloropromazine, a drug used in the treatment of prion disease,mepacrine possesses an aliphatic chain very similar to that of putrescine, a naturalpolyamine whose concentration in biological tissues is crucial for cells, but in particular forprions proliferation. Aim of our study was to investigate if mepacrine could antagonize theproliferative effect of putrescine. VERO and MCF-7 cells were cultured in MEM medium andtreated with increasing concentrations of putrescine, mepacrine and their association.MTT test was performed. Results show that putrescine significantly stimulated the growthof VERO and MCF-7 cells, whereas mepacrine confirmed to be a potent inhibitor. Whenused in association, sub-inhibitory concentrations of mepacrine antagonized the proliferativeeffect of putrescine on both cell cultures (inhibition >60%). The presence of thealiphatic chain in mepacrine, and putrescine, could explain this interaction. Finally, sinceit has been postulated by Prousiner (2) that the variation in the concentration ofpolyamines stimulates the proliferation of prions, on the basis of these preliminary data,we can assume that, similarly to chlorpromazine, mepacrine could be useful in thetreatment of prion diseases.

2004 - Interaction between saquinavir and antimycotic drugs on C-albicans and C-neoformans strains [Articolo su rivista]
Casolari, Chiara; Rossi, Tiziana; Baggio, Giosuè Gabriele; A., Coppi; G., Zandomeneghi; Ruberto, Ippazio Antonio; C., Farina; Fabio, Giuliana; A., Zanca; Castelli, Mario
abstract

Candidiasis and cryptococcosis are the most common fungal diseases among patients suffering from HIV infection. In the present work we assess whether the combined therapies, proteinase inhibitors and antimycotic drugs, could modify the therapeutic effect of antimycotics. An in vitro study to evaluate the antifungal effect of saquinavir and antimycotic drugs combination on yeast growth was performed. Strains of C. albicans and C. neoformans from HIV-seropositive patients were used. Susceptibility tests of yeasts to amphotericin B, 5-fluorocytosine, miconazole and fluconazole, singly and in combination with saquinavir, were performed in two different media. In the combinations the antimycotic agents and saquinavir were tested at sub-inhibitory concentrations: 0.1-10 mug ml(-1) and 12.50 mug ml(-1), respectively. The fractionary inhibitory concentration (FIC) index was also calculated. The results show that the interaction between saquinavir and all the antimycotic drugs never resulted in antagonism. Fluconazole acts in more synergistic way, no matter which medium is used. The combined therapy miconazole/saquinavir results in synergism, especially in Sabouraud. The total absence of antagonism and the presence of synergism suggest that a combined therapy could be proposed in the treatment of HIV-seropositive patients to reduce side effects, thanks to the use of lower doses of antimycotic drugs.

2004 - “In Vitro” investigation on the effects of some antimalarial drugs on MCF-7 and VERO cell replication. [Abstract in Rivista]
Rossi, T.; Coppi, Antonella; Zandomeneghi, G.; Lodi, S.; Ruberto, A; Baggio, G.
abstract

Mepacrine is no longer used as anti-malaria drug but rather as an antitumour drug. A previous in vivo study of ours showed that Primaquine speed up the development of Ehrlich ascites tumour and rapidly lead to the death of the infected rats. In the light of these observations, an in vitro study was undertaken to assess the response of human tumour cells to various anti-malarial drugs and consequently the safety of the anti-malarial therapy. Materials and Methods: MCF-7 cells and Vero cells (control line) were cultured in Eagle’s minimum Essential medium (EMEM) and then subjected to graded concentrations of different anti-malarial drugs. Trypan-blue exclusion, MTT and Western blotting tests were performed. Results: The findings showed that pyrimethamine (12.5 mg/L), chloroquine (12.5 mg/L) and primaquine (1.56 mg/L) stimulated MCF-7 cell growth. The proliferative effect was inhibited by doxorubicin only in cultures treated with chloroquine and primaquine. Results indicate that the combination with doxorubicine reduces the stimulating effects of chloroquine, while the combination doxorubicine/pirimethamine significantly improves MCF7 cell growth. We concluded that some anti-malarial drugs have a worrying tumour-promoting effect which should not be underestimated when undertaking anti-malarial prophylactic measures.

1992 - Effect of 18 B glycyrrhetinic acid on systolic blood pressure in normotensive rats [Articolo su rivista]
Rossi, T.; Vampa, G.; Benvenuti, S.; Ruberto, A.; Baggio, G.; Castelli, M.
abstract