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Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze sede Policlinico

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2024 - A network study to differentiate suicide attempt risk profiles in male and female patients with major depressive disorder [Articolo su rivista]
Sarti, P.; Colliva, C.; Varrasi, S.; Guerrera, C. S.; Platania, G. A.; Boccaccio, F. M.; Castellano, S.; Pirrone, C.; Pani, L.; Tascedda, F.; di Nuovo, S.; Caraci, F.; Blom, J. M. C.

Suicide attempts are a possible consequence of Major Depressive Disorder (MDD), although their prevalence varies across different epidemiological studies. Suicide attempt is a significant predictor of death by suicide, highlighting its importance in understanding and preventing tragic outcomes. Researchers are increasingly recognizing the need to study the differences between males and females, as several distinctions emerge in terms of the characteristics, types and motivations of suicide attempts. These differences emphasize the importance of considering gender-specific factors in the study of suicide attempts and developing tailored prevention strategies. We conducted a network analysis to represent and investigate which among multiple neurocognitive, psychosocial, demographic and affective variables may prove to be a reliable predictor for identifying the 'suicide attempt risk' (SAR) in a sample of 81 adults who met DSM-5 criteria for MDD. Network analysis resulted in differences between males and females regarding the variables that were going to interact and predict the SAR; in particular, for males, there is a stronger link toward psychosocial aspects, while for females, the neurocognitive domain is more relevant in its mnestic subcomponents. Network analysis allowed us to describe otherwise less obvious differences in the risk profiles of males and females that attempted to take their own lives. Different neurocognitive and psychosocial variables and different interactions between them predict the probability of suicide attempt unique to male and female patients.

2023 - Cluster Headache Genomewide Association Study and Meta-Analysis Identifies Eight Loci and Implicates Smoking as Causal Risk Factor [Articolo su rivista]
Winsvold, B. S.; Harder, A. V. E.; Ran, C.; Chalmer, M. A.; Dalmasso, M. C.; Ferkingstad, E.; Tripathi, K. P.; Bacchelli, E.; Børte, S.; Fourier, C.; Petersen, A. S.; Vijfhuizen, L. S.; Magnusson, S. H.; O'Connor, E.; Bjornsdottir, G.; Häppölä, P.; Wang, Y. F.; Callesen, I.; Kelderman, T.; Gallardo, V. J.; de Boer, I.; Olofsgård, F. J.; Heinze, K.; Lund, N.; Thomas, L. F.; Hsu, C. L.; Pirinen, M.; Hautakangas, H.; Ribasés, M.; Guerzoni, S.; Sivakumar, P.; Yip, J.; Heinze, A.; Küçükali, F.; Ostrowski, S. R.; Pedersen, O. B.; Kristoffersen, E. S.; Martinsen, A. E.; Artigas, M. S.; Lagrata, S.; Cainazzo, M. M.; Adebimpe, J.; Quinn, O.; Göbel, C.; Cirkel, A.; Volk, A. E.; Heilmann-Heimbach, S.; Skogholt, A. H.; Gabrielsen, M. E.; Wilbrink, L. A.; Danno, D.; Mehta, D.; Guðbjartsson, D. F.; Rosendaal, F. R.; Willems van Dijk, K.; Fronczek, R.; Wagner, M.; Scherer, M.; Göbel, H.; Sleegers, K.; Sveinsson, O. A.; Pani, L.; Zoli, M.; Ramos-Quiroga, J. A.; Dardiotis, E.; Steinberg, A.; Riedel-Heller, S.; Sjöstrand, C.; Thorgeirsson, T. E.; Stefansson, H.; Southgate, L.; Trembath, R. C.; Vandrovcova, J.; Noordam, R.; Paemeleire, K.; Stefansson, K.; Fann, C. S. J.; Waldenlind, E.; Tronvik, E.; Jensen, R. H.; Chen, S. P.; Houlden, H.; Terwindt, G. M.; Kubisch, C.; Maestrini, E.; Vikelis, M.; Pozo-Rosich, P.; Belin, A. C.; Matharu, M.; van den Maagdenberg, A. M. J. M.; Hansen, T. F.; Ramirez, A.; Zwart, J. A.

Objective: The objective of this study was to aggregate data for the first genomewide association study meta-analysis of cluster headache, to identify genetic risk variants, and gain biological insights. Methods: A total of 4,777 cases (3,348 men and 1,429 women) with clinically diagnosed cluster headache were recruited from 10 European and 1 East Asian cohorts. We first performed an inverse-variance genomewide association meta-analysis of 4,043 cases and 21,729 controls of European ancestry. In a secondary trans-ancestry meta-analysis, we included 734 cases and 9,846 controls of East Asian ancestry. Candidate causal genes were prioritized by 5 complementary methods: expression quantitative trait loci, transcriptome-wide association, fine-mapping of causal gene sets, genetically driven DNA methylation, and effects on protein structure. Gene set and tissue enrichment analyses, genetic correlation, genetic risk score analysis, and Mendelian randomization were part of the downstream analyses. Results: The estimated single nucleotide polymorphism (SNP)-based heritability of cluster headache was 14.5%. We identified 9 independent signals in 7 genomewide significant loci in the primary meta-analysis, and one additional locus in the trans-ethnic meta-analysis. Five of the loci were previously known. The 20 genes prioritized as potentially causal for cluster headache showed enrichment to artery and brain tissue. Cluster headache was genetically correlated with cigarette smoking, risk-taking behavior, attention deficit hyperactivity disorder (ADHD), depression, and musculoskeletal pain. Mendelian randomization analysis indicated a causal effect of cigarette smoking intensity on cluster headache. Three of the identified loci were shared with migraine. Interpretation: This first genomewide association study meta-analysis gives clues to the biological basis of cluster headache and indicates that smoking is a causal risk factor. ANN NEUROL 2023.

2023 - Dysbindin, D3 receptors, and their genetic interaction differently regulate neuroplasticity genes in the mouse brain [Abstract in Atti di Convegno]
Rivi, V.; Benatti, C.; Blom, J. M. C.; Pani, L.; Brunello, N.; Drago, F.; Papaleo, F.; Torrisi, S.; Leggio, G.; Tascedda, F.

2023 - Environmental enrichment influences mouse hippocampal neuroinflammatory response [Abstract in Rivista]
Rigillo, Giovanna; Benatti, Cristina; Toscano, Ylenia; Tascedda, Fabio; Pani, Luca; Brunello, Nicoletta; Alboni, Silvia

Background: Many studies have shown the positive effects of environmental enrichment on brain plasticity with significant implications for development, behavior, learning, memory, and recovery from brain damage (1). Experimental and clinical studies report that one’s living environment can modulate cellular and molecular responses in the brain, counteracting cognitive decline, alleviating anxiety, and depressive behaviours, as well as moderating the outcome of pharmacological treatments (2). Neuroinflammation has been well established as an important factor in the aetiopathogenesis and progression of brain disorders. It can affect neural development and alters hippocampal plasticity thus resulting in cognitive impairments. Neuroinflammation is characterized by a dysregulation of the NLRP3 inflammasome activation, an increase in the expression of inflammatory cytokines, and a decrease of neurotrophic factors (3). Behavioral and neurochemical changes, caused by neuroinflammation, have been most frequently investigated through peripheral administration of lipopolysaccharide (LPS), which can, directly and indirectly, affect the central nervous system (4). Based on these premises, the aim of this study was to explore the molecular effects of the quality of the living environment in modulating the LPS-induced neuroinflammatory response in the hippocampus of wild-type mice. Methods: Male C57BL6J mice (13 weeks-old) were randomly housed in Impoverished (IE) or Enriched Environment (EE) condition for 28 days, then exposed to LPS (0.830 mg/Kg, i.p.) or saline (SAL). Twenty-four hours after injection hippocampi were removed for gene expression analysis performed by means of qRT-PCR. Data from groups were analyzed by Two-way ANOVA followed by Tukey’s post hoc test. Results: The analysis of the environmental effects on the LPS signaling system highlighted the downregulation of the membrane-bound protein LBP, the receptor TLR4 and the co-receptor cluster CD14 expression levels in EE-housed animals compared to their counterparts. The exposure to an EE condition was able to attenuate the LPS-induced increase of TLR4 and NLRP3 inflammasome mRNA levels. Gene expression analysis revealed a significant downregulation of the pro-inflammatory cytokines IL-1β and TNFα levels in EE-housed mice while LPS exposure strongly increased IL-1β and TNFα mRNA levels irrespective of the housing conditions. Moreover, EE-exposed mice showed a significant upregulation of BDNF hippocampal mRNA levels, although no effects were observed after LPS treatment in both conditions. Conclusions: Our results displayed the beneficial effect of EE in regulating the expression of inflammatory mediators involved in the LPS-induced response in the hippocampus, a key area for learning, memory, and emotion. These data suggest that living environment may exert a positive and protective role on the brain by reducing susceptibility toward neurodegenerative or neuropsychiatric disorders.

2023 - Esmethadone-HCl (REL-1017): a promising rapid antidepressant [Articolo su rivista]
Fava, M.; Stahl, S. M.; De Martin, S.; Mattarei, A.; Bettini, E.; Comai, S.; Alimonti, A.; Bifari, F.; Pani, L.; Folli, F.; Guidetti, C.; Furlan, A.; Sgrignani, J.; Locatelli, P.; Cavalli, A.; O'Gorman, C.; Traversa, S.; Inturrisi, C. E.; Pappagallo, M.; Manfredi, P. L.

This review article presents select recent studies that form the basis for the development of esmethadone into a potential new drug. Esmethadone is a promising member of the pharmacological class of uncompetitive N-methyl-D-aspartate receptor (NMDAR) antagonists that have shown efficacy for major depressive disorder (MDD) and other diseases and disorders, such as Alzheimer’s dementia and pseudobulbar affect. The other drugs in the novel class of NMDAR antagonists with therapeutic uses that are discussed for comparative purposes in this review are esketamine, ketamine, dextromethorphan, and memantine. We present in silico, in vitro, in vivo, and clinical data for esmethadone and other uncompetitive NMDAR antagonists that may advance our understanding of the role of these receptors in neural plasticity in health and disease. The efficacy of NMDAR antagonists as rapid antidepressants may advance our understanding of the neurobiology of MDD and other neuropsychiatric diseases and disorders.

2023 - Farmaci antidepressivi parte II: SNRI e farmaci multimodali [Capitolo/Saggio]
Benatti, Cristina; Alboni, Silvia; Brunello, Nicoletta; Pani, Luca; Tascedda, Fabio

2023 - Galcanezumab for the treatment of chronic migraine and medication overuse headache: Real-world clinical evidence in a severely impaired patient population [Articolo su rivista]
Guerzoni, S.; Baraldi, C.; Castro, F. L.; Cainazzo, M. M.; Pani, L.

Background: Galcanezumab is a monoclonal antibody acting against the calcitonin gene-related peptide approved for the preventive treatment of migraine. The aim of this article is to explore its effectiveness and safety of galcanezumab in chronic migraine (CM) with medication overuse-headache (MOH). Methods: Seventy-eight patients were consecutively enrolled at the Modena headache center and followed up for 15 months. Visits were scheduled every 3 months, and the following variables were collected: the number of migraine days per month (MDM); the painkillers taken per month (PM); the number of days per month in which the patient took, at least, one painkiller; the six-item headache impact test; and the migraine disability assessment questionnaire (MIDAS) score. Demographic features of the analyzed sample were collected at the baseline and adverse events (AEs) were collected at every visit. Results: After 12 months, galcanezumab significantly reduced the MDM, the PM, the number of days on medication, the HIT-6 as well as the MIDAS scores (all p <.0001). The greatest amelioration was obtained in the first trimester of treatment. A higher MDM, a higher NRS score at the baseline, and a higher number of failed preventive treatments negatively predict the CM relief at the year of treatment. No serious AEs were registered and only one drop-out was due to AE. Conclusions: Galcanezumab is effective and safe for the treatment of patients affected by CM and MOH. Patients with a higher impairment at the baseline may found less benefits with galcanezumab.

2023 - Implications of Pediatric Initiatives on CNS Drug Development for All Ages—2020 and Beyond: Second of Three Sets of Expanded Proceedings from the 2020 ISCTM Autumn Conference on Pediatric Drug Development [Articolo su rivista]
Pandina, G.; Busner, J.; Horrigan, J. P.; Mcsherry, C.; Bateman-House, A.; Pani, L.; Kando, J.

This article expands upon a session, titled “Implications of Pediatric Initiatives on CNS Drug Development for All Ages—2020 and Beyond,” that was presented as part of a two-day meeting on pediatric drug development at the International Society for Central Nervous System (CNS) Clinical Trials and Methodology (ISCTM) Autumn Conference in Boston, Massachusetts, in October 2020. Speakers from various areas of pediatric drug development addressed a variety of implications of including children in drug development programs. The speakers wrote summaries of their talks, which are included here. The session’s lead chair was Dr. Gahan Pandina, who wrote introductory and closing comments. Dr. Joseph Horrigan addressed the current landscape of pediatric development programs. Dr. Gahan Pandina addressed how the approach to research in pediatric populations affects the drug development process and vice versa. Dr. Alison Bateman-House discussed the ethical implications of research in the pediatric population. Dr. Luca Pani discussed some of the global regulatory issues and challenges concerning research in pediatric patients. Dr. Judith Kando served as a discussant and posed new questions about means of facilitating pediatric research. Finally, Dr. Gahan Pandina provided closing comments and tied together the presented issues. This paper should serve as an expert-informed reference to those interested and involved in CNS drug development programs that are aimed at children and/or required, through regulations, to include children as part of the approval process.

2023 - OnabotulinumtoxinA: Still the Present for Chronic Migraine [Articolo su rivista]
Baraldi, C.; Lo Castro, F.; Ornello, R.; Sacco, S.; Pani, L.; Guerzoni, S.

OnabotulinumtoxinA (BT-A) is one of the few drugs approved for the preventive treatment of chronic migraine (CM). Despite this, some aspects of its mechanism of action are still a matter of debate, and the precise magnitude of BT-A effects needs to be completely elucidated. BT-A acts primarily upon trigeminal and cervical nerve endings, by inhibiting the release of inflammatory mediators such as calcitonin gene-related peptide, as well as reducing the insertion of ionotropic and metabotropic receptors into the neuronal membrane. These actions increase the depolarization threshold of trigeminal and cervical nerve fibers, thus reducing their activation. The central actions of BT-A are still a matter of debate: a retrograde axonal transport has been postulated, but not clearly assessed in humans. Clinically, the efficacy of BT-A in CM has been assessed by large, randomized placebo-controlled trials, such as the Phase 3 REsearch Evaluating Migraine Prophylaxis Therapy (PREEMPT) trials. Those results were also confirmed in a wide range of open-label studies, even for long-term periods. Recently, novel findings have led to a better understanding of its pharmacological actions and clinical usefulness in migraine prevention. This narrative review summarizes, updates and critically revises the available data on BT-A and its possible implementation in chronic migraine. Moreover, the current role of BT-A in CM treatment has been discussed.

2023 - Role of dopamine D3 receptors, dysbindin, and their functional interaction on the expression of key genes for neuroplasticity in the mouse brain [Abstract in Rivista]
Rivi, Veronica; Benatti, Cristina; Blom, Johanna; Pani, Luca; Brunello, Nicoletta; Drago, Filippo; Papaleo, Francesco; Torrisi, Sebastiano; Salomone, Salvatore; Leggio Gian, Marco; Tascedda, Fabio

2023 - Safety and Efficacy of Red Yeast Rice Phytocomplex and Lovastatin: a Comparative Analysis [Relazione in Atti di Convegno]
Rigillo, G; Baini, G; Miraldi, E; Pani, L; Tascedda, F; Biagi, M

In the last 20 years, the demand for red yeast rice-based food supplements has significantly increased as consumers have been looked for natural alternatives to manage blood cholesterol levels without the adverse effects associated with synthetic statins. Actually, monacolin K, derived from red yeast rice (RYR) and chemically identical to lovastatin, inhibits 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, offering a natural alternative to synthetic drugs. In 2018, despite the health claim approved in 2011, the European Food Safety Authority (EFSA) raised concerns about the safety of monacolin K consumption, especially for vulnerable populations, and highlighted the lack of knowledge on RYR phytocomplex. Consequently, in 2021, the European Commission established a new maximum intake limit < 2.99 mg/day for monacolins. This study aimed to investigate the safety and efficacy of RYR phytocomplex compared to the sole monacolin K (=lovastatin) by analyzing eight different RYR samples with different monacolin K and secondary monacolins content. Efficacy was evaluated through a validated cell-free enzymatic assay, while an integrated in vitro simulated digestion and in silico ADME prediction were employed to compare the pharmacokinetics of different samples and lovastatin. The safety of RYR was assessed by monitoring cytotoxicity in intestinal, hepatic, kidney, and skeletal muscle cells using cell viability assays. Furthermore, muscle damage-related targets and myokines were measured by qRT-PCR in myoblasts exposed to prolonged non-toxic stimuli. Results demonstrated that RYR samples have a large chemical variability not only related to monacolin K content, in lactone and hydroxy-acid form, but also as regards pigments, secondary monacolins, polyphenols, and triterpenes. The enzymatic test revealed that all samples were more effective in inhibiting HMG-CoA activity than lovastatin at equivalent monacolin K content. Although secondary monacolins had a weaker effect than monacolin K, a synergistic effect was observed within the phytocomplex. Additionally, the phytocomplex facilitated higher bioaccessibility of monacolin K in RYR compared to lovastatin. Regarding cell viability, lovastatin and RYR samples exhibited no toxic effects up to 150 g/mL on intestinal, hepatic, and kidney cells; however, variable effects were observed on skeletal muscle cells at medium-high concentrations. Nevertheless, at physiological non-toxic concentrations, RYR samples did not impact the expression levels of myokine IL-6, autophagy factors, and catabolic markers of protein balance, unlike synthetic statin. These results suggest that RYR and lovastatin share similar biological activities, but the phytocomplex's contribution from secondary monacolins, pigments, polyphenols, and triterpenes ensure a superior safety profile for this botanical food supplement. However, it is important to note that the composition of RYR phytocomplex can vary significantly between different products on the market. This variability highlights the need for standardization and quality control measures to ensure the safety and efficacy of this botanical food supplement. In conclusion, our study provides evidence supporting the enhanced efficacy, bioaccessibility, and safety profile of RYR phytocomplex compared to lovastatin demonstrating its potential as a natural and effective option for hypercholesterolemia management.

2023 - Subanalysis of subjective cognitive measures from a phase 2, double-blind, randomized trial of rel-1017 in patients with major depressive disorder [Articolo su rivista]
Guidetti, C.; Serra, G.; Pani, L.; Pappagallo, M.; Maglio, G.; De Martin, S.; Mattarei, A.; Folli, F.; Manfredi, P. L.; Fava, M.

Objective: Improvement of cognitive function in patients with major depressive disorder (MDD) is an important treatment outcome. REL-1017 (esmethadone HCl) is a novel N-methyl-d-aspartate receptor (NMDAR) channel blocker and a potentially rapidly acting antidepressant. The objective of this study was to define the effects of REL-1017 on subjective cognitive measures in patients with MDD. Methods: Post hoc analysis was conducted of subjective cognitive measures from the Montgomery-Asberg Depression Rating Scale (MADRS) and the Symptoms of Depression Questionnaire (SDQ) from a randomized, double-blind, placebo-controlled, Phase 2a study. The study, designed to assess the safety, tolerability, and efficacy of 2 dosages (25 mg and 50 mg) of REL-1017 as an adjunctive treatment in patients with MDD unresponsive to standard antidepressants, included 62 patients. We analyzed subjective cognitive measures derived from the MADRS and SDQ scales at baseline and up to day 14, 7 days after the last dose of study drug. We developed 2 composite indexes that included subjective cognitive measures selected from the MADRS and SDQ. Results: The subanalysis of single measures and the 2 composite indexes derived from the MADRS and SDQ measures showed clinically meaningful and statistically significant improvements in cognitive function (P <.05). Conclusions: In a Phase 2a clinical trial, REL-1017 improved subjective measures of cognitive impairment, in addition to improving total MADRS and SDQ scores. These results need to be confirmed in larger and longer studies in MDD that include objective measures of cognitive function. Phase 3 studies of REL-1017 for MDD are currently underway.

2023 - The Role of Dopamine D3 Receptors, Dysbindin, and Their Functional Interaction in the Expression of Key Genes for Neuroplasticity and Neuroinflammation in the Mouse Brain [Articolo su rivista]
Rivi, Veronica; Benatti, Cristina; Blom, Johanna; Pani, Luca; Brunello, Nicoletta; Drago, Filippo; Papaleo, Francesco; Caraci, Filippo; Geraci, Federica; Torrisi, Sebastiano Alfio; Leggio, Gian Marco; Tascedda, Fabio

2023 - The nexus of social alliances and diverse moral domains: a bedrock for participatory clinical research [Articolo su rivista]
Blom, Johanna; Rivi, Veronica; Tascedda, Fabio; Pani, Luca

Rigillo, G.; Ciani, M.; Benatti, C.; Blom, J. M. C.; Tascedda, F.; Pani, L.; Alboni, S.; Brunello, N.

Vortioxetine (VTX) is a multimodal antidepressant with an extensive pharmacological profile that includes modulation of various neurotransmitter systems, neuroprotective activity, and beneficial effects on cognitive functions. Recent research has revealed a novel aspect of VTX's activity - its antiinflammatory effects - that suggests an intriguing molecular mechanism may underpin its therapeutic benefits. Neuroinflammation, dysfunctional neurogenesis and neurotransmission, and dysregulation of the hypothalamus– pituitary–adrenal (HPA) axis are all pivotal in the onset and progression of depression. One particular immune-inflammatory pathway overactivated in brain disorders is the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, a multiprotein complex. This complex's activation is mediated by NF-kB and reactive oxygen species (ROS) signalling pathways, leading to caspase-1-dependent release of the proinflammatory cytokines, IL-1β and IL-18. Mounting evidence implicates NLRP3 inflammasome in neuroinflammation- related disorders, with its activation associated with cognitive function impairment. Of note, microglia, the resident immune cells crucial for brain plasticity, express high levels of the NLRP3-inflammasome components. Our initial findings indicate that VTX exerts a region-dependent modulatoryeffect on the NLRP3-inflammasome system in a LPS-induced memory impairment in vivo model. Furthermore, VTX's ability to modulate immune response suggests that microglia could be a direct target of the drug. AIM: In light of the compelling evidence surrounding the role of the NLRP3 inflammasome in cognitive dysfunctions and the recent discovery of VTX's anti-inflammatory activity, we aimed to investigate the molecular effects induced by VTX pre-treatment in the presence or absence of the inflammasome-inducer LPS in a well-established in vitro model of mouse microglia: BV2 cells. METHODS: To dissect the influence of VTX pre-treatment (24h) on the NLRP3 inflammasome signaling pathway and microglial polarization, we analyzed gene and protein expression in BV2 cells stimulated with LPS or vehicle for 6h. We also scrutinized the activation/translocation of NF-kB and ROS release under these conditions. We applied one-way or two-way ANOVA followed by Tukey’s post hoc test for statistical analysis based on the experimental design. RESULTS: Our data demonstrate that short-term exposure to LPS significantly induces the activation/translocation of NF-kB signaling and ROS release in BV2 cells. We observed a time-dependent transcriptional upregulation of the inflammasome complex, IL-1β and IL-18, and microglial pro-inflammatory targets post-LPS stimulation, alongside a downregulation of the anti-inflammatory factors. Interestingly, a pre-treatment with VTX (10 nM) for 24h effectively modulated the LPS-induced NF-kB translocation and ROS production compared to control cells. Cells pre-treated with VTX exhibited lower levels of LPS-induced NLRP3 inflammasome- and microglia pro-inflammatory-related targets. However, VTX did not influence the expression of anti-inflammatory factors in both unstimulated and LPS-stimulated BV2 cells. CONCLUSIONS: Our findings reinforce the emerging evidence that supports VTX's anti-inflammatory activity. This activity is mediated via modulation of the inflammasome signaling pathway, which plays a pivotal role in the inflammatory response of microglia cells.

2022 - A Phase 2a Double-Blind Randomized Trial of REL-1017 (Esmethadone) in Patients with MDD: Analysis of Subscales from the Symptoms of Depression Questionnaire [Articolo su rivista]
Guidetti, C.; Fava, M.; Pani, L.; Pappagallo, M.; Serra, G.; Demartin, S.; Mattarei, A.; Manfredi, P. L.

BACKGROUND: Major depressive disorder (MDD) is the second leading cause of disability and chronic disease burden in the United States. The importance of improving functional outcomes in MDD is increasingly recognized. The Symptoms of Depression Questionnaire (SDQ), a patient-reported measure, was developed to capture the heterogeneity of symptoms of MDD. REL-1017 (esmethadone HCl; d-methadone), is a novel N-methyl-d-aspartate receptor (NMDAR) channel blocker and potential rapid antidepressant currently in Phase 3 development. In a Phase 2a trial, REL-1017 showed robust, rapid, and sustained antidepressant efficacy as adjunctive treatment in patients with MDD. The objective of this study was to assess the effects of REL-1017 on SDQ subscales to better characterize the functional implications of its therapeutic effects. METHODS: A double-blind, placebo-controlled, inpatient, two-doses, 25 and 50 mg, three-arm, 1:1:1, randomized, phase 2a trial of REL-1017 was conducted at 10 centers in the United States. Least square (LS) mean scores and Cohen's effect sizes of the total score of a 44-item of SDQ and its 5 subscales: lassitude, mood, cognitive/social functioning (SDQ-1); anxiety, agitation, anger, and irritability (SDQ-2); desire to be dead (SDQ-3); disruptions in sleep quality (SDQ-4); changes in appetite and weight (SDQ-5) were compared between REL-1017 and placebo. RESULTS: A total of 62 adult male and female patients (18-65 years of age) diagnosed with MDD participated in the trial. On day 14, the last day of efficacy measurement, the difference from placebo of the LS mean (90% CI) for REL-1017 25 mg and REL-1017 50 mg groups, respectively, showed improvement for both tested doses on SDQ total score (-23.2; P = .0066 [effect size: 0.9]; -26.8 P = .0014 [effect size: 1.1]). Additionally, for SDQ subscales, REL-1017 25 mg and REL-1017 50 mg groups, respectively, showed significant improvement as compared with placebo: SDQ-1 (-13.9; P = .0025 [effect size: 1.0]; -15.0; P = .0009 [effect size: 1.1]), SDQ-2 (-4.6; P = .0398 [effect size: 0.7]; -7.2; P = .0012 [effect size: 1.1]) and SDQ-4 (-2.7; P = .0055 [effect size: 1.0]; -2.8; P = .0029 [effect size: 1.0]). No significant differences were observed between the treated groups and placebo in the SDQ-3 and SDQ-5 subscales. CONCLUSIONS: In patients with MDD, aside from improving the overall CFB compared to placebo in SDQ total score, REL-1017 resulted in clinically meaningful and statistically significant improvements in cognitive/motivational, anxiety/irritability, and sleep-specific domains. The robust, rapid, and sustained efficacy of REL-1017 for MDD is not limited to improving mood, but potentially extends to cognitive, motivational, sleep, and social functions, with potentially meaningful therapeutic and socioeconomic implications. These results may signal disease-modifying effects of esmethadone for MDD that may offer potential advantages over symptomatic treatment with standard antidepressants. FUNDING: Relmada Therapeutics, Inc.

2022 - Case Report: REL-1017 Reduces Abnormal Clinician Administered Dissociative States Scale Scores in Patients with Major Depressive Disorder [Articolo su rivista]
Guidetti, C.; Pani, L.; Serra, G.; Pappagallo, M.; Fava, M.; Manfredi, P. L.

BACKGROUND: Dissociative symptoms may be found in a subset of patients with major depressive disorders (MDD). The Clinician-Administered Dissociative States Scale (CADSS) is a 23-item scale for the measurement of present-state dissociative symptoms with good inter-rater reliability and construct validity that can discriminate patients with dissociative disorders. The total CADSS score is derived by adding the score for each of the 23 items. A score of 4 or more on the CADSS is considered abnormal and clinically meaningful. Uncompetitive N-methyl-d-aspartic acid receptor (NMDAR) channel blockers have been proposed as a treatment for post-traumatic stress disorder (PTSD). REL-1017 is a novel, low potency, NMDAR channel blocker currently in Phase 3 studies for MDD. METHODS: This retrospective case series describes a subset of patients from a double-blind, randomized, placebo-controlled, in-patient 7-day, phase 2 trial of oral, once daily, 25 mg (75 mg loading dose on day 1, first dose) and 50 mg REL-1017 (100 mg loading dose on day 1, first dose) as an adjunctive treatment for MDD. This subset of patients was selected based on abnormal CADSS score at baseline, pre-treatment with the study drug. As part of REL-1017 safety evaluation, the CADSS was administered at four timepoints to all study patients: (a) 30 to 60 minutes pre-treatment at baseline on day 1; (b) 2 hours post-treatment on day 1 (after the first dose of study drug); (c) 2 hours post-treatment on day 7 (after the last dose); and (d) prior to discharge on day 9 (2 days after the last dose). RESULTS: Among the 62 randomized patients, four patients had a CADSS score of at least 4 on day 1 before study drug administration (2 patients in the 25 mg arm [CADSS score 22 and 4]; 1 patient in the 50 mg arm [CADSS score 35]; 1 patient in the placebo arm [CADSS score 6]). Among these 4 patients, starting on day 1, 2 hours post-treatment, the 2 subjects in the 25 mg subgroup (75 mg loading dose) and 1 subject in the 50 mg subgroup (100 mg loading dose) showed a clinically meaningful decrease in their CADSS score, while the single patient in the placebo group showed no change. CADSS scores on Day 1 pre-treatment, day 1 post-treatment, day 7 post last treatment, and on day 9 prior to discharge were 22-2-6-0; 4-0-0-0; 35-14-9-0, and 6-6-n/a-n/a, for the two patients in the 25 mg REL-1017 subgroup, the single patient in the 50 mg REL-1017 subgroup, and the single patient in the placebo group, respectively. CONCLUSIONS: These retrospective case report data potentially signal that REL-1017 may determine rapid and sustained improvement in patients with MDD and concurrent clinically meaningful dissociative symptoms assessed by a CADSS score of 4 or above. Ongoing phase 3 trials with REL-1017 are expected to enroll a total of 1200 outpatients with MDD. These studies will potentially generate additional data that may support the initiation of controlled studies with REL-1017 for the treatment of PTSD. FUNDING: Relmada Therapeutics.

2022 - Chronic migraine evolution after 3 months from erenumab suspension: real-world-evidence-life data [Articolo su rivista]
Guerzoni, S.; Baraldi, C.; Pensato, U.; Favoni, V.; Lo Castro, F.; Cainazzo, M. M.; Cevoli, S.; Pani, L.

Background: Erenumab is a monoclonal antibody acting against calcitonin gene-related peptide receptor which has been found effective even for the treatment of chronic migraine (CM) complicated with medication overuse headache (MOH). According to the present guidelines, the treatment with erenumab should continue for up to 1 year. The aim of the present study is to explore the evolution of patients affected by CM and MOH at the baseline, after erenumab discontinuation. Methods: One hundred and eighty-five patients affected by CM and MOH were recruited and followed up after erenumab discontinuation. The number of migraine days per month, the number of painkillers taken per month, the number of days in which one medication was used for a month were collected every 30 days for the 3 months following erenumab suspension. Results: At the 3rd month after suspension, patients displayed a significantly higher number of migraine days per month, a significantly higher painkiller consumption, and a significantly higher migraine-related disability. A high body mass index and the presence of aura were positively correlated with the relapse of CM and MOH. Conclusion: Patients affected by CM and MOH at the baseline displayed a significant worsening of their headaches after erenumab discontinuation.

2022 - Depression and Catatonia Associated With Lansoprazole in an Adolescent With Phelan-McDermid Syndrome: A Case Report [Articolo su rivista]
Persico, A.; Ricciardello, A.; Alessandrini, S.; Viola, L.; Bergonzini, P.; Iughetti, L.; Pani, L.

2022 - Editorial: Digital biomarkers in testing the safety and efficacy of new drugs in mental health: A collaborative effort of patients, clinicians, researchers, and regulators [Articolo su rivista]
Blom, Johanna Maria Catharina; Benatti, Cristina; Mascalzoni, Deborah; Tascedda, Fabio; Pani, Luca

2022 - Esmethadone (REL-1017) and Other Uncompetitive NMDAR Channel Blockers May Improve Mood Disorders via Modulation of Synaptic Kinase-Mediated Signaling [Articolo su rivista]
Stahl, S. M.; De Martin, S.; Mattarei, A.; Bettini, E.; Pani, L.; Guidetti, C.; Folli, F.; de Somer, M.; Traversa, S.; Inturrisi, C. E.; Pappagallo, M.; Gentilucci, M.; Alimonti, A.; Fava, M.; Manfredi, P. L.

This article presents a mechanism of action hypothesis to explain the rapid antidepressant effects of esmethadone (REL-1017) and other uncompetitive N-methyl-D-aspartate receptor (NMDAR) antagonists and presents a corresponding mechanism of disease hypothesis for major depressive disorder (MDD). Esmethadone and other uncompetitive NMDAR antagonists may restore physiological neural plasticity in animal models of depressive-like behavior and in patients with MDD via preferential tonic block of pathologically hyperactive GluN2D subtypes. Tonic Ca2+ currents via GluN2D subtypes regulate the homeostatic availability of synaptic proteins. MDD and depressive behaviors may be determined by reduced homeostatic availability of synaptic proteins, due to upregulated tonic Ca2+ currents through GluN2D subtypes. The preferential activity of low-potency NMDAR antagonists for GluN2D subtypes may explain their rapid antidepressant effects in the absence of dissociative side effects.

2022 - Excellent Response to OnabotulinumtoxinA: Different Definitions, Different Predictors [Articolo su rivista]
Ornello, R.; Baraldi, C.; Ahmed, F.; Negro, A.; Miscio, A. M.; Santoro, A.; Alpuente, A.; Russo, A.; Silvestro, M.; Cevoli, S.; Brunelli, N.; Vernieri, F.; Grazzi, L.; Pani, L.; Andreou, A.; Lambru, G.; Frattale, I.; Kamm, K.; Ruscheweyh, R.; Russo, M.; Torelli, P.; Filatova, E.; Latysheva, N.; Gryglas-Dworak, A.; Straburzynski, M.; Butera, C.; Colombo, B.; Filippi, M.; Pozo-Rosich, P.; Martelletti, P.; Guerzoni, S.; Sacco, S.

The identification of patients who can benefit the most from the available preventive treatments is important in chronic migraine. We explored the rate of excellent responders to onabotulinumtoxinA in a multicenter European study and explored the predictors of such response, according to different definitions. A pooled analysis on chronic migraineurs treated with onabotulinumtoxinA and followed-up for, at least, 9 months was performed. Excellent responders were defined either as patients with a ≥75% decrease in monthly headache days (percent-based excellent responders) or as patients with <4 monthly headache days (frequency-based excellent responders). The characteristics of excellent responders at the baseline were compared with the ones of patients with a <30% decrease in monthly headache days. Percent-based excellent responders represented about 10% of the sample, whilst frequency-based excellent responders were about 5% of the sample. Compared with non-responders, percent-based excellent responders had a higher prevalence of medication overuse and a higher excellent response rate even after the 1st and the 2nd injection. Females were less like to be frequency-based excellent responders. Chronic migraine sufferers without medication overuse and of female sex may find fewer benefits with onabotulinumtoxinA. Additionally, the excellent response status is identifiable after the first cycle.

2022 - Identification and characterization of the kynurenine pathway in the pond snail Lymnaea stagnalis [Articolo su rivista]
Benatti, Cristina; Rivi, Veronica; Alboni, Silvia; Grilli, Andrea; Castellano, Sara; Pani, Luca; Brunello, Nicoletta; Blom, Johanna Maria Catharina; Bicciato, Silvio; Tascedda, Fabio

2022 - Occlusal splint therapy in patients with Ménière’s disease and temporomandibular joint disorder [Articolo su rivista]
Monzani, D.; Baraldi, C.; Apa, E.; Alicandri Ciufelli, M.; Bertoldi, C.; Roggla, E.; Guerzoni, S.; Marchioni, D.; Pani, L.

SUMMARY Objective. This retrospective study aimed to verify the outcomes of stabilising occlusal splint therapy prescribed to 22 patients with unilateral definite Ménière’s disease and comorbid temporomandibular joint disorder. Methods. The results of a battery of audiometric and vestibular tests were recorded before and after 6 months of treatment, as well as the scores of disease-specific questionnaires. Results. The average hearing threshold in the affected ear and the acoustic immittance were unchanged. No spontaneous and positional nystagmus were recorded. Caloric hyporesponsiveness and vestibular myogenic evoked responses did not vary. No changes of stabilometric body sway parameters in eyes opened condition and with optokinetic stimulation delivered to the unaffected labyrinth were observed. A significant reduction was recorded in eyes closed condition and with the optokinetic stimulation toward the affected ear. The Tinnitus Handicap Inventory, the Situational Vertigo Questionnaire and the Numeric Pain Rating Scale scores improved. The number of vertigo attacks was reduced. Conclusions. Occlusal splint therapy is a favourable option to reduce aural symptoms of Ménière’s disease and comorbid temporomandibular joint disorder, even if its pathophysiological mechanism remains elusive.

2022 - Oral Cannabinoid Preparations for the Treatment of Chronic Migraine: A Retrospective Study [Articolo su rivista]
Baraldi, Carlo; Lo Castro, Flavia; Negro, Andrea; Ferrari, Anna; Cainazzo, Maria Michela; Pani, Luca; Guerzoni, Simona

Objective: To explore the effectiveness and safety of 3 oral cannabinoid preparations (FM2®, Bedrocan® and Bediol®) in the treatment of chronic migraine. Design: Retrospective, cohort study. Subjects: Patients with chronic migraine who received FM2®, Bedrocan® or Bediol® daily for the off-label treatment of their headache, up to 6 months. Methods: The number of migraine days per month, pain intensity, the number of acute medications taken per month, the number of days per month when the patient took at least one acute medication, and adverse events were recorded at baseline, 3 months, and 6 months after the start of treatment with oral cannabinoid preparations. Results: The number of migraine days didn't change significantly after the 3rd and the 6th month when compared to baseline (P = 0.1182). The pain intensity (P = 0.0004), the acute medication consumption (P = 0.0006) and the number of days per month in which patients took, at least, one acute medication, (P = 0.0004) significantly decreased when compared to the baseline. No significant differences were found between patients who were still taking a preventive treatment for chronic migraine and those who weren't (all P > 0.05). Different oral cannabinoid preparations displayed similar effectiveness (all P > 0.05). The AEs were mostly mild and occurred in the 43.75% of patients. Conclusions: Oral cannabinoid preparations may have a role in reducing pain intensity and acute medication intake in patients with chronic migraine, but the magnitude of the effect seems modest; further studies are needed.

2022 - Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors [Articolo su rivista]
Bettini, E.; Stahl, S. M.; De Martin, S.; Mattarei, A.; Sgrignani, J.; Carignani, C.; Nola, S.; Locatelli, P.; Pappagallo, M.; Inturrisi, C. E.; Bifari, F.; Cavalli, A.; Alimonti, A.; Pani, L.; Fava, M.; Traversa, S.; Folli, F.; Manfredi, P. L.

Excessive Ca2+ currents via N-methyl-D-aspartate receptors (NMDARs) have been implicated in many disorders. Uncompetitive NMDAR channel blockers are an emerging class of drugs in clinical use for major depressive disorder (MDD) and other neuropsychiatric diseases. The pharmacological characterization of uncompetitive NMDAR blockers in clinical use may improve our understanding of NMDAR function in physiology and pathology. REL-1017 (esmethadone-HCl), a novel uncompetitive NMDAR channel blocker in Phase 3 trials for the treatment of MDD, was characterized together with dextromethorphan, memantine, (±)-ketamine, and MK-801 in cell lines over-expressing NMDAR subtypes using fluorometric imaging plate reader (FLIPR), automated patch-clamp, and manual patch-clamp electrophysiology. In the absence of Mg2+, NMDAR subtypes NR1-2D were most sensitive to low, sub-μM glutamate concentrations in FLIPR experiments. FLIPR Ca2+ determination demonstrated low μM affinity of REL-1017 at NMDARs with minimal subtype preference. In automated and manual patch-clamp electrophysiological experiments, REL-1017 exhibited preference for the NR1-2D NMDAR subtype in the presence of 1 mM Mg2+ and 1 μM L-glutamate. Tau off and trapping characteristics were similar for (±)-ketamine and REL-1017. Results of radioligand binding assays in rat cortical neurons correlated with the estimated affinities obtained in FLIPR assays and in automated and manual patch-clamp assays. In silico studies of NMDARs in closed and open conformation indicate that REL-1017 has a higher preference for docking and undocking the open-channel conformation compared to ketamine. In conclusion, the pharmacological characteristics of REL-1017 at NMDARs, including relatively low affinity at the NMDAR, NR1-2D subtype preference in the presence of 1 mM Mg2+, tau off and degree of trapping similar to (±)-ketamine, and preferential docking and undocking of the open NMDAR, could all be important variables for understanding the rapid-onset antidepressant effects of REL-1017 without psychotomimetic side effects.

2022 - REL-1017 (Esmethadone) May Rapidly Reduce Dissociative Symptoms in Adults With Major Depressive Disorder Unresponsive to Standard Antidepressants: A Report of 2 Cases [Articolo su rivista]
Guidetti, Clotilde; Serra, Giulia; Pani, Luca; Pappagallo, Marco; Maglio, Gino; Trasolini, Monia; De Martin, Sara; Mattarei, Andrea; Bifari, Francesco; Folli, Franco; Manfredi, Paolo L; Fava, Maurizio

2022 - REL-1017 (Esmethadone) as Adjunctive Treatment in Patients With Major Depressive Disorder: A Phase 2a Randomized Double-Blind Trial [Articolo su rivista]
Fava, M.; Stahl, S.; Pani, L.; De Martin, S.; Pappagallo, M.; Guidetti, C.; Alimonti, A.; Bettini, E.; Mangano, R. M.; Wessel, T.; de Somer, M.; Caron, J.; Vitolo, O. V.; DiGuglielmo, G. R.; Gilbert, A.; Mehta, H.; Kearney, M.; Mattarei, A.; Gentilucci, M.; Folli, F.; Traversa, S.; Inturrisi, C. E.; Manfredi, P. L.

Objective: The purpose of this study was to examine the effects of REL-1017 (esmethadone), a novel N-methyl-d-aspartate receptor (NMDAR) channel blocker, in patients with major depressive disorder who failed to benefit from one to three standard antidepressant treatments in their current major depressive episode. Methods: A 7-day phase 2 multicenter randomized double-blind placebo-controlled trial, comprising three arms, was conducted to assess the safety, tolerability, pharmacokinetics, and efficacy of two dosages of REL-1017 (25 mg or 50 mg orally once a day). Patients were randomly assigned in a 1:1:1 ratio to placebo (N=22), REL-1017 25 mg/day (N=19), or REL-1017 50 mg/day (N=21). Safety scales included the 4-item Positive Symptom Rating Scale for psychotomimetic symptoms, the Clinician-Administered Dissociative States Scale for dissociative symptoms, the Clinical Opiate Withdrawal Scale for withdrawal signs and symptoms, and the Columbia-Suicide Severity Rating Scale for suicidality. The primary efficacy endpoint was the Montgomery-Åsberg Depression Scale (MADRS) score. All 62 randomly assigned patients were included in the full analysis set population analysis. Results: Patients experienced mild or moderate transient adverse events and no evidence of dissociative or psychotomimetic effects, opioid effects, or withdrawal signs and symptoms. The improvement in MADRS score shown on day 4 in both of the REL-1017 dosage groups was sustained through day 7 (last dose) and day 14 (7 days after the last dose), with effect sizes from 0.7 to 1.0. Conclusions: This trial showed favorable safety, tolerability, and pharmacokinetic profiles and suggests that REL-1017 may have rapid and sustained antidepressant effects compared with placebo in patients with inadequate responses to antidepressant treatments. These results will need confirmation in larger and longer trials.

2022 - Respondents of health survey powered by the innovative NURO app exhibit correlations between exercise frequencies and diet habits, and between stress levels and sleep wellness [Articolo su rivista]
Gallucci, D.; Ho, E. C. Y.; Geraci, J.; Loren, J.; Pani, L.

Nurosene's NURO app ( is an innovative smartphone application that gathers and analyzes active self-report metrics from users, empowering them with data-driven health machine intelligence. We present the data collected and analyzed from the initial round of participants who responded to a 12-question survey on their life-style and health status. Exploratory results using a variational autoencoder (VAE) suggested that much of the variability of the 12 dimensional data could be accounted for by two approximately uncorrelated latent variables: one pertaining to stress and sleep, and the other pertaining to exercise and diet. Subsequent modeling of the data using exploratory and confirmatory factor analyses (EFAs and CFAs) found that optimal data fits consisted of four factors, namely exercise, diet, stress, and sleep. Covariance values were high between exercise and diet, and between stress and sleep, but much lower between other pairings of non-identical factors. Both EFAs and CFAs provided extra contexts to and quantified the more preliminary VAE observations. Overall, our results significantly reduce the apparent complexity of the response data. This reduction allows for more efficient future stratification and analyses of participants based on simpler latent variables. Our discovery of novel relationships between stress and sleep, and between exercise and diet suggests the possibility of applying predictive analytics in future efforts.

2022 - Systematic Data Monitoring and Analysis of Cardiovascular Off-label Prescriptions in Pediatrics: Focus on Angiotensin-Converting Enzyme Inhibitors (ACE-I) and Beta Blockers [Articolo su rivista]
Cammarata, S. M.; Capone, G.; Lombardi, N.; Pani, L.; Mugelli, A.

Introduction: Many efforts have been made to stimulate clinical trials (CTs) in pediatrics but most of the drugs are still authorized only in adults and used off-label in the pediatric population. Aim: To assess how widespread is the off-label prescription in Italy and to identify areas of unmet medical need by applying a model for the systematic collection and analysis of data. Methods: A study was performed using 2015 data from the Italian Medicines Utilization Monitoring Centre Health Database (OsMed). A study sample of 3,726,583 pediatric patients, was considered. Cardiovascular drugs were selected for this study. Assessment of the off-label use, the analysis of the pharmacovigilance signals, a bibliographic research and the analysis of ongoing CTs were carried out. Results: In 2015, 8,544 pediatric patients received treatment with a cardiovascular drug. Angiotensin converting enzyme inhibitors (ACE-I) followed by beta blockers agents are the most prescribed molecules. Eight molecules were selected and an in-depth analysis conducted. The PhV network showed only one record of adverse reaction as off-label in 2015. The results show several therapeutic areas of use in pediatrics. Conclusion: Off-label in pediatrics is largely widespread in Europe and US and our results show it is also present in Italy. Molecules selected are used off-label for therapeutic areas such as oncologic, hematological and rare diseases. Results of pharmacovigilance suggests underreporting. The analysis carried out in this study could be an open track for a systematic monitoring activity and of interest for prescribers, pediatricians and other healthcare professionals during the clinical practice.

2022 - The Use of Psychotropic Medication in Pediatric Oncology for Acute Psychological and Psychiatric Problems: Balancing Risks and Benefits [Articolo su rivista]
Blom, Johanna M C; Barisone, Elena; Bertolotti, Marina; Caprino, Daniela; Cellini, Monica; Clerici, Carlo Alfredo; Colliva, Chiara; Favara-Scacco, Cinzia; Di Giuseppe, Silvia; Jankovic, Momcilo; Pancaldi, Alessia; Pani, Luca; Poggi, Geraldina; Rivi, Veronica; Tascedda, Fabio; Torta, Riccardo; Scarponi, Dorella

Severe acute behavioral and emotional problems represent one of the most serious treatment-related adverse effects for children and adolescents who have cancer. The critical and severe nature of these symptoms often makes necessary the use of psychotropic drugs. A working group composed of experts in multiple disciplines had the task of creating an agreement regarding a management plan for severe acute behavioral and emotional problems (SABEPs) in children and adolescents treated for cancer. To obtain global information on the use of psychotropic drugs in pediatric oncology, the working group first developed and mailed a 15-item questionnaire to many Italian pediatric oncology centers. Overall, an evident lack of knowledge and education regarding the use of psychotropic medications for the treatment of SABEPs was found. Thus, by referring to an adapted version of the Delphi method of consensus and standard methods for the elaboration of clinical questions (PICOs), the working group elaborated evidence-based recommendations for psychotropic drugs in the pediatric oncology setting. Furthermore, based on a thorough multivariate analysis of needs and difficulties, a comprehensive management flow was developed to optimize therapeutic interventions, which allows more accurate and efficient matching of the acute needs of patients while guiding treatment options.

2022 - The association between onabotulinumtoxinA and anti-CGRP monoclonal antibodies: a reliable option for the optimal treatment of chronic migraine [Articolo su rivista]
Guerzoni, S.; Baraldi, C.; Pani, L.

Chronic migraine (CM) is a great challenge for physicians dealing with headaches. Despite the introduction of the monoclonal antibodies (mAbs) acting against the calcitonin gene-related peptide (CGRP) that has revolutionized the treatment of CM, some patients still experience an incomplete relief. So, the association of two preventive treatments may be a reliable option for these patients. So, onabotulinumtoxinA (BT-A) and anti-CGRP mAbs may be used together, and some pre-clinical and clinical evidence of an additive action of the 2 drugs is emerging. In particular, since BT-A acts mainly on C-fibers and anti-CGRP mAbs on Aδ ones, their association may prevent the wearing-off phenomenon of BT-A, thus giving an additional benefit in those patients experiencing an incomplete response to BT-A alone. Despite this, the clinical studies available in the literature have a small sample size, often a retrospective design, and are heterogeneous in terms of the outcomes chosen. Considering this, the evidence of a favorable effect of the association between BT-A and anti-CGRP mAbs is still scarce. Furthermore, this association is explicitly forbidden by many National regulatory agencies, due to the high costs of both treatments. Anyway, their association could help in reducing the burden associated with the most severe cases of CM, thus relieving the direct and indirect costs of this condition. More well-designed studies with big samples are needed to unveil the real therapeutic gain of this association. Moreover, pharmacoeconomics studies should be performed, to assess the economic suitability of this association.

2021 - A Delphi-method-based consensus guideline for definition of treatment-resistant depression for clinical trials [Articolo su rivista]
Sforzini, L.; Worrell, C.; Kose, M.; Anderson, I. M.; Aouizerate, B.; Arolt, V.; Bauer, M.; Baune, B. T.; Blier, P.; Cleare, A. J.; Cowen, P. J.; Dinan, T. G.; Fagiolini, A.; Ferrier, I. N.; Hegerl, U.; Krystal, A. D.; Leboyer, M.; McAllister-Williams, R. H.; Mcintyre, R. S.; Meyer-Lindenberg, A.; Miller, A. H.; Nemeroff, C. B.; Normann, C.; Nutt, D.; Pallanti, S.; Pani, L.; Penninx, B. W. J. H.; Schatzberg, A. F.; Shelton, R. C.; Yatham, L. N.; Young, A. H.; Zahn, R.; Aislaitner, G.; Butlen-Ducuing, F.; Fletcher, C.; Haberkamp, M.; Laughren, T.; Mantyla, F. -L.; Schruers, K.; Thomson, A.; Arteaga-Henriquez, G.; Benedetti, F.; Cash-Gibson, L.; Chae, W. R.; De Smedt, H.; Gold, S. M.; Hoogendijk, W. J. G.; Mondragon, V. J.; Maron, E.; Martynowicz, J.; Melloni, E.; Otte, C.; Perez-Fuentes, G.; Poletti, S.; Schmidt, M. E.; van de Ketterij, E.; Woo, K.; Flossbach, Y.; Ramos-Quiroga, J. A.; Savitz, A. J.; Pariante, C. M.

Criteria for treatment-resistant depression (TRD) and partially responsive depression (PRD) as subtypes of major depressive disorder (MDD) are not unequivocally defined. In the present document we used a Delphi-method-based consensus approach to define TRD and PRD and to serve as operational criteria for future clinical studies, especially if conducted for regulatory purposes. We reviewed the literature and brought together a group of international experts (including clinicians, academics, researchers, employees of pharmaceutical companies, regulatory bodies representatives, and one person with lived experience) to evaluate the state-of-the-art and main controversies regarding the current classification. We then provided recommendations on how to design clinical trials, and on how to guide research in unmet needs and knowledge gaps. This report will feed into one of the main objectives of the EUropean Patient-cEntric clinicAl tRial pLatforms, Innovative Medicines Initiative (EU-PEARL, IMI) MDD project, to design a protocol for platform trials of new medications for TRD/PRD.

2021 - Biomarkers and Surrogate End points in Multiple Sclerosis Trials: Regulatory Issues [Articolo su rivista]
Skeen, M. B.; Pani, L.

Summary:Evoked potentials have assisted in the diagnosis of multiple sclerosis for years, but the potential to demonstrate pathophysiologic change has prompted a reconsideration of their potential role as outcome measures in clinical trials of multiple sclerosis. The use of any surrogate end point or biomarker in clinical trials requires a thorough understanding of that end point's performance characteristics and utility in a particular setting. This article explores regulatory issues regarding the use of biomarkers and surrogate end points in clinical trials of multiple sclerosis with particular emphasis on challenges faced by evoked potential studies.

2021 - Chronic Migraine and Medication Overuse Headache Worsening After OnabotulinumtoxinA Withdrawn Due to the Severe Acute Respiratory Syndrome-Coronavirus-2 Pandemic [Articolo su rivista]
Baraldi, C.; Ornello, R.; Favoni, V.; Sacco, S.; Caponnetto, V.; Pierangeli, G.; Pani, L.; Cevoli, S.; Guerzoni, S.

Introduction: OnabotulinumtoxinA (BT-A) is a preventive treatment for chronic migraine (CM), which needs to be administered regularly by a trained clinician every 3 months. The spread of the severe acute respiratory syndrome coronavirus-2 pandemic has forced many patients to momentarily stop the scheduled BT-A injections. The goal of this study was to explore whether those patients experienced a worsening of their CM and, if any, the clinical predictors of migraine worsening after BT-A withdrawal. Methods: This was a retrospective, multicenter study. Patients' clinical data were obtained from their clinical documentation stored at each center. In particular, the following variables were collected: the mean number of headache days in the last month (NHD), the average number of painkillers taken in the last month (AC), the average number of days in which patients took, at least, one painkiller in the last month (NDM), the average intensity of migraine using the numeric rating scale (NRS) score in the last month, and the average score obtained at the six-item Headache Impact Test. The variables mentioned earlier were compared before and after BT-A withdrawal. Results: After BT-A suspension, there was a significant increase in the NHD (P = 0.0313, Kruskal–Wallis rank test), AC (P = 0.0421, Kruskal–Wallis rank test), NDM (P = 0.0394, paired t-test), NRS score (P = 0.0069, Kruskal–Wallis rank test), and six-item Headache Impact Test score (P = 0.0372, Kruskal–Wallis rank test). Patients who were not assuming other preventive treatments other than BT-A displayed similar results. Patients who experienced a >30% worsening in NHD after BT-A was withdrawn displayed a longer CM history (P = 0.001, Kruskal–Wallis rank test), a longer MOH duration (P = 0.0017, Kruskal–Wallis rank test), a higher AC value at the baseline (P = 0.0149, Kruskal–Wallis rank test), a higher NDM (P = 0.0024, t-test), and a higher average value of the NRS score (P = 0.0073, Kruskal–Wallis rank test). Conclusion: BT-A withdrawn during severe acute respiratory syndrome coronavirus-2 pandemic was associated with a general worsening in patients suffering from CM, hence the need to continue BT-A injection to avoid patients' worsening.

2021 - Digital Phenotyping and Dynamic Monitoring of Adolescents Treated for Cancer to Guide Intervention: Embracing a New Era [Articolo su rivista]
Blom, J. M. C.; Colliva, C.; Benatti, C.; Tascedda, F.; Pani, L.

2021 - Erenumab for the preventive treatment of chronic migraine complicated with medication overuse headache: an observational, retrospective, 12-month real-life study [Articolo su rivista]
Michela Cainazzo, Maria; Baraldi, Carlo; Ferrari, Anna; Lo Castro, Flavia; Pani, Luca; Guerzoni, Simona

Background Erenumab is a monoclonal antibody blocking the calcitonin gene–related peptide receptor, which has been approved for the preventive treatment of chronic migraine (CM). The aim of this study was to explore the safety and effectiveness of erenumab in patients suffering from CM and medication overuse headache (MOH) in a real-life setting, up to 1 year. Methods Data regarding 81 patients treated with erenumab were retrospectively analyzed. Every 3 months, the following variables were collected: the mean number of headache days per month (headache index (HI)), the average number of painkillers taken per month (analgesic consumption (AC)), the mean number of days with painkiller consumption (number of days on medication (NDM)), the headache intensity (numeric rating scale (NRS) score), the 6-item Headache Impact Test (HIT-6), and the Self-Reported Instrument to Assess Work-Related Difficulties in Patients With Migraine (HEADWORK) scores. Results The HI, AC, and NDM and the NRS, HIT-6, and HEADWORK scores were significantly lower at every time point from the 3rd month onward compared to baseline (all P < 0.0001). No significant differences were found between patients who underwent painkiller detoxification before starting erenumab and those who did not (all P > 0.05). No significant differences were found between patients taking erenumab in combination with other preventive treatments and the ones taking it alone (all P ≥ 0.05). Five patients dropped out because of adverse events, which resolved after stopping erenumab. Conclusion Erenumab was safe and effective for CM complicated with MOH. Painkiller withdrawal and the association with other preventive treatment(s) seem useless.

2021 - New Models for the Evaluation of Specialized Medicinal Products: Beyond Conventional Health Technology Assessment and Pricing [Articolo su rivista]
Pani, L.; Becker, K.

New specialized therapeutics coming to market, such as advanced therapy medicinal products (ATMPs) and orphan drugs, differ from traditional therapies in terms of how they are manufactured and administered, as well as the potentially transformative benefits they may provide. The current health technology assessment (HTA) process that has been used for traditional therapies, such as small molecule drugs and antibodies, does not work adequately for specialized therapeutics, with a key issue being the generation of sufficient evidence to adequately capture the full long-term benefits. The objectives of this article are to discuss why the current HTA process is inadequate for evaluating these new therapies, how evidence should be continuously generated and presented to regulators and payers to support their use, and to propose new approaches to pricing models. This will enable payers to have an affordable, risk-mitigated means of funding new therapies in a timely manner, thus guaranteeing patient access to new, potentially life-saving therapies, while providing manufacturers with a return on their investment. Without new approaches or adaptation of existing frameworks, certain ATMPs may not reach patients in some or all countries or be at risk of withdrawal from the market.

2021 - Predictors of response to erenumab after 12 months of treatment [Articolo su rivista]
Baraldi, C.; Castro, F. L.; Cainazzo, M. M.; Pani, L.; Guerzoni, S.

Objective: Erenumab is a monoclonal antibody acting against calcitonin gene-related peptide receptor and approved for the preventive treatment of chronic migraine. The aim of the present study is to identify clinical predictors of good response in patients with chronic migraine and medication overuse-headache. Material and methods: This was a retrospective single-center not funded study. Enrolled patients were affected by chronic migraine and medication overuse-headache treated with erenumab monthly, up to 1 year. At 1 year, patients were classified as good responders if they displayed a ≥50% reduction in the number of headache days per months compared to the baseline. Results: After 1 year, a significant improvement in the number of headache days per months, analgesic consumption, 6-items headache impact test, and migraine disability assessment questionnaire scores were obtained compared to the baseline. Patients who obtained a ≥50% reduction in the number of headache days per month compared to the baseline displayed a longer history of medication overuse-headache, a higher number of painkillers taken per month at the baseline and a higher number of failed preventive treatments in the past. Conclusions: Patients with longer medication overuse-headache duration, higher analgesic intake, and a higher number of previous preventive treatment failures may receive less benefit with erenumab.

2020 - A case-control study of visually evoked postural responses in childhood with primary headaches [Articolo su rivista]
Baraldi, C; Gherpelli, C; Alicandri Ciufelli, M.; Monzani, D.; Pini, L. A.; Pani, L.; Guerzoni, S.

Background: Disorientation, nausea, confusion, dizziness, and displacement are frequently complained by headache-suffering children. Anyhow, the cause of these symptoms is still unclear, and a dysfunction of vestibular pathways or their alteration due to central pain pathways hyper-activation, has been proposed. The aim of this study is to use posturography to explore the balance function of headache-suffering children during pain-free periods. Methods: Posturography was performed on 19 migraineurs, 11 tension-type headache sufferers, and 20 healthy controls. Posturographic measures were performed during headache-free periods under different conditions: with eyes opened, eyes closed, and during right and left optokinetic stimulation. The last 2 conditions were used to mimic unreliable visual signals that can confound vestibular system. Results: During eyes-closed conditions, headache-suffering children displayed higher displacements than healthy controls, since statokinesiogram surface was higher in tension-type headache sufferers and migraineurs compared with controls (P value = 0.0095). Romberg’s index, indicating the overall stability of the subject, was lower in healthy controls than in headache sufferers (P = 0.0139), thus suggesting a vestibular impairment in the seconds. Moreover, both during right and left optokinetic stimulation, the statokinesiogram length was higher in headache-suffering children (P < 0.0001). Thereafter, statokinesiogram surface was higher in migraineurs during right optokinetic stimulation (P = 0.0388) than in tension-type headache sufferers when stimulation was directed on the opposite side (P = 0.0249). Conclusions: These results suggest a central alteration of vestibular pathways in headache-suffering children, that makes balance function more dependent from visual inputs than healthy subjects, even in inter-ictal phases.

2020 - Impact of reimbursement limits on patient access to direct-acting antivirals in Italy: Analysis of data from national registries [Articolo su rivista]
Russo, P.; Pani, L.; Staniscia, T.; Romano, F.; Marzioni, M.

OBJECTIVE: Hepatitis C virus (HCV) infection is a global epidemic, still highly prevalent in Europe. Given efficacy and safety of HCV therapy by Direct Antiviral Agents (DAA), World Health Organization called for actions to eliminate HCV infection. A limit is represented by access to care, mostly due to the high costs of medicines. In Italy, in 2015, the access to DAA therapy was reimbursed for patients with advanced disease, whereas in 2017 universal access was granted. The aim of this study was to analyse changes in patient recruitment trends treated with DAA with or without limitations to access to therapy. PATIENTS AND METHODS: 165,105 patients treated with DAA in Italy from 2015 to December 2018 were analysed. Daily patient treatment rate was obtained by segmented regression of interrupted time series analysis. RESULTS: 74,199 patients with advanced disease (62% with cirrhosis) had access to the therapy during the time period from 2015 to 2017. Following the extension of reimbursement criteria, 90,906 additional patients were treated (43.2% with F0-F1 and 22.9% with F2), with an absolute reduction of 59.9% of patients with advanced disease (cirrhosis decreased to 18.5%). Segmented regression of interrupted time series analysis of daily patient treatment rate showed a progressive reduction of patients with advanced disease, offset by those with initial disease. Notably, elimination of restrictions to therapy did not change the overall treatment rate. CONCLUSIONS: This study showed that a no-limit reimbursement policy for DAAs prescriptions to HCV infected individuals in Italy widened the types of treated patients, but the process towards elimination of HCV infection was not significantly changed.

2020 - Should Patients Receiving ACE Inhibitors or Angiotensin Receptor Blockers be Switched to Other Antihypertensive Drugs to Prevent or Improve Prognosis of Novel Coronavirus Disease 2019 (COVID-19)? [Articolo su rivista]
Trifiro, G.; Crisafulli, S.; Ando, G.; Racagni, G.; Drago, F.; Berrino, L.; Re, M.; Bernardini, R.; Chiamulera, C.; D'Avolio, A.; Pani, L.; Clementi, E.; Capuano, A.; Scaglione, F.; Danesi, R.; Cirino, G.; Mugelli, A.; Bonanno, G.; Brunello, N.; Luca, A.; Hrelia, P.; Pistis, M.; Ghelardini, C.; Taglialatela, M.

2020 - The endless frontier? The recent increase of R&D productivity in pharmaceuticals [Articolo su rivista]
Pammolli, F.; Righetto, L.; Abrignani, S.; Pani, L.; Pelicci, P. G.; Rabosio, E.

Background: Studies on the early 2000s documented increasing attrition rates and duration of clinical trials, leading to a representation of a "productivity crisis" in pharmaceutical research and development (R&D). In this paper, we produce a new set of analyses for the last decade and report a recent increase of R&D productivity within the industry. Methods: We use an extensive data set on the development history of more than 50,000 projects between 1990 and 2017, which we integrate with data on sales, patents, and anagraphical information on each institution involved. We devise an indicator to quantify the novelty of each project, based on its set of mechanisms of action. Results: First, we investigate how R&D projects are allocated across therapeutic areas and find a polarization towards high uncertainty/high potential reward indications, with a strong focus on oncology. Second, we find that attrition rates have been decreasing at all stages of clinical research in recent years. In parallel, for each phase, we observe a significant reduction of time required to identify projects to be discontinued. Moreover, our analysis shows that more recent successful R&D projects are increasingly based on novel mechanisms of action and target novel indications, which are characterized by relatively small patient populations. Third, we find that the number of R&D projects on advanced therapies is also growing. Finally, we investigate the relative contribution to productivity variations of different types of institutions along the drug development process, with a specific focus on the distinction between the roles of Originators and Developers of R&D projects. We document that in the last decade Originator-Developer collaborations in which biotech companies act as Developers have been growing in importance. Moreover, we show that biotechnology companies have reached levels of productivity in project development that are equivalent to those of large pharmaceutical companies. Conclusions: Our study reports on the state of R&D productivity in the bio-pharmaceutical industry, finding several signals of an improving performance, with R&D projects becoming more targeted and novel in terms of indications and mechanisms of action.

2019 - Approaches to attenuated psychosis syndrome treatments: A perspective on the regulatory issues [Articolo su rivista]
Pani, L.; Keefe, R. S. E.

2019 - Microdosing psychedelics: More questions than answers? An overview and suggestions for future research [Articolo su rivista]
Kuypers, K. P. C.; Ng, L.; Erritzoe, D.; Knudsen, G. M.; Nichols, C. D.; Nichols, D. E.; Pani, L.; Soula, A.; Nutt, D.

Background: In the past few years, the issue of ‘microdosing’ psychedelics has been openly discussed in the public arena where claims have been made about their positive effect on mood state and cognitive processes such as concentration. However, there are very few scientific studies that have specifically addressed this issue, and there is no agreed scientific consensus on what microdosing is. Aim: This critique paper is designed to address questions that need to be answered by future scientific studies and to offer guidelines for these studies. Approach: Owing to its proximity for a possible approval in clinical use and short-lasting pharmacokinetics, our focus is predominantly on psilocybin. Psilocybin is allegedly, next to lysergic acid diethylamide (LSD), one of the two most frequently used psychedelics to microdose. Where relevant and available, data for other psychedelic drugs are also mentioned. Conclusion: It is concluded that while most anecdotal reports focus on the positive experiences with microdosing, future research should also focus on potential risks of (multiple) administrations of a psychedelic in low doses. To that end, (pre)clinical studies including biological (e.g. heart rate, receptor turnover and occupancy) as well as cognitive (e.g. memory, attention) parameters have to be conducted and will shed light on the potential negative consequences microdosing could have.

2018 - Comparing safety information of biosimilars with their originators: a cross-sectional analysis of European risk management plans [Articolo su rivista]
Lepelaars, Leroy R. A.; Renda, Francesca; Pani, Luca; Pimpinella, Giuseppe; Leufkens, Hubert G. M.; Trifirò, Gianluca; Tafuri, Giovanni; Mantel-Teeuwisse, Aukje K.; Trotta, Francesco

Background and aims: Biosimilars have been available in the European Union (EU) since 2006. However, their uptake in routine care is heterogeneous across countries. The aim of the present study was to compare the safety information of biosimilars and their originators based on the information in the European risk management plan (RMP). Methods: A cross-sectional analysis on publicly available regulatory documents (RMPs and Summaries of Product Characteristics) of biosimilars and corresponding originators up to 1 November 2015 was performed. The safety concerns were extracted and merged into general safety concerns, and clinical relevance was assessed. The frequency of safety concerns and the representation of these safety concerns per general safety concern were assessed by either comparing RMPs of biosimilars and originators (if available for both) or comparing RMPs with the Summary of Product Characteristics of the originator. Results: Nineteen biosimilars and six originators were included. Overall, 55 general safety concerns (12 low, 21 medium and 22 highly clinically relevant) were identified. For all active substances, except for infliximab, no or only one difference was found in the listed general safety concerns. Comparison of regulatory documents for infliximab identified three medium clinically relevant general safety concerns more for infliximab biosimilars and two general safety concerns more for its originator. Conclusion: Based on publicly available information filed for regulatory purposes, no substantial differences were observed in the reporting of safety information for biosimilars and related originators. A direct comparison between biosimilars and related originators through formal postmarketing studies is needed to evaluate specific safety issues emerging during the products’ life cycle.

2018 - Nutraceuticals: opening the debate for a regulatory framework [Articolo su rivista]
Santini, Antonello; Cammarata, Silvia Miriam; Capone, Giacomo; Ianaro, Angela; Tenore, Gian Carlo; Pani, Luca; Novellino, Ettore

Currently, nutraceuticals do not have a specific definition distinct from those of other food-derived categories, such as food supplements, herbal products, pre- and probiotics, functional foods, and fortified foods. Many studies have led to an understanding of the potential mechanisms of action of pharmaceutically active components contained in food that may improve health and reduce the risk of pathological conditions while enhancing overall well-being. Nevertheless, there is a lack of clear information and, often, the claimed health benefits may not be properly substantiated by safety and efficacy information or in vitro and in vivo data, which can induce false expectations and miss the target for a product to be effective, as claimed. An officially shared and accepted definition of nutraceuticals is still missing, as nutraceuticals are mostly referred to as pharma-foods, a powerful toolbox to be used beyond the diet but before the drugs to prevent and treat pathological conditions, such as in subjects who may not yet be eligible for conventional pharmaceutical therapy. Hence, it is of utmost importance to have a proper and unequivocal definition of nutraceuticals and shared regulations. It also seems wise to assess the safety, mechanism of action and efficacy of nutraceuticals with clinical data. A growing demand exists for nutraceuticals, which seem to reside in the grey area between pharmaceuticals and food. Nonetheless, given specific legislation from different countries, nutraceuticals are experiencing challenges with safety and health claim substantiation.

2018 - Safety and efficacy of ombitasvir/paritaprevir/ritonavir/dasabuvir plus ribavirin in patients over 65 years with HCV genotype 1 cirrhosis [Articolo su rivista]
Ascione, Antonio; De Luca, Massimo; Melazzini, Mario; Montilla, Simona; Trotta, Maria Paola; Petta, Salvatore; Puoti, Massimo; Sangiovanni, Vincenzo; Messina, Vincenzo; Bruno, Savino; Izzi, Antonio; Villa, Erica; Aghemo, Alessio; Zignego, Anna Linda; Orlandini, Alessandra; Fontanella, Luca; Gasbarrini, Antonio; Marzioni, Marco; Giannini, Edoardo G.; Craxì, Antonio; Abbati, Giuseppe; Alberti, Alfredo; Andreone, Pietro; Andreoni, Massimo; Angeli, Paolo; Angelico, Mario; Angarano, Gioacchino; Angrisani, Debora; Antinori, Andrea; Antonini, Cinzia; Avancini, Ivo; Barone, Michele; Bruno, Raffaele; Benedetti, Antonio; Bernabucci, Veronica; Blanc, Pier; Boarini, Chiara; Boffa, Nicola; Boglione, Lucio; Borghi, Vanni; Borgia, Guglielmo; Brancaccio, Giuseppina; Brunetto, Maurizia; Cacciola, Irene; Calabrese, Paolo; Calvaruso, Vincenza; Campagnolo, Davide; Canovari, Benedetta; Caporaso, Nicola; Capra, Franco; Carolo, Giada; Cassola, Giovanni; Castelli, Francesco; Cauda, Roberto; Silberstein, Francesca Ceccherini; Cecere, Roberto; Chessa, Luchino; Chiodera, Alessandro; Chirianni, Antonio; Ciancio, Alessia; Cima, Serena; Coco, Barbara; Colombo, Massimo; Coppola, Nicola; Corti, Giampaolo; Cosco, Lucio; Corradori, Silvia; Cozzolongo, Raffaele; Cristaudo, Antonio; Danieli, Elena; Monforte, Antonella D’Arminio; Monache, Marco delle; Del Poggio, Paolo; de Luca, Andrea; Dentone, Chiara; Di Biagio, Antonio; Di Leo, Alfredo; Di Perri, Giovanni; Di Stefano, Marco; D’Offizi, Giampiero; Donato, Francesca; Durante, Emanuele; Erne, Elke; Fagiuoli, Stefano; Falasca, Katia; Federico, Alessandro; Felder, Martina; Ferrari, Carlo; Gaeta, Giovanni Battista; Ganga, Roberto; Gatti, Pietro; Giacomet, Vania; Giacometti, Andrea; Gianstefani, Alice; Giordani, Maria; Giorgini, Alessia; Grieco, Antonio; Guerra, Michele; Gulminetti, Roberto; Ieluzzi, Donatella; Imparato, Michele; Iodice, Valentina; La Monica, Silvia; Lazzarin, Adriano; Lenzi, Marco; Levrero, Massimo; Lichtner, Myriam; Lionetti, Raffaella; Guercio, Carmela Lo; Madonna, Salvatore; Magnani, Silvia; Maida, Ivana; Marignani, Massimo; Marrone, Aldo; Marsetti, Fabio; Martini, Silvia; Masarone, Mario; Maserati, Renato; Mastroianni, Claudio Maria; Memoli, Massimo; Menzaghi, Barbara; Merli, Manuela; Miele, Luca; Milella, Michele; Mondelli, Mario; Montalbano, Marzia; Monti, Monica; Morelli, Olivia; Morisco, Filomena; Nardone, Gaetano; Novara, Sergio; Onnelli, Giovanna; Onofrio, Mirella; Paganin, Simona; Pani, Luca; Parisi, Maria Rita; Parruti, Giustino; Pasquazzi, Caterina; Pasulo, Luisa; Perno, Carlo Federico; Persico, Marcello; Piai, Guido; Picciotto, Antonino; Pigozzi, Grazielle Marie; Piovesan, Sara; Piras, Maria Chiara; Pirisi, Massimo; Piscaglia, Anna Maria; Ponti, Laura; Potenza, Domenico; Pravadelli, Cecilia; Quartini, Mariano; Quirino, Tiziana; Raimondo, Giovanni; Rapaccini, Gian Ludovico; Rendina, Maria; Rizzardini, Giuliano; Rizzetto, Mario; Rizzo, Salvatore; Romagnoli, Dante; Romano, Antonietta; Rossi, Cristina; Rumi, Maria Grazia; Russello, Maurizio; Russo, Francesca Paolo; Russo, Maria Luisa; Sansonno, Domenico Ettore; Santantonio, Teresa Antonia; Saracco, Giorgio; Schimizzi, Anna Maria; Serviddio, Gaetano; Simeone, Filomena; Solinas, Attilio; Soria, Alessandro; Tabone, Marco; Taliani, Gloria; Tarantino, Giuseppe; Tarquini, Pierluigi; Tavio, Marcello; Termite, Antonio; Teti, Elisabetta; Toniutto, Pierluigi; Torti, Carlo; Tundi, Paolo; Vecchiet, Giacomo; Verucchi, Gabriella; Gentilucci, Umberto Vespasiani; Vinci, Maria; Vullo, Vincenzo; Zolfino, Teresa; Zuin, Massimo

Purpose: To analyse safety and efficacy of treatment based on ombitasvir/paritaprevir/ritonavir/dasabuvir plus ribavirin in the sub-group of GT1 patients older than 65 years. Methods: We collected data extracted from the ABACUS compassionate-use nationwide Italian programme, in patients with cirrhosis due to hepatitis C virus (HCV) Genotype-1 (GT1) or 4 and at high risk of decompensation. GT1-HCV-infected patients received once-daily ombitasvir/paritaprevir, with the pharmacokinetic enhancer ritonavir (25/150/100 mg) and twice-daily dasabuvir (250 mg) plus Ribavirin (RBV) (OBV/PTV/r + DSV + RBV) for 12 (GT1b) or 24 (GT1a) weeks. Endpoints were to evaluate safety and efficacy, the latter defined as HCV RNA negative 12 weeks after the end of treatment (SVR12). Results: Patients who suffered any adverse event (AE) were 74/240 (30.8%); 13/240 (5.4%) discontinued the treatment. A multivariate analysis found albumin < 3.5 g/dL (OR 2.04: 95% CI 1.0–4.2, p < 0.05) and hypertension (OR 4.6: 95% CI 2.3–9.2, p < 0.001) as variables independently associated with AE occurrence. The SVR12 was 95% (228/240). Multivariate analysis identified baseline bilirubin < 2 mg/dL (OR 4.9: 95% CI 1.17–20.71, p = 0.029) as the only variable independently associated with SVR12. Conclusion: Our findings suggest that OBV/PTV/r + DSV + RBV is safe and effective in real-life use in patients with compensated cirrhosis, HCV-GT1 infection, and age over 65.

2018 - Take This Cognitive Training Efficacy Bar Fight Outside (to a Regulatory Agency) [Articolo su rivista]
Richard S. E., Keefe; Pani, Luca

Scientific and Regulatory Commentary to Harvey et al. Computerized cognitive training (CCT) to improve cognitive functioning is of enormous interest and has been applied in a broad range of populations with goals of improving both cognition and community functioning. Recent reviews presenting negative conclusions about CCT efficacy have inconsistent definitions of the treatment targets and cognitive improvement. They do not present an accurate representation of the typical process of CCT and cognitive remediation (CR), especially as delivered in major mental illnesses such as schizophrenia. This review provides guidance on the definitions of CCT and CR, the uses of CCT and CR, and the definitions and measurements of cognitive and functional gains. The review focuses on schizophrenia and healthy aging, with each population receiving unique CCT or CR approaches and substantial extant literature with which to elucidate fundamental CCT and CR concepts and research findings. It is our conclusion that CCT has been shown in most studies to improve cognitive performance on untrained tests in healthy older people and in people with schizophrenia. Functional gains in schizophrenia appear to be limited to CR studies. Clearly defining CCT, CR, and levels of treatment-related gains will be critical for understanding the benefits of these widely used treatment programs

2018 - The Italian compassionate use of sofosbuvir in HCV patients waitlisted for liver transplantation: A national real-life experience [Articolo su rivista]
Martini, Silvia; Donato, Maria Francesca; Mazzarelli, Chiara; Rendina, Maria; Visco-Comandini, Ubaldo; Filì, Daniela; Gianstefani, Alice; Fagiuoli, Stefano; Melazzini, Mario; Montilla, Simona; Pani, Luca; Petraglia, Sandra; Russo, Pierluigi; Trotta, Maria Paola; Carrai, Paola; Caraceni, Paolo; Angeli, Paolo; Ballardini, Giorgio; Bernabucci, Veronica; Bhoori, Sherrie; Burra, Patrizia; Civolani, Alberto; D'Offizi, Gianpiero; Felder, Martina; Gaeta, Giovanni Battista; Ganga, Roberto; Ginanni Corradini, Stefano; Iemmolo, Rosa Maria; Lenci, Ilaria; Lionetti, Raffaella; Montalbano, Marzia; Morelli, Maria Cristina; Picciotto, Antonino; Sapere, Cristina; Serviddio, Gaetano; Tamè, Mariarosa; Verucchi, Gabriella; Zignego, Anna Linda

Background & Aims: This study aimed to assess the real-life clinical and virological outcomes of HCV waitlisted patients for liver transplantation (LT) who received sofosbuvir/ribavirin (SOF/R) within the Italian compassionate use program. Methods: Clinical and virological data were collected in 224 patients with decompensated cirrhosis and/or hepatocellular carcinoma (HCC) receiving daily SOF/R until LT or up a maximum of 48 weeks. Results: Of 100 transplanted patients, 51 were HCV-RNA negative for >4 weeks before LT (SVR12: 88%) and 49 negative for <4 weeks or still viraemic at transplant: 34 patients continued treatment after LT (bridging therapy) (SVR12: 88%), while 15 stopped treatment (SVR12: 53%). 98 patients completed SOF/R without LT (SVR12: 73%). In patients with advanced decompensated cirrhosis (basal MELD ≥15 and/or C-P ≥B8), a marked improvement of the scores occurred in about 50% of cases and almost 20% of decompensated patients without HCC reached a condition suitable for inactivation and delisting. Conclusions: These real-life data indicate that in waitlisted patients: (i) bridging antiviral therapy can be an option for patients still viraemic or negative <4 weeks at LT; and (ii) clinical improvement to a condition suitable for delisting can occur even in patients with advanced decompensated cirrhosis.

2017 - Association of neovascular age-related macular degeneration with month and season of birth in Italy [Articolo su rivista]
Longo, Antonio; Casuccio, Alessandra; Pani, Luca; Avitabile, Teresio; Cillino, Salvatore; Uva, Maurizio G.; Bonfiglio, Vincenza; Russo, Andrea; Parisi, Guglielmo; Cennamo, Gilda; Furino, Claudio; Parravano, Mariacristina; Xoxi, Entela; Reibaldi, Michele

In order to investigate the influence of season and month of birth on the risk of neovascular age-related macular degeneration (n-AMD) in Italy, we evaluated the month birth and sex of all patients, recorded in the anti-vascular endothelial growth factor (VEGF) monitoring registry of the Italian Medicines Agency, born between 1925-1944, who received intravitreal anti-VEGF injections for n-AMD between January 1, 2013 and July 29, 2015. The numbers of all births in Italy in the same years, extracted from the Italian National Institute of Statistics, were used to calculate the expected number of n-AMD cases. Overall, 45,845 patients (19,207 men, 26,638 women) received intravitreal anti-VEGF for n-AMD; in the same years, 20,140,426 people (10,334,262 male, 9,806,164 female) were born in Italy. Comparing the observed number of n-AMD cases with the expected number of n- AMD cases in each season, we found that the season-specific risk for n-AMD was 2.5% higher for those born in summer (OR=1.03, Bonferroni-corrected P=0.008) and 3% lower for those born in winter (OR=0.96, Bonferroni-corrected P=0.0004). When considering the month of birth, the risk of n-AMD was 5.9% lower for people born in January (OR=0.93, Bonferroni-corrected P=0.0012). The factors causing such differences should be determined.

2017 - Italy: Post-marketing successful strategies to manage pharmaceutical innovation [Capitolo/Saggio]
Cicchetti, A.; Coretti, S.; Iacopino, V.; Montilla, S.; Xoxi, E.; Pani, L.

2017 - Ombitasvir, paritaprevir, and ritonavir, with or without dasabuvir, plus ribavirin for patients with hepatitis C virus genotype 1 or 4 infection with cirrhosis (ABACUS): a prospective observational study [Articolo su rivista]
Petta, Salvatore; Marzioni, Marco; Russo, Pierluigi; Aghemo, Alessio; Alberti, Alfredo; Ascione, Antonio; Antinori, Andrea; Bruno, Raffaele; Bruno, Savino; Chirianni, Antonio; Gaeta, Giovanni Battista; Giannini, Edoardo G; Merli, Manuela; Messina, Vincenzo; Montilla, Simona; Perno, Carlo Federico; Puoti, Massimo; Raimondo, Giovanni; Rendina, Maria; Silberstein, Francesca Ceccherini; Villa, Erica; Zignego, Anna Linda; Pani, Luca; Craxì, Antonio; Tabone, Marco; Andreoni, Massimo; Teti, Elisabetta; Angelico, Mario; Persico, Marcello; Masarone, Mario; Chiodera, Aledssandro; Solinas, Attilio; delle Monache, Marco; Cecere, Roberto; Maria Schimizzi, Anna; Piovesan, Sara; Campagnolo, Davide; Chiara Piras, Maria; Zolfino, Teresa; Paolo Russo, Francesca; Morelli, Olivia; Sangiovanni, Vincenzo; Onofrio, Mirella; Iodice, Valentina; Izzi, Antonio; Pirisi, Massimo; Danieli, Elena; Vinci, Maria; Rizzardini, Giuliano; Fagiuoli, Stefano; Pasulo, Luisa; D'Arminio Monforte, Antonella; Zuin, Massimo; Giorgini, Alessia; Simeone, Filomena; Piali, Guido; Lo Guercio, Carmela; Federico, Alessandro; Brancaccio, Giuseppina; Marrone, Aldo; Abbati, Giuseppe; Boarini, Chiara; Borghi, Vanni; Bernabucci, Veronica; Corti, Giampaolo; Monti, Monica; Rizzetto, Mario; Martini, Silvia; Andreone, Pietro; Gianstefani, Alice; Lenzi, Marco; Verucchi, Gabriella; Toniutto, Pierluigi; Borgia, Guglielmo; Caporaso, Nicola; Morisco, Filomena; Nardone, Gaetano; Angrisani, Debora; Giacometti, Andrea; Benedetti, Antonio; Tarantino, Giuseppe; Marsetti, Fabio; Tavio, Marcello; Novara, Sergio; Antonia Santantonio, Teresa; Serviddio, Gaetano; Brunetto, Maurizia; Coco, Barbara; Angarano, Gioacchino; Milella, Michele; Barone, Michele; Di Leo, Alfredo; Ettore Sansonno, Domenico; Cacciola, Irene; Boffa, Nicola; Saracco, Giorgio; Di Biagio, Antonio; Picciotto, Antonino; de Luca, Andrea; Calvaruso, Vincenza; Corradori, Silvia; Ferrari, Carlo; Orlandini, Alessandra; Maida, Ivana; Torti, Carlo; Chessa, Luchino; Felder, Martina; Vespasiani Gentilucci, Umberto; Angeli, Paolo; Romano, Antonietta; Ludovico Rapaccini, Gian; Miele, Luca; Cima, Serena; Luisa Russo, Maria; Cozzolongo, Raffaele; Onnelli, Giovanna; D'Offizi, Giampiero; Lionetti, Raffaella; Montalbano, Marzia; Guerra, Michele; Di Perri, Giovanni; Boglione, Lucio; Capra, Franco; Carolo, Giada; Ieluzzi, Donatella; Antonini, Cinzia; Termite, Antonio; Madonia, Salvatore; Tarquini, Pierluigi; Parruti, Giustino; Vecchiet, Giacomo; Falasca, Katia; Menzaghi, Barbara; Quirino, Tiziana; Dentone, Chiara; Maria Piscaglia, Anna; Rossi, Cristina; Giordani, Maria; Fontanella, Luca; Cassola, Giovanni; Russello, Maurizio; Cristaudo, Antonio; Giacomet, Vania; Colombo, Massimo; Donato, Francesca; Durante, Emanuele; Cosco, Lucio; Marignani, Massimo; Quartini, Mariano; Memoli, Massimo; Ganga, Roberto; Ponti, Laura; Soria, Alessandro; Grazia Rumi, Maria; Gulminetti, Roberto; Maserati, Renato; Mondelli, Mario; Lazzarin, Adriano; Rita Parisi, Maria; Canovari, Benedetta; Avancini, Ivo; Pravadelli, Cecilia; Blanc, Pier; Pasquazzi, Caterina; Maria Mastroianni, Claudio; Lichtner, Myriam; Distefano, Marco; Magnani, Silvia; Paganin, Simona; Erne, Elke; Gatti, Pietro; Tundi, Paolo; Calabrese, Paolo; Gasbarrini, Antonio; Grieco, Antonio; Coppola, Nicola; Del Poggio, Paolo; Levrero, Massimo; Talliani, Gloria; Vullo, Vincenzo; Cauda, Roberto; La Monica, Silvia; Potenza, Domenico; Rizzo, Salvatore; Castelli, Francesco; Marie Pigozzi, Grazielle; Ciancio, Alessia; Romagnoli, Dante; Barchetti, Federica; Ivanovic, Jelena; Longo, Olimpia; Petraglia, Sandra; Paola Trotta, Maria

Background We ran a compassionate use nationwide programme (ABACUS) to provide access to ombitasvir, paritaprevir, and ritonavir, with dasabuvir, plus ribavirin for hepatitis C virus (HCV) genotype 1 infection and ombitasvir, paritaprevir, and ritonavir, plus ribavirin for HCV genotype 4 infection in patients with cirrhosis at high risk of decompensation while approval of these regimens was pending in Italy. Methods In this prospective observational study, we collected data from a compassionate use nationwide programme from March 17, 2014, to May 28, 2015. Patients with HCV genotype 1 infection and cirrhosis at high risk of decompensation were given coformulated ombitasvir (25 mg), paritaprevir (150 mg), and ritonavir (100 mg) once daily and dasabuvir (250 mg) twice daily for 12 weeks (patients with HCV genotype 1b infection) or 24 weeks (patients with HCV genotype 1a infection). Patients with HCV genotype 4 infection were given coformulated ombitasvir (25 mg), paritaprevir (150 mg), and ritonavir (100 mg) once per day for 24 weeks. All patients were given weight-based ribavirin. The primary efficacy endpoint was sustained virological response at week 12 after the end of treatment (SVR12), analysed by intention-to-treat. Univariate and multivariate logistic regression analyses were used to identify baseline characteristics associated with SVR12. Adverse events were recorded throughout the study. Findings 728 (96%) of 762 patients with cirrhosis who were given ombitasvir, paritaprevir, and ritonavir, with or without dasabuvir, plus ribavirin therapy for 12 or 24 weeks achieved SVR12. Logistic regression analyses identified that bilirubin concentrations of less than 2 mg/dL were associated with SVR12 (odds ratio [OR] 4·76 [95% CI 1·83–12·3]; p=0·001). 166 (23%) of 734 patients included in safety analyses had an adverse event. 25 (3%) patients discontinued treatment because of adverse events. Asthenia was the most commonly reported adverse event, occurring in 36 (5%) patients. Interpretation Our findings suggest that the safety and effectiveness of ombitasvir, paritaprevir, and ritonavir, with or without dasabuvir, plus ribavirin in patients with HCV genotype 1 or 4 infection and cirrhosis at high risk of decompensation in a real-life setting are similar to those reported in clinical trials. The concordance with clinical trials provides reassurance that the reported efficacy of this treatment in clinical trials will translate to its use in routine clinical practice. Funding Dipartimento Biomedico di Medicina Interna e Specialistica dell'Universita di Palermo.

2017 - Safety of Antiplatelet Agents: Analysis of ‘Real-World’ Data from the Italian National Pharmacovigilance Network [Articolo su rivista]
Gozzo, Lucia; Navarria, Andrea; Benfatto, Giuseppe; Longo, Laura; Mansueto, Silvana; Sottosanti, Laura; Pani, Luca; Salomone, Salvatore; Drago, Filippo

Introduction: According to the Italian National Report on drug use, thienopyridines (ticlopidine, clopidogrel and prasugrel) and ticagrelor represent the most prescribed antiplatelet agents, beside aspirin. The aim of this study was to analyse the safety profile of these drugs using data from spontaneous reporting of suspected adverse reactions (ADRs). Methods: Suspected ADRs for ticlopidine, clopidogrel, prasugrel and ticagrelor, reported on the Italian National Pharmacovigilance Network between January 2009 and December 2016, were included in the analysis. All suspected ADRs were classified by frequency, seriousness, outcome, age and system organ class. Results: Clopidogrel showed the highest absolute number of suspected ADRs, followed by ticlopidine. However, these data need to be contextualized in view of the differences in marketing authorization dates, prescription rates and a characterization of the relative seriousness of ADRs per each drug. After the correction for prescription rate, ticagrelor showed the highest reporting trend and ticlopidine the lowest. Most ADRs occurred in the elderly, in particular for ticlopidine. Bleeding represents one of the most reported events (ticlopidine 40%, clopidogrel 26%, prasugrel 42%, ticagrelor 30%) and aspirin was the most frequently associated suspected drug. The majority of ADRs had complete recovery and were non-serious, except for ticlopidine (serious ADRs 53%). Prasugrel showed the highest percentage of ‘life-threatening’ events and ‘death’. Conclusions: Based on the analysis conducted on spontaneous ADRs reporting system in Italy, the safety profile of antiplatelet drugs seems favourable. However, the overall risk-benefit ratio of these drugs needs to be reassessed taking into account the appropriateness of use in particular populations at risk, such as the elderly. Based on this information, we believe that more attention from clinicians and/or an implementation of regulatory measures could be useful for clinical practice.

2017 - The Italian compassionate use of sofosbuvir observational cohort study for the treatment of recurrent hepatitis C: clinical and virological outcomes [Articolo su rivista]
Carrai, Paola; Morelli, Cristina; Cordone, Gabriella; Romano, Antonietta; Tamé, Mariarosa; Lionetti, Raffaella; Pietrosi, Giada; Lenci, Ilaria; Piai, Guido; Russo, Francesco Paolo; Coppola, Carmine; Melazzini, Mario; Montilla, Simona; Pani, Luca; Petraglia, Sandra; Russo, Pierluigi; Trotta, Maria Paola; Martini, Silvia; Toniutto, Pierluigi; Bandiera, Francesco; Bhoori, Sherrie; Brillanti, Stefano; Burra, Patrizia; Corsale, Sveva; De Luca, Andrea; Fagiuoli, Stefano; Fattovich, Giovanna; Fava, Gianmarco; Felder, Martina; Forte, Paolo; Galeota-Lanza, Alfonso; Gitto, Stefano; Grieco, Antonio; Grossi, Paolo; Ialungo, A. M.; Iemmolo, Rosa Maria; Loiacono, Laura; Mangia, Alessandra; Merli, Manuela; Piacentini, A.; Pellicelli, Adriano; Rigamonti, Cristina; Gabriella, Verucchi; Zignego, Anna Linda

Direct antivirals are available for treating recurrent hepatitis C (RHC). This study reported outcomes of 424 patients with METAVIR F3–F4 RHC who were treated for 24 weeks with sofosbuvir/ribavirin and followed for 12 weeks within the Italian sofosbuvir compassionate use program. In 55 patients, daclatasvir or simeprevir were added. Child–Pugh class and model of end stage liver disease (MELD) scores were evaluated at baseline and 36 weeks after the start of therapy. The sustained viral response (SVR) was 86.7% (316/365) in patients who received sofosbuvir/ribavirin and 98.3% (58/59) in patients who received a second antiviral (P < 0.01). In patients treated with sofosbuvir/ribavirin, a significant difference in SVR was observed between patients diagnosed with METAVIR F4 (211/250; 84.4%), METAVIR F3 (95/105; 90.5%) and fibrosing cholestatic hepatitis (10/10; 100%) (P = 0.049). A significant association was found between patients who worsened from Child–Pugh class A and who experienced viral relapse (4/26 vs. 8/189, P = 0.02). In patients with a baseline MELD score <15, a significant association was found between maintaining a final MELD score <15 and the achievement of SVR (187/219 vs. 6/10, P = 0.031). This real-world study indicates that sofosbuvir/ribavirin treatment for 24 weeks was effective, and the achievement of SVR was associated with a reduced probability of developing worsening liver function.

2016 - Adherence to selective serotonin and serotonin-norepinephrine reuptake inhibitor prescriptions affects overall medication adherence in older persons: Evidence from the Italian nationwide osmed health-db database [Articolo su rivista]
Marengoni, Alessandra; Onder, Graziano; Esposti, Luca Degli; Russo, Pierluigi; Sangiorgi, Diego; Buda, Stefano; Fini, Massimo; Marchionni, Niccol; Bonassi, Stefano; Mammarella, Federica; Marrocco, Walter; Pozzi, Giuseppe; Palmer, Katie; Monaco, Alessandro; Pecorelli, Sergio; Pani, Luca

Objective: This study aimed to evaluate prevalence of prescription of and adherence to selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs) and whether adherence to these classes of drugs affects overall medication adherence in older persons. Methods: In a cross-sectional analysis of administrative data comprehensive of all prescribed drugs reimbursed by the Italian national health care system, new prescriptions of SSRIs and SNRIs to persons aged 65 years or older were analyzed (n = 380,400 in 2011; 395,806 in 2012; 409,741 in 2013, from a total sample of 3,762,299 persons aged 65 years or older) as well as prescriptions of antihypertensives, statins, other psychiatric drugs, antidiabetics, antiplatelets, anticoagulants, drugs for chronic obstructive pulmonary disease, and antiosteoporotics. Adherence was estimated by calculating the proportion of days covered by drugs dispensed during a period of 365 days. Adherence was defined as a proportion of days covered of more than 80%. Results: Prevalence of SSRI and SNRI prescriptions varied from 11.4% in 2011 to 12.1% in 2013. Adherence to SSRI and SNRI prescriptions ranged from 31.2% in persons aged ≥ 95 years in 2011 to 41.8% in persons aged 75-84 years in 2013. Persons adherent to SSRI and SNRI prescriptions were more likely to be adherent to the other medications, after adjustment for age, gender, and number of drugs prescribed. The highest association was found for adherence to psychiatric drugs (OR = 1.9; 95% CI, 1.8-2.0). Conclusions: Adherence to SSRI and SNRI prescriptions is poor in older persons. However, people adherent to these classes of antidepressants are more likely to be adherent to the other medications they are prescribed. Studies are needed to evaluate the reasons for and the potential benefits of increasing adherence to antidepressants on overall adherence.

2016 - Advanced Age and Medication Prescription: More Years, Less Medications? A Nationwide Report From the Italian Medicines Agency [Articolo su rivista]
Onder, G.; Marengoni, A.; Russo, P.; Degli Esposti, L.; Fini, M.; Monaco, A.; Bonassi, S.; Palmer, K.; Marrocco, W.; Pozzi, G.; Sangiorgi, D.; Buda, S.; Marchionni, N.; Mammarella, F.; Bernabei, R.; Pani, L.; Pecorelli, S.

Background: In older adults co-occurrence of multiple diseases often leads to use of multiple medications (polypharmacy). The aim of the present study is to describe how prescription of medications varies across age groups, with specific focus on the oldest old. Methods: We performed a cross-sectional study using 2013 data from the OsMed Health-DB database (mean number of medicines and defined daily doses prescribed in 15,931,642 individuals). There were 3,378,725 individuals age 65 years or older (21.2% of the study sample). Results: The mean number of prescribed medications progressively rose from 1.9 in the age group <65 years to 7.4 in the age group 80-84 years and then declined, with a more marked reduction in the age group 95 years or older with a mean number of 2.8 medications. A similar pattern was observed for the mean number of defined daily doses. Among participants age ≥65 years, proton pump inhibitors were the most commonly prescribed medication (40.9% of individuals ≥65 years), followed by platelet aggregation inhibitors (32.8%) and hydroxy-methylglutaryl-coenzyme A reductase inhibitors (26.1%). A decline in prescription was observed among individuals age 90 years or older, but this reduction was less consistent for medications used to treat acute conditions (ie, antibiotics and glucocorticoids) rather than preventive medicines commonly used to treat chronic diseases (ie, antihypertensive medications and hydroxy-methylglutaryl-coenzyme A reductase inhibitors). Conclusions: The burden of medication treatment progressively increases till age 85 and substantially declines after age of 90 years. Patterns of medication prescription widely vary across age groups.

2016 - Balancing access to medicines and sustainability in Europe: An analysis from the network of competent authorities on pricing and reimbursement (CAPR) [Articolo su rivista]
Pani, L.; Montilla, S.; Nemeth, G.; Russo, P.; Viceconte, G.; Vogler, S.

2016 - Identification and validation of biomarkers for autism spectrum disorders [Articolo su rivista]
Loth, E.; Spooren, W.; Ham, L. M.; Isaac, M. B.; Auriche-Benichou, C.; Banaschewski, T.; Baron-Cohen, S.; Broich, K.; Bolte, S.; Bourgeron, T.; Charman, T.; Collier, D.; de Andres-Trelles, F.; Durston, S.; Ecker, C.; Elferink, A.; Haberkamp, M.; Hemmings, R.; Johnson, M. H.; Jones, E. J. H.; Khwaja, O. S.; Lenton, S.; Mason, L.; Mantua, V.; Meyer-Lindenberg, A.; Lombardo, M. V.; O'Dwyer, L.; Okamoto, K.; Pandina, G. J.; Pani, L.; Persico, A. M.; Simonoff, E.; Tauscher-Wisniewski, S.; Llinares-Garcia, J.; Vamvakas, S.; Williams, S.; Buitelaar, J. K.; Murphy, D. G. M.

2016 - Kounis Syndrome: An analysis of spontaneous reports from international pharmacovigilance database [Articolo su rivista]
Renda, F.; Landoni, G.; Trotta, F.; Piras, D.; Finco, G.; Felicetti, P.; Pimpinella, G.; Pani, L.

Introduction The coincidental occurrence of a cardiac symptomatology (e.g. an acute coronary syndrome or a myocardial infarction), during an anaphylactic or anaphylactoid episode is known as Kounis Syndrome. A variety of drugs, substances, food and environmental exposures are associated with this reaction. There is an exponential increase in the number of published scientific articles reports on this syndrome, but since it is rare, the largest case series published so far included only 10 and 6 patients. Methods We searched the global World Health Organization database called VigiBase™ to detect all cases of Kounis Syndrome ever reported (last update December 31st 2014). Results We identified 51 cases of Kounis Syndrome reported to International Pharmacovigilance Agency (VigiBase™). All these cases were reported in the period 2010-2014 and almost half cases (22 reports) belonged to the year 2014. Most cases occurred in the USA and non-steroidal anti-inflammatory drugs were the most frequent trigger drugs. Discussion We collected pharmacovigilance international data representing the largest case series ever published on the recently identified Kounis Syndrome.

2016 - Kounis syndrome due to antibiotics: A global overview from pharmacovigilance databases [Articolo su rivista]
Renda, F.; Marotta, E.; Landoni, G.; Belletti, A.; Cuconato, V.; Pani, L.

Background Kounis syndrome (KS) is characterized by concurrent presence of anaphylactic and cardiac components. Available evidence suggests that antibiotics are frequently associated to KS. We therefore analyzed KS cases associated with antibiotics use from the two largest pharmacovigilance databases. Methods Two pharmacovigilance databases, EudraVigilance and VigiLyze, were searched for cases reporting the adverse reaction “Kounis Syndrome” with antibiotics as suspected active substance. We analyzed the period from December 1st, 2001 to February 16th, 2016. For the most reported active substance, proportional reporting ratio (PRR) was calculated. Results A total of 10 cases of KS associated with antibiotic use were retrieved from EudraVigilance database. Mean patients' age was 58.2 years and 70% were male. The most frequently reported suspected antibiotic was the combination amoxicillin/clavulanic acid (four cases). VigiLyze database reported 13 KS cases associated to antibiotics. Mean age was 56 years and 61% of patients were male. The most frequently reported antibiotic was again the combination amoxicillin/clavulanic acid (five cases). Seven duplicate cases were identified, leaving a total of 16 cases of KS, with six of them associated to amoxicillin/clavulanic acid use. The PRR value for amoxicillin/clavulanic acid against other kinds of antibiotics was 2.62 considering EudraVigilance data and 1.61 considering VigiLyze data. Conclusions This analysis provided a complete picture of the cases of KS associated with antibiotic use and identified a possible association between amoxicillin/clavulanic acid and KS. Since the number of cases is low, especially considering its wide use, further analyses are needed to confirm the association.

2016 - Pricing and reimbursement experiences and insights in the European Union and the United States: Lessons learned to approach adaptive payer pathways [Articolo su rivista]
Faulkner, S. D.; Lee, M.; Qin, D.; Morrell, L.; Xoxi, E.; Sammarco, A.; Cammarata, S.; Russo, P.; Pani, L.; Barker, R.

Earlier patient access to beneficial therapeutics that addresses unmet need is one of the main requirements of innovation in global healthcare systems already burdened by unsustainable budgets. “Adaptive pathways” encompass earlier cross-stakeholder engagement, regulatory tools, and iterative evidence generation through the life cycle of the medicinal product. A key enabler of earlier patient access is through more flexible and adaptive payer approaches to pricing and reimbursement that reflect the emerging evidence generated.

2016 - Scaling up health knowledge at European level requires sharing integrated data: An approach for collection of database specification [Articolo su rivista]
Menditto, Enrica; Bolufer De Gea, Angela; Cahir, Caitriona; Marengoni, Alessandra; Riegler, Salvatore; Fico, Giuseppe; Costa, Elisio; Monaco, Alessandro; Pecorelli, Sergio; Pani, Luca; Prados-Torres, Alexandra

2015 - Alternative pricing strategies for cancer drugs [Articolo su rivista]
Messori, A.; De Rosa, M.; Pani, L.

2015 - An Integrated Approach for a Structural and Functional Evaluation of Biosimilars: Implications for Erythropoietin [Articolo su rivista]
Gianoncelli, A.; Bonini, S. A.; Bertuzzi, M.; Guarienti, M.; Vezzoli, S.; Kumar, R.; Delbarba, A.; Mastinu, A.; Sigala, S.; Spano, P.; Pani, L.; Pecorelli, S.; Memo, M.

Background: Authorization to market a biosimilar product by the appropriate institutions is expected based on biosimilarity with its originator product. The analogy between the originator and its biosimilar(s) is assessed through safety, purity, and potency analyses. Objective: In this study, we proposed a useful quality control system for rapid and economic primary screening of potential biosimilar drugs. For this purpose, chemical and functional characterization of the originator rhEPO alfa and two of its biosimilars was discussed. Methods: Qualitative and quantitative analyses of the originator rhEPO alfa and its biosimilars were performed using reversed-phase high-performance liquid chromatography (RP-HPLC). The identification of proteins and the separation of isoforms were studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and two-dimensional gel electrophoresis (2D-PAGE), respectively. Furthermore, the biological activity of these drugs was measured both in vitro, evaluating the TF-1 cell proliferation rate, and in vivo, using the innovative experimental animal model of the zebrafish embryos. Results: Chemical analyses showed that the quantitative concentrations of rhEPO alfa were in agreement with the labeled claims by the corresponding manufacturers. The qualitative analyses performed demonstrated that the three drugs were pure and that they had the same amino acid sequence. Chemical differences were found only at the level of isoforms containing N-glycosylation; however, functional in vitro and in vivo studies did not show any significant differences from a biosimilar point of view. Conclusion: These rapid and economic structural and functional analyses were effective in the evaluation of the biosimilarity between the originator rhEPO alfa and the biosimilars analyzed.

2015 - Cardiovascular effects of dipeptidyl peptidase-4 inhibitors in diabetic patients: A meta-analysis [Articolo su rivista]
Savarese, G.; Perrone-Filardi, P.; D'Amore, C.; Vitale, C.; Trimarco, B.; Pani, L.; Rosano, G. M. C.

Background Dipeptidyl peptidase-4 inhibitors (DPP-4is) improve glucose control in patients with type 2 diabetes mellitus (DM); however, only few studies were properly designed to evaluate their cardiovascular (CV) effects. The purpose of this study was to assess the impact of DPP-4i treatment on CV morbidity and mortality. Methods Randomized clinical trials enrolling more than 200 patients, comparing DPP-4 versus placebo or active treatments in patients with DM and reporting at least one event among all-cause and CV mortality, myocardial infarction (MI), stroke and new onset of heart failure (HF) were included in the analysis. Results Ninety-four trials enrolling 85,224 patients (median follow-up = 29 weeks) were included in the analysis. Compared to control, treatment with DPP-4i did not affect all-cause and CV mortality, as well as stroke, in the short and long terms (< and >= 29 weeks, respectively). DPP-4i reduced the risk of MI in the short (RR: 0.584 [95% CI: 0.361 to 0.943]; p = 0.028), but not in the long term. Additionally, long-term treatment with DPP-4 was associated with a 15.8% increased risk of HF (RR: 1.158 [CI: 1.011 to 1.326]; p = 0.034). No heterogeneity among studies or publication bias was detected. Conclusions DPP4is do not affect all cause-and CV-mortality and stroke in diabetic patients; the reduction in MI observed with short-term treatment does not persist in the long term. Long-term use of DPP-4i in diabetic patients is associated with increased risk of HF.

2015 - Changes and determination of dosing recommendations for medicinal products recently authorised in the European Union [Articolo su rivista]
Ehmann, F.; Papaluca, M.; Di Giuseppe, F.; Pani, L.; Eskova, A.; Manolis, E.; Herold, R.

Introduction: The quantity and quality of data for determining the dose and treatment schedule of medicinal products is directly related to how safe and efficacious these medicines are and how successful they can be used to treat patients.Areas covered: This review provides an analysis of dose-related label modifications of recently approved drugs. It shows which areas could benefit from a better dose-exposure-response understanding, both during initial assessment and after marketing authorisation. This analysis highlights regulators' considerations in dosage evaluations and provides reflections for drug developers on how to ensure best possible dose selection in the interest of the patients.Expert opinion: Using modelling and simulation, pharmacogenomics, population pharmacokinetics, physiologically based pharmacokinetic models and drug-drug interaction studies in conjunction with well-designed clinical trials will improve the understanding of the pharmacology of medicines, of the physiology of the disease and of the dose-exposure-response relationship during drug development. More focus should be given to the investigation of dose and regimens for special populations before applying for marketing authorisation. Consequently, regulators could review dose-exposure-response data with more certainty and better define dose recommendations in the label.

2015 - Computational and functional analysis of biopharmaceutical drugs in zebrafish: Erythropoietin as a test model [Articolo su rivista]
Guarienti, M.; Giacopuzzi, E.; Gianoncelli, A.; Sigala, S.; Spano, P.; Pecorelli, S.; Pani, L.; Memo, M.

The zebrafish (Danio rerio) is a very popular vertebrate model system, especially embryos represent a valuable tool for in vivo pharmacological assays. This is mainly due to the zebrafish advantages when compared to other animal models. Erythropoietin is a glycoprotein hormone that acts principally on erythroid progenitors, stimulating their survival, proliferation and differentiation. Recombinant human erythropoietin (rhEPO) has been widely used in medicine to treat anemia and it is one of the best-selling biotherapeutics worldwide. The recombinant molecule, industrially produced in CHO cells, has the same amino acid sequence of endogenous human erythropoietin, but differs in the glycosylation pattern. This may influence efficacy and safety, particularly immunogenicity, of the final product. We employed the zebrafish embryo as a vertebrate animal model to perform in vivo pharmacological assays. We conducted a functional analysis of rhEPO alpha Eprex® and two biosimilars, the erythropoietin alpha Binocrit® and zeta Retacrit®. By in silico analysis and 3D modeling we proved the interaction between recombinant human erythropoietin and zebrafish endogenous erythropoietin receptor. Then we treated zebrafish embryos with the 3 rhEPOs and we investigated their effect on erythrocytes production with different assays. By real time-PCR we observed the relative upregulation of gata1 (2.4 ± 0.3 fold), embryonic α-Hb (1.9 ± 0.2 fold) and β-Hb (1.6 ± 0.1 fold) transcripts. A significant increase in Stat5 phosphorylation was also assessed in embryos treated with rhEPOs when compared with the negative controls. Live imaging in tg (kdrl:EGFP; gata1:ds-red) embryos, o-dianisidine positive area quantification and cyanomethemoglobin content quantification revealed a 1.8 ± 0.3 fold increase of erythrocytes amount in embryos treated with rhEPOs when compared with the negative controls. Finally, we verified that recombinant human erythropoietins did not cause any inflammatory response in the treated embryos. Our data showed that zebrafish embryo can be a valuable tool to study in vivo effects of complex pharmacological compounds, such as recombinant human glycoproteins, allowing to perform fast and reproducible pharmacological assays with excellent results.

2015 - Daptomycin in paediatrics: Current knowledge and the need for future research [Articolo su rivista]
Principi, N.; Caironi, M.; Venturini, F.; Pani, L.; Esposito, S.

To overcome the problems stemming from antimicrobial resistance, there have been several attempts to develop new antimicrobials in recent years. Of the highly potent drugs targeting resistant Gram-positive bacteria, daptomycin has a number of attractive characteristics that suggest its possible use in the treatment of serious infections due to these organisms. Although several pharmacokinetic and clinical studies in adults have provided data to determine howthis drug should be prescribed to obtain the maximal clinical efficacy without significant risks of severe adverse events, we have not yet solved all of the problems related to the use of this antibiotic in paediatric patients. In this paper, the resolved and lingering problems of daptomycin treatment in newborns and children are reviewed and discussed. Studies have indicated that daptomycin is a promising therapeutic option for the treatment of paediatric diseases caused by MDR Gram-positive bacilli. However, before daptomycin can be licensed for use in newborns and children, further studies are needed to establish the appropriate dosages for paediatric patients of different ages. The data collected in adults can only be transferred to children older than 12 years, and the information available is not sufficient to determine the dosage that will assure the highest antimicrobial efficacy with only marginal risks of adverse events in younger patients. Thus, studies in neonates and younger infants are urgently needed to permit the use of daptomycin in the first months of life, a period in which infections due to MDR Gram-positive pathogens are increasing.

2015 - Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS): A National Analysis of Data from 10-Year Post-marketing Surveillance [Articolo su rivista]
Renda, F.; Landoni, G.; Bertini Malgarini, R.; Assisi, A.; Azzolini, M. L.; Mucchetti, M.; Pimpinella, G.; Pani, L.

Introduction: Drug reaction with eosinophilia and systemic symptoms (DRESS) is a rare, severe and potentially fatal cutaneous adverse drug reaction (the mortality rate is up to 10 %) associated with numerous and apparently heterogeneous drugs. The aetiology is unknown. Objective: To report Italian cases of DRESS over a 10-year period. Methods: We searched the National Pharmacovigilance Network (NPN) for the term ‘drug reaction with eosinophilia and systemic symptoms’ from 1 January 2004 to 1 January 2014, to identify all reports of DRESS. Each case was checked to avoid duplication. Results: In the NPN, we identified 91 serious cases of DRESS: 68 were spontaneous, still-unpublished reports, while 23 additional cases were derived from screening of the scientific literature, performed by marketing authorization holders. Notably, the single common element linking all cases of DRESS was intake of a drug containing an aromatic ring. Conclusion: Thanks to the largest national DRESS case series ever reported, we were able to hypothesize, for the first time, that there is an association between use of drugs containing an aromatic ring in their chemical structure and DRESS. This might aid understanding of the aetiology of DRESS and facilitate diagnosis.

2015 - Effectiveness and cost effectiveness of bevacizumab in metastatic colorectal cancer [Articolo su rivista]
Messori, A.; De Rosa, M.; Fadda, V.; Pani, L.

2015 - Generic and biosilimilar medicines use and expenditure: A report from the National Observatory on the Use of Medicines (OsMed) of the Italian Medicines Agency (AIFA) [Articolo su rivista]
Monaco, A.; Mammarella, F.; Cangini, A.; Marini, R.; Marengoni, A.; Russo, P.; Pecorelli, S.; Pani, L.

2015 - Monitoring registries at Italian medicines agency: Fostering access, guaranteeing sustainability [Articolo su rivista]
Montilla, S.; Xoxi, E.; Russo, P.; Cicchetti, A.; Pani, L.

Objectives: The AIFA (Agenzia Italiana del Farmaco - Italian Medicines Agency) Monitoring Registries track the eligibility of patients and the complete flow of treatments, guaranteeing appropriateness in use of pharmaceutical products, according to approved indications. Methods: This study describes the Italian pharmaceutical context and the aims and functioning of AIFA Monitoring Registries, focusing on the applications to the Managed Entry Agreements (MEAs) and HTA approaches. Results: The AIFA Monitoring Registries System has been operational in Italy since 2005. In 2012, the system became part of the NHS Information Technology system, aiming at enhancing appropriate use of pharmaceuticals and efficiency of the administrative activity. Currently, seventy-six medicines are monitored through the system, corresponding to fifty-eight therapeutic indications; individual treatments recorded are more than 515,000, for a population of approximately 505,000 patients. For each monitored product, patients eligible for treatment are registered in the specific therapeutic indication dynamic monitoring database to collect epidemiologic and clinical data, including data on the safety profile, and ex-post information missing at first evaluation stage. Conclusions: AIFA Monitoring Registries allow the evaluation of the pharmaceuticals' performance in clinical practice and may promote innovation and quicker access to medicines at affordable prices, for the benefit of patients.

2015 - Rare diseases and effective treatments: Are we delivering? [Articolo su rivista]
Luzzatto, L.; Hollak, C. E. M.; Cox, T. M.; Schieppati, A.; Licht, C.; Kaariainen, H.; Merlini, G.; Schaefer, F.; Simoens, S.; Pani, L.; Garattini, S.; Remuzzi, G.

2015 - Risk of hospitalization for heart failure in patients with type 2 diabetes newly treated with DPP-4 inhibitors or other oral glucose-lowering medications: A retrospective registry study on 127,555 patients from the Nationwide OsMed Health-DB Database [Articolo su rivista]
Fadini, G. P.; Avogaro, A.; Degli Esposti, L.; Russo, P.; Saragoni, S.; Buda, S.; Rosano, G.; Pecorelli, S.; Pani, L.; Martinetti, S.; Mero, P.; Raeli, L.; Migliazza, S.; Dellagiovanna, M.; Cerra, C.; Gambera, M.; Piccinelli, R.; Zambetti, M.; Atzeni, F.; Valsecchi, V.; Deluca, P.; Scopinaro, E.; Moltoni, D.; Pini, E.; Leoni, O.; Oria, C.; Papagni, M.; Nosetti, G.; Caldiroli, E.; Moser, V.; Roni, R.; Polverino, A.; Bovo, C.; Mezzalira, L.; Andretta, M.; Trentin, L.; Palcic, S.; Pettinelli, A.; Arbo, A.; Bertola, A.; Capparoni, G.; Cattaruzzi, C.; Marcuzzo, L.; Rosa, F. V.; Basso, B.; Saglietto, M.; Delucis, S.; Prioli, M.; Filippi, R.; Coccini, A.; Ghia, M.; Sanfelici, F.; Radici, S.; Scanavacca, P.; Campi, A.; Bianchi, S.; Verzola, A.; Morini, M.; Borsari, M.; Danielli, A.; Dal Maso, M.; Marsiglia, B.; Vujovic, B.; Pisani, M.; Bonini, P.; Lena, F.; Aletti, P.; Marcobelli, A.; Sagratella, S.; Fratini, S.; Bartolini, F.; Riccioni, G.; Meneghini, A.; Di Turi, R.; Fano, V.; Blasi, A.; Pagnozzi, E.; Quintavalle, G.; D'Avenia, P.; De Matthaeis, M. C.; Ferrante, F.; Crescenzi, S.; Marziale, L.; Venditti, P.; Bianchi, C.; Senesi, I.; Baci, R.; De Carlo, I.; Lavalle, A.; Trofa, G.; Marcello, G.; Pagliaro, C.; Troncone, C.; Farina, G.; Tari, M. G.; Motola, G.; De Luca, F.; Saltarelli, M. L.; Granieri, C.; Vulnera, M.; Palumbo, L.; La Viola, F.; Florio, L.; De Francesco, A. E.; Costantino, D.; De Francesco, A. E.; Rapisarda, F.; Lazzaro, P. L.; Pastorello, M.; Parlli, M.; Visconti, M.; Uomo, I.; Sanna, P.; Lombardo, F.

Aims Oral glucose-lowering medications are associated with excess risk of heart failure (HF). Given the absence of comparative data among drug classes, we performed a retrospective study in 32 Health Services of 16 Italian regions accounting for a population of 18 million individuals, to assess the association between HF risk and use of sulphonylureas, DPP-4i, and glitazones. Methods and results We extracted data on patients with type 2 diabetes who initiated treatment with DPP-4i, thiazolidinediones, or sulphonylureas alone or in combination with metformin during an accrual time of 2 years. The endpoint was hospitalization for HF (HHF) occurring after the first 6 months of therapy, and the observation was extended for up to 4 years. A total of 127 555 patients were included, of whom 14.3% were on DPP-4i, 72.5% on sulphonylurea, 13.2% on thiazolidinediones, with average 70.7% being on metformin as combination therapy. Patients in the three groups differed significantly for baseline characteristics: age, sex, Charlson index, concurrent medications, and previous cardiovascular events. During an average 2.6-year follow-up, after adjusting for measured confounders, use of DPP-4i was associated with a reduced risk of HHF compared with sulphonylureas [hazard ratio (HR) 0.78; 95% confidence interval (CI) 0.62-0.97; P = 0.026]. After propensity matching, the analysis was restricted to 39 465 patients, and the use of DPP-4i was still associated with a lower risk of HHF (HR 0.70; 95% CI 0.52-0.94; P = 0.018). Conclusion In a very large observational study, the use of DPP-4i was associated with a reduced risk of HHF when compared with sulphonylureas.

2015 - Steps forward in regulatory pathways for acute and chronic heart failure [Articolo su rivista]
Pani, Luca; Pecorelli, Sergio; Rosano, Giuseppe; Anker, Stefan D.; Peracino, Andrea; Fregonese, Laura; Prasad, Krishna; Rasi, Guido

A workshop was organized by the Agenzia Italiana del Farmaco (AIFA) to discuss unmet needs and ways forward in the development of medicines in heart failure, their rationale, and cost-effective use. An integrated, multidisciplinary approach, including patients' needs and perspectives, was advocated by all the participants as the way to the most effective treatment regimens. More work is needed for reaching consensus on clinical and functional endpoints, for validating patient reported outcomes and measurements of well-being. Similarly, the integration into the clinical programmes of the health technology assessment/payers perspective, in particular, the evaluation of 'real-life' treatment effectiveness and of health as a value, would help in shifting the development and authorization of medicines from the molecule paradigm to their evaluation in the context of the whole health care regimen. Through this kind of workshop, AIFA is trying to build a template for meetings devoted to debate unmet needs with all stakeholders towards tentative road maps for the future.

2015 - Taking stock: A multistakeholder perspective on improving the delivery of care and the development of treatments for Alzheimer's disease [Articolo su rivista]
Bradley, P.; Akehurst, R.; Ballard, C.; Banerjee, S.; Blennow, K.; Bremner, J.; Broich, K.; Cummings, J.; Dening, K.; Dubois, B.; Klipper, W.; Leibman, C.; Mantua, V.; Molinuevo, J. L.; Morgan, S.; Muscolo, L. A. A.; Nicolas, F.; Pani, L.; Robinson, L.; Siviero, P.; Van Dam, J.; Van Emelen, J.; Wimo, A.; Wortmann, M.; Goh, L.

Health-care stakeholders increasingly recognize that the scientific and economic challenges associated with Alzheimer's disease (AD) are simply too great for individual stakeholder groups to address solely from within their own silos. In the necessary spirit of collaboration, we present in this perspective a set of multicountry multistakeholder recommendations to improve the organization of existing AD and dementia care and the development of new treatments. In brief, the five recommendations are (1) health-care systems must make choices regarding the patient populations to be diagnosed and treated, (2) health-care systems should use an evidence-based standard of care, (3) increased collaboration between public and private institutions is needed to enhance research, (4) reimbursement end points need to be agreed on and validated, and (5) innovative business models should be used to spur the introduction of new medicines.

2015 - The European Medicines Agency's strategies to meet the challenges of Alzheimer disease [Articolo su rivista]
Haas, M.; Mantua, V.; Haberkamp, M.; Pani, L.; Isaac, M.; Butlen-Ducuing, F.; Vamvakas, S.; Broich, K.

Regulatory agencies have a key role in facilitating the development of new drugs for Alzheimer disease, particularly given the challenges associated with early intervention. Here, we highlight the strategies of the European Medicines Agency to help address such challenges.

2015 - The future is now: Model-based clinical trial design for Alzheimer's disease [Articolo su rivista]
Romero, K.; Ito, K.; Rogers, J. A.; Polhamus, D.; Qiu, R.; Stephenson, D.; Mohs, R.; Lalonde, R.; Sinha, V.; Wang, Y.; Brown, D.; Isaac, M.; Vamvakas, S.; Hemmings, R.; Pani, L.; Bain, L. J.; Corrigan, B.

Failures in trials for Alzheimer's disease (AD) may be attributable to inadequate dosing, population selection, drug inefficacy, or insufficient design optimization. The Coalition Against Major Diseases (CAMD) was formed in 2008 to develop drug development tools (DDT) to expedite drug development for AD and Parkinson's disease.1 CAMD led a process that successfully advanced a clinical trial simulation (CTS) tool for AD through the formal regulatory review process at the US Food and Drug Administration (FDA) and European Medicines Agency (EMA).

2015 - The international coalition of medicines regulatory authorities (ICMRA) [Articolo su rivista]
Skerritt, J.; De Oliveira, J. C. M.; Arora, A.; Rasi, G.; Rys, A.; O'Mahony, P.; Pani, L.; Kondo, T.; Hurts, H.; Ling, M. C. M.; Hela, M.; Hudson, I.; Hamburg, M.

A new global collaboration brings together senior leaders to provide coordinated, consistent, and strategic leadership in an increasingly globalized and complex regulatory environment. The International Coalition of Medicines Regulatory Authorities (ICMRA) is a voluntary, executive level entity that provides direction for a range of areas that are common to many regulatory authorities’ missions.

2015 - Transparency in medical research: Time for a paradigm shift [Articolo su rivista]
Pelliccia, F; Coats, Aj; Pani, L; Gaudio, C; Rosano, G

2015 - Transparency in medical research: Time for a paradigm shift [Articolo su rivista]
Pelliccia, F.; Coats, A. J. S.; Pani, L.; Gaudio, C.; Rosano, G.

2014 - Are we ready? What is missing and what is needed? A regulator's perspective [Articolo su rivista]
Bernardini, C.; Muscolo, L. A. A.; Siviero, P. D.; Montilla, S.; Pani, L.

2014 - Clinical trials and late-stage drug development for Alzheimer's disease: An appraisal from 1984 to 2014 [Articolo su rivista]
Schneider, L. S.; Mangialasche, F.; Andreasen, N.; Feldman, H.; Giacobini, E.; Jones, R.; Mantua, V.; Mecocci, P.; Pani, L.; Winblad, B.; Kivipelto, M.

The modern era of drug development for Alzheimer's disease began with the proposal of the cholinergic hypothesis of memory impairment and the 1984 research criteria for Alzheimer's disease. Since then, despite the evaluation of numerous potential treatments in clinical trials, only four cholinesterase inhibitors and memantine have shown sufficient safety and efficacy to allow marketing approval at an international level. Although this is probably because the other drugs tested were ineffective, inadequate clinical development methods have also been blamed for the failures. Here, we review the development of treatments for Alzheimer's disease during the past 30 years, considering the drugs, potential targets, late-stage clinical trials, development methods, emerging use of biomarkers and evolution of regulatory considerations in order to summarize advances and anticipate future developments. We have considered late-stage Alzheimer's disease drug development from 1984 to 2013, including individual clinical trials, systematic and qualitative reviews, meta-analyses, methods, commentaries, position papers and guidelines. We then review the evolution of drugs in late clinical development, methods, biomarkers and regulatory issues. Although a range of small molecules and biological products against many targets have been investigated in clinical trials, the predominant drug targets have been the cholinergic system and the amyloid cascade. Trial methods have evolved incrementally: inclusion criteria have largely remained focused on mild-to-moderate Alzheimer's disease criteria, recently extending to early or prodromal Alzheimer disease or 'mild cognitive impairment due to Alzheimer's disease', for drugs considered to be disease modifying. The duration of trials has remained at 6-12 months for drugs intended to improve symptoms; 18- to 24-month trials have been established for drugs expected to attenuate clinical course. Cognitive performance, activities of daily living, global change and severity ratings have persisted as the primary clinically relevant outcomes. Regulatory guidance and oversight have evolved to allow for enrichment of early-stage Alzheimer's disease trial samples using biomarkers and phase-specific outcomes. In conclusion, validated drug targets for Alzheimer's disease remain to be developed. Only drugs that affect an aspect of cholinergic function have shown consistent, but modest, clinical effects in late-phase trials. There is opportunity for substantial improvements in drug discovery and clinical development methods. © 2014 The Association for the Publication of the Journal of Internal Medicine.

2014 - Coalition Against Major Diseases/European Medicines Agency biomarker qualification of hippocampal volume for enrichment of clinical trials in predementia stages of Alzheimer's disease [Articolo su rivista]
Hill, D. L. G.; Schwarz, A. J.; Isaac, M.; Pani, L.; Vamvakas, S.; Hemmings, R.; Carrillo, M. C.; Yu, P.; Sun, J.; Beckett, L.; Boccardi, M.; Brewer, J.; Brumfield, M.; Cantillon, M.; Cole, P. E.; Fox, N.; Frisoni, G. B.; Jack, C.; Kelleher, T.; Luo, F.; Novak, G.; Maguire, P.; Meibach, R.; Patterson, P.; Bain, L.; Sampaio, C.; Raunig, D.; Soares, H.; Suhy, J.; Wang, H.; Wolz, R.; Stephenson, D.

Background Regulatory qualification of a biomarker for a defined context of use provides scientifically robust assurances to sponsors and regulators that accelerate appropriate adoption of biomarkers into drug development. Methods The Coalition Against Major Diseases submitted a dossier to the Scientific Advice Working Party of the European Medicines Agency requesting a qualification opinion on the use of hippocampal volume as a biomarker for enriching clinical trials in subjects with mild cognitive impairment, incorporating a scientific rationale, a literature review and a de novo analysis of Alzheimer's Disease Neuroimaging Initiative data. Results The literature review and de novo analysis were consistent with the proposed context of use, and the Committee for Medicinal Products for Human Use released an opinion in November 2011. Conclusions We summarize the scientific rationale and the data that supported the first qualification of an imaging biomarker by the European Medicines Agency. © 2014 The Alzheimer's Association. All rights reserved.

2014 - Drug utilization, safety, and effectiveness of exenatide, sitagliptin, and vildagliptin for type 2 diabetes in the real world: Data from the Italian AIFA Anti-diabetics Monitoring Registry [Articolo su rivista]
Montilla, S.; Marchesini, G.; Sammarco, A.; Trotta, M. P.; Siviero, P. D.; Tomino, C.; Melchiorri, D.; Pani, L.; Melchiorri, D.; Sbraccia, P.; Nicolucci, A.; Brignoli, O.; Coscelli, C.; Dell'Aera, M.; Mazzaglia, G.; Giustini, S. E.; De Rosa, M.; Covezzoli, A.; Rigazio, A.; Roncadori, A.

Background and aims: In Italy, the reimbursed use of incretin mimetics and incretin enhancers was subject to enrollment of patients into a web-based system recording the general demographic and clinical data of patients. We report the utilization data of glucagon-like peptide 1 (GLP1) receptor agonists and dipeptidylpeptidase-4 (DPP4) inhibitors in clinical practice as recorded by the Italian Medicines Agency (AIFA) Monitoring Registry. Methods and results: From February 2008 to August 2010, 75,283 patients with type 2 diabetes were entered into the registry and treated with exenatide, sitagliptin, or vildagliptin. The treatment was administered to patients in a wide range of ages (≥75 years, n=6125 cases), body mass index (BMI) (≥35kg/m2, n=22,015), and metabolic control (HbA1c≥11% ((96mmol/mol), n=3151). Overall, 1116 suspected adverse drug reactions were registered, including 12 cases of acute pancreatitis (six on exenatide). Hypoglycemic episodes mainly occurred in combination with sulfonylureas. Treatment discontinuation for the three drugs (logistic regression analysis) was negatively associated with the male gender and positively with baseline HbA1c, diabetes duration, and, limitedly to DPP-4 inhibitors, with BMI. Treatment discontinuation (including loss to follow-up, accounting for 21-26%) was frequent. Discontinuation for treatment failure occurred in 7.7% of cases (exenatide), 3.8% (sitagliptin), and 4.1% (vildagliptin), respectively, corresponding to 27-40% of all discontinuations, after excluding lost to follow-up. HbA1c decreased on average by 0.9-1.0% (9mmol/mol). Body weight decreased by 3.5% with exenatide and by 1.0-1.5% with DPP-4 inhibitors. Conclusions: In the real world of Italian diabetes centers, prescriptions of incretins have been made in many cases outside the regulatory limits. Nevertheless, when appropriately utilized, incretins may grant results at least in line with pivotal trials.

2014 - High prevalence of poor quality drug prescribing in older individuals: A nationwide report from the Italian Medicines Agency (AIFA) [Articolo su rivista]
Onder, G.; Bonassi, S.; Abbatecola, A. M.; Folino-Gallo, P.; Lapi, F.; Marchionni, N.; Pani, L.; Pecorelli, S.; Sancarlo, D.; Scuteri, A.; Trifiro, G.; Vitale, C.; Zuccaro, S. M.; Bernabei, R.; Fini, M.

Background.Poor quality of drug prescribing in older persons is often associated with increased drug-related adverse events, hospitalization, and mortality. The present study describes a set of prescribing quality indicators developed by the Geriatrics Working Group of the Italian Medicines Agency (AIFA) and estimates their prevalence in the entire elderly (≥65 years) population in Italy.Methods.We performed a cross-sectional study using 2011 data from the OsMed (Osservatorio dei Medicinali) database, which comprises all prescribed drugs that are reimbursed by the Italian National Healthcare System. Yearly prevalence of drug prescribing quality indicators in the Italian older population (n = 12,301,537) was determined.Results.Overall, 13 quality indicators addressing polypharmacy, adherence to treatment of chronic diseases, prescribing cascade, undertreatment, drug-drug interactions, and drugs to be avoided were identified. Polypharmacy was common, with more than 1.3 million individuals taking greater than or equal to 10 drugs (11.3% of the study population). The prevalence of low adherence and undertreatment was also elevated and increased with advancing age, with highest prevalence occurring in individuals aged 85 years and older. Prevalence was less than 3% for quality indicators assessing the prescribing cascade, drug-drug interactions, and drugs to be avoided.Conclusions.These results confirm the high frequency of suboptimal drug prescribing in older adults, using a database that covers the whole Italian population. In general, this descriptive study may help in prioritizing strategies aimed at improving the quality of prescribing in elderly population. © The Author 2013.

2014 - Insights Into the Decision Making of Advisory Groups to the Italian Medicines Agency [Articolo su rivista]
Marangi, M.; Cammarata, S. M.; Pani, L.

The Italian Medicines Agency (AIFA) is enhancing a strong transparency-oriented policy to improve information exchange and decision making with stakeholders. To this end, a questionnaire titled “Survey AIFA 2013” was sent to the 72 selected contacts on February 17, 2013 (closing date April 3, 2013), to assess influence on committees and secretariats’ opinions and decisions. The survey was divided into 2 sections (17 questions) with a 10-minute time limit. The results show that external resolutions have little influence on internal advisory groups, whereas internal ones carry more weight. So-called intellectual bias needs careful monitoring, as it can potentially condition decisions.

2014 - Licensing of generic medicines: Are there any challenges left? A pharmaceutical regulatory perspective [Articolo su rivista]
Borg, J. J.; Tomasi, P.; Pani, L.; Aislaitner, G.; Pirozynski, M.; Leufkens, H.; Melchiorri, D.

When an innovative product (innovator) is not covered anymore by intellectual property rights, cheaper equivalent medicinal products (generic products) may be marketed and used in clinical practice. The regulation of generic products is well-established, and is primarily based on standard rules for quality, therapeutic equivalence requirements (the latter in most instances proven through a bioequivalence study), and safety data for the innovator. The extensive experience from bringing generic products to the market over the last decades allows the conclusion that they are well-accepted and provide a useful alternative option for cost-effective pharmacotherapy. While supporting this conclusion, there are a number of issues to be considered during the assessment of a generic product application. Six scenarios are described in total, from an efficacy and a safety perspective, where potential concerns with the current regulatory standards could arise in the approval of generic products. We also propose solutions to these scenarios in order to foster debate on these issues.

2014 - Managed entry agreements [Articolo su rivista]
Muscolo, L. A. A.; Bernardini, C.; Siviero, P. D.; Montilla, S.; Pani, L.

2014 - Management of notifications of donors with Creutzfeldt-Jakob disease (post-donation information) [Articolo su rivista]
Calizzani, G.; Vaglio, S.; Vetrugno, V.; Delbo, M.; Pani, L.; Grazzini, G.

2014 - Novel epigenetic target therapy for prostate cancer: A preclinical study [Articolo su rivista]
Naldi, I.; Taranta, M.; Gherardini, L.; Pelosi, G.; Viglione, F.; Grimaldi, S.; Pani, L.; Cinti, C.

Epigenetic events are critical contributors to the pathogenesis of cancer, and targeting epigenetic mechanisms represents a novel strategy in anticancer therapy. Classic demethylating agents, such as 5-Aza-2′-deoxycytidine (Decitabine), hold the potential for reprograming somatic cancer cells demonstrating high therapeutic efficacy in haematological malignancies. On the other hand, epigenetic treatment of solid tumours often gives rise to undesired cytotoxic side effects. Appropriate delivery systems able to enrich Decitabine at the site of action and improve its bioavailability would reduce the incidence of toxicity on healthy tissues. In this work we provide preclinical evidences of a safe, versatile and efficient targeted epigenetic therapy to treat hormone sensitive (LNCap) and hormone refractory (DU145) prostate cancers. A novel Decitabine formulation, based on the use of engineered erythrocyte (Erythro-Magneto-Hemagglutinin Virosomes, EMHVs) drug delivery system (DDS) carrying this drug, has been refined. Inside the EMHVs, the drug was shielded from the environment and phosphorylated in its active form. The novel magnetic EMHV DDS, endowed with fusogenic protein, improved the stability of the carried drug and exhibited a high efficiency in confining its delivery at the site of action in vivo by applying an external static magnetic field. Here we show that Decitabine loaded into EMHVs induces a significant tumour mass reduction in prostate cancer xenograft models at a concentration, which is seven hundred times lower than the therapeutic dose, suggesting an improved pharmacokinetics/pharmacodynamics of drug. These results are relevant for and discussed in light of developing personalised autologous therapies and innovative clinical approach for the treatment of solid tumours. © 2014 Naldi et al.

2014 - Reflections on decisions made on the well-established use of medicinal products by EU regulators and the ECJ [Articolo su rivista]
Borg, J. J.; Laslop, A.; Pani, L.; Maciulaitis, R.; Melchiorri, D.

Background: In the European Union (EU), a medicinal product needs a marketing authorization (MA) to be placed on the market. The EU’s medicinal products’ legislative framework allows for a reduced application for medicines outside their data exclusivity. One such type of application is the well- established use (WEU) medicinal product application (i.e. bibliographic applications). Recently, these MA applications have been subject to arbitration procedures at the European Medicines Agency’s Committee for Medicinal Products for Human Use (CHMP) because of disagreements between member states during the authorisation process. This paper reflects on these cases and highlights their potential impact on future WEU applications. Methods: Decisions adopted by the European Commission on WEU applications between 2009 and 2012 were identified from the EU Community Register on medicinal products for human use. Subsequently, decisions were reviewed to understand the potential serious risk to public health (PSRPH) that EU regulators raised during MA application procedures. Results: Four decisions were adopted by the EU commission between 2009 and 2012. Three followed disagreements between member states on PSRPH grounds. One decision was the outcome of a centralised marketing authorisation application. Six key messages were identified from the four cases reviewed and presented. Conclusion: A guideline on WEU to implement the technical specifications to fulfil Annex I of Directive 2001/83/EC for MA applications is not available. Thus, reflections on recent decisions on WEU applications provide scientific direction to the industry as well as the medicinal product regulators on the documentation required to successfully file and obtain a WEU MA.

2014 - Stamina therapies: Let the record stand [Articolo su rivista]
Bianco, Paolo; Cattaneo, Elena; DE LUCA, Michele; Pani, Luca


2014 - Steps forward in regulatory pathways for acute and chronic heart failure [Articolo su rivista]
Pani, L.; Pecorelli, S.; Rosano, G.; Anker, S. D.; Peracino, A.; Fregonese, L.; Prasad, K.; Rasi, G.

A workshop was organized by the Agenzia Italiana del Farmaco (AIFA) to discuss unmet needs and ways forward in the development of medicines in heart failure, their rationale, and cost-effective use. An integrated, multidisciplinary approach, including patients' needs and perspectives, was advocated by all the participants as the way to the most effective treatment regimens. More work is needed for reaching consensus on clinical and functional endpoints, for validating patient reported outcomes and measurements of well-being. Similarly, the integration into the clinical programmes of the health technology assessment/payers perspective, in particular, the evaluation of ‘real-life’ treatment effectiveness and of health as a value, would help in shifting the development and authorization of medicines from the molecule paradigm to their evaluation in the context of the whole health care regimen. Through this kind of workshop, AIFA is trying to build a template for meetings devoted to debate unmet needs with all stakeholders towards tentative road maps for the future.

2014 - The effects of novel and newly approved antipsychotics on serum prolactin levels: A comprehensive review [Articolo su rivista]
Peuskens, J.; Pani, L.; Detraux, J.; De Hert, M.

Since the 1970s, clinicians have increasingly become more familiar with hyperprolactinemia (HPRL) as a common adverse effect of antipsychotic medication, which remains the cornerstone of pharmacological treatment for patients with schizophrenia. Although treatment with second-generation antipsychotics (SGAs) as a group is, compared with use of the first-generation antipsychotics, associated with lower prolactin (PRL) plasma levels, the detailed effects on plasma PRL levels for each of these compounds in reports often remain incomplete or inaccurate. Moreover, at this moment, no review has been published about the effect of the newly approved antipsychotics asenapine, iloperidone and lurasidone on PRL levels. The objective of this review is to describe PRL physiology; PRL measurement; diagnosis, causes, consequences and mechanisms of HPRL; incidence figures of (new-onset) HPRL with SGAs and newly approved antipsychotics in adolescent and adult patients; and revisit lingering questions regarding this hormone. A literature search, using the MEDLINE database (1966-December 2013), was conducted to identify relevant publications to report on the state of the art of HPRL and to summarize the available evidence with respect to the propensity of the SGAs and the newly approved antipsychotics to elevate PRL levels. Our review shows that although HPRL usually is defined as a sustained level of PRL above the laboratory upper limit of normal, limit values show some degree of variability in clinical reports, making the interpretation and comparison of data across studies difficult. Moreover, many reports do not provide much or any data detailing the measurement of PRL. Although the highest rates of HPRL are consistently reported in association with amisulpride, risperidone and paliperidone, while aripiprazole and quetiapine have the most favorable profile with respect to this outcome, all SGAs can induce PRL elevations, especially at the beginning of treatment, and have the potential to cause new-onset HPRL. Considering the PRL-elevating propensity of the newly approved antipsychotics, evidence seems to indicate these agents have a PRL profile comparable to that of clozapine (asenapine and iloperidone), ziprasidone and olanzapine (lurasidone). PRL elevations with antipsychotic medication generally are dose dependant. However, antipsychotics having a high potential for PRL elevation (amisulpride, risperidone and paliperidone) can have a profound impact on PRL levels even at relatively low doses, while PRL levels with antipsychotics having a minimal effect on PRL, in most cases, can remain unchanged (quetiapine) or reduce (aripiprazole) over all dosages. Although tolerance and decreases in PRL values after long-term administration of PRL-elevating antipsychotics can occur, the elevations, in most cases, remain above the upper limit of normal. PRL profiles of antipsychotics in children and adolescents seem to be the same as in adults. The hyperprolactinemic effects of antipsychotic medication are mostly correlated with their affinity for dopamine D2 receptors at the level of the anterior pituitary lactotrophs (and probably other neurotransmitter mechanisms) and their blood-brain barrier penetrating capability. Even though antipsychotics are the most common cause of pharmacologically induced HPRL, recent research has shown that HPRL can be pre-existing in a substantial portion of antipsychotic- naïve patients with first-episode psychosis or at-risk mental state. © 2014 The Author(s).

2014 - To the AIFA Director [Articolo su rivista]
Pani, L.

2014 - Towards a framework for treatment effectiveness in schizophrenia [Articolo su rivista]
Juckel, G.; de Bartolomeis, A.; Gorwood, P.; Mosolov, S.; Pani, L.; Rossi, A.; Sanjuan, J.

Introduction: Prompt administration of antipsychotic treatment that is adhered to is essential for the optimal treatment of schizophrenia. Many patients have benefited from the advent of second-generation antipsychotics, which can offer good symptomatic control with reduced incidence of extrapyramidal symptoms, although with higher risk of metabolic side effects. It is unsurprising that accounts as to whether first- and second-generation antipsychotics differ in their efficacy vary, since treatment effectiveness is a broad notion and difficult to define.Objectives: Numerous factors may be used to gauge treatment effectiveness and, while it has largely been defined in terms of improvements in four domains (symptoms of disease, treatment burden, disease burden, and health and wellness), the real-world clinical utility of this consensus is unclear. Therefore, this article aims to provide a framework that can aid psychiatrists in making assessments about treatment effectiveness.Methods and results: A panel of 12 psychiatrists and psychopharmacologists convened to develop and propose an accessible and globally-applicable framework for assessing the effectiveness of antipsychotic treatments in patients with schizophrenia. Following presentation of a preliminary proposal to a wider group of psychiatrists from across Europe, it was refined into a framework comprising five domains: symptomatic remission and retention of treatment; affective symptoms; cognitive functioning; treatment satisfaction; and personal and social functioning - each of which is discussed in this article.Conclusions: This article provides a framework that can aid psychiatrists in making assessments about treatment effectiveness. It is anticipated that the framework outlined here may contribute to improving clinical practice through the promotion of a patient-centered approach to the assessment of treatment effectiveness, using five specified domains, in patients with schizophrenia.

2014 - Use and reimbursement of off-label drugs in pediatric anesthesia: The Italian experience [Articolo su rivista]
Salvo, I.; Landoni, G.; Mucchetti, M.; Cabrini, L.; Pani, L.

Background Most of the drugs used in anesthesia are off-label in children even if they present solid clinical evidence in adults. This lack of authorization is caused by multiple factors including the difficulty in conducting research in this area (due to the ethical concerns and/or the low number of available participants, the high variability of the outcome measures) and the lack of economic interest of the pharmaceutical companies (due to the limited market). Objective Define a list of medicinal products commonly used off-label in pediatrics anesthesia to be reimbursed by Italian National Health System. Methods and results We hereby describe the methodological framework used to allow reimbursed use of a list of medicinal products, widely used off-label in pediatric patients, ensuring the best therapeutic results with the lowest possible risk for children. A task force of pediatric anesthesiologists from Italy petitioned the Italian Medicines Agency (AIFA) to allow a number of commonly utilized but off-label drugs for pediatric anesthesia to be reimbursed for specific indications. For each drug, both the supporting literature and expert opinion were used, and the resulting list of drugs allowed to be used/reimbursed officially by AIFA was significantly expanded. This paper documents one approach to the problem of off-label use of drugs for pediatric patients that can be a model for future efforts. Conclusion Continuous efforts are needed from government institutions and sponsors on drug development and on drug approval process in pediatrics, as research on drug effectiveness and safety is mandatory in children as in adults. At the same time, clinicians must become more familiar with the drug-approval process, participate to sponsored trials, and perform ztrials themselves. © 2014 John Wiley & Sons Ltd.

2014 - Validated method to determine quetiapine and norquetiapine in microsomal matrix by LC MS-MS: Implication in quetiapine metabolism [Articolo su rivista]
Peddio, G.; Pittau, B; Manca, I.; Salis, R.; Pani, L.; Pira, L.

2013 - Biosimilars: The paradox of sharing the same pharmacological action without full chemical identity [Articolo su rivista]
Pani, L.; Montilla, S.; Pimpinella, G.; Bertini Malgarini, R.

The use of biotech medicines is increasing, with consequent mounting expenses for National Health Systems (NHSs). Biosimilars should be considered an opportunity to improve access to care. On the other side, the general public might suspect to receive low-quality medicines to save money. Actually, no drugs with a lesser degree of pharmaceutical quality with respect to existing alternatives can be authorized on the ground of a lower price. Biosimilars can be authorized only if their quality is of the same level as that of the originator. There is no chemical identity between biosimilars and the originators: any differences in quality attributes must be justified and shown not to impact on the safety and efficacy of the biosimilar by scientific investigations including pre-approval nonclinical and/or clinical studies. The biosimilar safety profile may be different from the originator or change over time for the same product. Hence caveats limiting the widespread use of biosimilars yet exist and should be solved by education on the main biological issues of biotech medicines, and on continuous update of the rules set up by the Regulatory Authorities to assess biosimilarity and to monitor post-approval safety. © 2013 Informa UK, Ltd.

2013 - Disclosure of grounds of European withdrawn and refused applications: A step forward on regulatory transparency [Articolo su rivista]
Tafuri, G.; Trotta, F.; Leufkens, H. G. M.; Pani, L.

2013 - Is HCMV vaccine an unmet need? The state of art of vaccine development [Articolo su rivista]
Chiurchiu, S.; Calo Carducci, F. I.; Rocchi, F.; Simonetti, A.; Bonatti, G.; Salmaso, S.; Melchiorri, D.; Pani, L.; Rossi, P.

Congenital HCMV infection is the most frequent congenital infection, with an incidence of 0.2-2.5% among all live births. About 11% of infected newborns show symptoms at birth, including hepatosplenomegaly, thrombocytopenia, neurologic involvement, hearing impairment and visual deficit. Moreover, 5-25% of the asymptomatic congenital HCMV-infected neonates will develop sequelae over months or even years. The relevant social burden, the economic costs of pre-natal screening, post-natal diagnosis, follow-up and possible therapy, although still limited, are the major factors to be considered. Several types of vaccines have been explored in order to develop an effective and safe HCMV vaccine: live attenuated, subunit, vectored, peptide, DNA, and subviral ones, but none are available for use. This review illustrates the different vaccine types studied to date, focusing on the possible vaccination strategy to be implemented once the HCMV vaccine is available, in terms of target population. Copyright © by BIOLIFE, s.a.s.

2013 - Metformin-associated lactic acidosis requiring hospitalization. A national 10 year survey and a systematic literature review [Articolo su rivista]
Renda, F.; Mura, P.; Finco, G.; Ferrazin, F.; Pani, L.; Landoni, G.

Metformin is known to be rarely associated with lactic acidosis, a serious condition with a poor prognosis. To review the National Pharmacovigilance Network of the Italian Medicines Agency reporting cases of metformin-associated lactic acidosis. The National Pharmacovigilance Network of the Italian Medicines Agency, was searched for cases of lactic acidosis that occurred in a 10 years period (from November 2001 to October 2011). Data were analyzed, to identify associated clinical features. A systematic literature research was performed to identify other large case series on metformin associated lactic acidosis. Metformin was the antidiabetic drug most frequently associated with lactic acidosis in the assessed period. Metformin-associated lactic acidosis was the most frequent serious adverse reaction related to metformin reported to the national authority (18.2% of all 650 adverse drug reactions reported). There were 59 cases of metformin-associated lactic acidosis (mortality rate of 25.4%). In most patients (89.8%) there was at least one risk factor for the occurrence of lactic acidosis. The predictors of death were low arterial blood pH and absence of acute renal failure. The systematic research of the literature identified only six case-series with more than 30 patients. This is the second largest case series ever reported on metformin-associated lactic acidosis. We confirmed that this rare complication of metformin is frequently fatal. Death can be predicted when the patient arrive in the hospital with low pH and, not intuitively, if the patient has no acute kidney injury. Risk minimisation measures taken at national level to prevent this serious complication are described.

2013 - Regulatory evaluation of Glybera in Europe-two committees, one mission [Articolo su rivista]
Melchiorri, D; Pani, L; Gasparini, P; Cossu, G; Ancans, J; Borg, Jj; Drai, C; Fiedor, P; Flory, E; Hudson, I; Leufkens, Hg; Müller-Berghaus, J; Narayanan, G; Neugebauer, B; Pokrotnieks, J; Robert, Jl; Salmonson, T; Schneider, Ck.

2012 - DSM-5 and clinical trials in psychiatry: Challenges to come? [Articolo su rivista]
Butlen-Ducuing, F.; Haas, M.; Pani, L.; Van Zwieten-Boot, B.; Broich, K.

The publication of the fifth version of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) in 2013 raises important questions for clinical trials and associated regulatory decisions on new drugs for psychiatric disorders. © 2012 Macmillan Publishers Limited. All rights reserved.

2012 - Discriminative Hypomania Checklist-32 Factors in Unipolar and Bipolar Major Depressive Patients [Articolo su rivista]
Perugi, G.; Fornaro, M.; Maremmani, I.; Canonico, P. L.; Carbonatto, P.; Mencacci, C.; Muscettola, G.; Pani, L.; Torta, R.; Vampini, C.; Parazzini, F.; Dumitriu, A.; Angst, J.

Background: Although manic or hypomanic episodes define bipolar disorder (BD), most patients show a predominance of depressive symptomatology, often associated with delayed or disregarded BD diagnosis. The Hypomania Checklist-32 (HCL-32) has therefore been developed and tested internationally to facilitate BD recognition. Sampling andMethods: Five hundred seventy-one (563 eligible) patients diagnosed with a major depressive episode according to DSM-IV criteria were consecutively enrolled in a cross-sectional, multicenter, observational study (Come To Me). Lifetime manic or hypomanic features were assessed by the HCL-32, and severity of depressive and anxious symptomatology was assessed using the Zung's self-report questionnaires for depression and anxiety. Results: Among the patients diagnosed with BD (n = 119), either type I or type II, the occurrence of (hypo)manic symptoms was significantly higher compared to major depressive disorder (MDD) symptoms according to HCL-32 total and subscale scores obtained using a score of 14, which ensured an optimal discrimination between BD and MDD with a sensitivity of 0.85 and a specificity of 0.78. Conclusions: Although some false positives might occur, the HCL-32 was confirmed to be a useful instrument in the detection of past hypomania in MDD patients, finally contributing to proper therapeutic choices. Copyright © 2012 S. Karger AG, Basel.

2012 - European licensing of maintenance treatment in schizophrenia [Articolo su rivista]
Isaac, M.; Pani, L.; Gispen-De Wied, C.; Koch, A.

2012 - NESS038C6, a novel selective CB1 antagonist agent with anti-obesity activity and improved molecular profile [Articolo su rivista]
Mastinu, A.; Pira, M.; Pani, L.; Pinna, G. A.; Lazzari, P.

The present work aims to study the effects induced by a chronic treatment with a novel CB1 antagonist (NESS038C6) in C57BL/6N diet-induced obesity (DIO) mice. Mice treated with NESS038C6 and fed with a fat diet (NESS038C6 FD) were compared with the following three reference experimental groups: DIO mice fed with the same fat diet used for NESS038C6 and treated with vehicle or the reference CB1 antagonist/inverse agonist rimonabant, " VH FD" and " SR141716 FD" , respectively; DIO mice treated with vehicle and switched to a normal diet (VH ND).NESS038C6 chronic treatment (30. mg/kg/day for 31 days) determined a significant reduction in DIO mice weight relative to that of VH FD. The entity of the effect was comparable to that detected in both SR141716 FD and VH ND groups.Moreover, if compared to VH FD, NESS038C6 FD evidenced: (i) improvement of cardiovascular risk factors; (ii) significant decrease in adipose tissue leptin expression; (iii) increase in mRNA expression of hypothalamic orexigenic peptides and a decrease of anorexigenic peptides; (iv) expression increase of metabolic enzymes and peroxisome proliferator-activated receptor-α in the liver; (v) normalization of monoaminergic transporters and neurotrophic expression in mesolimbic area. However, in contrast to the case of rimonabant, the novel CB1 antagonist improved the disrupted expression profile of genes linked to the hunger-satiety circuit, without altering monoaminergic transmission.In conclusion, the novel CB1 antagonist compound NESS038C6 may represent a useful candidate agent for the treatment of obesity and its metabolic complications, without or with reduced side effects relative to those instead observed with rimonabant. © 2012 Elsevier B.V.

2012 - Subthreshold depression in older subjects: An unmet therapeutic need [Articolo su rivista]
Cherubini, A.; Nistico, G.; Rozzini, R.; Liperoti, R.; Di Bari, M.; Zampi, E.; Ferrannini, L.; Aguglia, E.; Pani, L.; Bernabei, R.; Marchionni, N.; Trabucchi, M.

Major depression, defined according to DSM IV TR criteria, is less common in older subjects, while other types of depression are two to three times more prevalent. This heterogeneous group of disturbances has received different names: depression not otherwise specified, minor depression, subthreshold or subsyndromal depression. Moreover, each condition has been defined using heterogeneous criteria by different authors. The term of subthreshold depression will be adopted in this position statement. Subthreshold depression has been associated with the same negative consequences of major depression, including reduced well being and quality of life, worsening health status, greater disability, increased morbidity and mortality. Nevertheless, there is a dearth of clinical trials in this area, and therefore older patients with subthreshold depression are either not treated or they are treated with the same non pharmacological and pharmacological therapies used for major depression, despite the lack of supporting scientific evidence. There is an urgent need to reach a consensus concerning the diagnostic criteria for subthreshold depression as well as to perform clinical trials to identify effective and safe therapies in this too long neglected patient group. © 2012 Serdi and Springer-Verlag France.

2012 - The placebo arm in clinical studies for treatment of Psychiatric Disorders: A Regulatory Dilemma [Articolo su rivista]
Gispen-de Wied, C.; Stoyanova, V.; Yu, Y.; Isaac, M.; Pani, L.; de Andres-Trelles, F.

Background: The use of placebo in clinical trials, and, related to this, ethical and feasibility aspects, are often debated. However, regulatory authorities must ensure that only new drugs with a positive benefit/risk would be granted a marketing authorization. It is therefore not surprising that they often put forward the need for placebo control in clinical trials in an area where many trials fail, and assay sensitivity is not self-evident. To illustrate the complexity that regulatory authorities encounter when faced with the registration dossier of products in the main psychiatric therapeutic areas, Major Depressive Disorder (MDD) and schizophrenia, the trial outcome for products receiving an opinion in the EU during the past 15 years were reviewed. Data source: European Public Assessment Reports and registration files. Results: A total of 45 studies qualified for analysis. For the indication MDD 38% of the studies (10/26) were recorded as failed, and another 15% (4/26) as negative. For schizophrenia, these figures were 16% (3/19) and 11% (2/19). Further exploration of the trials in MDD revealed an inconsistent pattern in terms of magnitude of placebo- and drug-mediated response (i.e. similar studies with consistent placebo response provided different treatment outcomes). Conclusion: From a regulatory perspective the dilemma of a priori exclusion of the placebo arm in clinical trials in the domains of depression or schizophrenia cannot be solved at this time as long as factors influencing trial variability are not better identified or understood. This counts in particular for MDD where the added drug effect is not consistent across trials with almost identical inclusion criteria. Unfortunately, this trend has not changed over the past 15 years. However, all efforts should be taken to optimize the clinical development of drugs in the psychiatric domain, and improve the intrinsic quality of the clinical trials in order to allow for a different viewpoint. © 2012 Elsevier B.V. and ECNP.

2012 - Tricyclic pyrazoles. Part 5. novel 1,4-Dihydroindeno[1,2-c]pyrazole CB2 ligands using molecular hybridization based on scaffold hopping [Articolo su rivista]
Murineddu, G; Asproni, B; Ruiu, S; Deligia, F; Falzoi, M; Pau, A; Thomas, Bf; Zhang, Y; Pinna, Ga; Pani, L; Lazzari, P.

2011 - Are "social drugs" (tobacco, coffee and chocolate) related to the bipolar spectrum? [Articolo su rivista]
Maremmani, I.; Perugi, G.; Rovai, L.; Maremmani, A. G. I.; Pacini, M.; Canonico, P. L.; Carbonato, P.; Mencacci, C.; Muscettola, G.; Pani, L.; Torta, R.; Vampini, C.; Akiskal, H. S.

Background: Across all ages and cultures, mankind has always used substances in order to induce pleasurable sensations or desirable psychophysical states. These substances, notably caffeine, tobacco, alcohol and chocolate, can be labeled 'social drugs'. Methods: We analyzed the social drug habits of 562 patients suffering from mood disorders, according to DSM-IV-R criteria (major depressive episode, recurrent depression, bipolar type I and II disorders and depression not otherwise specified). The sample was also divided into bipolar and non-bipolar according to Hypomania Check-list 32 (HCL-32), which proposes a broader concept of hypomania and soft bipolarity, comprising the spectrum of bipolar disorders proper, along with other, "softer" expressions of bipolarity intermediate between bipolar disorder and normality. Results: Using HCL-32 criteria, but DSM-IV-R criteria, a link was confirmed between bipolar spectrum and substance use including social drugs such as tobacco and coffee. Limitation: Observational correlational study. Conclusion: This study is in support of earlier theoretical formulations within the framework of the Pisa-San Diego collaboration. © 2011 Elsevier B.V. All rights reserved.

2011 - Novel approaches to drug-placebo difference calculation: Evidence from short-term antidepressant drug-trials [Articolo su rivista]
Rihmer, Z.; Gonda, X.; Dome, P.; Erdos, P.; Ormos, M.; Pani, L.

The calculation of antidepressant-placebo difference (50% - 30% = 20%) in drug trials is based on the postulate that all placebo responders should be 'automatically' antidepressant responders, an assumption that has been never been specifically investigated and substantiated. However, some studies show that a clinically significant part of placebo responders are also antidepressant nonresponders. The traditional calculation of antidepressant-placebo difference seems, therefore, to be wrong because of an inherent fundamental bias resulting in a marked overestimation of the placebo effect. If the mechanism of action of antidepressant and placebo are independent (unrelated) and the randomization results in two identical (homogenous) groups of patients, then the two basic principles by which the evaluation of potentially useful drugs are based on, such as the relationship between antidepressant response and placebo response rates, would also be independent. In this case, only 50% of placebo responders are antidepressant responders (and another 50% of them are antidepressant nonresponders) and the antidepressant-placebo difference would be 50% - 15% = 35%, instead of 50% - 30% = 20%, as calculated by the traditional method. For real interpretation, decisions that have been made on traditional drug-placebo difference evaluation should be recalculated and reviewed, not only in major depression but also in other psychiatric and medical disorders where the drug-placebo difference is in the same magnitude. © 2011 John Wiley & Sons, Ltd.

2011 - Prevalence and diagnostic distribution of medically unexplained painful somatic symptoms across 571 major depressed outpatients [Articolo su rivista]
Fornaro, M.; Maremmani, I.; Canonico, P. L.; Carbonatto, P.; Mencacci, C.; Muscettola, G.; Pani, L.; Torta, R.; Vampini, C.; Parazzini, F.; Dumitriu, A.; Perugi, G.

Objective: To assess the prevalence and distribution of medically unexplained painful somatic symptoms (PSSs) versus nonpainful somatic symptoms (NPSSs) in patients diagnosed with major depressive episode (MDE). Method: A total of 571 outpatients diagnosed with MDE according to DSM-IV-TR criteria were consecutively enrolled into a cross-sectional, multicentric, observational study over a period of 7 months. Subjects were evaluated by means of the ad hoc validated 30-item Somatic Symptoms Checklist (SSCL-30) and Zung's questionnaires for depression and anxiety. The 32-item Hypomania Checklist (HCL-32) was also administered in order to explore any eventual association of PSSs or NPSSs with sub-threshold (DSM-IV-TR [Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision] not recognized) bipolar disorder (BD). Results: In our sample, just 183 patients (32%) did not report painful somatic symptoms (NPSSs). Of these, 90 patients (15.76%) had no somatic symptoms at all. The remaining 388 (68%) had at least one PSS being subdivided as follows: 248 (43%) had one or two PSSs, while 140 (25%) experienced two or more. Patients with at least one PSS also reported a greater number of nonpainful somatic symptoms than NPSS. Bipolar patients (associated with higher HCL-32 scores) were less represented across PSS cases than NPSS subjects. Conversely, females were more prone to having a higher number of total somatic symptoms (and bipolar features). Conclusion: PSSs are common in patients with MDE, especially among those patients reporting fewer somatic symptoms in general as opposed to those patients who exhibit more somatic symptoms (both PSSs and NPSSs) with lower relative number of PSSs. A major therapeutic implication is that antidepressant monotherapy could be used with more confidence in unexplained PSS patients than in NPSS patients because of the latter group's lower frequency of (sub)-threshold bipolar features. © 2011 Fornaro et al.

2011 - Qualification opinion of novel methodologies in the predementia stage of Alzheimer's disease: Cerebro-spinal-fluid related biomarkers for drugs affecting amyloid burden - Regulatory considerations by European Medicines Agency focusing in improving benefit/risk in regulatory trials [Articolo su rivista]
Isaac, M.; Vamvakas, S.; Abadie, E.; Jonsson, B.; Gispen, C.; Pani, L.

The European Medicines Agency (EMA) in London is responsible for the Regulatory review of new medicinal products for Marketing Authorisation, through which pharmaceutical companies may obtain first Marketing Authorisation and subsequent Variations valid throughout the EU and EFTA.The qualification opinion of novel methodologies is a new procedure where applicants can obtain scientific advice on new methodologies for regulatory clinical trials of efficacy of new compounds. It will help benefit/risk assessment of the CHMP.The definition of prodromal AD is acceptable. The "Dubois Criteria" as criteria to define the population must be validated in full at the time of the submission of the dossiers.Including a positive CSF biomarker profile is considered predictive for the evaluation of the AD-dementia type. However, although high CSF tau and low CSF Aβ42 are predictive of Alzheimer's disease, the criterion "positive CSF tau/Aβ42 ratio" is not well defined.The qualification of biomarkers in the pre-dementia stage of Alzheimer's disease will allow better inclusion criteria of patients in pre-dementia trials in which the benefit/risk is higher for treatment with these novel compounds. © 2011 Elsevier B.V. and ECNP.

2011 - Unexplained somatic symptoms during major depression: Prevalence and clinical impact in a national sample of italian psychiatric outpatients [Articolo su rivista]
Perugi, G.; Canonico, P. L.; Carbonato, P.; Mencacci, C.; Muscettola, G.; Pani, L.; Torta, R.; Vampini, C.; Fornaro, M.; Parazzini, F.; Dumitriu, A.

Background: The aim of this study was to explore the prevalence and impact of unexplained somatic symptoms during major depression. Sampling and Methods: A total of 560 consecutive outpatients with a major depressive episode according to the DSM-IV (text revision) were evaluated in 30 psychiatric facilities throughout Italy. 'Unexplained' somatic symptoms were evaluated using the 30-item Somatic Symptoms Checklist (SSCL-30). Somatic symptoms were considered explained if they were best accounted for as coming from a concomitant physical illness or side effects. Patients evaluated their own mood symptomatology using the Zung questionnaires for depression and anxiety and the Hypomania Checklist-32. Results: According to the SSCL-30, only 90 subjects (16.1%) had no unexplained somatic symptoms, while 231 (41.3%) had 1-5 unexplained symptoms and 239 (42.7%) had more than 5. Asthenia was the most commonly observed unexplained somatic symptom (53% of patients). Unexplained somatic symptoms were more common in females and among those suffering from major depression and depression not otherwise specified rather than in patients with recurrent major depression and bipolar disorders. No relationship between unexplained somatic symptoms and hypomanic features was observed. Conclusions: The presence of a large number of unexplained somatic symptoms is associated with more severe depression and higher rates of misdiagnosis and inappropriate treatment. Copyright © 2011 S. Karger AG, Basel.

2011 - Weight loss induced by rimonabant is associated with an altered leptin expression and hypothalamic leptin signaling in diet-induced obese mice [Articolo su rivista]
Lazzari, Paolo; Sanna, Angela; Mastinu, Andrea; Cabasino, Simona; Manca, Ilaria; Pani, Luca

2010 - 1-(2′,4′-dichlorophenyl)-6-methyl-N-cyclohexylamine-1,4-dihy droindeno[1,2-c]pyrazole-3-carboxamide, a novel CB2 agonist, alleviates neuropathic pain through functional microglial changes in mice [Articolo su rivista]
Luongo, L.; Palazzo, E.; Tambaro, S.; Giordano, C.; Gatta, L.; Scafuro, M. A.; Rossi, F. s.; Lazzari, P.; Pani, L.; de Novellis, V.; Malcangio, M.; Maione, S.

Neuropathic pain is a devastating neurological disease that seriously affects quality of life in patients. The mechanisms leading to the development and maintenance of neuropathic pain are still poorly understood. However, recent evidence points towards a role of spinal microglia in the modulation of neuronal mechanisms. In this context, cannabinoids are thought to modulate synaptic plasticity as well as glial functions. Here, we have investigated the effect of chronic treatment with a selective agonist of cannabinoid type 2 receptor (CB2), 1-(2′,4′-dichlorophenyl)-6-methyl-N-cyclohexylamine-1,4-dihy droindeno[1,2-c]pyrazole-3 carboxamide (NESS400), on pain thresholds in the spared nerve injury (SNI) model in the mouse and on the distribution and activation of spinal microglia. Repeated treatment with NESS400 (4 mg/kg) significantly alleviated neuropathic mechanical allodynia and thermal hyperalgesia. In the dorsal horn (L4-L6) of neuropathic mice microglia activation (quantification of the length of microglial processes) and astrocytosis were associated with CB2 receptor over-expression on both cell types. Treatment with NESS400 significantly reduced the number of hypertrophic microglia while leaving microglial cell number unaffected and reduced astrogliosis. Moreover, prolonged administration of NESS400 reduced mRNA expression of pro-inflammatory markers and enhanced anti-inflammatory marker gene expression in dorsal horn extracts. In conclusion, we show that selective CB2 receptor stimulation prevents thermal hyperalgesia, alleviates mechanical allodynia and facilitates the proliferation of anti-inflammatory microglial phenotype in the ipsilateral dorsal horn of the spinal cord in SNI mice. © 2009 Elsevier Inc. All rights reserved.

2010 - A comparison of continuous subcutaneous paliperidone infusion and repeated subcutaneous injection of risperidone free-base in rats [Articolo su rivista]
Marchese, G.; Pittau, B.; Casu, G.; Peddio, G.; Spada, G. P.; Pira, M.; Deriu, A.; Portesani, F.; Pisu, C.; Lazzari, P.; Pani, L.

It is proposed that to achieve a therapeutic effect in schizophrenia patients, dopamine D2-receptor occupancy by antipsychotics within the striatum must exceed 60-65%. However, at high levels of D2-receptor occupancy, the risk of extrapyramidal symptoms (EPS) is increased. Following oral dosing of antipsychotics, peaks and troughs in plasma drug concentrations may be mirrored by fluctuations in D2-receptor occupancy. Paliperidone, a novel antipsychotic available as extended-release tablets (paliperidone ER), is the major active metabolite of risperidone and exhibits a plasma pharmacokinetic profile with reduced peak-trough fluctuations and consistent D2-receptor occupancy compared with conventional oral antipsychotic formulations. Using formulations that resemble those in clinical practice, this study provides a preclinical evaluation of the pharmacological properties of paliperidone ER and risperidone immediate-release formulation in terms of consistent antipsychotic efficacy over time and extrapyramidal symptom liability. Significant fluctuations in inhibition of d-amphetamine-induced hyperlocomotion were observed for repeated subcutaneous (SC) risperidone injections, whereas stable inhibitory efficacy was demonstrated during continuous SC paliperidone infusion. Similarly, significant fluctuations in latency on-bar were observed with repeated SC risperidone injections, whereas significantly lower latency on-bar was demonstrated following continuous SC paliperidone infusion. These results in an animal model suggest that although risperidone and paliperidone demonstrate similar pharmacologic effects, continuous administration of paliperidone achieves more stable antipsychotic efficacy with reduced motor impairment, akin to the effects observed with paliperidone ER in clinical studies. © 2009 Elsevier Masson SAS. All rights reserved.

2010 - Antinociceptive activity of Δ9-tetrahydrocannabinol non-ionic microemulsions [Articolo su rivista]
Lazzari, P.; Fadda, P.; Marchese, G.; Casu, G. L.; Pani, L.

Δ9-Tetrahydrocannabinol (Δ9-THC), the major psychoactive constituent of Cannabis sativa L., has been widely studied for its potential pharmaceutical application in the treatment of various diseases and disturbs. This sparingly soluble terpeno-phenolic compound is not easy to handle and to be formulated in pharmaceutical preparations.The aim of this work was to develop a stable aqueous Δ9-THC formulation acceptable for different ways of administration, and to evaluate the therapeutic properties of the new Δ9-THC based preparation for pain treatment. Due to the thermodynamic stability and advantages of microemulsion based systems, the study was focused on the identification of aqueous microemulsion based systems containing Δ9-THC.Oil in water Δ9-THC microemulsions were individuated through phase diagrams construction, using the non-ionic surfactant Solutol®HS15, being this surfactant acceptable for parenteral administration in human. A selected microemulsion samples containing 0.2wt% of Δ9-THC, stable up to 52°C, was successfully assayed on animal models of pain. Significant antinociceptive activity has been detected by both intraperitoneal and intragastric administration of the new Δ9-THC pharmaceutical preparation. The effect has been highlighted in shorter time if compared to a preparation of the same active principle based on previously reported conventional preparation. © 2010 Elsevier B.V.

2010 - Management of Comorbidity in Bipolar Disorder [Capitolo/Saggio]
Salloum, I. M.; Pani, L.; Cooke, T.

2010 - Multiplex genotyping of CYP3A4, CYP3A5, CYP2C9 and CYP2C19 SNPs using MALDI-TOF mass spectrometry [Articolo su rivista]
Falzoi, M.; Mossa, A.; Congeddu, E.; Saba, L.; Pani, L.

Background: Pharmacogenetics is the study of genetic variations that cause alterations in drug level, drug response and adverse drug reactions. SNPs found in CYP450 genes have the greatest genetic influences on interindividual variability in drug bioavailability. The polymorphic nature of these genes may modulate several enzyme levels that affect individual responses to pharmacological treatment. Among them, CYP3A4, CYP3A5, CYP2C9 and CYP2C19 isoforms of CYP450 enzymes are involved in the metabolism of many commonly prescribed drugs. Aims: In this study, we would like to develop a CYP450 genotyping platform that could lead a complete definition of a patients metabolic genotype in order to improve the clinical outcome of some drug treatments. Materials & methods: We used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (Sequenom®) to develop a SNP genotyping method. Results: This MALDI-TOF-based multiplexing system allows the simultaneous and efficient genotyping of a set of CYP450 gene polymorphisms. Conclusion: The multiple CYP450 gene testing achieved with this application can be used to develop diagnostic tests to predict drug responses and clinical outcomes. © 2010 Future Medicine Ltd.

2010 - Oct4 expression in in-vitro-produced sheep blastocysts and embryonic-stem-like cells [Articolo su rivista]
Sanna, D.; Sanna, A.; Mara, L.; Pilichi, S.; Mastinu, A.; Chessa, F.; Pani, L.; Dattena, M.

Transcription factor Oct4 (octamer-binding transcription factor-4) is important in early embryonic development and differentiation. It is also required for maintenance of pluripotency of the inner cell mass, and is used as a staminality marker of embryonic stem cells. Changes in Oct4 expression during the different stages of early embryo development have been reported, and therefore we have conducted a quantitative study of Oct4 gene expression of sheep blastocysts in vitro, and of embryonic-stem-like cells at the undifferentiated stage and in the course of differentiation. To characterize embryonicstem-like cells, alkaline phosphatase activity, stage-specific embryonic surface antigens SSEA-1, SSEA-3, SSEA-4 and three specific gene markers Nanog, Sox2 and Stat3 were assayed. cDNA produced by RT (reverse transcriptase)-PCR was synthesized and amplified by PCR; sequencing gave 98, 95 and 98% homology with the bovine sequences of Oct4, Nanog and Stat3 respectively. Using the ovine sequence of 290 bp, quantitative expression of Oct4 in the inner cell mass, trophoblast and embryonic-stem-like cells was performed by qRT-PCR (quantitative real-time PCR). Oct4 was expressed in the inner cell mass, trophoblast and embryonic-stem-like cells. Expression in the inner cell mass was significantly higher than in the trophoblast. This could be useful in defining the quality of embryos produced and makes it possible to use Oct4 to detect pluripotency. In addition, the different levels of Oct4 expression between undifferentiated and differentiating embryonic-stem-like cell cultures could be used to detect this gene as a staminality marker. © The Author(s) Journal compilation. © 2010 Portland Press Ltd.

2010 - Quetiapine anxiolytic-like effect in the Vogel conflict test is serotonin dependent [Articolo su rivista]
Pisu, C; Pira, L; Pani, L

2009 - Evaluation of amphetamine-induced hyperlocomotion and catalepsy following long-acting risperidone administration in rats [Articolo su rivista]
Marchese, G.; Casu, G.; Casti, P.; Spada, G. P.; Pani, L.

It has been proposed that long-acting risperidone could provide a constant antipsychotic efficacy associated with a reduced liability to induce extra-pyramidal symptoms. To ascertain this hypothesis, antagonism of amphetamine-induced hyperlocomotion and catalepsy were analyzed in rats for a period of 1-6 weeks following long-acting risperidone (20-60 mg/kg) injection. Long-acting risperidone reduced amphetamine-induced hyperlocomotion after 2-5 weeks from drug injection, without producing significant extra-pyramidal symptoms. Following the administration of long-acting risperidone a constant ability to antagonize amphetamine-induced hyperlocomotion was observed during the day, but not when the antipsychotic was chronically administered using a short-acting formulation. The pre-clinical results confirmed that long-acting risperidone may represent an advance in antipsychotic therapy. © 2009 Elsevier B.V. All rights reserved.

2009 - Expected clinical benefits of paliperidone extended-release formulation when compared with risperidone immediate-release [Articolo su rivista]
Pani, L.; Marchese, G.

Background: The development of paliperidone extended release (ER) may represent a new strategy to improve the pharmacological treatment of schizophrenia. The drug maintains the atypical antipsychotic profile of its parent compound risperidone, but it is associated with an innovative delivery system (OROS technology) that offers the possibility to obtain smooth drug plasma levels using an oral antipsychotic. Clinical trials confirmed that paliperidone ER is efficacious in the management of schizophrenia and well tolerated, however no direct clinical comparisons between paliperidone ER and immediate-release formulations of risperidone have been conducted to date. Objective: The present study evaluates possible differences between paliperidone ER and immediate-release formulations of risperidone due to structural/molecular and delivery system diversities, providing an estimation of their significance in the context of clinical results. Methods: A search of Medline and EMBASE was performed using the keywords 'Risperidone', 'Paliperidone' and 'OROS technology'. Results/conclusion: The analysis suggests that the chemical structure and pharmacokinetic profile of paliperidone ER might provide clinical benefits in terms of efficacy, tolerability and more consistent drug response among patients, when compared with the parent compound risperidone in its immediate release formulations. The relevance of these differences is discussed, taking into account several clinical aspects involved in the drug therapy of schizophrenia. © 2009 Informa UK Ltd All rights reserved.

2009 - Synthesis and enzymatic evaluation of novel partially fluorinated thiol dual ACE/NEP inhibitors [Articolo su rivista]
Olimpieri, F.; Tambaro, S.; Fustero, S.; Lazzari, P.; Sanchez-Rosello, M.; Pani, L.; Volonterio, A.; Zanda, M.

A novel family of peptidomimetics incorporating fluoroalkyl groups, mainly a trifluoromethyl, in α-position to a zinc(II)-binding thiol function, was synthesized in solution as well as in solid-phase. These compounds showed inhibitory potency in the nanomolar range against both angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP), whereas no inhibition of endothelin-converting enzyme-1 (ECE-1) was observed. The trifluoromethyl-derivatives were more potent than the parent unfluorinated analogues in the case of ACE, and less potent in the case of NEP. © 2009 Elsevier Ltd. All rights reserved.

2009 - Synthesis and in vitro evaluation of trifluoroethylamine analogues of enkephalins [Articolo su rivista]
Sinisi, R.; Ghilardi, A.; Ruiu, S.; Lazzari, P.; Malpezzi, L.; Sani, M.; Pani, L.; Zanda, M.

(Chemical Equation Presented) No gain with pain: At least when it comes to analgesics! Here we show that enkephalin analogues with the Gly 3-Phe 4 peptide bond replaced by a stereochemically defined trifluoroethylamine function display binding affinities in the nanomolar range for the - and -opioid receptors, only 30- to 80-fold lower than those of the natural compounds, whereas the Gly 2-Gly 3 trifluoroethylamine analogues have lower affinity. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.

2009 - The need for individualised antipsychotic drug therapy in patients with schizophrenia [Articolo su rivista]
Pani, L

2008 - Building a virtual archive using brain architecture and Web 3D to deliver neuropsychopharmacology content over the Internet [Articolo su rivista]
Mongeau, R.; Casu, M. A.; Pani, L.; Pillolla, G.; Lianas, L.; Giachetti, A.

The vast amount of heterogeneous data generated in various fields of neurosciences such as neuropsychopharmacology can hardly be classified using traditional databases. We present here the concept of a virtual archive, spatially referenced over a simplified 3D brain map and accessible over the Internet. A simple prototype (available at has been realized using current Web-based virtual reality standards and technologies. It illustrates how primary literature or summary information can easily be retrieved through hyperlinks mapped onto a 3D schema while navigating through neuroanatomy. Furthermore, 3D navigation and visualization techniques are used to enhance the representation of brain's neurotransmitters, pathways and the involvement of specific brain areas in any particular physiological or behavioral functions. The system proposed shows how the use of a schematic spatial organization of data, widely exploited in other fields (e.g. Geographical Information Systems) can be extremely useful to develop efficient tools for research and teaching in neurosciences. © 2007 Elsevier Ireland Ltd. All rights reserved.

2008 - Delta-9-tetrahydrocannabinol differently affects striatal c-Fos expression following haloperidol or clozapine administration [Articolo su rivista]
Marchese, G.; Sanna, A.; Casu, G.; Casti, P.; Spada, G. P.; Ruiu, S.; Pani, L.

It was previously shown that haloperidol, but not clozapine, induced intense rat catalepsy when co-administered with delta-9-tetrahydrocannabinol. The present study investigated whether similar alterations could be observed on striatal c-Fos immunoreactivity after administration of the same drug combinations. Western Blot and immunocytochemistry stereological analyses indicated that delta-9-tetrahydrocannabinol (0.5 mg/kg) increased striatal c-Fos immunoreactivity induced by haloperidol (0.1 mg/kg). Conversely, no significant alterations of striatal c-Fos immunoreactivity were observed after injections of clozapine (10 mg/kg) + vehicle, clozapine + delta-9-tetrahydrocannabinol or vehicle + delta-9-tetrahydrocannabinol. The present results indicate that the behavioral effects induced by delta-9-tetrahydrocannabinol in haloperidol- and clozapine-treated rats are associated with different striatal c-Fos expressions. © 2008 Elsevier B.V. All rights reserved.

2008 - Practical issues with amisulpride in the management of patients with schizophrenia [Articolo su rivista]
Pani, L.; Villagran, J. M.; Kontaxakis, V. P.; Alptekin, K.

Amisulpride is an atypical antipsychotic with a significantly greater effect size than first-generation, typical antipsychotics, and efficacy at least similar to that of olanzapine and risperidone in large-scale clinical trials in schizophrenia. Amisulpride provides greater improvement in positive and negative symptoms of schizophrenia, a better long-term outcome than typical antipsychotics, and distinct tolerability advantages over typical antipsychotics, which are reported to cause extrapyramidal symptoms (EPS) in 20-50% of patients. In addition, amisulpride is associated with significantly less weight gain than olanzapine and risperidone, does not increase body mass index, and favourably influences lipid profiles. In many patients with schizophrenia, adverse events impair adherence to treatment, and switching from typical or atypical antipsychotic therapy to amisulpride may be clinically appropriate. Observational drug-utilization studies suggest that many physicians switch to amisulpride because of fewer EPS and/or less weight gain and improved patient adherence. Cross-tapering (over 4 weeks), rather than abrupt cessation of pre-switch treatment, is preferred. Amisulpride has a low risk of drug-drug interactions, and, during cross-tapering, patients can remain on concurrent treatments (e.g. anticholinergics and antiparkinsonian agents) until the effective dosage has been reached. An appropriate amisulpride starting dose is 800 mg/day for patients with acute psychotic exacerbations, 400-800 mg/day for patients with predominantly positive symptoms, and 100-300 mg/day for predominantly negative symptoms. Amisulpride may be particularly suitable for clozapine-augmentation therapy in patients with refractory schizophrenia. Indeed, amisulpride is more effective than quetiapine as augmentation therapy in patients partially responsive to clozapine, and several prospective open-label studies and case series have reported promising results for amisulpride/ clozapine combination therapy. In three prospective studies, addition of amisulpride 200-800 mg/day to clozapine significantly reduced mean scores on the Brief Psychiatric Rating Scale (BPRS) total (-33% to -35%), Clinical Global Impression (CGI)-Severity scale (-31%), Positive and Negative Syndrome Scale total (-22%), and Scale for the Assessment of Negative Symptoms (-34%). The proportion of responders (CGI score ≥3 or BPRS improvement >20%) was 71-86%. Retrospective case-series analyses have also reported improved psychopathological state, reduced adverse events, and lower clozapine dosage requirement with use of this combination. The pharmacological and clinical profiles of amisulpride suggest that this agent is a viable clinical option when a change of antipsychotic therapy is required in patients with schizophrenia because of lack of efficacy, adverse events and poor adherence to treatment, or for augmentation of clozapine in treatment-resistant illness. © 2008 Adis Data Information BV. All rights reserved.

2008 - Remission in schizophrenia: Clinical and educational aspects. The Remission Project | La valutazione della remissione nella schizofrenia: Aspetti clinici ed educazionali. II progetto remissione [Articolo su rivista]
Aguglia, E; Bogetto, F; Cavallaro, R; Galderisi, S; Pani, L; Rossi, A; Sacchetti, E; Siracusano, A; Smeraldi, E.

2007 - 3-{2-[Bis-(4-fluorophenyl)methoxy]ethyl}-6-substituted-3,6-diazabicyclo[ 3.1.1]heptanes as novel potent dopamine uptake inhibitors [Articolo su rivista]
Loriga, G.; Ruiu, S.; Manca, I.; Murineddu, G.; Dessi, C.; Pani, L.; Pinna, G. A.

A series of analogues 2a-i related to 3-{2-[bis-(4-fluorophenyl)methoxy]ethyl}-8-(1H-indol-2-ylmethyl)-3,8-dia zabicyclo[3.2.1]octane (1) in which the 3,8-diazabicyclo[3.2.1]octane core was replaced by 3,6-diazabicyclo[3.1.1]heptane ring system has been synthesized and evaluated for their ability to inhibit DA reuptake into striatal nerve endings (synaptosomes). Biological data showed that compound 2a, the closest analogue of lead 1, possessed an increased reuptake inhibition activity over 1 (2a, Ki = 5.5 nM). Replacement of the indole ring with bioisosteric aromatic rings-benzothiophene (2b), benzofurane (2c), or indene (2d)-resulted, with the exception of 2d, in a double digit nanomolar activity. Changing the indenyl moiety of 2d with simplified aryl groups led to compounds 2e-h which displayed a similar or slightly decreased activity with respect to the ground term. Naphthalene derivative (2i) demonstrated a weaker activity than aromatic analogues. © 2007 Elsevier Ltd. All rights reserved.

2007 - Antipsychotic efficacy: Relationship to optimal D2-receptor occupancy [Articolo su rivista]
Pani, L.; Pira, L.; Marchese, G.

Clinically important differences exist between antipsychotic agents and formulations in terms of safety and tolerability. Features of the biochemical interaction between the antipsychotic and the D2-receptor may underlie these differences. This article reviews current information on the relationship between antipsychotic receptor occupancy and clinical response. A literature search was performed using the keywords 'antipsychotic or neuroleptic', 'receptor' and 'occupancy' and 'dopamine' and 'D2' supplemented by the authors' knowledge of the literature. Imaging and clinical data have generally supported the hypotheses that optimal D2-receptor occupancy in the striatum lies in a 'therapeutic window' between ∼65 and ∼80%, however, pharmacokinetic and pharmacodynamic properties of a drug should also be taken into account to fully evaluate its therapeutic effects. Additional research, perhaps in preclinical models, is needed to establish D2-receptor occupancy in various regions of the brain and the optimal duration of D2-receptor blockade in order to maximise efficacy and tolerability profiles of atypical antipsychotics and thereby improve treatment outcomes for patients with schizophrenia. © 2007 Elsevier Masson SAS. All rights reserved.

2007 - Contrasting effects of diazepam and repeated restraint stress on latent inhibition in mice [Articolo su rivista]
Mongeau, R.; Marcello, S.; Andersen, J. S.; Pani, L.

The effects on latent inhibition (LI; a delay in conditioning when a CS has been pre-exposed without consequences) of repeated restraint stress and the anxiolytic drug diazepam were examined in C57BL/6 mice to know whether previous aversive events or anxiolysis are factors determining the expression of LI. The LI model was optimized for this strain particularly sensitive to stress (using both the CER and the conditioned freezing procedures) and characterized with typical (haloperidol) and atypical (clozapine and olanzapine) antipsychotic drugs administered either during the conditioning or the pre-exposure phases. An acute challenge with amphetamine, a dopamine releaser, was done to verify the enhancement of hyperactivity in C57BL/6 mice after the restraint stress sensitization. At all doses tested, diazepam decreased latent inhibition when administered during the pre-exposure phase (similarly to atypical antipsychotic drugs). Repeated restraint stress enhanced LI by blocking the CS-induced freezing in pre-exposed mice. In contrast, pre-treatment with diazepam before pre-exposure allowed the expression of CS-induced freezing in stressed mice pre-exposed to the tone. It is suggested that stress and anxiolytic drugs can have opposite effects on attention or perseveration processes during learning of conflicting contingency responses. © 2007 Elsevier B.V. All rights reserved.

2007 - Effects of acute and chronic valproate treatments on p-CREB levels in the rat amygdala and nucleus accumbens [Articolo su rivista]
Casu, M. A; Sanna, A; Spada, G. P; Falzoi, M; Mongeau, R; Pani, L.

2006 - Bipolar depression and quetiapine: Results of a preliminary study [Articolo su rivista]
Mannu, P.; Minnai, G.; Pani, L.

Introduction. Although the most part of "bipolar life" is a "depressive life", literature data about the treatment of bipolar depression are very poor. Aim. Aim of this work is to test the efficacy and tolerability of quetiapine in the treatment of BD. Materials and Methods. Fourteen bipolar depressed patients (mean HAM-D 26.3±2.4) have been treated using quetiapine 450.0±75.0 mg/day in a naturalistic, flexible-dose setting. Results. Quetiapine has shown a "full" and fast effectiveness in all depressed bipolar patients. Conclusions. Although the clear methodological limitations of this study, the clinical activity and the good tolerability profile of quetiapine in depressive episodes associated to Bipolar Disorder (BD) have been proved.

2006 - Evolutionary psychopharmacology, mental disorders, and ethical behavior [Capitolo/Saggio]
Canali, S.; De Anna, G.; Pani, L.

The concept of pathology has a built-in normative character. An individual (or a state) is pathological if and only if it is not normal, that is, if it fails to be as an individual (or a state) of that sort ought to be. In other words, a pathological individual (or state) is an individual (or state) that does not conform to the standards to which all the individuals (or states) falling under its very sortal concept must conform. When applied to mental disorders, this view entails that the concept of psychopathology has a normative character. A human being is psychopathic if and only if he fails to have the mental capacities that are normal for humans, that is, the mental capacities that humans ought to have. Human specific mental capacities include emotional responses to the environment, as well as perceptual and cognitive abilities. Let us call this the “received view of psychopathology.” The received view has two main implications concerning psychiatry. The first implication is that psychiatry is ethically relevant, in two respects. First, the results of psychopathology may help to define what is normal for humans, and thus psychiatry may help to determine ethical norms. Second, the psychiatric treatment of mental disorders rests on various assumptions concerning what is normal for humans, and thus psychiatry has profound ethical bearings. The second implication of the received view of psychopathology is what we could call the “universal treatment thesis.”

2006 - Tricyclic pyrazoles. 4. Synthesis and biological evaluation of analogues of the robust and selective CB 2 cannabinoid ligand 1-(2′, 4′-dichlorophenyl)-6-methyl-N-piperidin-1-yl-1,4-dihydroindeno[1,2-c] pyrazole-3-carboxamide [Articolo su rivista]
Murineddu, G.; Lazzari, P.; Ruiu, S.; Sanna, A.; Loriga, G.; Manca, I.; Falzoi, M.; Dessi, C.; Curzu, M. M.; Chelucci, G.; Pani, L.; Pinna, G. A.

New analogues (2a-p) of the previously reported CB 2 ligands 6-methyl- and 6-chloro-1-(2′,4′-dichlorophenyl)-N-piperidin-1-yl-1, 4-dihydroindeno[1,2-c]pyrazole-3-carboxamides (1a,b) have been synthesized and evaluated for cannabinoid receptor affinity. One example, 1-(2′,4′- dichlorophenyl)-6-methyl-N-cyclohexyilamine-1,4-dihydroindeno[1,2-c] pyrazole-3-carboxamide (2a) was shown to have single digit nanomolar affinity for cannabinoid CB 2 receptors. Furthermore, compounds 2a and 2b, as well as lead structures 1a,b, were also shown to be agonist in an in vitro model based on human promyelocytic leukemia HL-60 cells. © 2006 American Chemical Society.

2005 - Effect of 1-butanol on the microstructure of lecithin/water/tripalmitin system [Articolo su rivista]
Caboi, F.; Lazzari, P.; Pani, L.; Monduzzi, M.

Warm microemulsions based on lipids characterized by a melting point over 50°C have been successfully used as starting matrix in a quenching process to obtain solid lipid nanoparticles (SLN). In this work, we have investigated the effect of 1-butanol (B) on the phase behavior of the lecithin (LCT)/water (W)/tripalmitin (TP) system at 70°C. The study has been carried out at LCT/B = 1 (weight ratio). Emulsion and liquid crystalline phase regions have been observed in the ternary phase diagram, while the presence of 1-butanol in the LCT/W/B/TP system allows the formation of a wide area of liquid isotropic phase from the whole (LCT + B)/TP binary axis up to 37 wt% of water. The microstructure of this isotropic phase has been investigated by means of 1H NMR PGSE technique. The self-diffusion coefficients of the different components along oil and water dilution lines indicate a microstructural organization characterized by a highly connected water in oil domains. © 2005 Elsevier Ireland Ltd. All rights reserved.

2005 - Effect of Δ9-tetrahydrocannabinol on phosphorylated CREB in rat cerebellum: An immunohistochemical study [Articolo su rivista]
Casu, M. A.; Pisu, C.; Sanna, A.; Tambaro, S.; Spada, G. P.; Mongeau, R.; Pani, L.

Several converging lines of evidence indicate that drugs of abuse may exert their long-term effects on the central nervous system by modulating signaling pathways controlling gene expression. Cannabinoids produce, beside locomotor effects, cognitive impairment through central CB1 cannabinoid receptors. Data clearly indicate that the cerebellum, an area enriched with CB1 receptors, has a role not only in motor function but also in cognition. This immunohistochemical study examines the effect of Δ9-tetrahydrocannabinol (Δ9-THC), the principal psychoactive component of marijuana, on the levels of phosphorylated CREB (p-CREB) in the rat cerebellum. Acute treatments with Δ9-THC at doses of 5 or 10 mg/kg induced a significant increase of p-CREB in the granule cell layer of the cerebellum, an effect blocked by the CB1 receptor antagonist SR 141716A. Following chronic Δ9-THC administration (10 mg/kg/day for 4 weeks), the density of p-CREB was markedly attenuated compared to controls, and this attenuation persisted 3 weeks after withdrawal from Δ9-THC. These data provide evidence for the involvement of cerebellar granule cells in the adaptive changes occurring during acute and chronic Δ9-THC exposure. This might be a mechanism by which Δ9-THC interferes with motor and cognitive functions. © 2005 Elsevier B.V. All rights reserved.

2005 - Evaluation of tamsulosin and alfuzosin activity in the rat vas deferens: Relevance to ejaculation delays [Articolo su rivista]
Tambaro, S.; Ruiu, S.; Dessi, C.; Mongeau, R.; Marchese, G.; Pani, L.

The effect of two α-adrenergic receptor antagonists widely employed in the therapy of benign prostatic hyperplasia, tamsulosin [(-)-(R)-5-[2-[[2-(0- ethoxyphenoxy) ethyl]amino]propyl]-2-methoxybenzenesulfonamide] and alfuzosin [(±)-N-[3-[(4-amino-6,7-dimethoxy-2-quinazolinyl) methylamino]propyl] tetrahydro-2-furancarboxamide], was investigated in the rat vas deferens. Because several clinical studies have shown that tamsulosin causes ejaculatory disorders, this study also evaluated the possible mechanisms implicated in these disorders by comparing the effect of tamsulosin with that of alfuzosin. Tamsulosin competitively antagonized the contractions induced by noradrenaline in vitro in the epididymal portion of the vas deferens with a potency pA 2 value of 9.2 ± 0.8. In the prostatic portion, tamsulosin increased the amplitude of intermittent spikes induced by exogenous noradrenaline (100-1000 μM). In both portions of the vas deferens, alfuzosin behaved as an α-adrenergic antagonist blocking the contractions induced by exogenous noradrenaline without altering spikes. The administration of tamsulosin (3 μg/kg i.v.) significantly reduced the contractions evoked by electrical pulses in the epididymal portion, whereas it increased those produced in the prostatic portion. Intravenous tamsulosin antagonized the contraction produced by exogenous noradrenaline, whereas alfuzosin administration (10 μg/kg i.v.) did not change the electrically induced contractions in both portions of the rat vas deferens and did not antagonize the contractions produced by exogenous noradrenaline. The fact that tamsulosin unusually enhances noradrenaline-induced intermittent spike contractions and nerve stimulation-induced twitches in the prostatic portions might be linked to its greater propensity to cause sexual dysfunctions.

2005 - Five mutations in the GABAA α6 gene 5′ flanking region are associated with a reduced basal and ethanol-induced α6 upregulation in mutated Sardinian alcohol non-preferring rats [Articolo su rivista]
Saba, L.; Porcella, A.; Sanna, A.; Congeddu, E.; Marziliano, N.; Mongeau, R.; Grayson, D.; Pani, L.

The presence of four nucleotide changes and a three base-pair deletion in the GABAA α6-subunit promoter is described in Sardinian alcohol non-preferring rats, selectively bred for their ethanol aversion. These mutations are associated with the R100Q α6 intragenic mutation that was previously characterized in the same animals. The possibility that these mutated nucleotides alter the ethanol-induced upregulation of the α6 gene was investigated by measuring cerebellar α6 mRNA levels after a chronic ethanol liquid diet in sNP rat. Real-time quantitative PCR showed an increased α6 gene expression after ethanol ingestion in normal and mutated rats. However, lower amounts of α6 mRNA levels were detected both in control and in ethanol-treated sNP rats carrying the five promoter and the intragenic mutations in a homozygous state. Using the electromobility shift assay, specific DNA binding sites were found in cerebellar extracts of the α6 regions comprising the five mutations. These results suggest that one or more of the mutated binding sites that were found in the 5′ flanking α6 region may be a consensus sequence for regulatory factors which are responsible for both basal and ethanol-induced α6 gene expression. © 2005 Elsevier B.V. All rights reserved.

2005 - Modulation of ATP-mediated contractions of the rat vas deferens through presynaptic cannabinoid receptors [Articolo su rivista]
Tambaro, S.; Mongeau, R.; Dessi, C.; Pani, L.; Ruiu, S.

The effect of R-(+)-[2,3-dihydro-5-methyl-3-[(morpholiny)methyl]pyrolol[1, 2,3-de]-1,4-benzoxazin-yl]-(1-naphthalenyl)methanone mesylate (WIN 55,212-2; a cannabinoid receptor agonist) was investigated on contractions of the bisected (epididymal and prostatic portions) rat vas deferens to assess the role of cannabinoid receptors in sympathetic ATP neurotransmission. WIN 55,212-2 inhibited the electrically induced contractions in both portions of the rat vas deferens. In the presence of the α1-adrenoreceptor antagonist prazosin, electrical stimulation produces a contraction mediated exclusively by ATP. In this condition, WIN 55,212-2 in the prostatic portion elicited a concentration-dependent inhibition that was antagonized by N-piperidinyl-[8- chloro-1-(2,4-dichlorophenyl)-1,4,5,6-tetrahydrobenzo[6,7]cyclohepta[1,2-c] pyrazole-3-carboxamide] (NESS 0327), a selective cannabinoid CB1 receptor antagonist. NESS 0327 caused a parallel dextral displacement of the WIN 55,212-2 concentration-response curve. It is suggested that activation of pre-junctional cannabinoid receptors on sympathetic nerves of the vas deferens modulates ATP neurotransmission. © 2005 Elsevier B.V. All rights reserved.

2005 - Tricyclic pyrazoles. 3. Synthesis, biological evaluation, and molecular modeling of analogues of the cannabinoid antagonist 8-chloro-1-(2′, 4′-dichlorophenyl)-N-piperidin-1-yl-1,4,5,6-tetrahydrobenzo[6,7] cyclohepta[1,2-c]pyrazole-3-carboxamide [Articolo su rivista]
Murineddu, G.; Ruiu, S.; Loriga, G.; Manca, I.; Lazzari, P.; Reali, R.; Pani, L.; Toma, L.; Pinna, G. A.

A series of analogues of 8-chloro-1-(2′,4′-dichlorophenyl)-AT- piperidin-1-yl-1,4,5,6-tetrahydrobenzo-[6,7]cyclohepta[1,2-c] pyrazole-3-carboxamide 4a (NESS 0327) (Ruiu, S.; Pinna, G. A.; Marchese, G.; Mussinu, J. M.; Saba, P.; Tambaro, S.; Casti, P.; Vargiu, R.; Pani, L. Synthesis and Characterization of NESS 0327: A Novel Putative Antagonist of CB 1 Cannabinoid Receptor. J. Pharmacol. Exp. Ther. 2003, 306, 363-370) was synthesized and evaluated for their affinity to cannabinoid receptors. Depending on the chemical modification of the lead structure that was chosen, compounds 4b, 4c, 4i, 4l, and 4m still proved to be potent binders of the CB1 receptor. Moreover, several analogues (4c, 4d, 4e, and 4m) demonstrated superior CB2 receptor binding affinities compared to the parent ligand. Compounds 4b, 4c, 4i, and 4l displayed the most promising pharmacological profiles, having the highest selectivity for CB1 receptors with Ki(CB2) to Ki(CB1) ratios of 11 250, 2000, 3330 and 4625, respectively. Compound 4c increased the intestinal propulsion in mice and antagonized the effect induced by the CB 1 receptor agonist WIN 55,212-2. Finally, molecular modeling studies were carried out on a set of tricyclic pyrazoles (2a-4a) and on rimonabant 1 (SR141716A), indicating that high CB1 receptors affinities were consistent for the tricyclic derivatives, both with a nonplanar geometry of the tricyclic cores and with a precise orientation of the substituent (chlorine) on this ring system. © 2005 American Chemical Society.

2005 - Tricyclic pyrazoles. Part 2: Synthesis and biological evaluation of novel 4,5-dihydro-1H-benzo[g]indazole-based ligands for cannabinoid receptors [Articolo su rivista]
Murineddu, G; Ruiu, S; Mussinu, J. -M; Loriga, G; Grella, G. E; Carai, M. A. M; Lazzari, P; Pani, L; Pinna, G. A.

2004 - Blockade of neurotensin receptors affects differently hypo-locomotion and catalepsy induced by haloperidol in mice [Articolo su rivista]
Casti, P.; Marchese, G.; Casu, G.; Ruiu, S.; Pani, L.

Antipsychotic drug treatment increases neurotensin (NT) neurotransmission, and the exogenous administration of NT produces antipsychotic-like effects in rodents. In order to investigate whether "endogenous" NT may act as a natural occurring antipsychotic or may mediate antipsychotic drug activity, the effects of the selective NT receptor antagonists SR 48692 and SR 142948A were analyzed in different behavioural tests of locomotor activity using vehicle, amphetamine, or haloperidol in mice. SR 48692 (0.1-1 mg/kg, i.p.) and SR 142948A (0.03-0.1 mg/kg, i.p.) failed to affect mouse spontaneous locomotor activity and amphetamine-induced (2.5 mg/kg, i.p.) hyper-locomotion. However, SR 48692 (0.1 and 0.3 mg/kg, i.p.) and SR 142948A (0.03 and 0.05 mg/kg, i.p.) significantly alleviated the reduction of locomotor activity elicited by haloperidol (0.01 and 0.04 mg/kg, s.c.) in vehicle- or amphetamine-treated mice. Finally, SR 48692 (0.3 mg/kg, i.p.) and SR 142948A (0.05 and 0.1 mg/kg, i.p.) increased mouse catalepsy produced by haloperidol (0.3 mg/kg, s.c.). The present results indicate that while endogenous NT is not involved in the modulation of either mouse spontaneous locomotor activity or amphetamine-induced hyper-locomotion, it might act by enhancing the therapeutic effects of haloperidol and by attenuating the extrapyramidal side effects elicited by this antipsychotic. © 2004 Elsevier Ltd. All rights reserved.

2004 - Consensus on the use of substituted benzamides in psychiatric patients [Articolo su rivista]
Racagni, G.; Canonico, P. L.; Ravizza, L.; Pani, L.; Amore, M.

The class of substituted benzamides includes compounds able to modulate dopaminergic neurons selectively and specifically. The first synthetic substituted benzamide was sulpiride, which has been replaced in the clinic by the more modern amisulpride. The compound is very selective for mesolimbic D2 and D3 receptors and, therefore, has a dual mechanism of action, which is associated with two different indications. At low doses (50 mg), amisulpride preferentially blocks presynaptic autoreceptors, producing an increase in dopamine release, and therefore acting as a dopaminergic compound able to resolve the dopaminergic hypoactivity that characterizes depression. At higher doses (400-1,200 mg), the drug exerts its activity on postsynaptic D 3/D2 receptors located in the limbic region and prefrontal areas, producing selective dopaminergic inhibition, eliciting antipsychotic effects. In the present review, the clinical use of amisulpride in depressive syndromes is discussed, in particular in dysthymia and in schizophrenia. Based on experimental data, amisulpride is a treatment of choice for dopaminergic transmission disorders, both in depression and in schizophrenia. Copyright © 2004 S. Karger AG, Basel.

2004 - Haloperidol versus risperidone on rat "early onset" vacuous chewing [Articolo su rivista]
Marchese, G.; Bartholini, F.; Casu, M. A.; Ruiu, S.; Casti, P.; Congeddu, E.; Tambaro, S.; Pani, L.

Similarly to acute rat catalepsy, "early onset" vacuous chewing movements (VCMs) induced by subchronic treatment with antipsychotic have recently been proposed as a model of human extrapyramidal symptoms. In the present study, the propensities of haloperidol and risperidone in inducing rat "early onset" VCMs were compared using doses of the two antipsychotics that acutely induce similar catalepsy. Comparable rat catalepsy states were observed when the effects produced by 0.1, 0.5, and 1mg/kg of haloperidol were compared with those induced by 1, 4, and 10mg/kg of risperidone, respectively. These doses of the two antipsychotics were then administered twice a day for 4 weeks and VCMs scored after 12h, 5 days, or 3 weeks of drug withdrawal. Among the haloperidol-treated groups, only those rats injected with 0.5 and 1mg/kg showed high levels of VCMs after 12h and 5 days of drug withdrawal when compared to vehicle-treated rats, while basal levels of VCMs were reached after 3 weeks from the last injection. High VCMs levels were observed in risperidone-treated rats only at the dose of 10mg/kg and after 12h of drug withdrawal, but not after 5 days or 3 weeks. The present results indicated that haloperidol possessed a much higher propensity to induce rat "early onset" VCMs than risperidone. © 2003 Elsevier B.V. All rights reserved.

2004 - Immunocytochemical study of the forebrain serotonergic innervation in Sardinian alcohol-preferring rats [Articolo su rivista]
Casu, M. A.; Pisu, C.; Lobina, C.; Pani, L.

Rationale: The anxiolytic effect of ethanol is generally considered to be causally related to the development of alcohol dependence, and serotonin (5-HT) has been involved in both alcohol abuse and anxiety disorders. Several lines of evidence suggest an inverse relationship between alcohol abuse and central serotonergic neurotransmission. Objectives: When tested in the elevated plus-maze, selectively bred Sardinian alcohol-preferring (sP) rats display a higher degree of anxiety than Sardinian alcohol-non-preferring rats (sNP); this behavior is reversed by voluntary ethanol intake. The present study examined whether sP rats differed with respect to the 5-HT innervation in different forebrain areas. Methods: We performed an immunohistochemistry study using an antibody raised against serotonin transporter (SERT), a marker for 5-HT fibers, coupled with an unbiased stereology, the method used to count the number of 5-HT neurons in the raphe nuclei. Results: The SERT-positive innervation density was found to be significantly lower in the medial-prefrontal cortex and in the shell of the nucleus accumbens of the ethanol-naive sP rats (sP-N) when compared with the sNP and unselected Wistar rats. No differences were found in the caudate putamen and hippocampus. The stereological analysis showed a significant difference in the number of 5-HT neurons in the dorsal but not in the median raphe of sP-N rats, compared with sNP and Wistar rats. Analysis of the cell body cross-sectional area revealed no differences among the three lines of rats either in the dorsal or in the median raphe. In sP rats that had voluntarily drunk ethanol for 14 consecutive days (sP-exp), no differences were found in the 5-HT innervation relative to sP-N animals. Conclusions: These results indicate a selective reduction of innervation in the medial portion of the mesocorticolimbic 5-HT system in sP rats, suggesting that this genetically determined difference may be involved in the contrasting alcohol preference and consumption of sP and sNP animals.

2004 - Practical application of pharmacotherapy with long-acting risperidone for patients with schizophrenia [Articolo su rivista]
Keith, S. J.; Pani, L.; Nick, B.; Emsley, R.; San, L.; Turner, M.; Conley, R.; Scully, P.; Chue, P. S.; Lachaux, B.

Objective: It is now generally accepted that the use of second-generation, or atypical, antipsychotics for schizophrenia represents an advance over conventional antipsychotic agents. However, adherence continues to be a problem, as with other medications for chronic disorders. Long-acting formulations of conventional antipsychotics partly address adherence problems, but their use is limited by tolerability issues. This article provides practical advice to physicians on the characteristics of patients who would benefit from treatment with long-acting atypical antipsychotic agents and offers suggestions on how to initiate treatment. Methods: A literature search for studies published between 1980 and 2003 that evaluated the treatment of patients with schizophrenia with long-acting atypical agents was conducted by using MEDLINE and EMBASE. The primary search parameters were "schizophrenia," "atypical," "antipsychotic," and "long-acting." As expected, long-acting risperidone was the only long-acting atypical agent identified; thus this article focuses on practical advice and suggestions on how to initiate therapy with long-acting risperidone. Results and discussion: From the results of the literature search and the discussion of a panel of experts at a meeting held in Dublin in 2003 and supported by Johnson & Johnson, it is possible to conclude that long-acting risperidone has demonstrated efficacy and tolerability, even among patients who are considered clinically stable on other antipsychotics. Most patients can switch safely and effectively to long-acting risperidone if appropriate strategies are applied. Long-acting risperidone provides a new and promising therapeutic option for the treatment of schizophrenia.

2004 - Ritanserin counteracts both rat vacuous chewing movements and nigro-striatal tyrosine hydroxylase-immunostaining alterations induced by haloperidol [Articolo su rivista]
Marchese, G.; Bartholini, F.; Ruiu, S.; Casti, P.; Casu, G. L.; Pani, L.

The effect of subchronic co-administration of ritanserin (1.5 mg/kg, i.p., twice a day) and haloperidol (1 mg/kg, i.p., twice a day) on rat vacuous chewing movements and on tyrosine hydroxylase-immunostaining was investigated. Ritanserin significantly reduced rat vacuous chewing movements observed following 2, 3 and 4 weeks of haloperidol administration and after 5 days of haloperidol withdrawal. Furthermore, ritanserin prevented the reduction of striatal tyrosine hydroxylase-immunostaining and the shrinkage of nigral dopaminergic cell bodies induced by haloperidol. The present results indicate that ritanserin may possess protective properties on both dopaminergic nigro-striatal neuron alterations and vacuous chewing movements induced by haloperidol, and provide further evidence indicating a possible association between these two haloperidol-induced effects. © 2003 Elsevier B.V. All rights reserved.

2004 - Synthesis and biological evaluation of 1,8-naphthyridin-4(1H)-on-3- carboxamide derivatives as new ligands of cannabinoid receptors [Articolo su rivista]
Ferrarini, P. L.; Calderone, V.; Cavallini, T.; Manera, C.; Saccomanni, G.; Pani, L.; Ruiu, S.; Gessa, G. L.

Cannabinoid receptors have been studied extensively in view of their potential functional role in several physiological and pathological processes. For this reason, the search for new potent, selective ligands for subtype CB receptors, CB1 and CB2, is still of great importance, in order to investigate their role in various physiological functions. The present study describes the synthesis and the biological properties of a series of 1,8-naphthyridine derivatives, characterised by the presence of some important structural requirements exhibited by other classes of cannabinoid ligands, such as an aliphatic or aromatic carboxamide group in position 3, and an alkyl or arylalkyl substituent in position 1. These compounds were assayed for binding both to the brain and to peripheral cannabinoid receptors (CB1 and CB2). The results obtained indicate that the naphthyridine derivatives examined possess a greater affinity for the CB2 receptor than for the CB1 receptor. In particular, derivatives 6a and 7a possess an appreciable affinity for the CB2 receptor, with K i values of 5.5 and 8.0 nM respectively; also compounds 4a, 5a and 8a exhibit a good CB2 affinity, with Ki values in the range of 10-44 nM. Furthermore, compounds 3g-i and 18 revealed a good CB 2 selectivity, with a CB1/CB2 ratio >20. © 2004 Elsevier Ltd. All rights reserved.

2004 - The atypical antipsychotic quetiapine increases both noradrenaline and dopamine release in the rat prefrontal cortex [Articolo su rivista]
Pira, L.; Mongeau, R.; Pani, L.

Quetiapine is a novel atypical antipsychotic drug with multi-receptorial affinity. Using in vivo microdialysis, we investigated if quetiapine modulates extracellular noradrenaline and dopamine in brain areas generally believed to be involved in the pathophysiology of schizophrenia and in the action of antipsychotic drugs. Quetiapine (5, 10 and 20 mg/kg, i.p.) increased levels of noradrenaline in both the prefrontal cortex and the caudate nucleus, while it increased dopamine levels mainly in the prefrontal cortex. It is argued that the marked increase of dopaminergic transmission in the prefrontal cortex induced by quetiapine might be relevant to its therapeutical action. © 2004 Elsevier B.V. All rights reserved.

2004 - The cerebellar GABAA α6 subunit is differentially modulated by chronic ethanol exposure in normal (R100R) and mutated (Q100Q) sNP rats [Articolo su rivista]
Sanna, A; Congeddu, E; Saba, L; Porcella, A; Marchese, G; Ruiu, S; Casti, P; Saba, P; Pani, L.

Sardinian alcohol non-preferring (sNP) rats carry a point mutation (R100Q) in the cerebellar expressed GABAA receptor α6 subunit gene, leading to a higher sensitivity to ethanol and diazepam. The role of the α6 subunit gene cluster in the ethanol non-preferring phenotype was here investigated by measuring the levels of α1, α6 and γ2 peptide in the cerebellum of normal (RR) and mutated (QQ) sNP rats after 2 weeks of chronic ethanol administration. Western blot analysis revealed that the α6 subunit is increased in RR sNP rats after chronic ethanol exposure (25.44%±8.69 versus control), while it remained unchanged in mutated QQ sNP rats. Interestingly, chronic ethanol administration decreased α1 peptide levels in the cerebellum of both rat lines to a similar extent (30.99%±6.74 and 27.12%±9.83 in RR and QQ rats, respectively), while γ2 peptide levels remained unchanged. To further correlate the genetic and biochemical difference of the normal and mutated sNP rats with their aversive phenotype, we exposed sNP rats to a protocol of acquisition and maintenance of ethanol drinking. QQ sNP rats drank less ethanol than RR rats during the acquisition phase, but such difference was lost during the maintenance phase. These data may contribute to elucidating the mechanisms of alcohol avoidance in rat lines selected for this behavior when exposed to ethanol solution. © 2003 Elsevier B.V. All rights reserved.

2003 - Characterization of wild-type (R100R) and mutated (Q100Q) GABAA α6 subunit in Sardinian alcohol non-preferring rats (sNP) [Articolo su rivista]
Sanna, A.; Congeddu, E.; Porcella, A.; Saba, L.; Pistis, M.; Peis, M.; Marchese, G.; Ruiu, S.; Lobina, C.; Grayson, D. R.; Gessa, G. L.; Pani, L.

Sardinian alcohol non-preferring (sNP) rats, selected for their low ethanol preference and consumption, carry a point mutation (R100Q) in the gene coding for GABAA receptor α6 subunit, which becomes more sensitive to diazepam-evoked GABA currents. We performed binding studies in the cerebellum of normal (RR) and mutated (QQ) sNP rats using [3H]Ro 15-4513, an inverse agonist for the benzodiazepine site which binds both diazepam insensitive and diazepam sensitive sites. Saturation curves performed on cerebellar membrane from genotyped rats indicated an higher affinity of [3H]Ro 15-4513 for GABAA receptors in QQ with respect to RR rats (Kd values 4.0±0.67 and 6.24±0.95 nM, respectively), with similar Bmax values (3.5±0.25 and 3.9±0.39 pmol/mg protein, respectively). Diazepam displacement curves showed a two component model for both genotypes, with similar Ki1 values for QQ and RR (3.6±0.62 and 4.9±0.33 nM, respectively). In QQ rats diazepam is able to completely displace [3H]Ro 15-4513 (Ki2=1.48±0.27 μM), while in RR rats the diazepam sensitive sites are still present (Ki2>10 μM). The basal mRNA and protein expression level of the α6 subunit were similar in RR and QQ rats. The electrophysiological profile of oocytes of Xenopus laevis injected with cerebellar synaptosomes showed that ethanol positively modulated GABA-evoked currents significantly more in QQ than in RR rats. These data contribute to the characterization of the function of GABAA α6 subunit and its involvement in determining alcohol related behavior. © 2002 Elsevier Science B.V. All rights reserved.

2003 - Co-release of noradrenaline and dopamine from noradrenergic neurons in the cerebral cortex induced by clozapine, the prototype atypical antipsychotic [Articolo su rivista]
Devoto, P.; Flore, G.; Vacca, G.; Pira, L.; Arca, A.; Casu, M. A.; Pani, L.; Gessa, G. L.

Rationale: Clozapine has been shown to increase extracellular dopamine (DA) and noradrenaline (NA) in the medial prefrontal cortex (mPFC). A recent study of ours suggested that extracellular DA in the PFC originates not only from dopaminergic but also from noradrenergic terminals, its release being controlled by α2-adrenoceptors. Objectives: Since clozapine binds to α2-adrenoceptors, the possibility that it might co-release DA and NA was studied. Methods: By means of microdialysis coupled to HPLC with electrochemical detection, the effect of clozapine on extracellular DA and NA in the mPFC, densely innervated by DA and NA, was compared to that in the occipital cortex, equally innervated by NA but receiving few DA projections. Results: Extracellular NA was found to be the same in the two cortices, consistent with homogeneous NA innervation. On the other hand, extracellular DA in the occipital cortex was only 29% lower than in the mPFC, in spite of the scarce dopaminergic innervation in the occipital cortex. Clozapine (10 mg/kg IP) increased extracellular DA and NA not only in the mPFC (by about 320% and 290%, respectively) but also in the occipital cortex (by 560% and 230%, respectively). Administration of the α2-agonist clonidine (0.15 mg/kg) reversed the effect of clozapine in both cortices, while the D2-agonist quinpirole (0.1 mg/kg IP) was ineffective. Conclusions: The results suggest that clozapine, by inhibiting α2-adrenoceptors, co-releases DA and NA from noradrenergic terminals in the occipital cortex and that the same mechanism might be responsible for the concomitant increase of the two monoamines in the mPFC.

2003 - Differential distribution of functional cannabinoid CB1 receptors in the mouse gastroenteric tract [Articolo su rivista]
Casu, M. A.; Porcella, A.; Ruiu, S.; Saba, P.; Marchese, G.; Carai, M. A. M.; Reali, R.; Gessa, G. L.; Pani, L.

Recently, the gastrointestinal pharmacology of cannabinoid CB1 receptors has been extensively explored. We employed western blotting and immunohistochemistry techniques to study the distribution of the cannabinoid CB1 receptor protein in the mouse gastroenteric tract. The cannabinoid CB1 receptor peptide was detected by western blotting only in its glycosylated form (63 kDa) with a significant differential distribution. The highest levels of expression were detected in the stomach and in the colon, while the pyloric valve was devoid of any cannabinoid CB1 receptor protein. The immunohistochemical study showed intense cannabinoid CB1 receptor immunoreactivity in ganglia subadjacent to the gastric epithelium and in the smooth muscle layers of both the small and large intestine. Only the small intestine showed (-)-3-[2-hydroxyl-4-(1,1-dimethylheptyl)-phenyl]-4-(3-hydroxylpropyl) cyclohexan-1-ol) ([3H]CP 55,940) specific binding (27%). These receptors mediated pharmacologically significant effects since the cannabinoid CB1 receptor agonist R(-)-7-hydroxy-delta-6-tetra-hydrocannabinol-dimethylheptyl (HU 210) dose dependently inhibited gastrointestinal transit up to 70%, while the cannabinoid CB1 receptor antagonist N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3- carboxamide (SR 141716A) increased gastrointestinal transit. Moreover, the dose of 0.3 μg/kg of HU 210, devoid per se of any activity on mouse intestinal propulsion, blocked the increased gastroenteric transit induced by the cannabinoid CB1 antagonist SR 141716A. © 2002 Elsevier Science B.V. All rights reserved.

2003 - Duration and stability of the rapid-cycling course: A long-term personal follow-up of 109 patients [Articolo su rivista]
Koukopoulos, A.; Sani, G.; Koukopoulos, A. E.; Minnai, G. P.; Girardi, P.; Pani, L.; Albert, M. J.; Reginaldi, D.

Background: Recognition by the DSM-IV of rapid cyclicity as a course specifier has raised the question of the stability and long-term outcome of rapid-cycling (RC) patients. Data on this topic is sparse and often inconsistent. To our knowledge, these are the first personally followed patients over the long term, dealing directly with the issue of the duration of the RC course. Methods: We examined the evolution of the course of 109 RC patients (68 women and 41 men) followed for a minimum of 2 years and up to 36 years, beginning with the index episode when the RC course was diagnosed by the authors (A.K., G.P.M., P.G., L.P., D.R.). Patients were included in the study if they met criteria for RC as defined by ≥4 affective episodes per year (Dunner and Fieve, 1974). The follow-up period varied from 2-5 years for 25 patients, 6-10 years for 24 patients, 11-15 years for 24 patients, 16-20 years for 19 patients, 21-25 years for 13 patients, 30-36 years for four patients. Results: In 13 patients (12%), RC emerged spontaneously and in 96 patients (88%), it was associated with antidepressant and other treatments. In 19 women (28% of all women) RC course started in perimenopausal age (45-54 years). The mean duration of RC during the follow-up period was 7.86 years (range 1-32) and its total duration (including RC course prior to the follow-up period) was 11 years (range 1-40). The total duration of the affective disorder, from the first episode to the end of the follow-up, was 21.78 years (range 1-70). At the end of the follow-up, 36 patients (33%) had complete remission for at least the past year, 44 (40%) stayed rapid cycling with severe episodes (six of this group committed suicide), while 15 (14%) were rapid cycling but with attenuated episodes. The other 14 patients (13%) became long cyclers, eight with severe episodes and six with milder ones. The main distinguishing features between those who remitted from and those who persisted in the RC course were: (1) the initial cycle pattern: patients with Depression-Hypomania(mania)-Free interval cycles (53 patients) had a worse outcome: 26.4% remitted and 52.8% persisted in the RC course through to the end of the follow up period. The Mania/Hypomania-Depression-Free interval cycles (22 patients) had a significantly better outcome, with 50% remitted and 27.2% persisting RC; and (2) the occurrence of the switch process from depression to hypomania/mania and the occurrence of agitated depressions made the prognosis worse. Continuous treatment was more effective against mania/hypomania than against depression, yet in all persisting RC cases the mania/hypomania remitted only partially. Limitations: These data derive from clinics known for their expertise in mood disorders, and they may have attracted and retained patients with a more severe course. Treatment was uncontrolled and consisted more of lithium than divalproex, lamotrigene and olanzapine, recently shown to be beneficial in subgroups of patients with rapid-cycling. Conclusions: Our findings suggest that rapid cyclicity, spontaneous or induced, once established, becomes for many years a stable rhythm in a substantial proportion of patients, linked to endogenous and environmental factors. The suggestion is made to consider as rapid-cyclers, at least for research purposes, those patients who have had a rapid cycling course for at least 2 years, borrowing the duration criterion currently employed for other chronic disorders such as Dysthymia and Cyclothymia. That our patients had poorer prognosis than some other cohorts in the literature is probably due to the shorter duration of "rapid-cycling" at entry in the latter cohorts. A true understanding of the nature of rapid-cycling will require a rigorous definition of not only duration, but also pole-switching and course patterns at entry into study. © 2002 Elsevier Science B.V. All rights reserved.

2003 - Haloperidol, but not clozapine, produces dramatic catalepsy in Δ 9-THC-treated rats: Possible clinical implications [Articolo su rivista]
Marchese, G.; Casti, P.; Ruiu, S.; Saba, P.; Sanna, A.; Casu, G.; Pani, L.

1. The effect on rat catalepsy induced by Δ 9- tetrahydrocannabinol (Δ 9-THC) in association with haloperidol (HP) or clozapine (CLOZ) administration was investigated. 2. Δ 9-THC dose-dependently increased HP (0.05 -1 mg kg -1, s.c.)-induced rat catalepsy, while no catalepsy was observed after CLOZ (1-20mg kg -1, s.c.) or Δ 9-THC + CLOZ administration. 3. The CB 1 antagonist SR141716A (0.5-5 mg kg -1, i.p.) reversed the increase mediated by Δ 9-THC on HP-induced catalepsy. 4. The D 2 agonist quinpirole completely reversed the catalepsy induced by both HP and HP + Δ 9-THC; however, higher doses of quinpirole were needed in the presence of Δ 9-THC. 5. The M 1 antagonist scopolamine and α 2 antagonist yohimbine were able to reduce the catalepsy induced by HP and HP + Δ 9-THC in a similar manner. 6. CLOZ and the 5-HT 2A/2C antagonists ritanserin, RS102221 and SB242084 were more effective in antagonizing HP than HP + Δ 9-THC-induced catalepsy. 7. HP and CLOZ failed to inhibit in vitro [ 3H]CP-55,940 binding, while Δ 9-THC and SR141716A did not show an appreciable affinity for the D 2 receptor. 8. It was suggested that the different effects on rat catalepsy induced by Δ 9-THC following HP or CLOZ administration may depend on the receptor-binding profiles of the two antipsychotics. 9. The preferential use of CLOZ rather than HP in the treatment of psychotic symptoms in cannabis abusers was discussed.

2003 - Hymenoptera Sting Anaphylactic Reactions in the Mediterranean Population of Albania [Articolo su rivista]
Mingomataj, E.; Ohri, D.; Dhimitri, V.; Priftanji, A.; Qirko, E.; Pani, L.; Fischer, T. C.; Dinh, Q. T.; Peiser, C.; Fischer, A.; Groneberg, D. A.

Background: Relatively few studies-have examined the relation of different hymenoptera sting reactions. Objective: To investigate the relation of anaphylactic reactions against stings of different hymenoptera subspecies in the Mediterranean population of Albania. Materials and Methods: A retrospective study was-conducted using the clinic files of 111 patients who were diagnosed for hymenoptera sting reactions from 1987 to 1996. Antigens used consisted of purified hymenoptera venom (bee, wasp, and paperwasp). The patients were diagnosed by intracutaneous tests in concentrations of 0.001 μg/ml, 0.01 μg/ml, 0.1 μg/ml, and 1 μg/ml. Results: The median age of the patients was 27 years. 57% of stings occurred between 20 to 40 years of age. The majority of anaphylactic reactions were recorded during the months of June to October. 81% of the patients were admitted to the hospital due to Mueller grade II to III reactions. In 26% of all cases, crossreactions (bee-wasp 16%, bee-wasp-paperwasp 7%, wasp-paperwasp 2%, bee-paperwasp 1%) were found. Of all anaphylactic reactions, 64% were attributed to bees, 24% to wasps, 8% to both bees and wasps, and 2% to paperwasps. Conclusions: In contrast to industrialized countries such as the United States or Western Europe where urban populations predominate, reactions to bee venom were more prevalent in the present study population.

2003 - In vitro evidence for the presence of [ 3H]-haloperidol uptake in rat brain [Articolo su rivista]
Ruiu, S.; Marchese, G.; Saba, P. L.; Satta, R.; Gessa, G.; Vaccari, A.; Pani, L.

1. The neuroleptic [ 3H]-haloperidol (HP) was taken up in synaptosomes prepared from rat brain, in a temperature-, sodium ion-, and energy-dependent process. 2. The highest concentration of uptake sites (V max=2.37 pmol mg -1 protein min -1) was in the striatum with the other brain areas displaying lower (by 50-70%) values. 3. The affinity values (K m≈40 nM) were similar in all brain areas considered. 4. The pharmacological characterization did not indicate a well-defined group of inhibitors, which suggested that HP might not use a transporter for recognized neurotransmitters. 5. The HP metabolites tested, including HPTP, were competitive inhibitors of [ 3H]-HP uptake, an indirect indication that they may actively enter the striatal nerve endings through the same carrier. 6. Since the uptake process was partially affected by the incubation of [ 3H]-HP in the presence of several antagonists of HP-transforming cytochrome P450 isoforms, the binding of HP at some enzyme sites inside the synaptosome cannot be excluded. 7. In conclusion, the present results suggest that HP may be actively transported in the rat brain.

2003 - Molecular characterization of new polymorphisms at the β2, α1, γ2 GABAA receptor subunit genes associated to a rat nonpreferring ethanol phenotype [Articolo su rivista]
Congeddu, E.; Saba, L.; Porcella, A.; Sanna, A.; Marchese, G.; Lobina, C.; Gessa, G. L.; Pani, L.

Recent preclinical and clinical studies have indicated a possible involvement of the genes encoding for the GABAA receptor subunits α6, β2, α1 and γ2 in the genetic susceptibility to alcohol abuse. We have recently found an (R) to (Q) mutation in codon 100 of the α6 GABAA subunit, that segregated in a rat line selectively bred for its voluntary ethanol aversion, Sardinian alcohol nonpreferring (sNP), but not in their Sardinian alcohol preferring (sP) counterpart, selected for its ethanol preference. In the present study the molecular composition of other GABAA subunits (β2, α1 and γ2) were analyzed in order to further investigate the involvement of the GABAA receptors in the genetic predisposition to voluntary alcohol intake. Automated sequencing analysis indicated the presence of six new silent substitutions (289 T→C in the β2 gene; 115 G→A in the α1 gene; 157 G→A, 174 C→T, 347 A→G and 385 A→T in the γ2 gene), in sNP but not in sP rats. These polymorphisms were linked to the α6 R100Q mutation previously described in sNP rats. The strict association between the α6 point mutation and the new polymorphisms found in the β2, α1 and γ2 genes, demonstrate that such genes belong to the same cluster and are inherited together in the rat. These results sustain the synteny for these clusters between the rodent and human genomes, and suggest that mutated GABAA β2, α6, α1 and γ2 subunit genes might contribute to the expression of an ethanol nonpreferring phenotype in a rat line that voluntarily avoids alcoholic solutions. © 2002 Elsevier Science B.V. All rights reserved.

2003 - Synthesis and characterization of NESS 0327: A novel putative antagonist of the CB1 cannabinoid receptor [Articolo su rivista]
Ruiu, S; Pinna, G. A; Marchese, G; Mussinu, J. -M; Saba, P; Tambaro, S; Casti, P; Vargiu, R; Pani, L.

2002 - Carmoxirole is able to reduce amisulpride-induced hyperprolactinemia without affecting its central effect [Articolo su rivista]
Marchese, G.; Ruiu, S.; Casti, P.; Bartholini, F.; Saba, P.; Gessa, G. L.; Pani, L.

Prolactin blood level and apomorphine-induced yawning were studied in rats treated with the substituted benzamide amisulpride in association with bromocriptine or carmoxirole; two dopamine D2 receptor agonists with high or low propensity to cross the brain-blood barrier, respectively. Administration of amisulpride produced a maximum increase in rat serum prolactin level (315±18%) vs. vehicle-treated animals (ED50=0.25±0.017 mg/kg, s.c.). The concurrent administration of carmoxirole or bromocriptine completely reversed the hyperprolactinemia induced by amisulpride (0.5 mg/kg, s.c.) (ID50=14.9±0.8 mg/kg and 0.81±0.03 mg/kg, respectively). Carmoxirole (15 mg/kg, i.p.) did not affect yawning induced by apomorphine (0.08 mg/kg, s.c.) nor amisulpride (0.5 mg/kg, s.c.) blockade of apomorphine-induced yawning. Conversely, a significant increase in the number of yawns was observed when bromocriptine (0.8 mg/kg, i.p.) was associated with apomorphine in the absence or presence of amisulpride. These results suggested that a peripheral dopamine D2 receptor agonists could be a useful tool in alleviating amisulpride-induced hyperprolactinemia without possibly affecting its central effect. © 2002 Elsevier Science B.V. All rights reserved.

2002 - Clinical implications of dopamine research in schizophrenia [Articolo su rivista]
Pani, L.

One of the most stimulating problems posed by second generation antipsychotics is the question of whether their ability to act on the negative, as well as the positive, symptoms of schizophrenia relies on the same neurochemical mechanisms as those responsible for their lack of extrapyramidal symptoms (EPS). Amisulpride is a substituted benzamide antipsychotic, which is known to be efficacious against both the positive and the negative symptoms of schizophrenia and to have a lower propensity to induce EPS than conventional antipsychotics. Amisulpride preferentially blocks the D2 and D3 subtypes of dopamine receptors, both presynaptically in the frontal cortex, enhancing dopaminergic transmission, and postsynaptically in subcortical areas such as the nucleus accumbens, so reducing dopaminergic transmission. Given that dopaminergic under-activity in the frontal cortex is thought to produce negative symptoms, and over-activity in the limbic system to produce positive symptoms, it is proposed that these are the mechanisms by which amisulpride produces its atypical profile. © 2002 Informa UK Ltd All rights reserved.

2002 - Dopaminergic deficit and mood disorders [Articolo su rivista]
Pani, L.; Gessa, G. L.

The roles of serotonin and noradrenaline in the pathogenesis of mood disorders have been elucidated by numerous studies, which support the therapeutic use of tricyclic antidepressants and selective serotonin re-uptake inhibitors. The same has not occurred for dopamine, notwithstanding the fact that the crucial role of the mesolimbic dopaminergic system in behaviour has been known to researchers for many years. The objective of this article is to provide an overview of the animal data that demonstrate the importance of dopamine in animal behaviour and suggest that dopaminergic deficiency may cause a number of psychiatric symptoms in man. © 2002 Lippincott Williams & Wilkins.

2002 - Effect of the amisulpride isomers on rat catalepsy [Articolo su rivista]
Marchese, G.; Bartholini, F.; Ruiu, S.; Casti, P.; Saba, P.; Gessa, G. L.; Pani, L.

The substituted benzamide amisulpride is currently administered in its racemic form. In the present study, the biochemical and cataleptogenic profiles of the two enantiomers (R+ and S-) were compared with those of the racemic mixture. Displacement binding studies showed that the (S-)-isomer possesses an higher affinity for dopamine D2-like receptor (Ki 5.2±0.4 nM) compared to (R+)-amisulpride (Ki 244±12 nM) and to (RS)-amisulpride (Ki 9.8±0.8 nM). In contrast, (S-)-amisulpride binds the α2-receptor with an affinity (Ki 1528±45 nM) lower than that of the (R+)-isomer (Ki 375±34 nM) and of (RS)-amisulpride (Ki 783±27 nM). The bar test was used to evaluate the catalepsy induced by each drug. (RS)-amisulpride induced catalepsy only at very high doses (>100 mg/kg, s.c.) whereas, (S-)-amisulpride produced a catalepsy at a lower dose (30 mg/kg, s.c.) and (R+)-amisulpride did not produce any catalepsy up to the dose of 75 mg/kg. Interestingly, (R+)-amisulpride reduced the catalepsy induced by (S-)-amisulpride (50 mg/kg, s.c.) or haloperidol (0.3 mg/kg, s.c.), at the doses of 50 or 30 mg/kg, respectively. These results indicate that the weak cataleptic properties of (RS)-amisulpride might partially rely on its (R+)-isomer and provide a further explanation for the atypical properties of amisulpride as an antipsychotic. © 2002 Elsevier Science B.V. All rights reserved.

2002 - Effect of the amisulpride isomers on rat prolactinemia [Articolo su rivista]
Marchese, G.; Ruiu, S.; Casti, P.; Saba, P.; Gessa, G. L.; Pani, L.

The effects on rat serum prolactin level of the two isomers constituting the racemic form of amisulpride were compared. (S-)-amisulpride induced hyperprolactinemia at lower doses (ED50=0.09±0.01 mg/kg) than racemic- (ED50=0.24±0.03 mg/kg) and (R+)-amisulpride (ED50=4.13±0.05 mg/kg), in accord with their affinities for pituitary dopamine D2 receptor (Ki=3.8±0.2, 6.4±0.2 and 143.3±2.3 nM, respectively). At doses twice the ED50, (S-)-amisulpride produced a maximal increase in prolactin level similar to that of the racemic form (403±21% and 425±15%, respectively), but higher than that of (R+)-amisulpride (198±8%). These results suggest that the hyperprolactinemia induced by the racemic-amisulpride is mostly due to its (S-)-isomer. © 2002 Elsevier Science B.V. All rights reserved.

2002 - Evidence for functional CB1 cannabinoid receptor expressed in the rat thyroid [Articolo su rivista]
Porcella, A.; Marchese, G.; Casu, M. A.; Rocchitta, A.; Lai, M. L.; Luigi, G.; Pani, L.

Objective: Previous reports have shown that the Δ9-tetrahydrocannabinol (Δ9TCH), the major psychoactive cannabinoid components of marijuana, is unable to inhibit thyroid hormonal activity. The aim of this study was to characterize the CB1 functional expression in the rat thyroid by a multi-methods approach. Methods and Results: RT-PCR was used to detect the mRNA expression of the CB1 cannabinoid receptor (17.8 2±4.0% of the normalizing reference gene β2 microglobulin), as well as the expression of the endocannabinoid hydrolyzing enzyme, fatty acid amide hydrolase (46.9±4.3% of β2 microglobulin), in the rat thyroid gland. The CB1-encoded protein was detected in its glycosylated form (63 kDa) by Western blot, employing a polyclonal antibody, while CB1 immunohistochemical localization showed an intracellular positive staining in both follicular and parafollicular cells. In addition, a 30% decrease in serum levels of both 3,5,3′ tri-iodothyronine (T3) and thyroxine (T4) was detected 4 h after the administration of the synthetic cannabinoid receptor agonist, WIN 55,212-2 (10mg/kg i.p.). These effects were antagonized by pretreatment with the CB1 antagonist SR 141716A (3 mg/kg i.p.); thyrotrophin levels were unaffected by both treatments. Conclusion: These data indicate that functional CB1 receptors which are able to modulate the release of T3 and T4 are expressed in the rat thyroid, and suggest a possible role of cannabinoids in the regulation of rat thyroid hormonal activity.

2002 - Reduced DAT- and DBH-immunostaining in the limbic system of Sardinian alcohol-preferring rats [Articolo su rivista]
Casu, M. A.; Dinucci, D.; Colombo, G.; Gessa, G. L.; Pani, L.

We have recently shown that tyrosine-hydroxylase immunostaining (TH-IM) is selectively decreased in the cingulate cortex and in the shell of the nucleus accumbens (nAcc) of Sardinian alcohol-preferring rats (sP) when compared with Sardinian alcohol-non preferring (sNP) and Wistar (W) rats. Since these regions contain both dopamine and noradrenaline (NA) fibers, clarification of the dopaminergic and noradrenergic contribution to the decreased TH-immunoreactivity was needed. To this aim, we carried out the present immunohistochemistry study using two antibodies raised against dopamine β-hydroxylase (DBH), the enzyme responsible for the conversion of dopamine into noradrenaline, and against the dopamine transporter (DAT), as markers for noradrenergic and dopaminergic fibers, respectively. The results show that DBH-immunostaining (DBH-IM) and DAT-immunostaining (DAT-IM) were both lower in the cingulate cortex of the sP rats with respect to sNP and W rats. In the shell of the nAcc a reduced DAT-IM in sP rats was found, while the DBH-IM did not differ between the three lines of rats. The analysis of the cell-body area of noradrenergic neurons in the locus coeruleus, revealed no differences between sP, sNP and W rats. These results indicate a selective reduction of the terminal innervation in the mesocorticolimbic dopamine and NA systems in sP rats. This genetically-determined difference may be involved in the opposite alcohol preference and consumption of sP and sNP rats. © 2002 Elsevier Science B.V. All rights reserved.

2002 - Reduced TH-immunoreactive fibers in the limbic system of Sardinian alcohol-preferring rats [Articolo su rivista]
Casu, M. A.; Colombo, G.; Gessa, G. L.; Pani, L.

The mesolimbic dopamine (DA) system has long been known to be involved in reward behaviors. As with other substances of abuse, it has been extensively reported that ethanol influences the dopaminergic system. The present study examined whether selectively bred Sardinian-alcohol-preferring (sP) and Sardinian alcohol non-preferring rats (sNP), differ in the DA innervation in structures of the forebrain that are related to rewarding behaviors. To this aim, we performed an immunohistochemistry study with an antibody raised against tyrosine hydroxylase (TH), the rate-limited step enzyme in the biosynthesis of monoamines. The TH-positive innervation density was found to be significantly lower in the cingulate cortex and in the shell of the nucleus accumbens of the sP when compared with the sNP and unselected Wistar rats. These anatomical structures both cluster in the medial aspect of the mesolimbic system. No differences in other major DA brain regions, such as the nigro-striatal pathway were found. The analysis of cell-body area revealed no differences between sP, sNP and Wistar rats in the ventral tegmental area and substantia nigra (pars compacta and reticulata) and the density of the TH-positive fibers was not different in the caudate-putamen. These results indicate a selective reduction of terminal innervation in the medial portion of the mesocorticolimbic DA system in sP rats and suggest that the latter may consume larger amounts of ethanol, when compared with sNP rats, to compensate for the deficiency of dopamine to produce an adequate level of reward. © 2002 Elsevier Science B.V. All rights reserved.

2002 - Sub-chronic treatment with classical but not atypical antipsychotics produces morphological changes in rat nigro-striatal dopaminergic neurons directly related to 'early onset' vacuous chewing [Articolo su rivista]
Marchese, G.; Casu, M. A.; Bartholini, F.; Ruiu, S.; Saba, P.; Gessa, G. L.; Pani, L.

In the present work, we investigated if an impairment of dopaminergic neurons after subchronic haloperidol treatment might be a possible physiopathologic substrate of the 'early onset' vacuous chewing movements (VCMs) in rats. For this purpose, different antipsychotics were used to analyse a possible relationship between VCMs development and morphological alterations of tyrosine-hydroxylase-immunostained (TH-IM) neurons. Rats treated twice a day with haloperidol displayed a significant increase of VCMs that was both time- (2-4 weeks) and dose (0.1-1 mg/kg) dependent. Immunocytochemical analysis showed a shrinkage of TH-IM cell bodies in substantia nigra pars compacta and reticulata and a reduction of TH-immunostaining in the striatum of haloperidol treated rats with the arising of VCMs. No differences were observed in TH-IM neurons of ventral tegmental area and nucleus accumbens vs. control rats. The atypical antipsychotics risperidone (2 mg/kg, twice a day), amisulpride (20 mg/kg, twice a day) and clozapine (10 mg/kg, twice a day) did not produce any nigro-striatal morphological changes or VCMs. TH-IM nigro-striatal neuron morphological alterations and VCMs were still present after three days of withdrawal in rats treated for four weeks with haloperidol (1 mg/kg). Both the main morphological changes and the behavioural correlate disappeared after three weeks of withdrawal. These results suggest that haloperidol induces a morphological impairment of the dopaminergic nigro-striatal neurons which is directly associated with the arising, permanency and disappearance of VCMs in rats.

2002 - The role of limbic and cortical regions in schizophrenia: Focus on dopamine [Articolo su rivista]
Marchese, G.; Pani, L.

Dopamine is implicated in the pathogenesis of both the positive and the negative symptoms of schizophrenia. Clinical efficacy of antipsychotic drugs, without the production of side-effects, may be achieved by a dose-response separation of pharmacological function, regional (i.e., anatomical) selectivity of action, or by the selective targeting of neuroreceptors. The atypical antipsychotics have many different ways of acting on receptors in the brain, but they have in common a decreased likelihood of producing extrapyramidal side-effects. Patients respond well to them by showing improvements of both positive and negative symptoms. The preclinical profile of amisulpride shows specificity for D2/D3 dopamine receptors and selective activity in the limbic system. There is evidence that amisulpride is effective in treating both the negative and positive symptoms of schizophrenia, and that it has a low propensity to induce motor side-effects. Therefore, both positive and negative symptoms can be treated, without inducing these side-effects, by selectively targeting dopamine receptors. © 2002 Éditions scientifiques et médicales Elsevier SAS.

2002 - The substituted benzamides and their clinical potential on dysthymia and on the negative symptoms of schizophrenia [Articolo su rivista]
Pani, Luca; L., and Gessa; G., L.

2001 - (-)S amisulpride binds with high affinity to cloned dopamine D3 and D2 receptors [Articolo su rivista]
Castelli M., Paola; Mocci, I.; Sanna, A. M.; Gessa, G. L.; Pani, L.

Amisulpride is a substituted benzamide antipsychotic with nanomolar affinity and high selectivity for dopamine D2 and dopamine D3 receptors. The interaction of racemic (+/-)RS amisulpride and its two enantiomers (+)R and (-)S with dopamine D2 and dopamine D3 receptors subtypes were compared with that of haloperidol. Binding studies were performed using either [3H]spiperone or [3H]nemonapride in baculovirus/Spodoptera frugiperda insect (Sf-9) cell system expressing either the human dopamine recombinant D2long (hD2L) or the rat dopamine recombinant D3 (rD3) receptors. Ki values at dopamine rD3 receptors were similar regardless of the radioligand used, whereas at hD2L receptors values were higher using [3H]spiperone than [3H]nemonapride. However, the rank order of compound potency against radiolabeled spiperone or nemonapride both at dopamine hD2L and at dopamine rD3 receptors was similar. (-)S amisulpride displaced [3H]spiperone or [3H]nemonapride binding from both dopamine hD2L or dopamine rD3 receptors, being twofold more potent than the racemic form and 38-19-fold more potent than (+)R enantiomer. Both racemic and the (-)S enantiomer exhibited 2-4 ([3H]spiperone)- and 3-4 ([3H]nemonapride)-fold higher affinity than haloperidol for dopamine rD3 receptor, respectively. The (+)R enantiomer has weaker affinity with respect to haloperidol for both dopamine hD2L and dopamine rD3 receptors. Our results show that (-)S amisulpride is the active enantiomer of amisulpride, showing high affinity for dopamine D3 and dopamine D2 receptors. © 2001 Elsevier Science B.V. All rights reserved.

2001 - Evidence for co-release of noradrenaline and dopamine from noradrenergic neurons in the cerebral cortex [Articolo su rivista]
Devoto, P.; Flore, G.; Pani, L.; Gessa, G. L.

The aim of this study was to determine whether extracellular dopamine (DA) in the prefrontal cortex (PFC) might originate other than from DA neurons, also from noradrenergic (NA) ones. To this aim, we compared the levels of DA and NA in the dialysates from the PFC, a cortical area innervated by NA and DA neurons, and cortices that receive NA but minor or no DA projections such as the primary motor, the occipital-retrosplenial, and the cerebellar cortex. Moreover, the effect of α2-ligands and D2-ligands that distinctly modify NA and DA neuronal activity on extracellular NA and DA in these areas was studied. Extracellular NA concentrations were found to be similar in the different cortices, as expected from the homogeneous NA innervation, however, unexpectedly, also DA concentrations in the PFC were not significantly different from those in the other cortices. The α2-adrenoceptor agonist clonidine, intraperitoneally (i.p.) injected or locally perfused into the PFC, reduced not only extracellular NA levels, as expected from its ability to inhibit NA neuron activity, but also markedly reduced extracellular DA levels. Conversely, the α2-adrenoceptor antagonist idazoxan, i.p. injected or locally perfused into the PFC, not only increased extracellular NA levels, in line with its ability to activate NA neuron activity, but also increased those of DA. Conversely, in contrast to its ability to inhibit DA neuronal activity, the D2 receptor agonist quinpirole only modestly and transiently reduced extracellular DA levels, while γ-butyrolactone failed to modify DA levels in the PFC; conversely, haloperidol, at variance from its ability to activate DA neurons, failed to significantly modify extracellular DA levels in the PFC. Both haloperidol and quinpirole were totally ineffective after local perfusion into the PFC. Systemically injected or locally perfused, clonidine and idazoxan also modified both DA and NA concentrations in dialysates from primary motor, occipital-retrosplenial and cerebellar cortices as observed in the PFC. Finally, i.p. injected or locally perfused, clonidine reduced and idazoxan increased extracellular NA levels in the caudate nucleus, but neither α2-ligand significantly modified extracellular DA levels. Our results suggest that extracellular DA in the PFC, as well as in the other cortices, may depend on NA rather than DA innervation and activity. They suggest that dialysate DA reflects the amine released from NA neurons as well, where DA acts not only as NA precursor but also as co-transmitter. The co-release of NA and DA seems to be controlled by α2-receptors located on NA nerve terminals.

2001 - Lead intoxication during intrauterine life and lactation but not during adulthood reduces nucleus accumbens dopamine release as studied by brain microdialysis [Articolo su rivista]
Devoto, P.; Flore, G.; Ibba, A.; Fratta, W.; Pani, L.

Environmentally relevant levels of lead (Pb) have been demonstrated to have a neurotoxic action, especially on children. In this study, Long-Evans rats were continuously exposed to Pb acetate in drinking water from early gestational days (2-6) or from 28 days of age. At the 13th week of age, the functional activity of the nucleus accumbens (NAC) dopaminergic system was studied by means of transversal microdialysis. Neither Pb treatment regimen modified dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) extracellular concentrations, with respect to control rats. However, neuronal depolarisation, induced by perfusion with 60 mM KCl, increased extracellular DA levels to a significantly minor degree in rats exposed to Pb during the intrauterine life, with respect to both control and adult Pb treated rats. The in utero treated rats also responded with a lower DA release to amphetamine (1 mg/kg ip) administration. On the other hand, no difference in NAC DA level was found amongst treatment groups in response to different concentrations of the D2-D3 dopaminergic agonist quinpirole, locally administered by means of inverse dialysis. These data indicate a preferential impairment of NAC DA synthesis and/or release in rats exposed to Pb acetate during their intrauterine life. © 2001 Elsevier Science Ireland Ltd.

2001 - The R100Q mutation of the GABAA α6 receptor subunit may contribute to voluntary aversion to ethanol in the sNP rat line [Articolo su rivista]
Saba, L.; Porcella, A.; Congeddu, E.; Colombo, G.; Peis, M.; Pistis, M.; Gessa, G. L.; Pani, L.

We have investigated the GABAA α6 subunit molecular composition in two rat lines selectively bred for high or low ethanol preference and consumption, namely Sardinian alcohol-preferring (sP) and Sardinian non-alcohol-preferring (sNP) rats, which have been bred at the University of Cagliari, Italy, since 1981. A total of 27 sP, 22 sNP and 25 control rats belonging to five other different strains, were studied by direct sequencing and amplification refractory mutation system analysis. Among the sNPs, only one was found to be normal, 11 heterozygotes, and 10 homozygotes for the G→A substitution in codon 100, the same R100Q point mutation previously described in Alcohol Non Tolerant rats, while no other animal showed any mutated allele. Pharmacological studies have extensively demonstrated that this substitution in the mature peptide changes the benzodiazepine-insensitive receptor to a sensitive one. In order to test the functional significance of this mutation in native cerebellar GABAA receptors, selective breeding from Q/R rats was employed to obtain a sufficient number of R/R homozygotes. Xenopus laevis oocytes were then injected with cerebellar synaptosomes extracted from Q/Q, R/Q and R/R sNP rats. Consistently, utilizing the two-electrode voltage-clamp technique, GABA-evoked currents mediated by GABAA receptors containing the mutated α6 subunit were potentiated by diazepam with about a two-fold increased potency, as compared to receptors containing the wild-type, benzodiazepine-insensitive α6 subunit. Our data show for the first time that a mutated GABAA α6 receptor subunit segregates in a rat line which voluntarily avoids alcohol consumption, and further support a possible involvement of the GABAA receptor containing a mutated α6 subunit in the genetic predisposition to alcohol preference. © 2001 Elsevier Science B.V.

2001 - The synthetic cannabinoid WIN55212-2 decreases the intraocular pressure in human glaucoma resistant to conventional therapies [Articolo su rivista]
Porcella, Anna; Maxia, Chiara; Luigi Gessa, Gian; Pani, Luca

2000 - Is there an evolutionary mismatch between the normal physiology of the human dopaminergic system and current environmental conditions in industrialized countries? [Articolo su rivista]
Pani, L.

A large body of evidence has recently defined a field theory known as 'evolutionary mismatch', which derives its attributes largely from the fact that current environmental conditions are completely different from those in which the human central nervous system evolved. Current views on the evolutionary mismatch theory lack, however, any attempts to define which brain areas or neuronal circuits should be mostly involved in coding such misevolved traits and to what extent our neurobiological knowledge can be applied to the topographical localization of a specific psychopathology. In this respect the mesocorticolimbic dopaminergic circuits have long been misconceptualized as simple reward or reinforcement systems. Instead, they motivate and coordinate the functions of the higher brain areas that mediate planning and foresight and direct finalized movement in both animals and humans. These systems make animals intensely interested in exploring the world around them, but by the same means they also make them susceptible to the environmental stimuli that have been sought and consumed. It is has been speculated that the cortical dopamine targets that developed most recently in phylogeny are of particular functional value, and that the mesocorticolimbic dopaminergic system is involved in more complex integrative functions than previously assumed. In the present paper I will argue that some mental disorders may have their deep roots in the evolutionary mismatch between the normal physiology of the mesocorticolimbic dopaminergic system and the current environmental conditions in affluent societies.

2000 - The 5-HT2 antagonist ritanserin blocks dopamine re-uptake in the rat frontal cortex [Articolo su rivista]
Ruiu, S.; Marchese, G.; Saba, P. L.; Gessa, G. L.; Pani, L.

The effect of ritanserin on dopamine (DA) re-uptake and efflux was studied in rat frontal cortex synaptosomes. When compared to other 5HT2 receptor antagonists such as ketanserin and risperidone or BA D2 receptor antagonists such as haloperidel and raclopride, the effect of ritanserin proved to be more potent. Ritanserin blocked the DA transporter with e K1 of 0.18 ± 0.06 μM, similar to cocaine (0.11 ± 0.005 μM), while ketanserin had a K1 of 0.93 ± 0.045; haloperidol of 2.07 ± 0.12; risperidone of 18.01 ± 0.62 and raclopride of 24.01 ± 1.55. In addition, 15 min from its local application to the synaptosomes, ritanserin potently released [3]H-DA leaving only 29.6 ± 1.6% of DA content, while ketanserin effect was equal to 45.5 ± 0.9%; haloperidol to 70.4 ± 2.2% and risperidone to 73.9 ± 1.5%, all tested at the dose of 10 μM. Cocaine had no effect on DA efflux. These results suggest that ritanserin has a intrinsic dopaminergic effect which may help to explain its reported improvement on mood, cognition and negative symptoms of schizophrenia.

2000 - The human eye expresses high levels of CB1 cannabinoid receptor mRNA and protein [Articolo su rivista]
Porcella, A.; Maxia, C.; Gessa, G. L.; Pani, L.

We used reverse transcriptase polymerase chain reaction to detect the expression of the central and peripheral cannabinoid receptors (CB1 and CB2, respectively) mRNA, and Western blotting to show the presence of the CB1 protein in subregions of the human eye. CB2 mRNA transcripts were undetectable, while levels of CB1 mRNA were significantly expressed in the human retina (25.8 ± 2.46%), ciliary body (210 ± 11.55%) and iris (62.7 ± 5.94%) when compared with those of the normalizing reference gene β2 microglobulin. The CB1 gene encodes a functional protein which is detected in its glycosylated (63 kDa) and unglycosylated (54 kDa) form in the same areas by a specific purified antibody raised against the amino terminus (residues 1-77) of the CB1 receptor. These results further support the proposed role of the CB1 receptor in controlling intraocular pressure, helping to explain the antiglaucoma properties of marijuana.

2000 - The role of stress in the pathophysiology of the dopaminergic system [Articolo su rivista]
Pani, Luca; L., and Porcella; A., and Gessa; G., L.

1998 - Cannabinoid receptor CB1 mRNA is highly expressed in the rat ciliary body: Implications for the antiglaucoma properties of marihuana [Articolo su rivista]
Porcella, A.; Casellas, P.; Gessa, G. L.; Pani, L.

We used RT-PCR to measure relative differences in cannabinoid receptor (CB) mRNAs in the rat eye, comparing CB1 or CB2 transcripts to that of the normalizing reference gene β2 microglobulin (β2m). Significantly higher levels of CB1 mRNA levels were found in the ciliary body (0.84 ± 0.05% of β2m) than in the iris, (0.34 ± 0.04% of β2m), retina (0.07 ± 0.005% of β2m) and choroid (0.06 ± 0.005% of β2m). CB2 mRNA was undetectable. This expression pattern supports a specific role for the CB1 receptor in controlling intraocular pressure, helping to explain the antiglaucoma property of cannabinoids.

1998 - The role of dopamine in the biological basis of dysthymia [Articolo su rivista]
Pani, L.; Gessa, G. L.

This paper reviews the recent literature supporting the hypothesis that a reduced neurotransmission in the mesolimbic dopamine (DA) system may sustain some of the core and subsidiary symptoms of dysthymia and other depressive conditions, namely anhedonia, lack of interest, lack of drive, lack of concentration, and psychomotor retardation. Experimental evidence indicates that mesolimbic DA mediates the rewarding; motivating and incentive effect of natural and artificial stimuli such as sex, foods, liquids, intracranial self-stimulation, and the drugs such as cocaine and amphetamine. Conversely, it has been shown that withdrawal from chronic treatment with these drugs is associated with depressive symptomatology and reduced release of DA in the ventral striatum. Similarly, different models of depression such as 'behavioral despair', 'learned helplessness' and 'chronic mild stress' are associated with reduced DA activity in the limbic system, which is reversed by chronic treatment with antidepressants. Various antidepressants, irrespective of their acute action on the uptake of specific neurotransmitters such as noradrenaline, serotonin and DA, when given chronically have the common property of potentiating the behavioral responses to psychostimulants that are mediated by DA receptors in the limbic areas. The DA hypothesis of depression offers a logical explanation for the antidepressive effect of drugs such as sulpiride and amisulpride, which block DA autoreceptors and thus increase DA output.

1998 - Δ9-Tetrahydrpcannabinol increases sequence-specific AP-1 DNA-binding activity and Fos-related antigens in the rat brain [Articolo su rivista]
A, Porcella; G, L Gessa; Pani, L.

Delta-9-tetrahydrocannabinol (Δ9-THC), the psychoactive principle of marijuana, has been shown to upregulate the mRNA levels of immediate-early genes in the rat brain. Using electrophoretic mobility-shift assay and onedimensional Western blot, we here report that Δ9-THC increases Activator protein-1 (AP-1) DNA-binding and Fos-related antigen activity in discrete areas of the rat brain. One hour after the intraperitoneal administration of Δ9-THC at a dose of 10 or 15 mg/kg, AP-1 DNA-binding activity in the nucleus accumbens increased by 33 and 49%, respectively, while Western blot showed an increase in both c-Fos, FosB, Fra-1 (Fos-related antigen) and Fra-2. In the cingulate cortex and caudate-putamen, Δ9-THC significantly increased AP-1 DNA-binding activity only at the highest dose used (57 and 71 %, respectively). While in the caudate-putamen the increase in AP-1 DNA binding was mainly due to an elevation of the c-Fos and FosB proteins, the same phenomenon depended on the FosB, Fra-1 and Fra-2 peptides in the cingulate cortex. The effect of Δ9-THC on the AP-1 DNA binding and the Fos-related antigens in the nucleus accumbens was blocked by the specific cannabinoid antagonist SR141716 A (3 mg/kg i.p.). Δ9-THC failed to modify Specificity protein 1 (Sp1) DNA-binding activity. The results indicate that Δ9-THC activates gene coding for AP-1 DNA-binding proteins by acting on cannabinoid receptors, and induces a different transcriptional program on the early-immediate gene of the Fos family, in different areas in the rat brain, suggesting that this mechanism might be involved in the central actions of cannabinoids. © European Neuroscience Association.

1997 - Evolution of the dopaminergic system and its relationships with the psychopathology of pleasure [Articolo su rivista]
Pani, L.; Gessa, G. L.

This paper summarizes the fundamental steps in the evolution of the dopaminergic system. A rudimentary dopaminergic system is present in primitive creatures, already able to select information processing, modulate 'emotional' behaviours and react to perturbations in environmental conditions. Pharmacological manipulations of the dopaminergic transmission are able to modify basic behaviours present in all animals from fishes to lizards to mammals. The ability to put the organism in motion and the hedonic capacity of giving pleasure, would justify the conservation through evolution of such a neuronal system. The fact however that the dopaminergic system has remained identical for the last several centuries, while many external conditions which interfere with its physiology have dramatically changed, may contribute to explain the transition from the original vital advantages of the dopaminergic system to its crucial role in the psychopathology of pleasure.

1997 - Fluoxetine-induced conditioned place preference: A preliminary study [Articolo su rivista]
Collu, M.; Poggiu, A. S.; Pani, L.; Serra, G.

1997 - Rat tyrosine hydroxylase gene polymorphisms [Articolo su rivista]
Murru, S; Pani, L; Poddie, D; Gessa, G; Cao, A; Pirastu, M.

1996 - Circulating levels of anticonvulsant metabolites of progesterone in women with partial epilepsy in the intercritical phase [Articolo su rivista]
Galli, R.; Michelini, S.; Bartalena, L.; Massetani, R.; Pani, L.; Grasso, L.; Cassano, G. B.; Martino, E.; Purdy, R. H.; Murri, L.

The Salpha-hydroxy metabolites of progesterone (P), 3alpha-hydroxy-5alpha-pregnan-20-one (allopregnanolone, A-PREG) and 3alpha-hydroxy-5beta-pregnan-20-one (pregnanolone, PREG), have been found to be among the most potent ligands of gamma-aminobutyric (GABA) eceptors; in experimental animals, they have been found to have anxiolytic, hypnotic and anticonvulsant effects 'ilar to those of the benzodiazepines and barbiturates that interact with GABA-A receptors. The present study was iertaken to determine plasma A-PREG and PREG concentrations in the luteal phase in women with partial epilep;, , in order to determine if an impaired metabolism of P occurs in this convulsive disorder. We measured plasma P, A-PREG and PREG levels in 75 women with partial epilepsy in the intercritical phase, and in 15 age-matched healthy women, during the luteal phase of the ovarian cycle (22nd24th day). The mean plasma ±S.E. A-PREG levels (three blood samples) were 0.7±0.6 ng/ml in the epileptic women and 0.5±0.2 ng/ml in controls, with no significant difference beteen the two groups (p = NS); the PREG levels were also similar (J.4±J ng/ml and I ±1.1 ng/ml, respectively: p = NS). A significant correlation was found between P levels and both A-PREG and PREG levels (r=0.72, p<0.001 and r = 0.79, p<0.00l, respectively). There were no significant differences between the two groups in terms of serum adrenocorticotropic hormone, Cortisol, dihydroepiandrosterone-sulfate, follicle stimulating hormone, prolactin, luteinizing hormone or estradiol levels.

1996 - Correlation between increased AP-1NGF binding activity and induction of nerve growth factor transcription by multiple signal transduction pathways in C6-2B glioma cells [Articolo su rivista]
Colangelo, A. M.; Pani, L.; Mocchetti, I.

Transcription mechanisms regulating nerve growth factor (NGF) gene expression in the CNS are yet to be thoroughly understood. We have used C6-2B rat glioma cells to characterize the signal transduction pathways that contribute to transcriptional and posttranscriptional regulation of NGF mRNA. Because the NGF promoter contains an AP-1 consensus sequence, we have investigated whether increases in AP-1 binding activity correlate with enhanced NGF mRNA expression. Gel mobility shift assays using an oligonucleotide homologous to the AP-1 responsive element of the rat NGF gene (AP-1NGF) revealed that 12-O-tetradecanoyl phorbol-13-acetate (TPA) and, to a lesser extent, isoproterenol (ISO) and thapsigargin, a microsomal Ca2+-ATPase inhibitor, stimulated binding to AP-1NGF within 2 h. All of these stimuli increased NGF mRNA levels within 3 h. Cycloheximide pretreatment blocked the TPA and ISO-mediated binding to AP-1NGF suggesting that de novo synthesis of c-Fos/c-Jun may be required for the transcriptional regulation of NGF gene. Nuclear run-on assays and NGF mRNA decay studies revealed that TPA increases NGF transcription whereas ISO affects both transcription and mRNA stabilization. We propose that (i) different signal transduction mechanisms regulate the expression of the NGF gene in cells derived from the CNS, and (ii) both mRNA transcription and stability account for the cAMP-mediated increase in NGF mRNA levels.

1996 - Dysthymia in neurological disorders [Articolo su rivista]
Akiskal, H. S.; Bolis, C. L.; Cazzullo, C.; Costa, E Silva; J. A., Gentil; V., Lecrubier; Y., Licinio; Qemail, Author; Linden, M.; Lopez-Ibor, J. J.; Ndiaye, I. P.; Pani, L.; Prilipko, L.; Robertson, M. M.; Robinson, R. G.; Starkstein, S. E.; Thomas, P.; Wang, Y.; Wong, M.

1995 - Animal models of mania [Articolo su rivista]
Gessa, G. L.; Pani, L.; Serra, G.; Fratta, W.

1995 - Anxious-excited depression. A mixed affective syndrome [Articolo su rivista]
Koukopoulos, A.; Pani, L.; Serra, G.; Minnai, G.; Reginaldi, D.

A mixed affective syndrome is described which meets the criteria for major depression but not those of the DSM III-R for a mixed state. The clinical picture is characterized by lack of motor retardation and fluent verbalization; the facial expression is animated and sometimes dramatic. Patients suffer considerably and are often tearful. They complain of inner tension and restlessness, racing thoughts and despair. Emotional lability and momentary irritability are observed. Insomnia occurs initially or with frequent early waking. Suicidal ideation occurs and makes the syndrome of concern in view of its impulsive nature. Antidepressants increase restlessness, insomnia, aggressiveness and the impulsiveness of suicidal ideation. Low-dose neuroleptics, lithium and anticonvulsivants are highly effective. A few sessions of ECT offer rapid improvement.

1995 - Chronic imipramine, l-sulpiride and mianserin decrease corticotropin releasing factor levels in the rat brain [Articolo su rivista]
Fadda, P.; Pani, L.; Porcella, A.; Fratta, W.

Among clinically effective antidepressant drugs, the action mechanism of mianserin has recently been related to variations in corticotropin releasing factor (CRF) levels in the rat locus coeruleus. We describe a specific effect on CRF levels after chronic treatment with different antidepressants: mianserin (10 mg/kg), imipramine (20 mg/kg), both for 21 days, or l-sulpiride (1 mg/kg) for 15 days. While all antidepressants used greatly decreased CRF concentratrions in the hypothalamus, only mianserin decreased CRF concentrations by 40% in extrahypothalamic sites. Acute treatments failed to modify CRF levels. Chronic treatment with mianserin did not affect CRF density either in the hypothalamus or the extrahypothalamic areas. This new finding may add another facet to the therapeutic action of certain antidepressants and in particular to the atypical profile of mianserin. © 1995.

1995 - Cytokine regulation of the liver transcription factor hepatocyte nuclear factor-3β is mediated by the C/EBP family and interferon regulatory factor 1 [Articolo su rivista]
Samadani, U.; Porcella, A.; Pani, L.; Johnson, P. F.; Burch, J. B. E.; Pine, R.; Costa, R. H.

Three distinct hepatocyte nuclear factor-3 (HNF-3) proteins (α, β, and γ) regulate the transcription of numerous liver-enriched genes. The HNF-3 proteins bind DNA via a homologous winged helix motif common to a number of proteins known to be critical for determination events in embryogenesis. We have demonstrated previously that two binding sites in the -184 HNF-3β promoter are recognized by widely distributed factors and that there is also a critical autoregulatory site present that is recognized by members of the HNF-3 family. Adjacent to the autoregulatory site, we identified a binding site for a cell-specific factor, LF-H3β, that may function in restricting HNF-3β gene expression to hepatocytes. Our present study demonstrates that members of the C/EBP and proline and acidic amino acid-rich subfamilies of basic region leucine zipper transcription factors bind the LF-H3β site, and cotransfection of HepG2 cells shows that these factors are able to activate an HNF-3β promoter reporter construct. The LF-H3β-C/EBP binding sequence also confers HNF-3β promoter stimulation in response to interleukin (IL)-1 and IL-6. Upstream of this HNF-3β proximal promoter region, an IFN- stimulated response element core sequence (-231 to -210) was found that mediates transcriptional induction by IFN-γ but not IFN-α. Gel mobility supershift assay demonstrates that an IFN-γ-induced protein-DNA complex is disrupted by an antibody specific for interferon regulatory factor- 1/interferon-stimulated gene factor-2. Consistent with this finding, we observed that IFN-γ induction requires ongoing protein synthesis. Surprisingly, the effect of the three cytokines (IL-1, IL-6, and IFN-γ) in combination as assayed by the same model is not synergistic. HNF-3β joins the C/EBP family on the list of liver-enriched transcription factors, the expression of which is modulated by cytokines.

1995 - Dysthymia biochemistry: The role of dopamine [Articolo su rivista]
Gessa, G. L.; Serra, G.; Pani, L.

1995 - Sleep deprivation in the rat: an animal model of mania [Articolo su rivista]
Gessa, G. L.; Pani, L.; Fadda, P.; Fratta, W.

The model of sleep deprivation in rats by the platform method has been extensively studied in our laboratory as a possible animal model of mania. At the end of the period of sleep deprivation, the rat does not fall asleep as soon as it is returned to its home cage, but shows a period of wakefulness of about 30 min, during which the animal presents a cohort of symptoms that appear to mimic those present in idiopathic mania. In particular, during this period the animal displays insomnia, a high degree of hyperactivity, irritability, aggressiveness, hypersexuality and stereotypy. Haloperidol (0.2 mg/kg) was effective in reducing latency to sleep, while l-sulpiride was much weaker (< 50 mg/kg). The dopamine D1 receptor antagonist SCH 23390 exhibited an extremely high potency and efficacy in reducing sleep latency, a significant effect being observed with 3 μg/kg. The administration of the specific D1 receptor agonist SKF 38393 markedly prolonged the period of insomnia with the correlated behavioral syndrome. When lithium was added to the diet and consumed during the sleep deprivation period in adequate amounts to produce serum lithium levels of 0.7-1.0 mEq/1, sleep latency and locomotor activity were significantly reduced. The administration of naloxone (1-10 mg/kg) reduced the latency to sleep in a dose-related manner. By contrast, morphine (1 and 5 mg/kg, i.p.), β-endorphin and [d-Ala2,d-Leus5]enkephalin (i.c.v., 2 and 1 μg, respectively) markedly prolonged the insomnia. The model not only represents a confirmation in the rat that sleep loss often precedes and may trigger a manic episode in man, but suggests that an opioid-dopamine interaction may play a pathogenetic role in mania. © 1995.

1995 - The long term prophylaxis of affective disorders [Articolo su rivista]
Koukopoulos, A.; Reginaldi, D.; Minnai, G.; Serra, G.; Pani, L.; Johnson, F. N.

1994 - Antidepressant-like effect of selective dopamine D1 receptor agonists in the behavioural despair animal model of depression [Articolo su rivista]
D'Aquila, P. S.; Collu, M.; Pani, L.; Gessa, G. L.; Serra, G.

We compared the effect of two selective dopamine D1 receptor agonists, SKF 38393 ((±)-1-phenyl-2,3,4,5-tetrahydro-(1H)- 3-benzazepine-7,8-diol · HCl) and A68930 ((1R,3S)-1-aminomethyl-5,6-dihydroxy-3-phenylisochroman · HCl), and that of imipramine in the behavioural despair model of depression. The dopamine D1 receptor agonists and impramine showed an anti-immobility effect. Moreover we found that the 'antidepressant' effect of imipramine in the behavioural despair test was antagonized by SCH 23390, a selective dopamine D1 receptor blocker. The results further support the hypothesis that dopamine D1 receptor stimulation plays an important role in the mechanism of action of amtidepressants and suggest that dopamine D1 receptor agonists might be considered as potential antidepressant drugs. © 1994.

1994 - Pharmacology of neurosteroid biosynthesis. Role of the mitochondrial DBI receptor (MDR) complex [Articolo su rivista]
Costa, E; Cheney, D. L; Grayson, D. R; Korneyev, A; Longone, P; Pani, L; Romeo, E; Zivkovich, E; Guidotti, A.

1993 - ACTH-induced mitochondrial DBI receptor (MDR) and diazepam binding inhibitor (DBI) expression in adrenals of hypophysectomized rats is not cause-effect related to its immediate steroidogenic action [Articolo su rivista]
Cavallaro, S.; Pani, L.; Guidotti, A.; Costa, E.

Diazepan binding inhibitor (DBI) is a 10-kDa polypeptide that is enriched in steroidogenic cells such as adrenocortical, Leydig, and glial cells. In these cells, DBI and some of its processing products bind to the mitochondrial DBI receptor (MDR), located on the outer mitochondrial membrane, and stimulate pregnenolone formation by facilitating cholesterol access to the inner mitochondrial membrane where the cytochrome P-450 side chain cleavage enzyme is located. To determine whether the ACTH-induced increase in adrenal steroidogenesis occurs via changes in DBI and MDR expression the adrenal content of DBI-like immunoreactivity (DBI-LI), the MDR density, and the expression of mRNAs encoding for DBI and MDR were studied in hypophysectomized rats treated with vehicle or ACTH. After 9 days from the hypophysectomy, the levels of DBI-like immunoreactivity (DBI-LI) and DBI-mRNA declined to approximately 20% of their normal value; in contrast MDR-density and MDR-mRNA levels were reduced by 50-60% and were associated to a similar decrease in the activity of type A monoamine oxidase, a marker for mitochondrial proteins. Prolonged administration of ACTH-R (ACTH in saline containing 16% gelatin, 15 U/kg/day, from day 7 after surgery) to hypophysectomized rats, completely restored DBI and MDR adrenal expression to values similar to those of sham-operated rats. Our results indicate that ACTH, probably acting at the transcriptional level, is required for the normal expression of DBI and MDR in adrenal cortex. Changes in DBI and MDR expression after ACTH administration were not temporally related to the immediate steroidogenesis induced by ACTH, and may reflect its long-term trophic action on adrenocortical cells. © 1993.

1993 - Cloning and tissue-specific functional characterization of the promoter of the rat diazepam binding inhibitor, a peptide with multiple biological actions [Articolo su rivista]
Kolmer, M.; Alho, H.; Costa, E.; Pani, L.

Diazepam binding inhibitor (DBI) is a 10-kDa polypeptide that regulates mitochondrial steroidogenesis, glucose-induced insulin secretion, metabolism of acyl-CoA esters, and the action of γ-aminobutyrate on GABAA receptors. To investigate the regulation of DBI gene expression, three positive clones were isolated from a rat genomic library. One of them contained a DBI genomic DNA fragment encompassing 4 kb of the 5′ untranslated region, the first two exons, and part of the second intron of the DBI gene. Two other overlapping clones contained a processed DBI pseudogene. Several transcription initiation sites were detected by RNase protection and primer extension assays. Different tissues exhibited clear differences in the efficiencies of transcription startpoint usage. Transient expression experiments using DNA fragments of different length from the 5′ untranslated region of the DBI gene showed that basal promoter activity required 146 bp of the proximal DBI sequence, whereas full activation was achieved with 423 bp of the 5′ untranslated region. DNase I protection experiments with liver nuclear proteins demonstrated three protected regions at nt -387 to -333, -295 to -271, and -176 to -139 relative to the ATG initiation codon; in other tissues the pattern of protection was different. In gel shift assays the most proximal region (-176 to -139) was found to bind several general transcription factors as well as cell type-restricted nuclear proteins which may be related to specific regulatory patterns in different tissues. Thus, the DBI gene possesses some features of a housekeeping gene but also includes a variable regulation which appears to change with the function that it subserves in different cell types.

1993 - Identification of nine tissue-specific transcription factors of the hepatocyte nuclear factor 3/forkhead DNA-binding-domain family [Articolo su rivista]
Clevidence, D. E; Overdier, D. G; Tao, W; Qian, X; Pani, L; Lai, E; Costa, R. H.

1992 - Hepatocyte nuclear factor 3β contains two transcriptional activation domains, one of which is novel and conserved with the Drosophila fork head protein [Articolo su rivista]
Pani, L.; Overdier, D. G.; Porcella, A.; Qian, X.; Lai, E.; Costa, R. H.

The hepatocyte nuclear factor 3 (HNF-3) gene family is composed of three proteins (α, β, and γ) that are transcription factors involved in the coordinate expression of several liver genes. All three proteins share strong homology in their DNA binding domains (region I) and are able to recognize the same DNA sequence. They also possess two similar stretches of amino acids at the carboxyl terminus (regions II and III) and a fourth segment of homology at the amino terminus (region IV). Furthermore, the HNF-3 proteins demonstrate homology with the Drosophila homeotic gene fork head in regions I, II, and III, suggesting that HNF-3 may be its mammalian homolog. In order to define HNF-3β protein domains involved in transcriptional activation, we have used a reporter gene, whose transcription is dependent on HNF-3 binding, for hepatoma cell cotransfection assays with expression vectors that produced different truncated HNF-3β proteins. A position-independent activation domain which contained conserved regions II and III was identified at the carboxyl terminus of the HNF-3β protein (amino acids 361 to 458). Moreover, site-directed mutations that altered the sequences within regions II and III demonstrated their importance to transactivation. The region II-III domain does not possess amino acid sequences in common with other transcription factors and may define a novel activation motif. HNF-3β amino-terminal sequences defined by conserved region IV also contributed to transactivation, but region IV activity required the participation of the region II-III domain. Region IV is abundant in serine amino acids and contains two putative casein kinase I phosphorylation sites, a feature similar to protein motifs described for the transcription factors Pit-1/GHF-1 and HNF-1.

1992 - The restricted promoter activity of the liver transcription factor hepatocyte nuclear factor 3β involves a cell-specific factor and positive autoactivation [Articolo su rivista]
Pani, L.; Qian, X.; Clevidence, D.; Costa, R. H.

The transcription factor hepatocyte nuclear factor 3 (HNF-3) is involved in the coordinate expression of several liver genes. HNF-3 DNA binding activity is composed of three different liver proteins which recognize the same DNA site. The HNF-3 proteins (designated α, β, and γ) possess homology in the DNA binding domain and in several additional regions. To understand the cell-type-specific expression of HNF-3β, we have defined the regulatory sequences that elicit hepatoma-specific expression. Promoter activity requires -134 bp of HNF-3β proximal sequences and binds four nuclear proteins, including two ubiquitous factors. One of these promoter sites interacts with a novel cell-specific factor, LF-H3β, whose binding activity correlates with the HNF-3β tissue expression pattern. Furthermore, there is a binding site for the HNF-3 protein within its own promoter, suggesting that an autoactivation mechanism is involved in the establishment of HNF-3β expression. We propose that both the LF-H3β and HNF-3 sites play an important role in the cell-type-specific expression of the HNF-3β transcription factor.

1991 - Flunarizine attenuates cocaine-induced inhibition of A9 dopaminergic neurons [Articolo su rivista]
Diana, M.; Pani, L.; Rossetti, Z.; Passino, N.; Gessa, G. L.

The intravenous administration of cocaine (0.5-8 mg/kg) produced a dose-related inhibition of the firing rate of substantia nigra (A9) dopaminergic (DA) neurons. Pretreatment with the calcium antagonist flunarizine (5 mg/kg i.v.) prevented cocaine-induced effects but failed to antagonize the inhibition of firing induced by a low dose of apomorphine (10μg/kg i.v.). This finding rules out the possibility that flunarizine antagonism of cocaine effect might depend on DA autoreceptors blockade by flunarizine. It is suggested that flunarizine, by blocking calcium influx into DA neurons, prevents DA release from dendrites, thereby counteracting the main mechanism by which cocaine inhibits dopaminergic neuronal activity. © 1991 The Italian Pharmacological Society.

1991 - Inhibition of [3H]dopamine uptake by flunarizine [Articolo su rivista]
Devoto, P.; Pani, L.; Kuzmin, A.; Montis, G. D.

The effect of different calcium channel blockers was studied on basal and cocaine-inhibited [3H]dopamine uptake in rat striatal synaptosomes. Isradipine (dihydropyridine calcium antagonist) and diltiazem (L-type calaium antagonist) were devoid of effect on [3H]dopamine uptake, while flunarizine (T-type calcium antagonist) inhibited [3H]dopamine uptake. Flunarizine inhibition was competitive and the inhibitory curve was biphasic with a Hill coefficient of 2.1. The high Hill number suggested a mechanism of positive cooperativity between two sites. Flunarizine inhibition showed a complex interaction with cocaine inhibition. While flunarizine at low concentrations interacts with a distinct site, at higher concentrations it interacts with the same site as cocaine. The relevance of this finding for the potentiation by flunarizine of cocaine-induced dopamine release in vivo is discussed. © 1991.

1991 - The calcium antagonist PN 200-110 inhibits the reinforcing properties of cocaine [Articolo su rivista]
Pani, Luca; L., and Kuzmin; A., and Martellotta; M. C., and Gessa; G. L., and Fratta

1990 - Brain dialysis and dopamine: does the extracellular concentration of dopamine reflect synaptic release? [Articolo su rivista]
Pani, L.; Gessa, G. L.; Carboni, S.; Portas, C. M.; Rossetti, Z. L.

We considered the drug-induced circling behaviour of rats monolaterally lesioned with kainic acid (KA) as a marker of the dopamine (DA) concentration in the synaptic space. D-Amphetamine produced a dose-related increase in the DA concentration of the dialysate from an intact striatum and a proportional number of ipsilateral circlings. Pargyline or L-dihydroxyphenylalanine (L-Dopa), alone or in combination with benserazide, increased the concentration of DA to a similar or even higher level than d-amphetamine, but failed to elicit a circling response. Apomorphine given after these drugs at the peak of DA accumulation always induced circling behaviour. The results suggest that released DA may undergo different inactivation processes before reaching the dialysis probe, and that these processes may be differentlly affected by drug treatments. Alternatively, it may be suggested that DA can be released into the synaptic space, in a functional manner, or into the interstitial fluid, from where it cannot reach the synaptic receptors. © 1990.

1990 - Calcium receptor antagonists modify cocaine effects in the central nervous system differently [Articolo su rivista]
Pani, L.; Kuzmin, A.; Diana, M.; De Montis, G.; Gessa, G. L.; Rossetti, Z. L.

The effect of different calcium antagonists on cocaine-induced dopamine (DA) release in the striatum, as measured by brain microdialysis in freely moving rats, and on cocaine-induced motor stimulation was studied. While two dihydropyridine calcium antagonists, nimodipine (20 mg/kg) and isradipine (2.5 mg/kg), prevented cocaine-induced DA release and motor stimulation, the diphenylalkylamine-type calcium antagonist flunarizine (20 mg/kg) strongly potentiated both effects of cocaine. Moreover, two calcium antagonists, veraparnil (20 mg/kg) and diltiazem (20 mg/kg), were ineffective. The results indicate that various classes of calcium antagonists differ in their interaction with the effects of cocaine in the CNS and suggest that dihydropyridine calcium channel antagonists might be clinically useful for the treatment of cocaine abuse. © 1990.

1990 - Conditioned place preference induced by ethanol in a rat line selected for ethanol preference [Articolo su rivista]
Colombo, G.; Kuzmin, A.; Fadda, F.; Pani, L.; Luigi Gessa, G.

1990 - Dihydropyridine calcium antagonists prevent cocaine-, but not amphetamine-, induced dopamine release and motor activity in rats [Articolo su rivista]
Rossetti, Z. L.; Pani, L.; Kuzmin, A.; Carboni, S.; Gessa, G. L.

1990 - Flunarizine potentiates cocaine-induced doparaine release and motor stimulation in rats [Articolo su rivista]
Pani, L.; Kuzmin, A.; Stefanini, E.; Gessa, G. L.; Rossetti, Z. L.

Pretreatment with fiunarizine (20 mg/kg), a diphenylalkylamine calcium channel antagonist, markedly potentiated cocaine (10 mg/kg) -induced dopamine (DA) output from the ventral striatum, as measured by microdialysis in freely moving rats. Moreover, fiunarizine enhanced cocaine-induced motor stimulation. Pretreatment with the D2 receptor antagonist (-)-sulpiride (25 mg/kg) potentiated cocaine-induced DA output but, unlike fiunarizine, inhibited cocaine-induced motor stimulation. When added to striatal membrane preparations, flunanzine displaced [3H]spiperone binding with a Ki of about 100 nM. Since recent evidence indicates that the effects of cocaine are calcium-dependent and are prevented by dihydropyridine calcium antagonists, these findings suggest that the paradoxical potentiating effect of fiunarizine is probably due to an interaction with the DA system and is not due to its calcium antagonist property. © 1990.

1990 - Nimodipine inhibits cocaine-induced dopamine release and motor stimulation [Articolo su rivista]
Pani, L.; Carboni, S.; Kusmin, A.; Gessa, G. L.; Rossetti, Z. L.

1990 - Stress increases noradrenaline release in the rat frontal cortex: Prevention by diazepam [Articolo su rivista]
Rossetti, Z. L.; Portas, C.; Pani, L.; Carboni, S.; Gessa, G. L.

Foot-shock produced a more than 2-fold increase in noradrenaline (NA) release from the frontal cortex of freely moving rats. The effect of acute stress was almost completely prevented by the administration of diazepam (5 mg/kg i.p.). Diazepam alone inhibited cortical NA release, the maximal inhibition (-57%) being observed 90 min after the injection. Cortical NA release therefore appears to be a reliable index of central noradrenergic activity in response to stressful conditions. © 1990.

1990 - The non-competitive NMDA-receptor antagonist MK-801 prevents the massive release of glutamate and aspartate from rat striatum induced by 1-methyl-4-phenylpyridinium (MPP+) [Articolo su rivista]
Carboni, S.; Melis, F.; Pani, L.; Hadjiconstantinou, M.; Rossetti, Z. L.

The concentrations of dopamine (DA) and of the excitatory amino acids (EAAs) glutamate (Glu) and aspartate (Asp) were measured in dialysates from the striatum of awake rats in order to study the link between the release of DA and of EAAs induced by the infusion of 1-methyl-4-phenylpyridinium ion (MPP+). DA and EAAs were detected simultaneously by HPLC-EC. The infusion of MPP+ at the concentration of 1 mM elevated DA levels in the perfusates, but did not affect EAA release. However, MPP+ at 10 mM maximally stimulated Glu and Asp release to 230- and 68-fold of baseline, respectively. In this condition, pretreatment with the N-methyl-d-aspartate (NMDA) receptor antagonist MK-801 (5 mg/kg, i.p.), prevented the MPP+-induced EAA release. In contrast, MK-801 had no effect on DA release induced either by 1 or 10 mM MPP+. These results suggest that MPP+-induced DA and EAA release are independently regulated processes. In addition, the finding that MK-801 inhibits MPP+-induced EAA release suggests that EAAs may act on NMDA receptors to stimulate their own release through a positive-feedback mechanism. © 1990.

1989 - Brain dialysis provides evidence for D2-dopamine receptors modulating noradrenaline release in the rat frontal cortex [Articolo su rivista]
Rossetti, Z. L.; Pani, L.; Portas, C.; Gessa, G.

1989 - Differential postmortem release of noradrenaline and dopamine from rat brain [Articolo su rivista]
Rossetti, Z. L; Pani, L; Gessa, G. L.

1988 - Are D1 dopamine receptor agonists potential antidepressants? [Articolo su rivista]
Serra, G; Collu, M; D'Aquila, P; Pani, L; Gessa, G. L.