L’insorgenza di fenomeni coinvolti nello sviluppo della farmacoresistenza costituisce al momento la principale causa di mancata risposta al trattamento chemioterapico nell’osteosarcoma. Questo è in parte dovuto ad una sovraespressione di diversi trasportatori ABC nelle cellule tumorali che causano un aumento dell’efflusso extracellulare del chemioterapico e pertanto una ridotta risposta al trattamento farmacologico. L'oncogene C-MYC è coinvolto nella resistenza al metothrexate, alla doxorubicina e al cisplatino ed è un fattore prognostico avverso, se sovraespresso al momento della diagnosi, in pazienti affetti da osteosarcoma. C-MYC è in grado di regolare l'espressione di diversi trasportatori ABC, probabilmente coinvolti nella resistenza ai farmaci nell’osteosarcoma, e questo potrebbe spiegare l’impatto prognostico avverso dell’oncogene in questo tumore. L’espressione genica di C-MYC e di 16 trasportatori ABC, regolati da C-MYC e / o responsabili dell'efflusso di diversi chemioterapici, è stata valutata su due diverse casistiche cliniche e su un pannello di linee cellulari di osteosarcoma umano mediante real-time PCR. L'espressione della proteina è stata valutata per i 9 trasportatori ABC risultati più rilevanti.Infine l'efficacia in vitro di un inibitore, specifico per ABCB1 e ABCC1, è stata valutata su linee cellulari di osteosarcoma. ABCB1 e ABCC1 sono i trasportatori più espressi nelle linee cellulari di osteosarcoma. ABCB1 è sovraespresso al momento della diagnosi in circa il 40-45% dei pazienti affetti da osteosarcoma e si conferma essere un fattore prognostico avverso se sovraespresso al momento della diagnosi. Pertanto ABCB1 diventa il bersaglio di elezione per lo sviluppo di strategie terapeutiche alternative, nel trattamento dell’osteosarcoma, atte al superamento della farmacoresistenza. L’inibizione dell'attività di tale trasportatore causa un aumento della sensibilità al trattamento chemioterapico nelle linee cellulari di osteosarcoma farmacoresistenti, indicando questo approccio come una possibile strategia per superare il problema della mancata risposta al trattamento farmacologico nei pazienti con osteosarcoma che sovraesprimono ABCB1. / Resistance to chemoterapeutic agents is a major obstacle for successful treatment of high grade osteosarcoma. This is in part caused by disregulation of the activity of ATP-binding cassette transporters (ABC) responsible for the efflux of chemotherapeutic agents in cancer cells. Studying how these transporters are regulated at transcription level is a crucial step to understanding multidrug resistance in cancer. The C-MYC oncogene has been identified to be related to resistance against methotrexate, doxorubicin and cisplatin and to be a prognostic factor for high grade osteosarcoma. C-MYC can regulate the expression of several ABC transporter genes, possibly related to drug resistance in osteosarcoma, and this may explain its adverse prognostic impact in this tumor. We have analyzed C-MYC and 16 ABC transporters, which are regulated by C-MYC and/or have been found to be responsible for the efflux of conventional drugs used in the treatment of human tumors on two different series of clinical samples and in a panel of human osteosarcoma cell lines by real-time PCR. The protein expression was evaluated for the 9 ABC transporters resulted as the most relevant from gene espression experiments. Finally the efficacy of a specific ABCB1 and ABCC1 inhibitor, was evaluated on osteosarcoma cell lines. ABCB1 and ABCC1 were the most expressed transporters in osteosarcoma cell lines. ABCB1 overexpression at diagnosis was found in about 40-45% of osteosarcoma patients and it was confirmed to be associated with adverse prognosis, being involved in drug resistance of osteosarcoma cells. Because of this, ABCB1 can be considered as a candidate therapeutic target for improving the treatment response of high-grade osteosarcoma patients. Inhibition of ABCB1 activity resulted to increase chemotherapeutics sensitivity of drug resistant human osteosarcoma cell lines, indicating this approach as a possible strategy to overcome treatment unresponsiveness in ABCB1-overexpressing osteosarcoma patients.

The transcribed ultraconserved regions (T-UCRs) are a group of long non-coding RNAs involved in human carcinogenesis. The factors regulating the expression of T-UCRs and their mechanism of action in human cancers are unknown. In this work it was shown that high expression of uc.339 associates with lower survival in 204 non-small cell lung cancer (NSCLC) patients. Moreover, it was shown that uc.339 found up-regulated in archival NSCLC samples, acts as a decoy RNA for miR-339-3p, -663-3p and -95-5p. So, Cyclin E2, a direct target of three microRNAs is up-regulated, inducing cancer growth and migration. Evidence of this mechanism was provided from cell lines and primary samples confirming that TP53 directly regulates uc.339. These results support a key role for uc.339 in lung cancer.

Autism Spectrum Disorder (ASD) is a range of early-onset conditions classified as neurodevelopmental disorders, characterized by deficits in social interactions and communication, as well as by restricted interest and repetitive behaviors. Among the proteins associated with this spectrum of disease there are Caspr2, α-NRXN1, NLGN1-4. Caspr2 is involved in the clustering of K+ channels at the juxtaparanodes, where it is proposed to bind TAG-1. Recent works reported a synaptic localization of Caspr2, but little is know on its role in this compartment. NRXNs and their ligand NLGNs, instead, have a well-defined role in the formation and maintenance of synapses. Among the neuroligins, NLGN2 binds NRXNs with the lowest affinity, suggesting that it could have other not yet characterized ligands. The aim of this work was to better characterize the binding of Caspr2 to TAG-1 and to identify new potential binding partner for Caspr2 and NLGN2. Unexpectedly, using Isothermal Titration Calorimetry and co-immunoprecipitation experiments the direct association of the first two proteins could not be verified and the results indicate that the first evidences reporting it were biased by false-positive artifacts. These findings, together with the uncharacterized synaptic localization of Caspr2, made the identification of new potential binding partners for this protein necessary. To find new proteins that associate with Caspr2 and NLGN2, affinity chromatography in tandem with mass spectrometry experiments were performed. Interestingly, about 25 new potential partners were found for these two proteins and NLGN1, that was originally included as a control: 5 of those, namely SFRP1, CLU, APOE, CNTN1 and TNR, were selected for further investigations. Only the association of CLU to NLGN2 was confirmed. In the future, screenings of the remaining candidates have to be carried out and the functional role for the proposed NLGN2-CLU complex has to be studied.

Il presente studio si concentra sull’analisi degli aspetti traslazionali nella ricerca farmacologica applicata alla Gastroenterologia. La trattazione si articola in due parti: una prima elaborazione teorica, che permette di inquadrare nel contesto della ricerca traslazionale il razionale scientifico ed etico alla base delle attività sperimentali eseguite durante il triennio; una seconda parte, nella quale si riportano i metodi, i risultati e le osservazioni conclusive derivanti dallo studio sperimentale. Nella prima parte vengono analizzate alcune caratteristiche delle procedure, adottate nella ricerca in ambito farmacologico gastrointestinale, che permettono di ottenere un dato verosimile derivabile da modelli diversi rispetto all’organismo umano. Sono inclusi nella trattazione gli aspetti etici dell’utilizzo di alcuni modelli animali di patologie intestinali organiche e funzionali in relazione al loro grado di predittività rispetto alla realtà sperimentale clinica. Nella seconda parte della trattazione, viene presentato uno studio esplorativo tissutale multicentrico sul ruolo del sistema oppioide e cannabinoide nella sindrome dell’intestino irritabile (IBS). Obiettivo dello studio è la valutazione dell’espressione e la localizzazione del recettore oppioide µ (µOR), del suo ligando β endorfina (β-END) e del recettore cannabinoide 2 (CB2) nei pazienti con IBS ad alvo costipato (IBS-C) e diarroico (IBS-D), ed in soggetti sani (HC). I dati ottenuti indicano un’implicazione del sistema oppioide e cannabinoide nella risposta immune alterata riscontrata nei pazienti con IBS ed in particolare nel sottogruppo IBS-C. La presente trattazione suggerisce come la creazione di nuovi sistemi di indagine sempre più validi da un punto di vista traslazionale possa dipendere, almeno in parte, dalla capacità di integrare realtà disciplinari, tecnologie ed esperienze metodologiche diverse nel contesto della ricerca in campo biomedico e farmacologico ed in particolare tramite un mutuo scambio di informazioni tra realtà clinica e ricerca di base / The present study focuses on the analysis of translational issues of pharmacological research applied to Gastroenterology. The dissertation is divided in two parts: the first one consists in a theroethical elaboration allowing to contextualize - within the framework of translational research - the scientific and ethical rationale underlying the experimental activity carried out over the last three years. The second part deals with the methods, the results and the conclusive observations deriving from the experimental study presented. In the first part some features of the procedures adopted in the field of pharmacology applied to gastroenterology are presented. These allow to obtain a reliable data deriving from models other than human one. This section also addresses the ethical issues concerning the use of animal models of both organic and functional intestinal pathologies. Such issues are put in relation to the level of predicitivity peculiar to the model and related to the research. In the second part of the dissertation the author presents a multicenter study about the role of the opioid and cannabinoid system in the irritable bowel syndrome (IBS). The aim of the study is the assessment of the expression of µ opioid receptor (µOR), of its ligand β-endorphin (β-END) and the cannabinoid receptor 2 (CB2) in IBS patients with either constipation (IBS-C) or diarrhea (IBS-D) compared to healthy controls (HC). The data obtained suggest that the opioid and cannabinoid systems contribute to the abnormal immune response detected in IBS patients, particularly in IBS-C. The present dissertation suggests how the creation of innovative research methodologies ever more effective from a translational standpoint may depend, at least in part, from the ability to integrate different research areas, technologies and methodological expertise in the context of biomedical and pharmacological research, especially through the mutual exchange of information between clinical and basic research.

Colorectal cancer (CRC) is a multistep process that goes through adenoma-carcinoma sequence. Many forms of CRC may be prevented by routine control, which can detect precancerous neoplasm before they undergo malignant transformation (123). For this reasons we hypothesized that a combination of simple faecal tests, may help to identify patients with higher risk of adenomas and/or CRC. The aim of this study is to clarify whether FOBT, enzyme Tumor M2-PK and endocannabinoid system molecules (CB1, CB2, FAAH), could represent diagnostic non-invasive markers, alone or in combination, for early diagnosis of CRC and its precancerous lesions. In a pivotal study we analyzed a selected population, using i-FOBT and quantitative ELISA stool test for t-M2-PK detection. i-FOBT showed the highest specificity and PPV (88.8% and 52.7% respectively); M2-PK had the best sensitivity (87.2%); the best results it obtained with combination tests; in fact if our patients had been subjected only to the i-FOBT test, 33 high risk adenoma and 14 CRC would not have been diagnosed. Supported by these findings, we analyzed a consecutive population: patients with both positive tests have only 31.6% risk of developing CRC; in contrast, patients negative to both markers, cancer risk was less than 2% (VPN 98.5%). We confirmed, also with immuoistochemistry, that increase of tumour M2-PK in patients with CRC, as well as in stool samples, is correlated with pre neoplastic stages. To investigate the expression of endocannabinoid system in CRC, CB1, CB2 and FAAH markers were studied immunochemically in different stages and normal tissue. CB receptors and their ligands, as well as FAAH inhibitions, showed to have a protective role in colorectal cancer. They, in combination with other markers (such as t-M2-PK and FOBT), could be indicated to develop a non-invasive test for an early diagnosis of cancerous and pre-cancerous colorectal lesions.

Nella presente tesi, abbiamo caratterizzato un modello di ratto trattato con dieta iperlipidica e streptozotocina (D), utilizzato per valutare i potenziali effetti anti-steatotici di una miscela contenente 5 estratti vegetali (caigua, soia, erba medica, carciofo e riso rosso fermentato) somministrata a due differenti dosaggi (0,3 [A] e 1 mg/Kg [B]) e confrontata con il controllo positivo eugenolo (E) ed il veicolo di somministrazione latte di soia (M). I valori concernenti colesterolo, LDL, glucosio, ALT ed il contenuto di lipidi nel tessuto epatico sono risultati aumentati in D, A ed M, rispetto ai CTR, e ripristinati significativamente in B ed E. Le attività marcatrici del CYP2E1 (p-nitrofenolo idrossilasi ed anilina-idrossilasi) e le analisi di Western blotting per le proteine CYP2E1 e CYP4A hanno evidenziato una significativa induzione su D, A ed M ed un parziale ripristino dei valori di controllo in B. Dal dosaggio del GSH e della carbonilazione delle proteine è stata evidenziata la presenza di stress ossidativo nei ratti iperlipidici, significativamente ridotta nel B. La presenza del colesterolo nella dieta iperlipidica è stata determinante per la significativa riduzione dei geni HMGCR ed LDLr. Dall’analisi di geni coinvolti nell’infiammazione e nello stress mitocondriale si è evidenziato un significativo aumento nei gruppi D, A e M rispetto al CTR, mentre una significativa riduzione si è osservata in B ed E. Per estendere la caratterizzazione del ratto HFD/STZ, in un nuovo esperimento abbiamo analizzato geni chiave del metabolismo del colesterolo e del trasporto degli acidi biliari. Abbiamo osservato un’induzione dei geni responsabili della formazione dell’acido colico CYP7A1 e CYP8B1, ma non di quelli dell’acido chenodeossicolico CYP27A1 e CYP7B1. Inoltre è emersa una diminuita espressione di SHP (gene chiave per l’inibizione feedback del CYP7A1 e del CYP8B1) e dei geni responsabili del trasporto degli acidi biliari, del colesterolo, e della fosfatidilcolina. / In this thesis, we characterized a rat model treated with a high fat diet and low dose of streptozotocin (HFD/STZ, D), to evaluate the anti-steatotic effects of a 5 plant extracts mixture (caigua, soybean, alfalfa, artichoke and fermented red rice) administered at two different doses (0.3 [A] and 1 mg/kg [B]) and compared to the positive control eugenol (E) and the vehicle soy milk (M) administration. Cholesterol, LDL, glucose and ALT blood parameters values and liver lipids content were increased in D, A and M, with the respect to CTR, and significantly recovered in B and E. CYP2E1 specific activities (p-nitrophenol hydroxylase and aniline hydroxylase) and Western blot analysis for CYP2E1 and CYP4A proteins were induced in D, A and M, and partially restored in B. GSH and protein carbonylation analysis highlighted, compared to CTR, the presence of oxidative stress in HFD/STZ rats, which was significantly reduced in B. The analysis of genes involved in inflammation (IL6, TNFα) and mitochondrial stress revealed a significant increase in A, D, M groups, but not in E and B. The presence of 2% of cholesterol in the high fat diet determined a significant reduction of HMGCR and LDLr transcripts. To extend the characterization of our steatotic rat model, we analyzed key genes of cholesterol metabolism and bile acids transporters in a new experiment. It was observed an induction of CYP7A1 and CYP8B1 genes responsible for the formation of cholic acid (CA), but not of CYP27A1 and CYP7B1, which lead to chenodeoxycholic acid (CDCA). In addition, we observed a decreased expression of FXR-regulated SHP (the key gene for the feedback CYP7A1 and CYP8B1 inhibition) and of the genes responsible for the transport of bile acids (NTCP, MRP2/4, OATP1A1, OATP1B2, BSEP), cholesterol (ABCG5), and phosphatidylcholine (MDR3).

Negli ultimi decenni, in virtù delle eccellenti proprietà antiossidanti della vitamina E (VE), la possibile associazione tra questa e la riduzione dell’incidenza di patologie neoplastiche è stata studiata mediante numerosi trial clinici. I risultati sono però ambigui e l’impiego della VE come agente chemiopreventivo su larga scala è oggi più che mai al centro del dibattito scientifico. Il Selenium and Vitamin E Cancer Prevention Trial (SELECT) ha addirittura evidenziato un rischio d’incidenza più alto per il cancro alla prostata nel gruppo d’intervento in cui è stata somministrata la VE. Tuttavia, il meccanismo d’azione non è noto. Poiché la VE induce l’espressione di alcune isoforme del P450 nel fegato e poichè tale induzione è associata ad un aumento della produzione di specie reattive centrate sull’ossigeno, ci siamo posti il problema di come un’eventuale up-regulation a livello prostatico avrebbe potuto generare uno stress ossidativo responsabile del fenomeno di cui sopra. Il presente lavoro ha mostrato come la VE provochi una marcata induzione delle isoforme CY1A1, CYP1A2, e CYP1B1/2 nel rene e nella prostata di ratto trattato i.p. con VE (100 o 200 mg/kg p.c. per sette o quattordici giorni consecutivi), e nel rene una generale inattivazione degli enzimi post-ossidativi GST e UDPGT ed una riduzione della potenzialità antiossidante. La spettroscopia EPR abbinata alla tecnica radical trapping ha rilevato una generazione anomala di radicali liberi in entrambi i tessuti. I risultati sono stati confermati in vitro in cellule epiteliali di prostata umana RWPE-1 esposte alla VE. Insieme ad un aumento dell’espressione genica (mRNA) di differenti CYPs, è stato osservato un incremento di radicali liberi e della prostaglandina E2 (PGE2) rispetto al controllo. Lo studio indica che la VE induce la superfamiglia CYP e uno stress ossidativo a livello prostatico (co-cancerogenesi) e può contribuire a spiegare i risultati inaspettati del trial SELECT. / Several meta-analysis and randomized clinical trials have seriously questioned chemoprevention based on vitamins including vitamin E (VE). Recently, the Selenium and Vitamin E Cancer Prevention Trial (SELECT) has pointed out an increased risk for prostate cancer among VE long–term users. However, to date, the mechanism underlying these findings still remain unknown. Evidence from both in vitro and in vivo models reported how VE might increase the expression of hepatic cytochrome P450 (CYP). Induction may increase the biotransformation of ubiquitous pre-carcinogens and trigger an over-production of oxygen centred radicals (ROS) in the target tissue. We apotheosized that if such phenomenon occured also in the prostate, it could contribute to explain the SELECT unexpected data. Male Sprague-Dawley rats were daily treated i.p. with either 100 or 200 mg/kg b.w. for 7 or 14 consecutive days. A powerful booster effect of various CYP isoforms such as CY1A1, CYP1A2, and CYP1B1/2, coupled with a marked free radical over-generation were recorded in renal and prostate tissues. VE treatment led to a wide down-regulation of antioxidant (catalase, NAD(P)H:quinone reductase) and phase II enzymes (glutathione S-transferase, UDP-glucuronosyl transferase capability. Results observed in the in vivo study were consistent with those obtained by the use of a RWPE-1 human prostate cell based model. Compared to the control RWPE-1, cells exposed to VE reported a general CYP up-regulation associated with a higher content of free radicals. Interestingly, VE treatment also induced the cyclooxygenase (COX-2) expression with a consequently increased of the prostaglandin E2 levels. The present study suggests that VE can act as a co-carcinogen and pro-oxidant agent. If such epigenetic mechanisms occur in human, may contribute to explain the harmful outcomes raised up from the SELECT study.

REST (RE-1 silencing transcription factor, or NRSF –neuron-restrictive silencing factor) binds to a conserved RE-1 motif present in the promoter region of regulated genes and represses their transcription in neuronal and non-neuronal cells (Bruce et al., 2004). REST recruits corepressors (CoREST, mSin3a) and multiple chromatin modifying enzymes (HDAC1/2, demethylase LSD1 and methyltransferase G9a), causing chromatin compaction and altering gene expression by changing epigenetic tagets (Ballas et al., 2005). REST contributes to orchestrate the epigenetic regulation of target genes through several miRNAs including miR-9/9*, miR-29a, miR-124a, miR-218 and others (Wu and Xie, 2006). My thesis has ascertained an anti-tumor properties of transcription factor REST on a model of cervical adenocarcinoma where class I histone deacetylase (HDAC) inhibitor: valproic acid (VPA) down-regulates REST and its corepressors CoREST and HDAC1 at mRNA and protein level. These effects are related to a potent effect on cell apoptosis, possibly mediated by miR-9 overexpression as consequence of REST and CoREST down-regulation. I report the presence of a double-negative feedback loop between REST and miR-9 in HeLa cell line: in absence of REST, miR-9 levels substantially increase while miR-9 overexpression promotes REST down-regulation. Interestingly, I have observed that REST is sufficient to induce a noteworthy chromatin remodeling in HeLa cells. HeLa cell apoptosis induced by these events, involves mitochondrial control of apoptosis signaling pathways, particularly Bcl-2 family gene BAX. In conclusion, the present study aims to contribute to a more accurate comprehension of the processes responsible for REST activity in a model of epithelial cervical adenocarcinoma, and relevant for a detailed knowledge of important events causing oncogenesis. Moreover, considering the crucial role of epigenetic regulation of gene transcription in the etiology of many pathological conditions, any further knowledge in this field could find important and innovative pharmacological applications.

Chronic obstructive pulmonary disease (COPD) is a multifactorial disease characterized by airflow obstruction that is usually progressive and associated with an abnormal inflammatory response of the lungs to noxious particles and gases. Cigarette smoke (CS) is the main risk factor, but only a small proportion of smokers (15-20%) develop symptomatic disease, this suggests that there are individual susceptibility factors involved in disease onset and progression. Considering the impact of environmental and genetic risk factors in COPD, this dissertation sought to uncover genetic susceptibility biomarkers in a population affected by COPD and explore tissue metabolic alterations induced by chronic CS exposure in a mouse model. A case-control study was carried on in a COPD population, aiming to investigate whether polymorphisms of microsomal epoxide hydrolase (EPHX1) gene, and related phenotypes, had any bearing on individual susceptibility to COPD onset and severity. DNA of COPD patients and controls was genotyped for functional polymorphisms of EPHX1 gene (exon 3 Tyr113His, exon 4 His139Arg), and haplotype analysis was performed using PCR-RFLP and PCR-RT techniques. The statistical analysis did not show any significant result about the potential relationship between analyzed SNPs, related phenotypes, and COPD risk and severity. Stable isotope-resolved metabolomics approach was used to study glycolytic pathway alterations induced by chronic CS exposure in lung and liver tissue of an emphysema mouse model. C57BL/6J mice, after CS exposure, were injected via IP injection with glucose tracer containing carbon stable isotope - 13C6-glucose – then, tissues were collected and the incorporation of 13C into metabolites was monitored by mass spectrometry and nuclear magnetic resonance spettroscopy. Lung tissue analyses did not reveal any significant alteration in lung tissue glycolysis of mice exposed to CS. On the other hand, CS may contribute to dysregulated glycolysis, PPP, glycogen synthesis and utilization, in emphysema mouse model liver tissue.

Obiettivo Analisi di consumi e costi degli antibiotici sistemici negli ospedali dell’Emilia-Romagna dal 2004 al 2011, con attenzione alla variabilità interaziendale e al significato, in termini di resistenza batterica, dell’aumento di alcuni gruppi terapeutici; Sottoanalisi nei reparti pediatrici, individuando i gruppi terapeutici critici, e valutazione delle reazioni avverse pediatriche da antibiotici segnalate, per il periodo in esame. Metodi I dati di consumo e spesa degli antibiotici sistemici per il periodo 2004-2011 sono stati ottenuti dal database regionale AFO e le giornate di degenza per ogni reparto dal database regionale di dimissione ospedaliera SDO. Le segnalazioni di sospette reazioni avverse da antibiotici tra gennaio 2004 e dicembre 2011 sono state estratte dal database nazionale VigiSegn. Risultati Negli otto anni, il consumo di antibiotici negli ospedali dell’Emilia-Romagna è aumentato del 27% e la spesa del 3%. Il consumo è apparso nettamente superiore nei reparti chirurgici che medici. La prima classe per consumo e spesa sono le penicilline/inibitori delle beta lattamasi. Nei reparti pediatrici, sono stati utilizzati 65 principi attivi diversi e amoxicillina/acido clavulanico è stato il più usato (26% del totale del 2011). Tra gli antibiotici critici, le cefalosporine di terza generazione sono state le più consumate in tutti i reparti pediatrici nel 2011. Tra le molecole il cui uso ospedaliero è vincolato, spiccano il linezolid e la teicoplanina che, comunque, hanno inciso più di tutte nella spesa del 2011 (18% e 15%, rispettivamente). Per la farmacovigilanza, i bambini (3-13 anni) sono stati coinvolti in 23 casi, mentre gli infanti (≤2 anni) solo in 4. L’associazione amoxicillina/acido clavulanico è stata più frequentemente segnalata (n=7), e soltanto 2 casi erano gravi. Conclusioni I risultati mostrano un quadro critico sul massiccio uso delle cefalosporine di terza generazione e sull’incremento del linezolid, da approfondire se per inappropriatezza d’uso oppure per aumento delle resistenze batteriche. / Aim To assess antibiotic consumption and expenditure in all hospitals of Emilia-Romagna from 2004 to 2011, highlighting on variability and meaning, in terms of bacterial resistance, of the rise in some therapeutic groups; To analyze antibiotic use among the pediatric wards, identifying the critical therapeutic groups, and evaluation of antibiotic-associated adverse reactions in pediatric inpatients, in the period considered. Methods Consumption and expenditure data of antibacterial for systemic use in the period 2004-2011 were obtained from the regional database (AFO) and the number of bed-days was available for the single wards from the regional hospital discharge database (SDO). Adverse reaction reports associated to antibiotics and recorded between January 2004 and December 2011 have been extracted from the national database VigiSegn. Results In the eight years analyzed, the overall antibiotic consumption increased by 27% and expenditure by 3% in all hospitals of Emilia-Romagna. Antibiotic consumption appeared markedly higher in the wards of surgical wards than medical ones. Penicillins/beta-lactamase inhibitors ranked first in consumption and expenditure. Sixty-five different antibiotics were used among the pediatric wards and amoxicillin/clavulanic acid was the most used (26% of the total in 2011). Among the critical antibiotics identified, the third-generation cephalosporins were the most used in all pediatric wards in 2011. Among the antibiotics with a defined limited use, the data highlighted that linezolid and teicoplanin affected most of all expenditure in 2011 (18% and 15%, respectively). About pharmacovigilance data, children (3-13 years) were involved in 23 cases, whereas infants (≤2 years) just in 4. Amoxicillin/clavulanic acid was the antibiotic most frequently reported (n = 7), and 2 cases of them were serious. Conclusions Results show a critical picture on the marked use of third-generation cephalosporins and on increase of linezolid consumption, because of inappropriateness of use or rise of bacterial resistance.