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Rossella AVALLONE

Ricercatore Universitario presso: Dipartimento di Scienze della Vita sede ex-Scienze Biomediche


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Pubblicazioni

2020 - BV-2 Microglial Cells Respond to Rotenone Toxic Insult by Modifying Pregnenolone, 5alpha-Dihydroprogesterone and Pregnanolone Level [Articolo su rivista]
Avallone, Rossella; Lucchi, Chiara; Puja, Giulia; Codeluppi, Alessandro; Filaferro, Monica; Vitale, Giovanni; Rustichelli, Cecilia; Biagini, Giuseppe
abstract

Neuroinflammation, whose distinctive sign is the activation of microglia, is supposed to play a key role in the development and progression of neurodegenerative diseases. The aim of this investigation was to determine levels of neurosteroids produced by resting and injured BV-2 microglial cells. BV-2 cells were exposed to increasing concentrations of rotenone to progressively reduce their viability by increasing reactive oxygen species (ROS) production. BV-2 cell viability was significantly reduced 24, 48 and 72 h after rotenone (50–1000 nM) exposure. Concomitantly, rotenone (50–100 nM) determined a dose-independent augmentation of ROS production. Then, BV-2 cells were exposed to a single, threshold dose of rotenone (75 nM) to evaluate the overtime release of neurosteroids. In particular, pregnenolone, pregnenolone sulfate, progesterone, 5alpha-dihydroprogesterone (5-DHP), allopregnanolone, and pregnanolone, were quantified in the culture medium by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. BV-2 cells synthesized all the investigated neurosteroids and, after exposure to rotenone, 5DHP and pregnanolone production was remarkably increased. In conclusion, we found that BV-2 cells not only synthesize several neurosteroids, but further increase this production following oxidative damage. Pregnanolone and 5alpha-DHP may play a role in modifying the progression of neuroinflammation in neurodegenerative diseases.


2020 - Protective Effects of Borago officinalis (Borago) on Cold Restraint Stress-Induced Gastric Ulcers in Rats: A Pilot Study [Articolo su rivista]
Di Cerbo, Alessandro; Carnevale, Gianluca; Avallone, Rossella; Zavatti, Manuela; Corsi, Lorenzo
abstract

Stress is a typical body's natural defense to a generic physical or psychic change. A specific linking mechanism between ulcer onset and psycho-physical stress prolonged exposure has been reported. We decided to investigate the possible effects of Borago officinalis L. (Borago) in preventing physical (stress)-induced gastric ulcers in a rat model. Eighty male Sprague-Dawley rats were randomly divided into 16 groups, pretreated with a control solution, omeprazole (20 mg/kg), Borago methanolic extract (25, 50, 100, 250, and 500 mg/kg), Borago organic extract (50, 100, 250, and 500 mg/kg), Borago aqueous extract (5, 10, 20, 30, and 40 mg/kg), and D(-)-2-Amino-5-phosphonovaleric acid (AP5) (25 mg/kg) and kept in stressful conditions such as water immersion and restraint-induced stress ulcers. The animals were sacrificed and their stomach scored for the severity and the number of gastric ulcers. Methanolic extract (500 mg/kg) significantly reduced both ulcer parameters (***p < 0.001 and **p < 0.01, respectively). Aqueous and organic extract significantly decreased severity score at 5 and 10 mg/kg (**p < 0.01 and ***p < 0.001, respectively), and at 250 and 500 mg/kg (***p < 0.001), respectively, while gastric ulcers' resulted number significantly reduced only at 10 mg/kg (*p < 0.05) and at 500 mg/kg (**p < 0.01), respectively. On the other hand, aqueous extract significantly increased the mucosal gastric content of cAMP (*p < 0.05) and NR2A and NR2B subunits (*p < 0.05 and **p < 0.01, respectively) at 5 mg/kg. Organic extract showed also a significant cytotoxic effect at 500 and 1,000 mg/kg with a 3T3 cell viability reduction of 43.6% (**p < 0.01) and 92.1% (***p < 0.001), respectively. Borago aqueous extract at 10 mg/kg could be considered as a potential protective agent against stress-induced ulcers, and it is reasonable to possibly ascribe such protective activity to a modulation of the NR2A and NR2B subunit expression.


2019 - Omega-3 Fatty Acids and Neurodegenerative Diseases: New Evidence in Clinical Trials [Articolo su rivista]
AVALLONE, Rossella; VITALE, Giovanni; BERTOLOTTI, Marco
abstract

A nutritional approach could be a promising strategy to prevent or slow the progression of neurodegenerative diseases such as Parkinson’s and Alzheimer’s disease, since there is no eectivetherapy for these diseases so far. The beneficial effects of omega-3 fatty acids are now well established by a plethora of studies through their involvement in multiple biochemical functions, including synthesis of anti-inflammatory mediators, cell membrane fluidity, intracellular signaling, and gene expression. This systematic review will consider epidemiological studies and clinical trials that assessed the impact of supplementation or dietary intake of omega-3 polyunsaturated fatty acids on neurodegenerative diseases such as Parkinson’s and Alzheimer’s diseases. Indeed, treatment with omega-3 fatty acids, being safe and well tolerated, represents a valuable and biologically plausible tool in the management of neurodegenerative diseases in their early stages.


2019 - Structure Model and Toxicity of the Product of Biodissolution of Chrysotile Asbestos in the Lungs [Articolo su rivista]
Gualtieri, A. F.; Lusvardi, G.; Pedone, A.; Di Giuseppe, D.; Zoboli, A.; Mucci, A.; Zambon, A.; Filaferro, M.; Vitale, G.; Benassi, M.; Avallone, R.; Pasquali, L.; Lassinantti Gualtieri, M.
abstract

Asbestos is a commercial term indicating six natural silicates with asbestiform crystal habit. Of these, five are double-chain silicates (amphibole) and one is a layer silicate (serpentine asbestos or chrysotile). Although all species are classified as human carcinogens, their degree of toxicity is still a matter of debate. Amphibole asbestos species are biopersistent in the human lungs and exert their chronic toxic action for decades, whereas chrysotile is not biopersistent and transforms into an amorphous silica structure prone to chemical/physical clearance when exposed to the acidic environment created by the alveolar macrophages. There is evidence in the literature of the toxicity of chrysotile, but its limited biopersistence is thought to explain the difference in toxicity with respect to amphibole asbestos. To date, no comprehensive model describing the toxic action of chrysotile in the lungs is available, as the structure and toxic action of the product formed by the biodissolution of chrysotile are unknown. This work is aimed at fulfilling this gap and explaining the toxic action in terms of structural, chemical, and physical properties. We show that chrysotile's fibrous structure induces cellular damage, mainly through physical interactions. Based on our previous work and novel findings, we propose the following toxicity model: inhaled chrysotile fibers exert their toxicity in the alveolar space by physical and biochemical action. The fibers are soon leached by the intracellular acid environment into a product with residual toxicity, and the dissolution process liberates toxic metals in the intracellular and extracellular environment.


2015 - Idiopathic Recurrent Stupor, Still an Unsolved Issue [Articolo su rivista]
Fois, Chiara; Murgia, Bastianino; Avallone, Rossella; Murrighile, Raffaela Maria; Sechi, Gianpietro
abstract

We report a case of unexplained recurrent stupor for which, given the current knowledge and the lack of easily available diagnostic tools, we were unable to reach a definite diagnosis. In the patient the syndrome was associated with an ominous prognosis. Worldwide accepted and easily available proceeding method is needed to manage patients with recurrent stupor for which a toxic origin may be supposed


2015 - Modulatory effects of neurosteroids and thyroid hormones on GABA-evoked currents in cultured dorsal root ganglion cells [Abstract in Atti di Convegno]
Puja, Giulia; Ravegnani, Laura; Ravazzini, Federica; Avallone, Rossella; Bardoni, Rita
abstract

Neurosteroids (NSs) and Thyroid hormones (THs, T3, triiodothyronine and T4, thyroxine) are important endogenous modulators of GABAA receptor (GABAAR) function (Puia and Losi, 2011). The involvement of NSs in several physiological and pathological processes has been largely acknowledged, among them pain transmission. Several studies revealed the antinociceptive properties of some NSs and demonstrated that they can induce a potent peripheral analgesia via a direct GABAAR allosteric modulation (Poisbeau et al., 2014). Very little is known instead of how THs affect synaptic transmission in structures devoted to pain transmission. Pain sensitivity is related to the thyroid status, indeed hyperthyroidism confers greater sensitivity to thermal noxious stimuli (Edmondson et al., 1990) and alters the nociceptive responses in rats (Bruno et al., 2005). Dorsal root ganglion (DRG) cells are primary sensory neurons playing important roles in pain transmission between periphery and CNS. By using the patch clamp technique in the whole cell configuration we analyzed the effect of THs and of some NSs (Pregnenolone Sulfate, PS, and Allopregnenolone, ALLO) on GABA-evoked currents in rat DRG cells grown in primary cultures T3, T4 and PS (from 500 nM to 50 µM) reduce GABA-evoked currents with an IC50 of 0.8±0.3μM for T3 (eff max =-42±9%), of 1.4±0.7 μM for T4 (eff max =-41±6%) and of 4.3±1.2μM (eff max =-60±8%) for PS. ALLO potentiates GABA-evoked current in DRG neurons with an IC50 of 1.3±0.8 μM and a maximal effect of 110±20%. To investigate the mechanism of action of THs and PS we applied increasing concentrations of GABA (5, 10 and 50 µM) to the same concentration of modulator (10 µM). The effect of T3, T4 and PS was not dependent on the GABA concentration used suggesting that they act in a non-competitive way. The modulatory activity of THs and PS on sIPSCs amplitude and frequency measured in lamina II neurons of acutely dissociated spinal cord slices was also investigated. In conclusion, since DRG neurosteroidogenesis is a physiologically relevant process (Schaeffer et al., 2010), our findings suggest that NSs modulation of GABAAR in this cells could play an important role in pain transmission from periphery to spinal cord. Furthermore the decreased GABAAR activity induced by T3 and T4 result in a reduced inhibitory neurotransmission that could contribute to the increased pain sensitivity detected in hyperthyroid animals.


2015 - Supplementation of omega 3 fatty acids improves oxidative stress in activated BV2 microglial cell line [Articolo su rivista]
Corsi, Lorenzo; MOMO DONGMO, BENDJ EDITH; Avallone, Rossella
abstract

Abstract Many reports have shown promising beneficial effects of long-chain polyunsaturated fatty acids (L-PUFAs) of the omega 3 series in several brain diseases. In the present study, we tested the hypothesis that omega 3 fatty acids supplement reduced pro-inflammatory functions in vitro and in vivo. We demonstrated that a supplement rich in PUFAs (SRP) increased cell viability in a dose-dependent manner suggesting its protective role against lipopolysaccharide (LPS)-induced cell death in BV2 microglial cell line. In the same cultures, the supplement rich in PUFAs reduced the reactive oxygen species (ROS) and nitric oxide (NO) production. A most prominent target for ROS management is the family of peroxisome proliferator-activated receptors (PPARs). The co-treatment with SRP and LPS increased significantly the nuclear immunoreactivity of PPAR-γwhen compared the LPS treatment alone. Moreover, the chronic administration of the SRP in rats, increased the immunoreactivity of the PPAR-γ1 protein confirming its potential neuroprotective effect.


2012 - Polyunsaturated fatty acid levels in rat tissues after chronic treatment with dietetic oils [Articolo su rivista]
Rustichelli, Cecilia; Avallone, Rossella; Campioli, Enrico; Braghiroli, Daniela; Baraldi, Mario
abstract

BACKGROUND: The essential fatty acids can be helpful in the prevention of several pathologies. The purpose of this study was to quantify the major n-3 and n-6 fatty acids in tissues of rats fed with flaxseed oil and with a dietetic oil in order to evaluate how their chronic supplementation could influence the correspondent in vivo levels and to study the effectiveness of the dietetic oil compared to flaxseed oil. RESULTS: Fatty acids were successfully extracted from biological samples, subjected to derivatization procedure and analysed by high-performance liquid chromatography with UV detection under gradient elution mode. The developed method showed good linearity, precision and accuracy with recoveries ranging from 89% to 92%. Animals treated with flaxseed and dietetic oils showed enhanced levels of n-3 fatty acids compared to control groups, with a significant higher levels of EPA and DHA in the brain and in the adipose tissue of the dietetic groups compared to the flaxseed groups. CONCLUSION: The obtained data underline that the tested oils can effectively enhance the tissue levels of n-3 fatty acids and therefore they could be successfully used in the dietetic treatment of lipid related diseases.


2012 - n-3 Dietary supplementation and lipid metabolism: Differences between vegetable- and fish-derived oils [Articolo su rivista]
Campioli, Enrico; Rustichelli, Cecilia; Avallone, Rossella
abstract

The effect of flaxseed oil rich in linolenic acid (ALA), and a mixed oil (flaxseed oil and fish oil) rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on the lipid clearance and peroxisome proliferator activated receptors (PPARs) in liver and adipose tissue of rats fed for 30 days with the two oils was evaluated. The results showed that after treatment with the mixed oil the hematic triacylglycerol content was significantly decreased compared to control animals. Regarding the tissue distribution of the major omega-3 fatty acids, both oils were able to increase ALA, EPA, docosapentaenoic acid (DPA) in liver and adipose tissue; and DHA solely in the adipose tissue. Finally the treatment with either flaxseed or mixed oil increased hepatic PPAR-γ expression but only the mixed oil enhanced the hepatic expression of PPAR-α. No effect on adipose tissue PPAR-γ expression was observed with both oils’ treatment.


2011 - Morphological and Receptorial Changes in the Epididymal Adipose Tissue of Rats Subjected to a Stressful Stimulus. [Articolo su rivista]
Campioli, Enrico; Carnevale, Gianluca; Avallone, Rossella; Guerra, Deanna; Baraldi, Mario
abstract

Obesity is nowadays related to other pathological conditions such as inflammation, insulin resistance, and diabetes, but little is known about the relationship between psychological stress and adipocytes. We decided to study the expression of the translocator protein (TSPO) 18-kDa, peroxisome proliferator-activated receptor-γ (PPAR-γ), mitochondrial uncoupling protein-1 (UCP-1), and adipocyte morphology in the adipose tissue of rats subjected to stress conditions. In our model of stress, rats fasted for 24 h were placed in a restraint cage and then immersed vertically to the level of the xiphoid process in a water bath at 23 °C for 7 h. After that period, we removed the epididymal adipose tissues for the subsequent analysis. The optical and electron microscopy revealed that adipocytes of control rats formed a continuous epithelial-like cell layer; on the contrary in the adipocytes of stressed rats some cells have merged together and the number of vessels formed seems to increase. Stressed adipocytes presented unilocular cells with numerous mitochondria with a morphology ranging between that of brown adipose tissue (BAT) and white adipose tissue (WAT). Interestingly, when we investigated the subcellular distribution of UCP-1 by immunogold electron microscopy, the adipose tissue of stressed rats was positive for UCP-1. From the immunoblot analysis with anti-PPAR-γ antibody, we observed an increased expression of PPAR-γ in the adipocytes of stressed group compared with control group (P < 0.05). Stress induced the expression of TSPO 18-kDa receptor (B(max) = 106.45 ± 5.87 fmol/mg proteins), which is undetectable by saturation-binding assay with [(3)H]PK 11195 in the control group.


2010 - EFFETTI DELLA SUPPLEMENTAZIONE DIETETICA CON OLIO DI LINO SULLA SINTESI DELLE KINURENINE IN ANIMALI CON IL MORBO DI HUNTINGTON. [Abstract in Atti di Convegno]
Avallone, Rossella; Rustichelli, Cecilia; M. F., Notarangelo; Campioli, Enrico
abstract

E’ stato ipotizzato che il pathway delle kinurenine abbia un ruolo importante nel morbo di Huntington, una patologia neurodegenerativa causata dall’espansione del tratto di poliglutamina della proteina huntingtina. Durante la fase iniziale della patologia, i livelli cerebrali di due metaboliti della via delle kinurenine, l’acido chinolinico e il suo precursore la 3-idrossi chinurenina sono risultati elevati. Inoltre in uno studio clinico è stato dimostrato che il trattamento con acido eicosapentenoico aveva effetti benefici sulla corea e sui sintomi motori.Lo scopo della ricerca è stato quello di valutare gli effetti della supplementazione di acidi grassi della serie omega 3 nell’Huntington studiando in particolare il pathway delle kinurenine.Topi transgenici con morbo di Huntington (R6/2 - HD) e topi wild-type (WT) sono stati alimentati cronicamente per un mese con olio di oliva (utilizzato come controllo) e con olio dietetico costituito da un’associazione di olio di lino, ricco in acido linolenico (ALA), olio di girasole e olio di pesce ricco in acido eicosapentenoico (EPA) e docosaesanoico (DHA).Nel cervello è stato determinato un significativo aumento (p<0,05) dei livelli di ALA, EPA e DHA nel topo WT trattato con olio dietetico rispetto al corrispondente trattato col olio di oliva: lo stesso andamento è stato determinato nel topo HD trattato con olio dietetico con un aumento significativo in EPA e DHA (p<0,01). Non è stata determinata nessuna variazione nel livelli di acido linoleico e di acido arachidonico né nei topi WT né in quelli HD. Nei topi HD è stato determinato un minore livello di DHA (p<0,01) e un maggiore livello di acido arachidonico (p<0,01) rispetto ai topi WT. Il profilo degli acidi grassi nel tessuto adiposo riflette quello cerebrale mentre nel fegato è stato misurato un aumento significativo di EPA e DHA sia nel topo WT che nel topo HD dopo trattamento cronico con olio dietetico. EPA e DHA sono maggiori nel topo HD (p<0,001) al WT (p<0,05).Per quanto riguarda gli esperimenti sul pathway delle kinurenine, l’3H-kinurenina è stata iniettata nello striato e si sono misurati i metaboliti prodotti mediante analisi HPLC. Non è stata determinata alcuna variazione nelle concentrazioni di acido chinurenico, acido chinolinico e 3-idrossi chinurenina.Concludendo il trattamento dietetico con olio arricchito in acidi grassi delle serie omega 3 ha mostrato benefici nella patologia di Huntington.


2010 - Evidence that isopropylthioxanthone (ITX) is devoid of anxiolytic and sedative effect [Articolo su rivista]
Campioli, Enrico; Zavatti, Manuela; Avallone, Rossella; Puja, Giulia; Losi, Gabriele; Baraldi, Mario
abstract

Isopropylthioxanthone (ITX) is a well-known photo-initiator in ultraviolet light-cured inks frequently used in milk packaging materials, yoghurt, ready-to-feed infant formula, and other drinks. Traces of ITX have beenfound in milk and, as a consequence, there was considerable interest in studying the biological activity of this molecule and its potential hazard for the human health. Although the ITX genotoxic effects have been excluded 10 by the European Food Safety Authority (EFSA), the US Environmental Protection Agency (USEPA) is still examining its possible toxic potential depending on a dose–effect ratio. Little is known about the ITX activity on the function of the central nervous system and cerebral neurotransmitters. Using behavioural, biochemical, andelectrophysiological tests, the authors have found that: (1) ITX did not exert an in vivo anxiolytic or sedativeeffect when administered orally to rats; (2) ITX did not affect the binding characteristics of central and peripheral15 benzodiazepine receptors studied in vitro; and (3) ITX did not influence the ability of GABA to increase thechloride channel permeability studied by patch clamp technique in a single neuron of cultured cerebellargranule cells.


2009 - Alpha-Linolenic acid and Linoleic acid in serum and tissues after flaxseed (Linum usitatissimum) oil in vivo administration [Articolo su rivista]
Avallone, Rossella; Rustichelli, Cecilia; Campioli, Enrico; Notarangelo, Francesca Maria Pia; Braghiroli, Daniela; Baraldi, Mario
abstract

Essential fatty acids can be helpful in the prevention of several pathologies. The bioavailability of acute supplementation of different doses of flaxseed oil was studied by analyzing the level of alpha-linolenic acid (ALA) and linoleic acid (LA) in serum and tissues (adipose, liver) of rats tested at 2, 4, 8 and 16 h after the administration. The amount of flaxseed oil administered at increasing doses corresponded to 1, 2.5 and 5 g ALA/kg of body weight. The corresponding fatty acid methyl esters obtained via direct methylation were quantified using gas chromatography/mass spectrometry. Serum ALA level increased after 1 or 2.5 g/kg. ALA was increased in both adipose and liver tissue 4 h after the administration of 1 g/kg of flaxseed oil. There was no further increase by using a higher oil dosage. LA did not change in serum at the doses used.


2009 - Experimental evidence of the anaphrodisiac activity of Humulus lupulus L. in naïve male rats [Articolo su rivista]
Zanoli, Paola; Zavatti, Manuela; Rivasi, Marianna; Benelli, Augusta; Avallone, Rossella; Baraldi, Mario
abstract

Ethnopharmacological relevance: In the folk medicine Humulus lupulus L. (hops) is mainly recommendedas a mild sedative with antispasmodic and digestive properties. It is also reputed to exert an anaphrodisiaceffect but it is still lacking the experimental evidence of this activity.Aim of the study: To evaluate the influence of Humulus lupulus extract on sexual behavior of both naïveand sexually potent male rats; thereafter to investigate the role of 8-prenylnarigenin (8-PN) in the effectdisplayed by the hop extract.Materials and methods: Sprague–Dawley male rats both naïve and sexually potent were acutely administeredwith the hop extract dosed at 5, 10, 25 and 50 mg/kg. In addition the extractwas administered dailyfor 10 consecutive days at the dose of 0.25 mg/kg/day in sexually potent animals. The pure compound8-PN was acutely administered in naïve rats at the dosages of 5, 12.5 and 25g/kg. All the animals werescreened for their sexual behavior manifestation during the mating test.Results: In naïve rats the acute administration of Humulus lupulus extract at the doses of 25 and 50 mg/kgsignificantly reduced the percentage of mounting and ejaculating animals, in comparison to vehiclecontrols. The other parameters recorded during the mating test were not affected by the hop extract.In sexually potent rats nor the acute neither the repeated administration of the extract modified theircopulatory behavior.The pure compound 8-PN failed to influence male sexual behavior of naïve rats.Conclusion: Humulus lupulus extract exerted an anaphrodisiac effect only in naïve rats by inhibiting theirmounting and ejaculating behavior. The presence of 8-PN in the extract could be only partially involvedin the observed anaphrodisiac effect.


2009 - HPLC analysis of n-3 and n-6 fatty acid levels in rat serum after chronic treatment with dietetic oils [Articolo su rivista]
Rustichelli, Cecilia; Avallone, Rossella; Campioli, Enrico; Braghiroli, Daniela; Parenti, Carlo; Baraldi, Mario
abstract

High-performance liquid chromatography HPLC analysis of the major fatty acids of the n-3 family and n-6 family in rat serum was carried out. To perform sample cleanup, existing techniques of lipid extraction were tested and modified to achieve maximal free fatty acids recovery in a reasonable time. Concerning chromatographic analyses, p-bromophenacyl bromide was used as ultraviolet (UV)-derivatizing agent followed by reversed-phase HPLC/UV under mobile phase gradient conditions. The optimized and validated procedure was then applied to rats fed with flaxseed oil and a combination of flaxseed, fish and sunflower oils, defined as "dietetic oil", in order to evaluate how their chronic supplementation can influence serum levels of n-3 and n-6 fatty acids.


2009 - HPLC determination of fatty acid levels in rats after chronic supplementation of flaxseed oil and dietetic oil. [Abstract in Atti di Convegno]
Rustichelli, Cecilia; Avallone, Rossella; Campioli, Enrico; Braghiroli, Daniela; Baraldi, Mario
abstract

Essential fatty acids (EFAs) belonging to -3 and -6 families play an important role in vivo, supporting the cardiovascular, reproductive, immune and nervous system functions; an inappropriate balance of these EFAs contributes to the development of diseases, while a proper balance helps maintain and even improve health.The purpose of this research was to study the levels of the principal -3 (ALA: n-lino-lenic acid; EPA: eicosapentaenoic acid; DPA: docosapentaenoic acid; DHA: docosa-hexaenoic acid) and -6 (LA: linoleic acid; AA: arachidonic acid) fatty acids in rat serum and tissues (liver, brain, adipose tissue) after a chronic treatment with flaxseed oil and with a dietetic oil, containing flaxseed oil, fish oil and sunflowers oil. The treatment was planned in order to give to rats the same ALA amount.The biological samples were subjected to liquid-liquid extraction, followed by derivatisation with p-bromophenacyl bromide and then analysed by HPLC/UV on a C18 reversed-phase column under gradient mobile phase conditions.Rats treated with flaxseed oil and dietetic oil showed enhanced levels of n-3 fatty acids compared to the correspondent control group. In serum, ALA levels were slightly higher in the dietetic oil group than in the flaxseed oil group; this fact is probably due to an enhanced absorption and also a decreased metabolism of ALA. The EPA, DPA and DHA serum levels, higher in the dietetic than in the flaxseed group, could derive both from ALA through its metabolism and also from EPA and DHA contained in the fish oil. Brain and adipose tissue showed EPA and DHA levels higher in the dietetic than in the flaxseed group, as occurred for serum samples. Concerning liver samples, the treatment with flaxseed or dietetic oil caused an enhancement for ALA, EPA and DPA levels, while no variations were found for DHA compared to control groups.The data obtained underline that the tested oils can enhance the levels of -3 fatty acids and therefore they could be successfully used in the dietetic treatment of lipid related diseases.


2009 - Natural endogenous ligands for benzodiazepine receptors in hepatic encephalopathy. [Articolo su rivista]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; Venturini, I; Baraldi, Claudia; Zeneroli, Maria Luisa
abstract

Benzodiazepines of natural origin (NBZDs) have been found in human blood and brains as well as in medicinal plants and foods. In plasma and brain tissue there are i.e. diazepam and nordiazepam equal to commercial drugs but there are also other benzodiazepine-like compounds termed “endozepines”, which act as agonists at the benzodiazepine receptors of central type (CBR). A synthetic pathway for the production of NBZDs has not yet been found, but it has been suggested that micro-organisms may synthesize molecules with benzodiazepine-like structures. Hence NBZDs could be of both endogenous and exogenous source and be considered as natural anxyolitic and sedative. Interestingly there are also natural compounds, such as the polypeptide Diazepam Binding Inhibitor (DBI) acting as an “inversive agonist” implicated in fair and panic disorders. It has been suggested that NBZDs may play a role in the pathogenesis of hepatic encephalopathy (HE). Multidirectional studies evaluated NBZDs levels (1) in the blood of normal subjects, of cirrhotic with or without HE and in commercial benzodiazepine consumers; (2) in the blood of cirrhotic treated or not with a non-absorbable antibiotic; (3) in several constituents of our diet. In conclusion, NBZDs increase sometime in cirrhotics with or without HE but they reach concentrations not higher than those found in commercial benzodiazepines consumers. Hence NBZDs must be considered as occasional precipitating factor of HE and benzodiazepine antagonists only symptomatic drugs. The finding that NBZDs may be in part synthesized by intestinal bacterial flora and in part constituent of our diet underlines the importance to feed cirrhotic patients with selected food.


2008 - Flaxseed oil: acute and chronic supplementation increases serum and tissue concentrations of omega fatty acids in rats [Abstract in Atti di Convegno]
Campioli, Enrico; Avallone, Rossella; Rustichelli, Cecilia; Notarangelo, Francesca Maria Pia; Braghiroli, Daniela; Baraldi, Mario
abstract

We studied the bioavailability of acute supplementation of scalar doses of flaxseed oil by analysing the level of Linolenic acid (ALA, -3) and Linoleic acid (-6) in serum and tissues (adipose, liver and brain) of rats tested at 2-4-8-16 h after the administration. The amount of flaxseed oil administered by oral rate was 1.9, 4.7, 9.5 mL/kg corresponding to 1, 2.5, 5 g ALA/kg. Two techniques of lipid extraction were investigated to achieve maximal free fatty acids recovery in a reasonably short time. The corresponding fatty acid methyl esters obtained with direct methylation with MeOH/HCl, were quantified by gas chromatography/mass spectrometry (GC/MS) technique. GC-MS analyses were performed on a Gas-Chromatograph Varian 3400 on a HP-INNOWAX column (30 m x 0.25 mm; 0.25 m film thickness). Mass spectra were acquired on a Finnigan MAT SSQ 710A mass spectrometer in the electron impact (EI) mode.Serum ALA levels at 1 g/kg after 2h in the flaxseed oil group increased by 70% from 0.067 ± 0.007 to 0.096 ± 0.008 mg/mL (P<0.001 Anova) whereas no significant increase occurred in the flaxseed oil group at 2.5 g/kg (0.142 ± 0.009) or at 5 g/kg after 2 h (0.140 ± 0.008) when compared with the value obtained after 1 g/kg. A statistically significant increase of ALA was found in adipose tissue and in liver 4 h after the administration of 1 g/kg of ALA whereas higher doses (2.5-5 g/kg) did not produce any significant changes. Concerning linoleic acid (-6) no significant increased concentrations were found in serum at the three doses studied confirming that flaxseed oil is mainly a source of -3 fatty acids.


2008 - La supplementazione di olio di lino aumenta la concentrazione plasmatica e tissutale dell'acido linolenico e cambia l'espressione dei recettori attivanti la proliferazione dei perossisomi (PPAR) [Abstract in Atti di Convegno]
Avallone, Rossella; Rustichelli, Cecilia; Campioli, Enrico; Baraldi, Mario
abstract

E’ stata studiata la biodisponibilità dell’olio di lino dopo supplementazione acuta analizzando i livelli di acido linolenico (ALA), acido linoleico, EPA, DPA e DHA nel siero e nei tessuti (adiposo, epatico e cerebrale) di ratti dopo 2-4-8-16 ore dalla somministrazione. La quantità di olio di lino somministrata è stata corrispondente a 1-2,5-5 g di ALA/kg. Gli esteri metilici degli acidi grassi ottenuti mediante metilazione diretta con MeOH/HCl, sono stati quantificati mediante GC/MS (GC Varian 3400/ Finnigan MAT SSQ 710 A) ad impatto elettronico a –70 eV su colonna capillare HP-INNOWAX (30 m x 0,25 mm DI x 0,25 m).I livelli serici di ALA nel gruppo trattato con olio di lino alla dose di 1 g/kg, dopo 2h aumentano del 70% andando da 0,067 ± 0,007 a 0,096 ± 0,008 mg/mL (P<0.001 Anova, post test Bonferroni) mentre nessuna variazione significativa è stata riscontrata nel gruppo trattato con 2,5 g/kg (0,142 ± 0,009) o con 5 g/kg dopo 2 h (0,140 ± 0,008). Un aumento significativo di ALA è stato determinato nel tessuto adiposo e nel fegato dopo 4 ore dalla somministrazione di 1 g/kg mentre le dosi maggiori (2,5-5 g/kg) non producono nessun cambiamento significativo. Per quanto riguarda l’acido linoleico non è stato determinato nessun aumento significativo confermando che l’olio di lino è soprattutto una fonte di acidi grassi della serie -3.Alla luce dei risultati ottenuti dopo trattamento acuto, si è reso necessario valutare la biodisponibilità dell’olio di lino dopo trattamento cronico di un mese alla dose ottimale di 1 g di ALA/kg. In questo caso la quantificazione degli acidi grassi è stata effettuata mediante HPLC/UV con una colonna a fase inversa C18 previa derivatizzazione con p-bromofenacil bromuro [1]. Si è potuto notare un significativo incremento nel siero e nel fegato dei valori di ALA, EPA e DPA in conformità con studi precedenti [2]. Non ci sono significative variazioni dei livelli serici di colesterolo totale, HDL, LDL mentre si nota una diminuzione significativa della trigliceridemia.E’ stata inoltre valutata mediante tecnica di Western blot l’espressione dei recettori PPAR nelle due isoforme α, e γ rispettivamente nel fegato e nel tessuto adiposo dopo trattamento cronico con olio di lino.Le bande relative a PPAR  e  e -actina sono state quantificate e l’espressione di PPAR è stata valutata come rapporto PPAR- e  /-actina.[1] Metha et al. J. Chromat. B 1998, 719: 9-23[2] Harper C.R. et al. J. Nutr. 2006, 136: 2844-2848.


2008 - Variazione degli acidi grassi W3 e W6 e plasticità del recettore attivante la proliferazione dei perossisomi (PPAR-a) dopo trattamento cronico con olio di lino e olio dietetico nel fegato. [Abstract in Atti di Convegno]
Campioli, Enrico; Avallone, Rossella; Rustichelli, Cecilia; Baraldi, Mario
abstract

Lo scopo del presente lavoro è stato di quantificare l’acido α-linolenico (ALA), eicosapentaenoico (EPA), docosapentaenoico (DPA), docosaesaenoico (DHA), linoleico e arachidonico nel fegato di ratti trattati cronicamente con olio di lino e con un’associazione di oli vegetali e pesce. Si è valutata inoltre, la plasticità dell’isoforma α del recettore PPAR nel fegato.I PPAR sono dei recettori nucleari, eterodimerizzati con il recettore per l’acido 9-cis retinoico [1]. Questi agiscono come fattori di trascrizione per geni coinvolti nel metabolismo. Molti composti si legano con alta affinità ai recettori PPARs, tra questi troviamo gli acidi grassi insaturi a lunga catena (acido linoleico e linolenico), acidi grassi ramificati, acidi grassi ossidati o coniugati, ed infine eicosanoidi, leucotrieni e fibrati.Il tessuto epatico degli animali trattati per un mese con i suddetti oli è stato omogeneizzato con una miscela estraente costituita da ispropanolo/eptano/acido solforico secondo la metodica descritta da Mehta et al. [2]. Dopo aver aggiunto uno standard interno costituito dai 6 acidi grassi da analizzare, i campioni sono stati portati a secco in corrente di azoto e derivatizzati secondo la metodica descritta da Puttmann et al. [3] per ottenere i corrispondenti p-bromofenacil bromuri derivati. I campioni così ottenuti sono stati analizzati mediante HPLC a 35°C, colonna RP-18 endcapped (125 x 4 mm; 5 μm), detector UV a 254 nm.Successivamente il tessuto è stato omogeneizzato in buffer di lisi RIPA, addizionato di inibitori delle proteasi e centrifugato. I campioni così ottenuti sono stati caricati su gel di poliacrilamide bis-tris 4-12% e sottoposti ad analisi di Western blot.Dall’analisi HPLC si è riscontrato per quanto riguarda l’olio di lino un aumento significativo di ALA (p<0,001; one way Anova, Bonferroni’s post test), EPA (p<0,05) e DPA (p<0,05). Per quanto riguarda l’olio dietetico si è avuto un aumento significativo di ALA ed EPA, mentre si è avuto una diminuzione significativa dell’acido linoleico (p< 0,01) e dell’acido arachidonico (p<0,001). I risultati dei Western blot condotti sul tessuto epatico non evidenziano un aumento dell’espressione recettoriale del PPAR-α nei ratti trattati con olio di lino ed olio dietetico.


2007 - DENTIFICATION OF COMPOUNDS OF BIOLOGICAL INTEREST IN TOBACCO EXTRACTS BY LIQUID CHROMATOGRAPHY COUPLED TO MASS SPECTROMETRY [Abstract in Atti di Convegno]
Avallone, Rossella; A., Raffaelli2; J. P., Schaller3; M., Baraldi1
abstract

One of the challenging questions in benzodiazepine (BDZ) research is the identification of endogenous BDZ ligands in the brain, which could modulate -aminobutyric acid neurotrasmission. It has been established that endogenous substances with benzodiazepine-like activity play a role in the pathogenesis of different pathologies like hepatic encephalopathy or idiopathic recurring stupor , Several substances with benzodiazepine-like activity were also found in food and in several officinal plants used in folk medicine as hypnotic or tranquillisers such as and Matricaria chamomilla. The presence of molecules with high affinity for central and peripheral benzodiazepinereceptors was also determined in the pod and leaves of Ceratonia siliqua.The aim of the present work was to determine the possible presence of BDZ-like compounds in leaves of Nicotiana tabacum and to test their ability to bind central or peripheral BDZ receptors.Tobacco leaves have been extracted with methanol and the crude extract was chromatographed at 0.8 ml/min on a LiChrospher 100 RP-18 column (250x4.0 mm; 5 m) equilibrated with 80% water/1% FA and 20% acetonitrile. The sample was analysed using a water/1%FA and acetonitrile gradient at 0.5% from 20 to 58% acetonitrile. 75 fraction were collected, lyophilised and tested for their ability to inhibit [3H]RO 15-1788 specific binding to central BDZ binding site or [3H]PK 11195 specific binding to peripheral BDZ binding site (PBR). The crude extract, as well as the separated HPLC fractions, have been analyzed by HPLC-MS and HPLC-MS-MS in order to identify any substances related to the observed biological activity. Collision Induced Decomposition (CID) spectra allowed to identify some possible compounds, such as steroids and glucosides of different nature. The analysis of the separated fractions showed that these components are spread along several fractions, suggesting a quite complex mixture and high concentration, which tend to affect the chromatographic behaviour of the purification step.Future work will focus on MS-MS scan analysis directed at families of compounds and will combine HPLC-MS-MS to biological activity data in order to identify ions strictly related to the activity.


2007 - Endogenous benzodiazepines and intestinal bacterial flora in normal and pathological conditions [Abstract in Atti di Convegno]
Campioli, Enrico; Avallone, Rossella; A., Ferrieri; M., Baraldi
abstract

Evidence has been provided that benzodiazepine (BZD)-like substances are increased in the brains of animals with experimental HE, as well as in serum and in cerebrospinal fluid of patients with HE due to liver cirrhosis and fulminant hepatic failure. Thus, BDZ-like compounds can be regarded as precipitating factors of hepatic encephalopathy requiring the presence of a cerebral supersensitivity to these compounds. The endogenous biosynthetic pathway for the production of natural benzodiazepines has not yet been found while their presence in food has been already described. Furthermore it has been demonstrated by in vitro experiments that microorganims can synthesise molecules with BDZ structures and by in vivo studies that intestinal bacterial flora contributed by 40% to level of total natural benzodiazepines in serum of cirrhotic patients. The aim of the present study was to: 1) confirm the partial contribution of intestine to the level of circulating natural BDZ in commercial BDZ free patients, 2) to study the influence of the chronic administration of a symbiotic compound (Zyrfos-Alfa Wassermann) on the serum concentration of BDZ-like compounds in rats.The serum samples obtained by acid precipitation was chromatographed at 0.8 ml/min on a LiChrospher 100 RP-18 column (250x4.0 mm; 5 m) equilibrated with 80% water/0.1% TFA and 20% acetonitrile. The sample was analysed using a water/0.1% TFA and acetonitrile gradient at 0.5% from 20 to 58% acetonitrile. 75 fraction were collected, lyophilised and tested for their ability to inhibit [3H]RO 15-1788 specific binding to central BDZ binding site.The concentration of BDZ-like substances in portal vein of cirrhotic patients (n=10) reach levels of 1024.24 ± 22.78 ng diazepam equivalent (DE)/mL in comparison with the circulating value of 162.44 ± 10.89 ng DE/mL. These data further demonstrate that there is a intestinal source of the above mentioned compounds. As far as regard the influence of the symbiotic which is composed by probiotic, prebiotic and B vitamins, herein we report that after 14 days of chronic administration the level of BDZ-like compounds decreased from level of 173.5 ± 43.76 ng DE/mL present in controls to level under the detection limit (0.5 ng DE/mL).The reported experiments and data seem to further demonstrate that the intestinal bacterial flora contribute to the amount of circulating natural BDZs both in normal and pathological conditions. Moreover it might be possible to modulate the level of these compounds by the administration of dietary supplements.


2007 - Evidence that the β-acid fraction of hops reduces central GABAergic neurotransmission [Articolo su rivista]
Zanoli, Paola; Zavatti, Manuela; Rivasi, Marianna; F., Brusiani; Losi, Gabriele; Puja, Giulia; Avallone, Rossella; Baraldi, Mario
abstract

Humulus lupulus (hops) is traditionally used as a tranquilizing herbal remedy. Here, we investigated the in vivo and in vitro effect of hop β-acids on central nervous system function. Oral administration of β-acids (5–10 mg/kg) in rats produced an increased exploratory activity in the open field, a reduction in the pentobarbital hypnotic activity and a worsening of picrotoxin-induced seizures. When dosed at 10 mg/kg, β-acids increased, in the elevated plus maze, open arm entries reducing in parallel those in closed arms. In the forced swimming test, we observed a reduction in the immobility time that could suggest an antidepressant-like activity. Electrophysiological studies performed on cerebellar granule cells in culture showed that the β-acids fraction decreased GABA-evoked current in a dose-dependent way. The effect was not inhibited by the benzodiazepine antagonist Ro 15-1788. Benzodiazepine receptors involvement was also excluded by [3H]-Ro 15-1788 binding assay. In conclusion, the behavioral effects of β-acids fraction could be explained by a reduction in the GABAergic activity although we cannot rule out the involvement of other neurotransmitter systems


2007 - Flaxseed oil supplementation increases plasma and tissue concentrations of W3 fatty acids in rats. [Abstract in Atti di Convegno]
Notarangelo, Francesca Maria Pia; Avallone, Rossella; Rustichelli, Cecilia; Campioli, Enrico; Braghiroli, Daniela; Baraldi, Mario
abstract

-Linolenic acid (ALA) is a major dietary (-3) fatty acid. The essential fatty acids must be absorbed by food intake and play a very important role in the coagulation (inhibition of platelets aggregation) and in the inflammatory reaction (anti-inflammatory effects). In cardiovascular diseases, particularly in coronary diseases, studies demonstrated a decreased mortality in populations who eat an omega-3 rich diet or who take an omega-3 supplement.We studied the bioavailability of acute supplementation of scalar doses of flaxseed oil (Organic Oils–Perugia) by analysing the level of ALA (-3) and Linoleic acid (-6) in serum and tissues (adipose, liver and brain) of rats tested at 2-4-8-16 h after the administration. The amount of flaxseed oil administered by oral rate was 1.9, 4.7, 9.5 mL/kg corresponding to 1, 2.5, 5 g ALA/kg. Two techniques of lipid extraction were investigated to achieve maximal free fatty acids recovery in a reasonably short time. The corresponding fatty acid methyl esters obtained with direct methylation with MeOH/HCl, were quantified by gas chromatography/mass spectrometry (GC/MS) technique. GC-MS analyses were performed on a Gas-Chromatograph Varian 3400 on a HP-INNOWAX column (30 m x 0.25 mm; 0.25 m film thickness). Mass spectra were acquired on a Finnigan MAT SSQ 710A mass spectrometer in the electron impact (EI) mode with an ionization energy of 70 eV; the ion source temperature was 250°C, the filament current was 200 A, the conversion dynode power was –15.0 kV and electron multiplier voltage was 1500 V.Serum ALA levels at 1 g/kg after 2h in the flaxseed oil group (n=25) increased by 70% from 0.067 ± 0.007 to 0.096 ± 0.082 mg/mL (P<0.001 Anova) whereas no significant increase occurred in the flaxseed oil group at 2.5 g/kg (0.142 ± 0.071) or at 5 g/kg after 2 h (0.140 ± 0.106) when compared with the value obtained with 1 g/kg. ALA (1g/kg) significantly increased after 4 h in adipose tissue and in liver but also in this case at higher doses (2.5-5 g/kg) the concentration wasn’t increased. Concerning linoleic acid (-6) no significant increased concentrations were found in serum at the three doses studied confirming that flaxseed oil is a source of -3 fatty acids. These data suggested that there is a limiting step in the adsorption of these fatty acids and that there is no advantage to take more than 1 g/kg of ALA supplementation.


2007 - HPLC analysis of essential fatty acids in rat plasma [Abstract in Atti di Convegno]
Rustichelli, Cecilia; Avallone, Rossella; Notarangelo, Francesca Maria Pia; Campioli, Enrico; Braghiroli, Daniela; Baraldi, Mario
abstract

Linoleic acid (n-6 family) and -Linolenic acid (n-3 family) are two polyunsaturated fatty acids (PUFAs) that are known as essential fatty acids and must be provided by diet.They play an important role in coagulation and inflammatory reactions; concerning cardiovascular diseases studies demonstrated a decreased mortality in populations who eat an omega-3 rich diet or who take an omega-3 supplement.This paper deals with HPLC analysis of linoleic acid and linolenic acid in rat plasma in order to investigate the effect of flaxseed oil supplementation on plasma levels of these essential fatty acids.HPLC procedures are recommended for accurately quantifying of thermally labile moieties like polyunsaturated fatty acids; nevertheless a derivatization step is required as these analytes are lacking both in chromophore and fluorophore group and cannot therefore be detected at sufficiently low concentrations.In this work, p-bromophenacyl bromide is used as UV-derivatizing agent followed by HPLC/UV on C18 reversed phase column. The mobile phase gradient was studied in order to obtain a complete separation for the anaytes and the internal standard from matrix components.Concerning plasma sample clean-up, existing techniques of lipid extraction were tested and modified to achieve maximal free fatty acids recovery in a reasonable time.Studies on rats fed with flaxseed oil are currently in progress in order to evaluate how the chronic supplementation can increase plasma levels of n-3 and n-6 fatty acids.


2006 - Analisi GC-EI-MS di acidi grassi omega-3 e omega-6 in matrici complesse [Abstract in Atti di Convegno]
Avallone, Rossella; Braghiroli, Daniela; Rustichelli, Cecilia; Manzini, Daniela; Campioli, Enrico; Baraldi, Mario
abstract

Lo scopo del presente lavoro è stato quello di sviluppare un metodo semplice e rapido per l'analisi degli acidi grassi nel siero e nel tessuto dopo somministrazione acuta o cronica di olio di semi di lino.


2006 - Evidence that natural benzodiazepine-like compounds increase during spontaneous labour [Articolo su rivista]
Facchinetti, Fabio; Avallone, Rossella; Modugno, Giuseppe; Baraldi, Mario
abstract

Natural benzodiazepine-like compounds (NBDZ) are present in the blood of normal people free of commercial benzodiazepine medication. In this work, we evaluated the Levels of NBDZ in maternal/foetal serum during delivery after spontaneous tabour (VD) or caesarean section (CS). For both the VD (n=11) and the CS (n=11) groups (VD+CS=22), three blood samples were collected at three different times: the first was collected three days before tabour, the second immediately after delivery or at fetal abdominal extraction and the third one was obtained at second day post-partum. NBDZ were measured by radioreceptor binding assay after HPLC extraction and purification white cortisol was measured through radioimmunoassay. In the VD group, a significant increase of NBDZ levels occurred at tabour in comparison with the levels found in pre- and post-partum periods. By the contrary, no differences in NBDZ Levels were found in the CS group at the three different times. The levels of cortisol in the VD group were found to be higher at tabour than that determined at pre- and post-partum. Again no significant changes were found in the CS group. These findings suggest for the first time that tabour is associated with a marked increase of NBDZ which could be envisaged as a stress-retated event.


2005 - 2,1,3-Benzothiadiazine derivatives: synthesis and screening versus PDE4 enzyme [Articolo su rivista]
Tait, Annalisa; Luppi, Amedeo; Avallone, Rossella; Baraldi, Mario
abstract

A series of N-1,3 disubstituted 2,1,3-benzothiadiazine derivatives (BTDs) were synthesized and evaluated for their inhibitory activity versus enzymatic isoform PDE4 extracted from U937 cell line. Some of the tested compounds showed a high PDE4 inhibitory activity at 100 μM and the IC50 value of the most interesting terms were evaluated. The structure–activity relationships of these compounds showed that the 3,5-di-tert-butyl-4-hydroxybenzyl moiety at N-1 position is important to obtain activity at micromolar level as previously reported. For the same compounds the antioxidant activity were evaluated highlighting 14 as the most significative one. The introduction of other bulky substituents in N-1 position is detrimental


2005 - Endozepines in recurrent stupor [Articolo su rivista]
P., Cortelli; Avallone, Rossella; Baraldi, Mario; Zeneroli, Maria Luisa; J., Mandrioli; Corsi, Lorenzo; R., Riva; P., Tuniper; E., Lugaresi; A., Baruzzi; P., Montagna
abstract

Stupor is a condition from which the subject can be aroused only by vigorous stimuli. Most patients with stupor have a diffuse organic cerebral dysfunction. Rarely stupor is recurrent and no specific causes can be found. Patients with idiopathic recurrent stupor were awakened by i.v. administration of an antagonist (flumazenil) of the benzodiazepine recognition site located in the GABA(A) receptor. Since no exogenous benzodiazepines were detected in plasma and cerebrospinal fluid by high performance liquid chromatography, an excess of endogenous benzodiazepine-like compounds (endozepines) was proposed as the cause of stupor. The existence of endozepines, their widespread distribution in the CNS and their involvement in hepatic encephalopathy are established. However, the origin of these compounds, how biosynthesis occurs and the mechanisms and causes through which they alter brain functions are poorly understood. The fact that a number of synthetic benzodiazepines are difficult to detect using conventional techniques and the discovery that some cases of recurrent stupor were caused by fraudulent administration of lorazepam question whether the concept of endozepine recurrent stupor can be sustained. This review summarizes the state of endozepine physiology and pharmacology and the clinical syndromes attributed to their involvement. A diagnostic work-up to define endozepine-induced recurrent stupor is suggested.


2005 - Evaluation of rifaximin, placebo and lactulose in reducing the levels of benzodiazepine-like compounds in patients with liver cirrhosis: a pilot study [Articolo su rivista]
I., Venturini; A., Ferrieri; Farina, Franco; Cosenza, Francesca; Avallone, Rossella; Corsi, Lorenzo; Baraldi, Mario; Zeneroli, Maria Luisa
abstract

Benzodiazepine-like compounds (BZDs), either taken with the diet or synthesized by intestinal bacterial flora, may represent a precipitating factor for hepatic encephalopathy (HE) in cirrhotic patients. We evaluated whether a diet and/or treatment with rifaximin or lactulose can reduce serum concentrations of BZDs in 18 cirrhotic patients without HE. Patients were given a standard diet for 7 days to keep the dietary intake of BZDs constant and were then randomized to a 7-day treatment with rifaximin 1,200 mg/day, lactulose 10-20 g three times daily, or placebo. Blood samples were collected at enrollment, at the end of the diet and drug treatment periods, and 7 days after the drug was stopped (follow-up). Serum concentrations of BZDs were measured by a radioligand binding technique after high-performance liquid chromatography extraction and purification and were expressed as diazepam equivalents (DE). No change in serum BZD concentrations was observed during the diet, while a statistically significant decrease from 105.6 +/- 66.5 to 63.5 +/- 49.5 pmol DE/ml was achieved in rifaximin-treated patients (p < 0.05) but not in patients treated with lactulose or placebo. During the followup, serum BZD concentrations returned to 104.5 +/- 74.0 pmol DE/ml in rifaximin-treated patients (p < 0.05 vs. end-treatment values), while no significant change was observed in the lactulose- and placebo-treated patients. These data indicate that control of bacterial flora with cyclic administration of rifaximin plays a pivotal role in avoiding increased plasma concentrations of BZDs, which represent a precipitating factor for HE inpatients with severe liver disease.


2005 - Management of hepatic encephalopathy: Role of rifaximin [Articolo su rivista]
Zeneroli, Maria Luisa; Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Baraldi, Claudia; Baraldi, Mario
abstract

Hepatic encephalopathy (HE) is a neuropsychiatric syndrome, which develops in patients with acute or chronic liver failure. It is widely accepted to be due to impairment of hepatic clearance of toxic products from the gut such as ammonia. Accumulation of ammonia induces a glutamate neurotoxicity leading to an increased tone of the gamma-aminobutyric acid A (GABA-A) receptor system in the brain which results in HE. Factors either increasing the ammonia levels (protein load, constipation, sepsis, or gastrointestinal bleeding) or potentiating the functional activity of the GABAergic system [natural benzodiazepine-like compounds (NBZDs) or exogenous benzodiazepines] may act as precipitating factors of HE. NBZDs are present in trace amounts in the blood of normal subjects and have been found to be increased in the blood of patients with liver cirrhosis, with or without HE. These compounds may derive either from the diet since they have been found in plants, vegetables and animals or from gut bacteria. The observation that intestinal bacterial flora is involved in the production of both primary agent of HE (ammonia) and precipitating factors (NBZDs) suggests that the use of nonabsorbable antibiotics such as rifaximin may be useful in preventing episodes of HE in patients with liver cirrhosis. Copyright


2005 - Nuclear location-dependent role of peripheral benzodiazepine receptor (PBR) in hepatic tumoral cell lines proliferation [Articolo su rivista]
Corsi, Lorenzo; Geminiani, Elisa; Avallone, Rossella; Baraldi, Mario
abstract

PBR is involved in numerous biological functions, including steroid biosynthesis, mitochondrial oxidative phosphorylation and cell proliferation. The presence of PBR at the perinuclear/nuclear subcellular level has been demonstrated in aggressive breast cancer cell lines and human glioma cells where it seems to be involved in cell proliferation. In our study we investigated the presence of perinuclear/nuclear PBR in different hepatic tumor cell lines with regard to binding to [H-3] PK 11195 and protein analysis. The results obtained by saturation binding experiments and scatchard analysis of perinuclear/nuclear PBR density in parallel with the results on the growth curves of the cell lines tested, indicate that the perinuclear/nuclear PBR density correlates inversely with cell doubling time. Moreover, the cell line with high perinuclear/nuclear PBR proliferated in response to PBR ligand, whereas that with low perinuclear/nuclear PBR did not. Our results reinforce the idea that the subcellular localisation of PBR defines its function and that this receptor could be a possible target for new strategies against cancer.


2005 - Nuclear location-dependent role of peripheral benzodiazepine receptor (PBR) in hepatic tumoral cell lines proliferation [Abstract in Atti di Convegno]
Geminiani, Elisa; Corsi, Lorenzo; Avallone, Rossella; Baraldi, Mario
abstract

The peripharal typo benzodiazepine receptor (PBR) was described as another class of binding sites for benzodiazepines distinct from the central benzodiazepine receptor (CBR). In our study we investigated the presence of nuclear PBR in different hepatic tumor cell lines with ragards to binding to [3H]PK11195 and protein analysis. The results obtained indicated that the nuclear PBR density correlates inversely with cell doubling time.


2005 - Pharmacological characterisation of beta-acids fraction of hops [Abstract in Atti di Convegno]
Zavatti, Manuela; Zanoli, Paola; Rivasi, Marianna; Avallone, Rossella; Baraldi, Mario
abstract

Humulus lupulus (hops) is traditionally used as a tranquilizing herbal remedy. Here, we investigated the in vivo and in vitro effect of hop -acids on central nervous system function. Oral administration of -acids (5–10 mg/kg) in rats produced an increased exploratory activity in the open field, a reduction in the pentobarbital hypnotic activity and a worsening of picrotoxin-induced seizures. When dosed at 10 mg/kg, -acids increased, in the elevated plus maze, open arm entries reducing in parallel those in closed arms. In the forced swimming test, we observed a reduction in the immobility time that could suggest an antidepressant-like activity. Electrophysiological studies performed on cerebellar granule cells in culture showed that the -acids fraction decreased GABA-evoked current in a dose-dependent way. The effect was not inhibited by the benzodiazepine antagonist Ro 15-1788. Benzodiazepine receptors involvement was also excluded by [3H]-Ro 15-1788 binding assay. In conclusion, the behavioraleffects of -acids fraction could be explained by a reduction in the GABAergic activity although we cannot rule out the involvement of other neurotransmitter systems.


2004 - Peripheral benzodiazepine receptors in potatoes (Solanum tuberosum) [Articolo su rivista]
Corsi, Lorenzo; Avallone, Rossella; Geminiani, Elisa; Cosenza, Francesca; I., Venturini; Baraldi, Mario
abstract

The peripheral benzodiazepine receptor (PBR), an internal protein of the mammalian mitochondrial membrane, is involved in several metabolic functions such as steroidogenesis, oxidative phosphorylation, and regulation of cell proliferation. Here we report the presence of PBRs in parenchymal and meristematic tissues of potato (Solanum tuberosum). PBRs are heterogeneously distributed in potato and are highly expressed in meristematic cells. In particular the receptor protein is mainly localised in the meristematic nuclear subcellular preparation. This 30-36 kDa protein, which corresponds to PBR, is increased, indeed, in meristematic compared to the parenchymal tissue. This suggests an involvement of this receptor in the regulation of cell plant growth. In addition, the demonstration that PBRs are also present in vegetables supports the hypothesis of a highly conserved receptor system during phylogenesis. Published by Elsevier Inc.


2003 - Evidence that endogenous benzdiazepine-like compounds increase during spontaneous delivery [Abstract in Atti di Convegno]
Avallone, Rossella; Modugno, Giuseppe; Facchinetti, Fabio; Baraldi, Mario
abstract

It has been established that endogenous benzodiazepines (EBDZ) play a role in the pathogenesis of different pathologies like hepatic encephalopathy or idiopathic recurring stupor(1-2). Evidences have not been provided about the role of EBDZ during pregnancy and delivery. In this work we evaluated a possible changes of EBDZ in maternal/foetal serum during delivery after spontaneous labour (VD) or caesarean section (CS). For both VD and CS groups (n=24) 3 blood samples were collected at 3 different times: the first was collected at least 3 days before labour and delivery, the second was collected at the moment of the delivery or at foetal abdominal extraction; the third sample was obtained at 2nd day post-partum. Determination of EBDZ was performed as already described (1) and cortisol was measured through commercially available radioimmunoassay kit. For the statistical analysis of EBDZ Wilcoxon test was used for within-subjects comparison and Mann-Whitney tests for between subjects comparison. For the statistical analysis of cortisol levels the analysis of variance and paired t-test was used. In the VD group a significant increase of EBDZ occurred at delivery respect with both baseline (P=0.008) and postpartum (P=0.028) values. Interestingly in CS group were found no differences in the 3 different analysed samples. The cortisol levels in VD group at delivery (560 ± 156 ng/ml) were found to be higher than that at baseline (252 ± 93, t= 5.47, P<0.0001) and post-partum (241 ± 44, t=6.11, P<0.0001) while in CS group no significant changes were found. These findings demonstrated for the first time that parturition is associated with a marked increase of EBDZ which could be attributed to stress present at delivery time.


2003 - Presenza di composti ad attivita’ ansiolitico-sedativa nella filera di produzione del parmigiano reggiano. [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; Puja, Giulia; Losi, Gabriele; Baraldi, Mario
abstract

Da tempo è stata descritta la presenza di sostanze ad attività sedativa nel latte tuttavia la loro struttura chimica è ancora oggetto di studio. Recentemente è stato isolato dall’s1-caseina un peptide denominato s1-CN-(f91-100) o s1-casozepina [1] mentre il nostro gruppo ha identificato nel latte altre sostanze non peptidiche ad attività benzodiazepino-simile [2]. Non è ancora noto, comunque, quali e quante sostanze presenti nel latte si ritrovano nei vari formaggi dopo i processi di produzione. Nel presente lavoro è stata valutata la presenza di composti ad attività benzodiazepino-simile nel parmigiano reggiano. In particolare è stata analizzata la filiera di produzione del parmigiano reggiano prelevando i seguenti campioni presso un’unica azienda in modo da escludere variabilità legate a fattori di alimentazione, ambiente, procedimento di produzione: fieno, latte intero, latte magro, siero, siero innesto, panna, cagliata, tosone e parmigiano reggiano. I campioni estratti con metanolo sono stati purificati mediante analisi HPLC a 0.8 ml/min utilizzando una colonna LiChrospher 100 RP-18 column (250x4.0 mm; 5 m) equilibrata con 80% di acqua/0.1% TFA e 20% di acetonitrile. I campioni sono stati analizzati a gradiente (0.5%) di acqua/0.1% TFA and acetonitrile dal 20 al 58% di acetonitrile. Per ogni campione sono state raccolte 75 frazioni, liofilizzate e testate per valutare la loro capacità di legarsi specificatamente al sito di riconoscimento centrale benzodiazepinico mediante dosaggio radioisotopici utilizzando come ligando marcato il [3H]RO 15-1788, antagonista benzodiazepinco. La concentrazione ottenuta per estrapolazione da una curva di competizione generata con autentico diazepam, viene espressa in ng diazepam equivalenti /g di campione. Nel tosone e nel parmigiano reggiano sono state rilevate piccole quantità di sostanze BDZ-simili (0,78 ± 0,12 e 0,86 ± 0,17 ng DE/g rispettivamente) mentre nel latte e nel siero la loro concentrazione risulta significativamente maggiore (117,75 ± 9,2 e 215 ± 15,7 ng DE/g rispettivamente). E’ interessante sottolineare che la concentrazione di tali sostanze nel fieno (14,8 ± 1,64 DE ng/g) è inferiore rispetto al latte, considerando che l’alimentazione della mucca è esclusivamente basata sul fieno si può ipotizzare che il contenuto che si ritrova nel latte sia anche il contributo di una sintesi endogena. L’attività ansiolitico-sedativa delle frazione isolate dal siero di latte è stata valutata sia in vitro su neuroni corticali con la tecnica elettrofisiologia del patch-clamp che in vivo.Bibliografia1. L. Miclo, E. Perrin, A. Driou, V. Papadopoulos et al. Characterization of a-casozepina, a tryptic peptide from bovine as1-caseina with benzodiazepine-like activity. FASEB J: 15, 1780-1782, 20012. R. Avallone, L. Corsi, M.L. Zeneroli and M. Baraldi. Presence of benzodiazepine-like molecules in food and their implication in the nutrition of cirrhotic patients. Innovative Food Science and Technologies: 2/3, 193-198, 2001


2003 - Presenza di sostanze Benzodiazepino-simili in estratti di foglie di tabacco (Nicotiana tabacum) [Relazione in Atti di Convegno]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; Geminiani, Elisa
abstract

Le finalità della ricerca sono rappresentate dal desiderio di chiarire le possibili utilizzazioni a fini salutistici dei singoli principi attivi presenti nelle foglie di tabacco avendo ben presenti gli efftti prodotti dai composti benzodiazepino-simili a secondo della loro capacità di interferire sui recettori centrali (attività anziolitico-sedativa e miorilassante) e su quelli periferici (steroidogenesi, proliferazione cellulare ed attività sui processi infiammatori).


2003 - Recettore periferico delle benzodiazpine e diazeoma binding inhibitor nel carcinoma epatocellulare [Abstract in Atti di Convegno]
Corsi, Lorenzo; I., Venturini; Avallone, Rossella; Cosenza, Francesca; Baraldi, Mario; Zeneroli, Maria Luisa
abstract

L'elevata espressione del recettore periferico delle benzodiazepine e del suo ligando endogeno DBI in tessuti tumorali potrebbe fortemente indicare un'implicazione di questo soistema proteico nel metabolismo delle cellule neoplastiche.


2002 - Antiproliferative effects of Ceratonia siliqua L. on mouse hepatocellular carcinoma cell line [Articolo su rivista]
Corsi, Lorenzo; Avallone, Rossella; F., Cosenza; F., Farina; Baraldi, Claudia; Baraldi, Mario
abstract

Extracts from pods and leaves of carob (Ceratonia siliqua L.) were tested for their ability to inhibit cell proliferation of mouse hepatocellular carcinoma cell line (T1). The two extracts showed a marked alteration of T1 cell proliferation in a dose-related fashion reaching the maximal effect at 1 mg/ml. Moreover, we demonstrated that leaf and pod extracts were able to induce apoptosis in T1 cell lines after 24-h treatment mediating a direct activation of the caspase 3 pathway. HPLC analysis revealed the presence of gallic acid, epigallocatechin-3-gallate and (-) epi catechin - 3 -gallate in pod and leaf extracts, compounds well known to exert antiproliferative effects. Their concentration reached 6.28 mg/g in carob leaves and 1.36 mg/g in carob pods extract. The discovery that carob pod and leaf extracts contained antiproliferative agents could be of practical importance in the development of functional foods and/or chemopreventive drugs.


2002 - Extraction and purification from Ceratonia siliqua of compounds acting on central andperipheral benzodiazepine receptors [Articolo su rivista]
Avallone, Rossella; Cosenza, Francesca; Farina, Franco; Baraldi, Claudia; Baraldi, Mario
abstract

The presence of molecules with high affinity for central and peripheral benzodiazepine receptors was determined in the pod and leaves of Ceratonia siliqua carob. The amount of the substances able to selectively bind the central benzodiazepine receptor recovered fromcarob pods and leaves was respectively 12.17 and 18.7 ng diazepam equivalentg. The amount of compounds active on peripheral benzodiazepine receptor in both pods and leaves was higher in comparison with the central one, being 49.83 and 40.00 PK 11195 equivalentg, respectively. In particular the compounds acting on peripheral benzodiazepine receptors were found to be extremely concentrated in the young leaves Ž2572.57 ng PK 11195equivalentg. The presence of substances with central benzodiazepine activity in carob extracts seems of great importance in view of the possibility to use carob extract as potential natural products with anxiolytic-sedative effects. Moreover, the prevalence in leaves ofsubstances acting on peripheral benzodiazepine receptor suggests the possible utilisation of leave extracts as chemopreventive agents.


2002 - Vegetable and mammalian milk as a source of benzodiazepine-like molecules [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; Baraldi, Mario
abstract

One of the challenging questions till facing benzodiazepine (BDZ) researchers is the identification of endogenous BDZ ligands in the brain, which could modulate -aminobutyric acid (GABAA) neurotrasmission. Several substances with benzodiazepine-like activity were found in food. In particular their amount was particularly high in milk. Different type of milk of animal or vegetal origin were analysed by HPLC analysis. The serum obtained by acid precipitation of milk was chromatographed at 0.8 ml/min on a LiChrospher 100 RP-18 column (250x4.0 mm; 5 m) equilibrated with 80% water/0.1% TFA and 20% acetonitrile. The sample was analysed using a water/0.1% TFA and acetonitrile gradient at 0.5% from 20 to 58% acetonitrile. 75 fraction were collected, lyophilised and tested for their ability to inhibit [3H]RO 15-1788 specific binding to central BDZ binding site or [3H]PK 11195 specific binding to peripheral BDZ binding site (PBR). The results are reported in figure. Thermic treatments, like UHT, or the biological procedures do not change the BDZ-like molecules concentration. Since the PBR is involved in cell proliferation the fractions active on PBR were tested on the ability to modulate cell proliferation of hepatocellular carcinoma cell lines (T1). The chronic treatment with the active fraction showed a significant dose-dependent inhibition of T1 cell proliferation indicating a potential use of milk as functional food.


2001 - Ammonia and endogenous benzodiazepine-like compounds in the pathogenesis of hepatic encephalopathy [Articolo su rivista]
Venturini, I.; Corsi, L.; Avallone, R.; Farina, F.; Bedogni, G.; Baraldi, C.; Baraldi, M.; Zeneroli, M. L.
abstract

Background: Ammonia and endogenous benzodiazepines (BDZs) are two of the most important agents among those taken into consideration in the pathogenesis of hepatic encephalopathy (HE). Methods: Venous ammonia and endogenous BDZs sera levels were assayed in 58 liver cirrhosis patients (34 male, 24 female) free of commercial BDZs. Endogenous BDZs were measured by binding assay after highperformance liquid chromatography purification. Ammonia was assessed by colorimetric test. Results: Endogenous BDZs and ammonia were significantly higher in Child-Pugh class C than in class B and class A (P &lt; 0.05), correlating to the severity of the liver dysfunction but not with the degree of HE. A significant difference, in fact, was noted between degree 0 (no HE) versus III-IV of HE (P &lt; 0.05), but not between degrees I-II versus III-IV. Regression analysis performed to find a correlation between the ammonia and BDZ levels in HE resulted negative. Conclusion: Clinical evidence is provided in cirrhotic patients that ammonia and endogenous BDZ levels do not correlate with each other in the outcome of HE.


2001 - Effect of epicatechin-3-gallate (ECG) and epigallocatechin-3-gallate (EGCG) on cel proliferation, role of peripheral benzodiazepine receptor (PBR) [Abstract in Atti di Convegno]
Corsi, Lorenzo; R., Guagliumi; Avallone, Rossella; Baraldi, Mario
abstract

The polyphenolic compounds showed cancer chemopreventive effect in animal tumor model. Peripheral benzodiazepine receptor i involved in cell proliferation and malignancy. We investigated and compared the effect og epigallocatechin-3-gallate and epicatechin-3-gallate on the proliferation of mouse hepatocellular carcinoma cel line.


2001 - Natural Modulators of Anxiety and Sleep Disturbances in Food [Relazione in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; G., Lugli; Baraldi, Mario
abstract

HPLC analysis of foos extracts revealed the presence of several fractions that exhibited diaplacing activity on benzodiazepine-binding assay to rat cerebellar membranes.


2001 - Presence of benzodiazepine-like compound molecules in food and their implication in the nutrition of cirrhotic patients [Articolo su rivista]
Avallone, Rossella; Corsi, Lorenzo; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Benzodiazepine (BDZ)-like compounds, present in trace amounts in normal subjects increase in the blood of liver cirrhotic patients. The origin of these compounds is still unknown but they are present in medicinal plants and foods. Herein we report the detection of BDZ-like molecules in fruits, vegetables, cereals, meat, milk and cheeses and in different cultivars of potatoes, tomatoes and carrots. The extracted food was separated by HPLC purification and the collected fractions were tested by radioreceptor binding assay in order to evaluate their ability to selectively bind the central benzodiazepine receptors. The mean value was 14.80 ng of diazepam equivalent (DE)/g in fruits, 4.34 ng DE/g in vegetables, 6.35 in cereals and 4.09 in meat. BDZ-like compounds are poorly present in cheeses and completely absent in olive and seeds oil. From these findings it is possible to select food with low amount of BDZ-like molecules useful for cirrhotic diet in order to prevent hepatic encephalopathy.


2001 - Sedative and anxiolytic properties od chrysin, a constituent of Passiflora incarnata [Abstract in Atti di Convegno]
Rivasi, Marianna; Muscatello, Cinzia; Zanoli, Paola; Avallone, Rossella; Baraldi, Mario
abstract

The behavioral effects of acute administration of chrysin, contained in Passiflora incarnata, were studied in rats. The data demonstrate that in our experimental conditions, chrysin was able to reduce locomotor activity when injected in rats at a minimal effective dose of 25 mgrkg. However, chrysin exhibited a clear anxiolytic effect when injected at the dose of 1 mgrkg,. The sedative effect of the flavonoid cannot be ascribed to an interaction with GABA]benzodiazepine receptors, since it was not counteracted by the benzodiazepine antagonist Flumazenil. To the contrary, the anxiolytic effect of chrysin, which was blocked by the injection of Flumazenil, could be linked to an activation of the GABAA receptor unit.


2000 - Behavioral characterisation of the flavonoids apigenin and chrysin [Articolo su rivista]
Zanoli, Paola; Avallone, Rossella; Baraldi, Mario
abstract

The behavioral effects of acute administration of two flavonoids, apigenin and chrysin, contained in Matricaria chamomilla and in Passiflora incarnata, respectively, were studied in rats. The data demonstrate that in our experimental conditions, the two flavonoids were equally able to reduce locomotor activity when injected in rats at a minimal effective dose of 25 mgrkg. However, while chrysin exhibited a clear anxiolytic effect when injected at the dose of 1 mgrkg, apigenin failed to exert this activity. The sedative effect of these flavonoids cannot be ascribed to an interaction with GABA]benzodiazepine receptors, since it was not counteracted by the benzodiazepine antagonist Flumazenil. To the contrary, the anxiolyticeffect of chrysin, which was blocked by the injection of Flumazenil, could be linked to an activation of the GABAA receptor unit.


2000 - Benzodiazepine esogene ed endogene: iplicazioni in taluni stati patologici [Capitolo/Saggio]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; Zeneroli, Maria Luisa
abstract

Le benzodiazepine che si trovano nel plasma dell'uomo potrebbero derivare dagli alimenti, dall'azione della flora batterica intestinale e probabilmente da una sintesi endogena ancora sconosciuta.


2000 - Caratterizzazione dell'attività farmacologica dell'iperforina in confronto all'estratto di Hypericum perforatum [Abstract in Atti di Convegno]
Muscatello, Cinzia; Baraldi, Claudia; Rivasi, Marianna; Avallone, Rossella; Zanoli, Paola
abstract

Allo scopo di chiarire il meccanismo con cui l'iperforina svolge la sua azione ansiolitica, sono stati eseguiti studi preliminari con antagonisti famcaologici che hanno suggerito un possibile coinvolgimento del sistema serotoninergico e una scarsa implicazione del sistema recettoriale benzodiazepinico.


2000 - Characterisation of pharmacoligical activity of hyperforin in comparison with Hypericum extract [Abstract in Atti di Convegno]
Zanoli, Paola; Muscatello, Cinzia; Baraldi, Claudia; Rivasi, Marianna; Avallone, Rossella
abstract

In our experiments we studied the influence of hyperforin on locomotor behavior of rats and evaluated its effect in two experimental models of anxiety in comparison with the administration of hypericum extract.


2000 - Definizione delle attività farmacologiche dell'iperforina e relativi meccanismi d'azione [Abstract in Atti di Convegno]
Zanoli, Paola; Muscatello, Cinzia; Baraldi, Claudia; Rivasi, Marianna; Avallone, Rossella
abstract

nel nostro studio abbiamo preso in considerazione l'effetto dell'iperforina sul comportamento motorio del ratto e la sua influenza in modelli sperimentali di ansia. In tali esprimenti abbiamo messo a confronto l'attività dell'iperforina acetato con quella di un estratto totale di iperico ottenuto con la tecnica della CO2 supercritica.


2000 - Endogenous and exogenous benzodiazepines [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Herein we report the detection of benzodiazepine-like molecules in food suc as carrot, maize, soy-bean, peam wheat, salad and different cultivars of potatoes and tomatoes. The extracted food exhibit after HPLC separation a series of substances able to inhibit the binding of [3H]RO 15-1788 to rat cerebellar membranes hence indicating a benzodiazepine-like activity.


2000 - Endogenous and exogenous benzodiazepines [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

It is clear that there is a level of benzodiazepines in the serum healthy man free of benzodiazepines medication and that alteration of these levels could influence the state of vigilance and anxiety.


2000 - Endogenous benzodiazepines [Articolo su rivista]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; I., Venturini; C., Baraldi; Zeneroli, Maria Luisa
abstract

The existence of endogenous benzodiazepines such as diazepam and nordiazepam has been provided in human blood and brains as well as in medicinal plants and foods. It must be stressed, however, that in plasma and brain tissue there are also other benzodiazepine-like compounds termed 'endozepines' which are not halogenated. A synthetic pathway for the production of benzodiazepine-like compounds and endozepines has not yet been found, hence it may be surmised that these compounds could be of exogenous source. Changes in the level of endogenous circulating benzodiazepines due to food or drug ingestion could be responsible for pathological conditions. Clinical experiments were designed in order to study the levels of the endogenous benzodiazepines in vegetables and in the blood of control subjects and of cirrhotic patients. These patients accumulate benzodiazepines because of decreased liver metabolization capacity and impaired renal secretion, reaching plasma concentrations similar to those recorded in commercial benzodiazepine consumers.


2000 - Endogenous benzodiazepines: from physiology to patology [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Endogenous benzodiazepines were measured by the radioligand binding technique after high performance liquidchromatography (HPLC) purification. The presence of diazepam andN-desmethyldiazepam was assayed by HPLC-electrospray tandem mass spectrometry. Diazepam binding inhibitor was studied in serum by radioimmunoassay. Results—Endogenous benzodiazepineswere below the limit of detection in 7% of patients with encephalopathy. When detectable, their levels were at least comparable with those of benzodiazepine consumers and correlated with the liver dysfunction but not the stage of encephalopathy.Serum levels of diazepam binding inhibitor tended to decrease when endogenous benzodiazepines levels increased. Conclusions—Endogenous benzodiazepines may accumulate in patients withliver cirrhosis during the course of the disease, and the phenomenon appears to be independent of the presence or absenceof encephalopathy.


2000 - Le benzodiazepine dalla molecola alla pratica clinica [Monografia/Trattato scientifico]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; Baraldi, Claudia; Zeneroli, Maria Luisa
abstract

La ricerca di molecola endogene ad attività benzodiazepino-simile ha portato alla scoperta di molecole sia negli alimenti che nell'uomo.


2000 - Le benzodiazepine naturali [Capitolo/Saggio]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; Baraldi, Claudia; Ml, Zeneroli
abstract

Sull’origine delle benzodiazepine naturali trovate nei tessuti e nel sangue di mammiferi sono state formulate due ipotesi: a) origine esogena; b) origine endogena. Entrambe sono state prese in considerazione.Per quanto riguarda l’origine esogena, un nostro studio sistematico sul contenuto di sostanze benzodiazepino-simili negli alimenti (verdure, cereali, frutta, carne, latte, formaggio e condimenti) utilizzati nella dieta mediterranea, ha messo in evidenza una differenza significativa nel contenuto di tali composti fra le varietà dei vegetali diverse sia per genotipo sia per fenotipo [2]. La nozione che le benzodiazepine naturali sono presenti in molti alimenti ha suggerito la possibilità di interferire sullo stato di vigilanza dell’uomo, attraverso la somministrazione di diete particolari capaci di modificarne l’omeostasi nel sangue. E’ possibile pertanto mettere a punto diete a basso contenuto di tali sostanze nei pazienti a rischio di sviluppare disturbi neurologici come nel caso della cirrosi epatica o utilizzare diete ricche di benzodiazepine naturali quali il latte nei soggetti ansiosi o con disturbi del sonno.Per quanto riguarda l’origine endogena, pazienti affetti da cirrosi epatica di media gravità sono stati in primo luogo sottoposti ad una dieta in modo da controllare l’apporto benzodiazepinico alimentare [3]. Dopo 3 giorni di dieta essi sono stati trattati per via orale con rifaximina (800 mg/dì per os per 7 giorni), antibiotico non assorbibile capace di modificare la flora batterica intestinale. L’azione antibatterica della rifaximina ha prodotto in questo breve periodo di terapia una riduzione media del 40 % dei livelli di sostanze benzodiazepino-simili nel sangue. Questa osservazione sembra indicare che la flora batterica intestinale sia capace di influenzare i livelli di sostanze benzodiazepino-simili nel sangue e suggerisce la possibilità di utilizzare nella dieta probiotici e prebiotici e relative associazioni nel tentativo di aumentare la flora batterica intestinale utile alla produzione di benzodiazepine endogene capaci di prevenire eccessive lesioni psichiche e disturbi del sonno.


2000 - Pharmacological profile of apigenin, a flavonoid isolated from Matricaria chamomilla [Articolo su rivista]
Avallone, R.; Zanoli, P.; Puia, G.; Kleinschnitz, M.; Schreier, P.; Baraldi, M.
abstract

Dried flowers of Matricaria chamomilla L. are largely used to provide sedative as well as spasmolytic effects. In the present study, we examined in particular the pharmacological property of a fraction isolated from a methanolic extract of M. chamomilla, which was identified by HPLC-MS-MS analysis as apigenin. By radioreceptor binding assays, we demonstrated the ability of the flavone to displace a specific radioligand, [3H]Ro 15-1788, from the central benzodiazepine binding site. Electrophysiological studies performed on cultured cerebellar granule cells showed that apigenin reduced GABA (gamma-aminobutyric acid)-activated Cl-currents in a dose-dependent fashion. The effect was blocked by co-application of Ro 15-1788, a specific benzodiazepine receptor antagonist. Accordingly, apigenin reduced the latency in the onset of picrotoxin-induced convulsions. Moreover, apigenin injected i.p. in rats reduced locomotor activity, but did not demonstrate anxiolytic, myorelaxant, or anticonvulsant activities. The present results seem to suggest that the inhibitory activity of apigenin on locomotor behaviour in rats cannot be ascribed to an interaction with GABA(A)-benzodiazepine receptor but to other neurotransmission systems, since it is not blocked by Ro 15-1788. (C) 2000 Elsevier Science Inc.


2000 - Presence of benzodiazepines-like molecules in food and theri implication in the nutrition of cirrhotic patients. [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; I., Ventu; Farina, Franco; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

The data presented indicate that it is possible to select vegetables with low amount of benzodiazepines-like molecules which could be more appropriate for cirrhotic diet in order to avoid the occurence of episodes of encephalopathy in patients with severe liver disease.


2000 - Presenza di sostanze ad attività benzodiazepino-simile negli alimenti: implicazione nella nutrizione di pazienti cirrotici [Abstract in Atti di Convegno]
Avallone, Rossella; F., Begnozzi; Farina, Franco; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Sulla tavola di ogni italiano ci sono tranquillanti naturali che in alcuni casi, come il latte fresco, contengono quantità farmacologiche di principi attivi alla base di noti medicinali come il tavor. Il latte fresco, umano, vaccino e caprino, possiede un'attività tranquillante intrinseca paragonabile a quella di farmaci molto impiegati come il diazepam (principio attivo del valium) poiché contiene naturalmente le stesse molecole benzodiazepiniche vendute in farmacia. Oltre al latte, hanno effetti tranquillanti sull'uomo anche alcuni frutti, come pere, arance e mele, i cerali e la soia. Alimenti ad elevato contenuto di queste sostanze, potrebbero essere impiegati nella prevenzione di stati patologici ansiosi, mentre alimenti a ridotto contenuto di tranquillanti potrebbero trovare una giusta applicazione nelle linee dietetiche per pazienti cirrotici e encefalopatici che non possono assumere composti ad azione sedativa.


2000 - Prresenza di sostanze attive sia sul recettore centrale che periferico delle benzodiazepine in estratti di farina e foglie di carruba (Certonia siliqua) [Abstract in Atti di Convegno]
Baraldi, Claudia; Avallone, Rossella; Cosenza, Francesca; A., Trovato; Baraldi, Mario
abstract

Nell'ambito di numerose indagini dedicat alla determinazione di composti benzodiazepino-simili in diversi prodotti alimentari, parte delle ricerche hanno riguardato la presenza di tali composti nel bacello e nelle foglie di carruba (Ceratonia siliqua), pianta sempreverde ampiamentediffusa in tutta l'area mediterranea. Se i semi trovano una grande utilizzazione nell'indusria alimentare per l'alta concentrazione di gomme, l'utilizzo dell'intero bacello ha invece delle strette limitazioni per il prevalente carattere astringente dovuto alla presenza dei tannini.


1999 - Attività sedativa dei flavonoidi contenuti in piante medicinali: Proprietà farmacologiche a confronto [Abstract in Atti di Convegno]
Zanoli, Paola; Avallone, Rossella; Baraldi, Mario
abstract

Nella presente ricerca abbiamo evidenziato che flavponoidi come la crisina e l'apigenina contenuti in particolare nella camomilla e nella passiflora rispettivamente, sono dotati di proprietà sedative. Infatti quando vengono somminstrate a ratti per via intraperitoneale, a partire da una dose minima attiva di 25 mg/Kg sono in grado di ridurre, in modo dose-dipendente, l'attività locomotoria degli animali sottoposti al test dell'open field.


1999 - Endogenous benzodiazepines [Abstract in Atti di Convegno]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Zeneroli, Maria Luisa
abstract

The presence of specific receptors for benzodiazepines (BDZ) suggested that these recognition sites could subserve a physiological role in the regulation of anxiety and sleep only if stimulated by endoigenous ligands. A polypeptide nmed DBI could be one putative endogenous ligands with diazepma binding inhibitory properties and with anxiogenic activity. Clinical experiments were designed in order to study the levels of the endogenous BDZs and in particular the relationship between BDZs and DBI in control.


1998 - A possible mechanism of resistance to death during cell proliferation in hepatocellular carcinoma: the mitochondrial benzodiazepine receptor system. [Abstract in Atti di Convegno]
I., Venturini; Zeneroli, Maria Luisa; Corsi, Lorenzo; R., Pellicci; G., Ardizzone; A., Arrigo; Farina, Franco; C., Ferrarese; N., Pecora; M., Frigo; Avallone, Rossella; Baraldi, Mario
abstract

The aims of the present research w to detremine the role of the mitocondrial benzodiazepine receptors (MBRs) in hepatocellular carcinoma (HCC). The densities of MBRs and the level of DBI in PPIK were tested in tumoral and non tumoral liver tissue of patients with HCC.


1998 - Ammonia and 1,4-benzodiazepines in encephalopathic patients with fulminatn hepatic failure [Abstract in Atti di Convegno]
Zeneroli, Maria Luisa; I., Venturini; Corsi, Lorenzo; Avallone, Rossella; Farina, Franco; G., Ardizzone; M., Centenaro; A., Arrigo; L., Miglioli; P., Schreier; M., Kleinschnitz; Baraldi, Mario
abstract

Recently it has been surmised that both ammonia and benzodiazepine-like compounds (BDZs) may sinergistically contribute to hepatic encephalopathy (HE). In order to clarify this hypothesis, the levels of venous ammonia and BDZs were assayed in patients wuth HE due to fulminant hepatic failure.


1998 - Ammonia and 1,4-benzodiazepines in patients with fulminat hepatic failure [Abstract in Atti di Convegno]
Zeneroli, Maria Luisa; I., Venturini; Corsi, Lorenzo; Avallone, Rossella; Farina, Franco; G., Ardizzone; M., Centenaro; A., Arrigo; A., Caronna; P., Schreier; M., Kleinschnitz; Baraldi, Mario
abstract

The inconsistent presence of benzodiazepines in hepatic encephalopathy (HE) due to fulminat hepatic failure and the presence at the awakening from coma, indicate that HE i note strictly depoendent on the levels of these compounds.


1998 - Benzodiazepine endogene [Abstract in Atti di Convegno]
Avallone, Rossella; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

La reale importanz di questi studi è rappresentata dal fatto che tali sostanze presenti nella dieta sono sorgenti di benzodiazepine per l'organismo umano. il riconoscimento che gli alimenti sono una sorgente di benzodiazepine, suggerisce la possibilità di selezionare diete a basso od alto contenuto di benzodiazepine in base alla patologia presente.


1998 - Benzodiazepine-like compound in foods and plants [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; Baraldi, Mario
abstract

Extracts of potatoes and tomatoes exhibited after HPLC separation a series of substances able to inhibit the binding of [3H]RO 15-1788 to rat cerebellar membranes. The level ranged from 0.03 ng diazepam equivalent (DE)/g to 3.56 ng DE/g inthe potatoes and from 0.01 to 1.55 DE/g in the tomatoes depending from the selected cultivars.


1998 - Benzodiazepine-like compounds in plasma of patients with fulminant hepatic failure [Articolo su rivista]
Zeneroli, Maria Luisa; I., Venturinii; Corsi, Lorenzo; Avallone, Rossella; Farina, Franco; G., Ardizzone; M., Centanaro; A., Arrigo; P., Schreier; M., Kleinschnitz; Baraldi, Mario
abstract

Background: Benzodiazepine-like compounds have been implicated in the pathogenesis of encephalopathy after fulminant hepatic failure. Methods: The levels and the nature of benzodiazepine-like compounds were determined in six cases of fulminant hepatic failure during the course of the disease. Blood samples were collected on admission and a few days later, when the neurologic status had improved in five cases and immediately before death in one case. The compounds were measured in sera with a binding technique after high-performance liquid chromatography purification and analyzed with mass spectrometry;, Results: Their levels were highly variable in those with severe encephalopathy and were still Increased on awakening in some cases. Diazepam and N-desmethyldiazepam were inconsistently present. Conclusions: The inconsistent presence of benzodiazepine-like compounds in encephalopathy after fulminant hepatic failure and their persistence, in some cases, at high levels on awakening from coma seem to Indicate that the encephalopathy is not strictly dependent on the levels of these compounds.


1998 - Endogenous benzodiazepine-like compounds and diazepam binding inhibitor in serum of patients with liver cirrhosis with and without overt encephalopathy [Articolo su rivista]
Avallone, Rossella; Zeneroli, Maria Luisa; I., Venturini; Corsi, Lorenzo; P., Schreier; M., Kleinschnitz; C., Ferrarese; Farina, Franco; Baraldi, Claudia; N., Pecora; M., Frigo; Baraldi, Mario
abstract

Background/Aim—Despite some controversy, it has been suggested that endogenous benzodiazepine plays a role in the pathogenesis of hepatic encephalopathy. The aim of the present study was to evaluatethe concentrations of endogenous benzodiazepines and the peptide, diazepam binding inhibitor, in the blood of patients with liver cirrhosis with and without overt encephalopathy, and to compare theselevels with those of consumers of commercial benzodiazepines.Subjects—Normal subjects (90), benzodiazepine consumers (14), and cirrhotic patients (113) were studied. Methods—Endogenous benzodiazepines were measured by the radioligand bindingtechnique after high performance liquid chromatography (HPLC) purification. The presence of diazepam and N-desmethyldiazepam was assayed by HPLC-electrospray tandem mass spectrometry.Diazepam binding inhibitor was studied in serum by radioimmunoassay. Results—Endogenous benzodiazepineswere below the limit of detection in 7% of patients with encephalopathy. When detectable, their levels were at least comparable with those of benzodiazepine consumers and correlated with the liver dysfunction but not the stage of encephalopathy.Serum levels of diazepam binding inhibitor tended to decrease when endogenous benzodiazepines levels increased. Conclusions—Endogenous benzodiazepines may accumulate in patients withliver cirrhosis during the course of the disease, and the phenomenon appears to be independent of the presence or absence of encephalopathy.


1998 - Endogenous benzodiazepine-like compounds in serum of normal subjects and in liver cirrhosis patients. [Abstract in Atti di Convegno]
I., Venturini; Avallone, Rossella; Corsi, Lorenzo; Zeneroli, Maria Luisa; P., Schreier; M., Kleinschnitz; Farina, Franco; Baraldi, Mario
abstract

The concentration of benzodiazepines (BDZs) in serum from normal subject resulted under the limit of detection (lower tha 2 nmol DE/l) in 46 cases (51%). In the remaining 44 normal subjects the total amount binding ranged between 6 and 20 nmol DE/l. In the liver cirrhosis BDZs were under the detection limit in 7% of patients withencephalopathy. When detectable, tehi levels were at least comparable with those of benzodiazepines consumers and correlated with the liver dysfuntion but not with the stages of encephlopathy.


1998 - Endogenous benzodiazepines sources and role in liver cirrhosis and in fulminat hepatic failure [Abstract in Atti di Convegno]
Baraldi, Mario; Zeneroli, Maria Luisa; I., Venturini; Corsi, Lorenzo; Avallone, Rossella; Farina, Franco
abstract

The source and the role on endogenous benzodiazepines (BZDs)in the induction of hepatic encephalopathy (HE) are still unknown. BDZs are measured by radioligand binding thecnique after HPLC purificationin normnal subjects (n.90), benzodiazepine consumers (BZC) (n.14), cirrhotic patients (LC) (n.133), fluminat hepatic failure (FHF) (n.6).


1998 - Evidence that plasma levels of natural benzodiazepines are regulated by intestinal bacterial flora. [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Farina, Franco; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

The source od natural benzodiazepines is still unknown but they have been described in medicinal plants, vegetables and milk. In order to establish the content of BDZs in our diet, we measured natural BDZs by HPLC followed by radioligand binding assay in potatoes and tomatoes.


1998 - Food and bacterial flora are source of benzodiazepine-like compounds [Abstract in Atti di Convegno]
Zeneroli, Maria Luisa; I., Venturini; S., Stefanelli; Farina, Franco; A., Ferrieri; Corsi, Lorenzo; Avallone, Rossella; R., Cosenza; Baraldi, Mario
abstract

The content of natural benzodiazepines (BDZs) in diet was assayed by HPLC followed by radioligand binding technique in potatoes and tomatoes. We assayed the levels of BDZs in the plasma of 10 liver cirrhosis patients in child B class free of commercial benzodiazepines medication and under a constant diet. The patients were treated with rifaximin (800 mg/die x 7 days) in order to modify the bacterial flora.


1998 - Mitochondrial benzodiazepine receptor system in hepatocellular carcinoma: a possible mechanims of resistance to death during cell proliferation. [Abstract in Atti di Convegno]
I., Venturini; Zeneroli, Maria Luisa; Corsi, Lorenzo; R., Pellicci; G., Ardizzone; A., Arrigo; Farina, Franco; C., Ferrarese; N., Pecora; M., Frigo; Avallone, Rossella; Baraldi, Mario
abstract

These data demonstrated that in HCC patients there is an increased availability of the substrate for steroid synthesis, associated with an increase functional status of MBRs in HCC tissue. This suggest that in HCC tissue there is an incresed steroidogenesis and a resistance of tumoral against radical damage.


1998 - Peripheral benzodiazepine receptors in liver tumors and presence of thei ligands in vegetbale and officinal plants. [Abstract in Atti di Convegno]
Corsi, Lorenzo; Avallone, Rossella; I., Venturini; Zeneroli, Maria Luisa; H., Alho; Baraldi, Mario
abstract

in this study we provided evidence that during degenerative liver disease as hepatocellular carcinoma (HCC) the peripheral benzodiazepine receptor (PBR) binding sites increse drammatically. Extract of tomatoes, mentha piperita, lavadula officinalis, citrus aurantium and melissa officinalis after HPLC separation gave rise to a series of compounds able to inhibit the binding of [3H]PK 11195, a specific binding for PBRs to rat adrenal glands.


1998 - Pharmacological profile of flavonoids present in medicinal plants with sedative and axiolytic properties [Abstract in Atti di Convegno]
Zanoli, Paola; Avallone, Rossella; Truzzi, Cristina; Vezzalini, Francesca; Baraldi, Mario
abstract

In the present study we investigated the pharmacological effect of two flavonoids, chrysin and apigenin, contained in Passiflora incarnata and Matricaria chamomilla respetively.


1998 - Presence o fbenzodiazepine-like molecules in vegetables and thei implication in the nutrition of cirrhotic pateints. [Abstract in Atti di Convegno]
Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

The findings indicate that it is possible to select vgetables with low amount of benzodiazepine-like molecules which appear to be more appropriate in order to avoid the occurence of episodes of encephalopathy in patients with severe liver disease.


1998 - Sedative and Hypothermic Effects Induced by b-Asarone, a Main Component of Acorus calamus [Articolo su rivista]
Zanoli, Paola; Avallone, Rossella; Baraldi, Mario
abstract

In the present study we investigate the behavioural effects of b-asarone, the main compound of the essential oil obtained from Acorus calamus. b-asarone, when administered intraperitoneally in rats, exerts sedative and hypothermic but not analgesic effects. b-asarone, however, when administered in association with the cannabinomimetic drug WIN 55,212-2, was shown to potentiate some of the typical behavioural activities induced in animals by cannabinoids. Binding assays, performed on cortical synapticmembrane preparations using a specific cannabinoid radioligand ([3H]CP-55,940), excluded the ability of b-asarone to exert a direct agonistic activity on CB1 receptors. Hence, b-asarone cannot be considered a pure cannabinomimetic agent but it can be envisaged at least as an allosteric modulatory agent.


1998 - Sedative and hypothermic effects induced by β-asarone, a main component of Acorus calamus [Articolo su rivista]
Zanoli, P.; Avallone, R.; Baraldi, M.
abstract

In the present study we investigate the behavioural effects of β- asarone, the main compound of the essential oil obtained from Acorus calamus. β-asarone, when administered intraperitoneally in rats, exerts sedative and hypothermic but not analgesic effects. β-asarone, however, when administered in association with the cannabinomimetic drug WIN 55,212-2, was shown to potentiate some of the typical behavioural activities induced in animals by cannabinoids. Binding assays, performed on cortical synaptic membrane preparations using a specific cannabinoid radioligand ([3H]CP-55,940), excluded the ability of β-asarone to exert a direct agonistic activity on CB1 receptors. Hence, β-asarone cannot be considered a pure cannabinomimetic agent but it can be envisaged at least as an allosteric modulatory agent.


1998 - Sostanze ad attività benzodiazepino-simile in alimenti e droghe vegetali [Abstract in Atti di Convegno]
Baraldi, Mario; Avallone, Rossella; Corsi, Lorenzo; I., Venturini; Zanoli, Paola; M. L., Zeneroli
abstract

Per chierire l'otigine delle benzodiazepine trovate nei tessuti, sono stati analizzati diversi alimenti fra cui patata e pomodoro. Mediante analisi HPLC sono state separate da estratti di patata e pomodoro diverse sostanze capaci di inibire il legame del [3H]RO 15-1788 alle membrane cerebellari e del [3H]PK 11195 alle membrane surrenali di ratto. La quantità di queste sostanze varia da 0,03 ng/g diazepam equivalente (DE) a 3,56 ng/g di patata e da 0,01 a 1,55 ng/g di pomodoro a seocnda del cultivar analizzato.


1998 - Sources od benzodiazepine-like compounds: food and bacterial flora [Abstract in Atti di Convegno]
Avallone, Rossella; I., Venturini; Corsi, Lorenzo; Farina, Franco; DI BELLA, Maria; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

The observation indicates that intestinal bacterila flora is involved in the production of these compounds. In conclusion, even if cannot excludean endogenous synthesis, these observations seems to prove that the sources of benzodiazepine-like compounds in human could be the result of a combination of the food ingestion and bacterial flora production.


1998 - Up-regulation of peripheral benzodiazepine receptors system in hepatocellular carcinoma. [Articolo su rivista]
Venturini, I; Zeneroli, Maria Luisa; Corsi, Lorenzo; Avallone, Rossella; Farina, Franco; Alho, H; Baraldi, C; Pecora, N; Frigo, M; Ardizzone, G; Arrigo, A; Pellicci, R; Baraldi, Mario
abstract

Increased number of peripheral benzodiazepine receptors (PBRs) have been found in some tumors outside the liver. The present study was to verify whether the PBR system is altered in hepatocellular carcinoma (HCC). The levels of endogenous benzodiazepine-like compounds (BZDs). measured by radioreceptor binding technique after HPLC purification and the endogenous ligand for PBRs. termed diazepam binding inhibitor (DBI). measured by radioimmunoassay utilizing a specific antibody for human DBI, were studied in the blood of 15 normal subjects, 12 liver cirrhosis and 10 patients with HCC. The levels of BZDs in serum were increased hundred fold in liver cirrhosis patients and slightly elevated in HCC patients. DBI was found to be increased in HCC patients. The binding recognition sites for PBRs (Bmax) were increased 4 to 7 fold in HCC tissue in comparison with that found in non-tumoral liver tissue (NTLT). On the contrary the concentrations of DBI were found to be significantly decreased in HCC tissue in comparison with the respective NTLT. These results seem to suggest an implication of PBRs and of their putative endogenous ligands in the metabolism of these neoplastic cells and possibly in their proliferation. The up-regulation of PBRs found in HCC tissue seems to indicate an increased functional activity of these receptors and opens up the possibility of new pharmacological and diagnostic approaches while the changes in the circulating endogenous ligands for the above receptors might be envisaged as early markers of tumorigenesis in liver cirrhosis.


1997 - ANTIBACTERIAL ACTIVITY OF RIFAXIMIN REDUCES THE LEVELS OFBENZODIAZEPINE-LIKE COMPOUNDS IN PATIENTS WITH LIVERCIRRHOSIS [Articolo su rivista]
Zeneroli, Maria Luisa; I., Venturini; S., Stefanelli; Farina, Franco; R., Cosenza; L., Miglioli; E., Minelli; R., Amedei; A., Ferrieri; Avallone, Rossella; Baraldi, Mario
abstract

Benzodiazepine-like compounds are present in trace amounts in the blood of normal subjects and increase in liver cirrhotic patients with or without encephalopathy. Their increased presence may, however, represent an occasional precipitating factor of hepatic encephalopathy. The source of these compounds is still unknown, but they are constituents of our diet since benzodiazepine receptor ligands have been described in plants, vegetablesand in animals. They may also be synthesized, at least in part, by intestinal bacterial flora. In this article we report that the level of these compounds in the blood decreased by 40%after therapy with rifaximin, which reduces the aerobic and anaerobic intestinal bacterial flora. This observation indicates that intestinal bacterial flora is involved in the production of these compounds and that repeated short-term medications with this non-absorbableantibiotic may be useful in reducing the levels of benzodiazepine-like compounds in patients with liver cirrhosis.


1997 - Asymmetric synthesis of (R)- and (S)-2-pyrrolidinemethanesulfonic acid [Articolo su rivista]
Braghiroli, Daniela; Avallone, Rossella; DI BELLA, Maria
abstract

(R)- and (S)-2-Pyrrolidinemethanesulfonic acid, 3a and 3b, were synthesized from the corresponding N-Boc-2-(hydroxymethyl)-1-pyrrolidine, 6a and 6b. This asymmetric synthesis proceeds in mild conditions, with good overall yields and high enantiomeric purities (>99% ee).


1997 - Benzodiazepine-like compounds in plasma of patients with fulminant hepatic failure. [Abstract in Atti di Convegno]
I., Venturini; R., Cosenza; L., Miglioli; Farina, Franco; Corsi, Lorenzo; Avallone, Rossella; R., Amdedei; G., Ardizzone; E., Minelli; S., Stefanelli; Baraldi, Mario; P., Schreier; M., Kleinschnitz; Zeneroli, Maria Luisa
abstract

In conclusion the inconsistent presence od benzodizepines in hepatic encephalopathy (HE) due to fulminat hepatic failure and the persistence at high levels at the awakening from coma indicate that HE is not strickly dependent on the level of these compounds


1997 - Changes in endogenous benzodiazepine-like compound levels during the course of fulminant hepatic failure: potential effects of decreased renal function [Articolo su rivista]
Zeneroli, Maria Luisa; I., Venturini; Avallone, Rossella; Farina, Franco; L., Miglioli; R., Cosenza; R., Amedei; G., Ardizzone; M., Centenaro; A., Arrigo; Baraldi, Mario
abstract

The pathogenetic agents which cause encephalopathy due to fulminant hepatic failure are still under debate. Ammonia and benzodiazepine-like compounds are two of the most important agents considered, so far. Herein, we report the levels of benzodiazepine-like compounds in serum and in urine and of venous ammonia measured during the course of the disease (30 days). The patient rapidly developed stage IV encephalopathy with high levels of ammonia and with only a slight increase of benzodiazepine-like compounds. At that moment, the levels of these compounds were similar to those recorded in the blood when the patient regained full consciousness 28 days later. During the course of the disease, there was a 10-fold increase of benzodiazepine-like compounds in serum which was recorded in parallel with an impaired excretion due to oliguria. This observation seems to indicate that encephalopathy may develop in the absence of significantly increased levels of these compounds and that their episodic increase during fulminant hepatic failure may be an epiphenomenon linked with several factors such as impaired renal function


1997 - Determination of chemical composition of Carob (Ceratonia Siliqua): protein, fat, carbohydrates and tannins [Articolo su rivista]
Avallone, Rossella; Plessi, Maria; Baraldi, Mario; Monzani, Agar
abstract

Carob pod, germ, and seed were analyzed for moisture, ash, protein, fat, carbohydrates and particularly for their tannins content. Recovery of tannins as affected by various solvent extraction systems was investigated. Carob pod meal contained high levels of carbohydrates, appreciable amounts of protein and low levels of fat. Germ and seed contained more fat and less carbohydrates. Seventy percent acetone was the most effective solvent for the exctration and recovery of tanniins.


1997 - Endogenous benzodiazepine-like compounds in a patients with fulminatn hepatic failure: influence of a decreased renal excretion [Abstract in Atti di Convegno]
R., Cosenza; I., Venturini; L., Miglioli; Farina, Franco; Corsi, Lorenzo; Avallone, Rossella; R., Amedei; G., Ardizzone; A., Arrigo; E., Minelli; S., Stefanelli; Baraldi, Mario; Zeneroli, Maria Luisa
abstract

These observations suggest that encephalopathy due to fulminant hepatic faliure (FHF) may precipitate in the presence of low level of endogenous benzodiazepine and that these compounds may accumulate in the blood during the course of FHF without inducing encephalopath. One of the mechanisms of the increse of these compounds in the blood seems to be decreased urinary excretion due to diuresis.


1997 - Peripheral benzodiazepine receptors in hepatocellular carcinoma. [Capitolo/Saggio]
M. L., Zeneroli; R., Pellicci; I., Venturini; G., Ardizzone; A., Arrigo; Corsi, Lorenzo; Avallone, Rossella; Baraldi, Mario
abstract

Peripheral benzodiazepine receptor system (PBRs) triggers intracellular metabolic events and has been associated with cell proliferation. Its endogenous ligand, the diazepam binding inhibitor (DBI), contributes to steroidogenesis by promoting cholesterol delivery to the inner mitochondrial membrane. Since an increased number of PBRs have been found in several tumour tissues and cell lines, the present study was performed in order to verify if the expression of PBRs and DBI were altered in liver tumours. Using immunocytochemestry and in situ hybridisation we studied PBRs end DBI expression in 6 human tumours sited in the liver, in 1 liver hyperplasia, in 1 cirrhotic nodular regeneration, in 1 intestinal adenocarcionma and in correspondent surrounding non tumoral liver tissue (NTLT). Immunocytochemical study and in situ hybridisation revealed that PBRs and DBI were prominently expressed in neoplastic tissues than in NTLT and that they were colocalised in the same cells.


1997 - Sostanze ad attività benzodiazepino-simile in alimenti e in droghe vegetali: implicazioni fisiologiche e patologiche [Articolo su rivista]
Avallone, Rossella; Corsi, Lorenzo; Zanoli, Paola; DI BELLA, Maria; Monzani, Agar; I., Venturini; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Nel presente lavoro sono state estratte, purificate cromatograficamente e catratterizzate mediante dosaggi radioisotopici sostanze ad attività benzodiazepino-simile in 32 varietà di papata (Solanum tuberosum L.), 12 varietà di pomodoro (Solanum lycopersium L.) e nella camomilla (Matricaria chamomilla L.).


1996 - Benzodiazepine-like compounds and GABA in flower heads of Matricaria Chamomilla [Relazione in Atti di Convegno]
Avallone, Rossella; Zanoli, Paola; Corsi, Lorenzo; Cannazza, Giuseppe; Baraldi, Mario
abstract

Extracted and purified benzodiazepine like compounds from dried flwer heads of Matricaria chamomilla was investigated through radioligand binding assay on rat cerebellar membrane. Moreover intracerbroventrivular injection of purified active fraction produced a significant decrease of locomotor activity in rats.


1996 - Hepatic Encephalopathy in Liver Transplant Recipients Precipitated By Benzodiazepines Present in Transfused Blood [Articolo su rivista]
Zeneroli, Maria Luisa; I., Venturini; Avallone, Rossella; Farina, Franco; Corsi, Lorenzo; Baraldi, Claudia; G., Ardizzone; M., Centanaro; A., Arrigo; Baraldi, Mario
abstract

The observation that there are episodes of encephalopathy in liver cirrhosis patients after orthotopic liver transplantation, despite a well functioning graft and despite the lack of cerebral complications, prompted us to investigate the potential role of circulating benzodiazepine-like compounds in these episodes. The plasma levels of benzodiazepines were examined in 14 liver cirrhotic patients before and after transplantation. The benzodiazepines in the fluids infused during surgery and in individual bags of blood administered after surgery to 4 of these patients were also assayed. Herein we report that benzodiazepines accumulating in the blood of some transplanted patients appear to derive from blood transfusions utilized during surgery. The analysis of the types of benzodiazepines present in the blood utilized for transfusions suggests the use of commercial benzodiazepines by the donors. These compounds seem to be able to precipitate hepatic encephalopathy in patients with preexisting encephalopathy. Hence we suggest not using benzodiazepine consumers as blood donors, at least for patients with encephalopathy undergoing to liver transplantation.


1996 - Hepatic encephalopathy in patients submitted to liver transplantation precipitated by benzodiazepines present in transfused blood. [Articolo su rivista]
Zeneroli, Maria Luisa; I., Venturini; Avallone, Rossella; Farina, Franco; Corsi, Lorenzo; G., Ardizzone; M., Centanaro; A., Arrigo; Baraldi, Mario
abstract

Abstract: The observation that there are episodes of encephalopathy in liver cirrhosis patients after orthotopic liver transplantation, despite a well functioning graft and despite the lack of cerebral complications, prompted us to investigate the potential role of circulating benzodiazepine-like compounds in these episodes. The plasma levels of benzodiazepines were examined in 14 liver cirrhotic patients before and after transplantation. The benzodiazepines in the fluids infused during surgery and in individual bags of blood administered after surgery to 4 of these patients were also assayed. Herein we report that benzodiazepines accumulating in the blood of some transplanted patients appear to derive from blood transfusions utilized during surgery. The analysis of the types of benzodiazepines present in the blood utilized for transfusions suggests the use of commercial benzodiazepines by the donors. These compounds seem to be able to precipitate hepatic encephalopathy in patients with preexisting encephalopathy. Hence we suggest not using benzodiazepine consumers as blood donors, at least for patients with encephalopathy undergoing to liver transplantation.


1996 - Studio analitico della fibra dietetica in frutti dell'appennino modenese [Abstract in Atti di Convegno]
Plessi, Maria; Avallone, Rossella; Bertelli, Davide; Monzani, Agar
abstract

Nuts and chestnuts were analized for SDF and IDF using AOAC official method. Pectins, cellulose, hemicellulose and lignin were also evaluated.


1995 - Studio analitico della componente lipidica e di componenti minori (fibra dietetica e tannini) nella castagna (Castanea Sativa Mill.). . , [Relazione in Atti di Convegno]
Plessi, Maria; M. S., Simonetti; L., Cossignani; F., Santinelli; P., Damiani; Avallone, Rossella; Monzani, Agar
abstract

Caratterizzazione nutrizooinale e contenuto in metalli di castagne


1994 - The pathophysiology of hepatic encephalopathy. [Capitolo/Saggio]
Venturini, I; Avallone, Rossella; Avanzato, S; Zeneroli, Maria Luisa; Baraldi, Mario
abstract

Despite several clinical and expeimental studies, the pathogenesis of hepatic encephalopathy (HE) is still under debate. This is certainly due to the complexity of the issue but, at least in part, the lack of clarity resides on the method of approach utilised in the discussion of this theme. In order to reach a clear cut understanding of the cause-effect ratio the primary peripheral factors accumulating in the blood must be distinghuished from the neurochemical events. Moreover, since there are nowe clinical adn experimental data which demostrate that the patogenesis of HE is multifactorial, each toxin "per se" may not correlate with HE and may not fully explainal the aspects of the syndrome.


1992 - Effetti di deficit ed accumulo di metalli pesanti sulla funzionalità del sistema nervoso centrale [Capitolo/Saggio]
Zanoli, Paola; Rossi, Tiziana; Vaccari, G; Giacobazzi, A; Truzzi, C; Avallone, Rossella; Ventura, A.
abstract

Vengono descritti gli effetti da accumulo di metlli pesanti come lo zinco, il magnesio, il manganese, il piombo, il metilmercurio, l'alluminio sul sistema nervoso centrale


1991 - Effects of chronic treatment with flunitrazepam on GABAA, adenosine and glutamate receptor plasticity in rats [Relazione in Atti di Convegno]
Baraldi, Mario; Zanoli, Paola; A., Giacobazzi; G., Vaccari; Avallone, Rossella
abstract

A crhonic exposure to an anxyolitic dose of flunitrazepam induces an increased funactional activity of the GABAA receptors without affecting the purinergic system.


1990 - Decreased presence of zinc in brain areas of chronically morphine-exposed rats [Articolo su rivista]
Avallone, R.; Vaccari, G.; Giacobazzi, A.; Zanoli, P.; Baraldi, M.
abstract


1990 - Dence that a chronic treatment with RO 15-3505, a partial negative allosteric modulator of GABAA receptor, affects the glutamatergic system and improve learning in rats [Articolo su rivista]
Giacobazzi, A.; Vaccari, G.; Avallone, R.; Ventura, A.; Baraldi, M.
abstract


1990 - Effect of chronic treatment with flunitrazepam on the cerebral purinergic system in rats [Articolo su rivista]
Vaccari, G.; Giacobazzi, A.; Avallone, R.; Ventura, A.; Baraldi, M.
abstract


1990 - Urine retention indeced in rats by intra-spinal cord injection of colchicine as a tool to find new pharmacological treatment [Relazione in Atti di Convegno]
Baraldi, Mario; Zanoli, Paola; C., Truzzi; G., Vaccari; A., Giacobazzi; Avallone, Rossella
abstract

we report thet the administration of 30 mg/Kg/day of monosialoganglioside in combination with a single i.s.c. administration of NGF (0.8 microgram/rat) improved the finctional recovery of the micturition function judging from urine emission and content in the bladder at day 3 and 7.