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Professore Associato presso: Dipartimento di Scienze della Vita sede ex-Biologia (Orto Botanico)

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2020 - Carte verdi nell’Archivio di Stato di Modena: l’Erbario Estense, foglie tra i fogli, un rebus, un progetto. Parte II [Articolo su rivista]
Vicentini, C. B.; Buldrini, F.; Bosi, G.; Bonazza, M.; Romagnoli, C.

The Este Herbarium, preserved in the Este Secret Archives of the Modena State Ar- chives, is one of the few surviving Renaissance herbaria to date. It is the most fascinating and mysterious. It is probably the sixth volume of a lost series. In the present article, we discuss the results of a joint work (still in progress) among the Universities of Ferrara, Modena and Reggio Emilia, and Bologna. All the correspondence between simplists and botanists of that epoch (primarily Ulisse Aldrovandi from Bologna) was thoroughly analysed, to search the solution of the rebus: which author (or authors), which temporal location, which context of production for this herbarium? The documents consulted allow to understand that living plants, seeds and illustra- tions were commonly exchanged, but herbarium sheets never. The consultations on ta- xonomic attributions were frequent. The term herbario/herbarj, that is recurring in the letters written by Alfonso Cattaneo to Ulisse Aldrovandi, is also mysterious (thing or person?).

2020 - Tobacco in the Erbario Estense and other Renaissance evidence of the Columbian taxon in Italy [Articolo su rivista]
Vicentini, C. B.; Buldrini, F.; Romagnoli, C.; Bosi, G.

The Erbario Estense, preserved in the Archivio Segreto Estense (Modena State Archives, Italy), is one of the very few sixteenth century herbaria still existing today. Among its exsiccata are a dozen species coming from the Americas, one of which is tobacco. The author of the specimen calls the plant Tabacho, ouer Herba Regina; Camus and Penzig, in the late nineteenth century, identify it as Nicotiana tabacum and affirm that it could be the most ancient direct proof of the presence of this plant in Italy. Today, attribution of the specimen to the above-mentioned species is certain and, according to the studies carried out for the present research, only three other sixteenth century exsiccata of N. tabacum still exist, all of them preserved in the Erbario Aldrovandi in Bologna. Therefore, the specimen of the Erbario Estense is extremely precious from a historical and scientific viewpoint. Tobacco was certainly known by the simplists who were working at that epoch in the lively scientific and medical environment of Ferrara, even if, according to documentary sources, real pharmacological use of the plant seems to have taken place only in successive phases.

2018 - Melissa officinalis L. subsp. altissima (Sibth. & Sm.) Arcang. essential oil: Chemical composition and preliminary antimicrobial investigation of samples obtained at different harvesting periods and by fractionated extractions [Articolo su rivista]
Bozovic, M.; Garzoli, S.; Baldisserotto, A.; Romagnoli, C.; Pepi, F.; Cesa, S.; Vertuani, S.; Manfredini, S.; Ragno, R.

A comprehensive study on essential oils extracted from different samples of Melissa officinalis L. subsp. altissima (Sibth. & Sm.) Arcang. (Lamiaceae) collected in Tarquinia (Italy) is reported. In this study, the 24-h steam distillation procedure for essential oil preparation, in terms of different harvesting periods and extraction times was applied. The GC/MS analysis showed cis-caryophyllene and its oxide as the major essential oil's constituents. A great impact of phenological stage on chemical profile of essential oil was observed. Regarding the duration of the extraction process, prolonged distillation led to chemically more diverse oil samples. Preliminary microbiological evaluations of the essential oils samples revealed the lack of significant efficacy against Candida albicans.

2018 - “Spigo nardo”: from the Erbario Estense a possible solution for its taxonomical attribution [Articolo su rivista]
Vicentini, Chiara Beatrice; Buldrini, Fabrizio; Bosi, Giovanna; Romagnoli, Carlo

The spikenard is a plant mentioned since early antiquity, mostly known for its intense and pungent aroma that made it a precious ingredient of many cosmetic recipes for luxury perfumes. The nature of the spikenard has been controversial since the ancient era and, still today, there is the need for a correct attribution of such a name to a currently accepted botanical species. Thanks to a specimen from the Este Herbarium (Erbario Estense), preserved in the Modena State Archives (Italy), we tried to bring our contribution to the debate concerning this topic. We checked all the Italian ancient herbaria in search of samples of spikenard, analysed past literary and medical–pharmaceutical sources from classical antiquity to the XIX century, and searched former and present ethno-botanical uses. Our investigation allowed us to ascertain that the spikenard samples preserved in the Renaissance Italian herbaria belong to Nardostachys jatamansi (D. Don) DC., a herbaceous species belonging to the Valerianaceae, native to alpine regions of the north-western Himalaya, still used in Ayurveda medicine and as a component of cosmetic products.

2017 - Essential oil extraction, chemical analysis and anti-candida activity of calamintha nepeta (L.) Savi subsp. glandulosa (Req.) ball-new approaches [Articolo su rivista]
Božović, Mijat; Garzoli, Stefania; Sabatino, Manuela; Pepi, Federico; Baldisserotto, Anna; Andreotti, Elisa; Romagnoli, Carlo; Mai, Antonello; Manfredini, Stefano; Ragno, Rino

A comprehensive study on essential oils extracted from different Calamintha nepeta (L.) Savi subsp. glandulosa (Req.) Ball samples from Tarquinia (Italy) is reported. In this study, the 24-h steam distillation procedure for essential oil preparation, in terms of different harvesting and extraction times, was applied. The Gas chromatography-mass spectrometry (GC/MS) analysis showed that C. nepeta (L.) Savi subsp. glandulosa (Req.) Ball essential oils from Tarquinia belong to the pulegone-rich chemotype. The analysis of 44 samples revealed that along with pulegone, some other chemicals may participate in exerting the related antifungal activity. The results indicated that for higher activity, the essential oils should be produced with at least a 6-h steam distillation process. Even though it is not so dependent on the period of harvesting, it could be recommended not to harvest the plant in the fruiting stage, since no significant antifungal effect was shown. The maximum essential oil yield was obtained in August, with the highest pulegone percentage. To obtain the oil with a higher content of menthone, September and October should be considered as the optimal periods. Regarding the extraction duration, vegetative stage material gives the oil in the first 3 h, while material from the reproductive phase should be extracted at least at 6 or even 12 h.

2016 - Antidermatophytic action of resorcinol derivatives: Ultrastructural evidence of the activity of phenylethyl resorcinol against microsporum gypseum [Articolo su rivista]
Romagnoli, Carlo; Baldisserotto, Anna; Vicentini, Chiara B.; Mares, Donatella; Andreotti, Elisa; Vertuani, Silvia; Manfredini, Stefano

In this work, we evaluated the antidermatophytic activities of three resorcinol derivatives that have a history of use in dermo-cosmetic applications to discover molecules with multiple dermatological activities (i.e., multi-Target drugs), thereby reducing the cost and time necessary for new drug development. The antidermatophytic activities of the three skin lighteners were evaluated relative to the known antifungal drug fluconazole on nine dermatophytes responsible for the most common dermatomycoses: Microsporum gypseum, Microsporum canis, Trichophyton violaceum, Arthroderma cajetani, Trichophyton mentagrophytes, Epidermophyton floccosum, Nannizzia gypsea, Trichophyton rubrum and Trichophyton tonsurans. Among the three tested resorcinols, only two showed promising properties, with the ability to inhibit the growth of all tested dermatophytes; additionally, the IC50 values of these two resorcinols against the nine dermatophytes confirmed their good antifungal activity, particularly for phenylethyl resorcinol against M. gypseum. Ultrastructural alterations exhibited by the fungus were observed using scanning electron microscopy and transmission electron microscopy and reflected a dose-dependent response to treatment with the activation of defence and self-preservation strategies.

2015 - A Multi-Target Approach toward the Development of Novel Candidates for Antidermatophytic Activity: Ultrastructural Evidence on α-Bisabolol-Treated Microsporum gypseum [Articolo su rivista]
Romagnoli, Carlo; Baldisserotto, Anna; Malisardi, Gemma; Vicentini, Chiara Beatrice; Mares, Donatella; Andreotti, Elisa; Vertuani, Silvia; Manfredini, Stefano

Multi-target strategies are directed toward targets that are unrelated (or distantly related) and can create opportunities to address different pathologies. The antidermatophytic activities of nine natural skin lighteners: α-bisabolol, kojic acid, β-arbutin, azelaic acid, hydroquinone, nicotinamide, glycine, glutathione and ascorbyl tetraisopalmitate, were evaluated, in comparison with the known antifungal drug fluconazole, on nine dermatophytes responsible for the most common dermatomycoses: Microsporum gypseum, Microsporum canis, Trichophyton violaceum, Nannizzia cajetani, Trichophyton mentagrophytes, Epidermophyton floccosum, Arthroderma gypseum, Trichophyton rubrum and Trichophyton tonsurans. α-Bisabolol showed the best antifungal activity against all fungi and in particular; against M. gypseum. Further investigations were conducted on this fungus to evaluate the inhibition of spore germination and morphological changes induced by α-bisabolol by TEM.

2013 - A multivariate analysis approach to the study of chimica and functional properties of chemodiverse plant derivatives: lavender essential oil [Articolo su rivista]
Maietti, Silvia; Rossi, Damiano; Guerrini, Alessandra; Useli, Chiara; Romagnoli, Carlo; Poli, Ferruccio; Bruni, Renato; Sacchetti, Gianni

Six lavender essential oils, L. angustifolia and five Lavandula x hybrida cultivars (Super Z, Abrialis, R.C., Alardii and Ordinario), were evaluated from a phytochemical and biological standpoint, and the results were computed by using multivariate data analysis. The essential oils were analysed by gas chromatography, gas chromatography– mass spectrometry and headspace gas-chromatography. Multivariate analyses (principal component analysis) identified three main phytochemical clusters among lavender essential oils, represented by 1,8-cineole, linalyl acetate and linalool. Functional properties of the essential oils were checked by (1) estimating cytotoxicity and genotoxicity using the Saccharomyces cerevisiae D7 strain; (2) determining antifungal activity against three common phytopathogens (Pythium ultimum, Magnaporthe grisea and Botrytis cinerea), by performing an agar vapour bioassay; and (3) calculating the antioxidant capacity by using the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and b-carotene bleaching tests. No mutagenic effects were detected, but multivariate analyses (PLS, partial least squares regression) showed that the essential oils belonging to the linalool cluster were the most cytotoxic. Antifungal activity against phytopathogens confirmed the predictive results of PLS. The differences among lavender essential oils regarding weak antioxidant capacity showed a positive relationship between the high polarity compounds and the DPPH method, as determined by PLS. The opposite effect was shown for the same type of compound and b-carotene. Ketones and esters did not exert any significant antioxidant activity. In conclusion, taking lavender essential oils as a model and computing multivariate data of a reduced number of parameters, the proposed approach assured the description of the relationship between a phytocomplex, its constituents and bioactivities, and allowed a comprehensive, predictive approach to be defined, for which the chemical profile provides a possible synergic overall effort in terms of applicative perspectives.

2012 - Synthesis, Antioxidant and Antimicrobial Activity of a New Phloridzin Derivative for Dermo-cosmetic Applications [Articolo su rivista]
A., Baldisserotto; G., Malisardi; E., Scalambra; Andreotti, Elisa; Romagnoli, Carlo; C. B., Vicentini; S., Manfredini; S., Vertuani

The phenolic compound phloridzin (phloretin 2′-O-glucoside, variously named phlorizin, phlorrhizin, phlorhizin or phlorizoside) is a prominent member of the chemical class of dihydrochalcones, which are phenylpropanoids. Phloridzin is specifically found in apple and apple juice and known for its biological properties. In particular we were attracted by potential dermo-cosmetic applications. Here we report the synthesis, stability studies and antimicrobial activity of compound F2, a new semi-synthetic derivative of phloridzin. The new derivative was also included in finished formulations to evaluate its stability with a view to a potential topical use. Stability studies were performed by HPLC; PCL assay and ORAC tests were used to determine the antioxidant activity. F2 presented an antioxidant activity very close to that of the parent phloridzin, but, unlike the latter, was more stable in formulations. To further explore potential health claims, antifungal activity of phloridzin and its derivative F2 were determined; the results, however, were rather low; the highest value was 31,6% of inhibition reached by F2 on Microsporum canis at the highest dose.

2011 - Antidermatophytic activity of pyrazolo[3,4-c]isothiazoles: a preliminary approach on 4-chlorophenyl derivative for evaluation of mutagenic and clastogenic effects on bacteria and human chromosomes in vitro [Articolo su rivista]
D., Rossi; D., Mares; Romagnoli, Carlo; Andreotti, Elisa; S., Manfredini; C. B., Vicentini

The antifungal activity of eight pyrazolo[3,4-c]isothiazole derivatives was evaluated on five 21 dermatophytes: three are anthropophilic species, i.e. Epidermophyton floccosum, Trichophyton 22 rubrum and Trichophyton tonsurans, and two are geophilic species, Microsporum gypseum and 23 Nannizia cajetani. The new compounds proved to be unlikely effective in inhibiting the growth of 24 the different strains. In general the fungi parasitic on man were more sensitive than the geophilic25 species. This fact can be positive for a possible practical-therapeutic utilization of these class of 26 compounds. To verify their possible use against fungi of medical interest, the most interesting 27 substance at low doses, 6-(4-chlorophenyl)-4-methyl-6H-pyrazolo[3,4-c]isothiazol-3-amine, was 28 chosen to perform in vitro genotoxicity tests using: Salmonella/microsome test (SAL), Sister 29 Chromatid Excange test (SCE), Cytokinesis-Blocked Micronucleus test (CBMN) and its 30 improvement (Ara-C/CBMN). The compound showed no mutagenic activity at low doses, whereas 31 at the highest dose (100 μg/mL) it causes a generalized cytotoxic effect. The high growth inhibition32 exerted on fungi at the lowest dose and the concomitant lack of genotoxicity, at least until the dose 33 of 50 μg/mL, might suggest the compound as a safe candidate as antidermatophytic substance.

2011 - Chemical characterization (GC-MS and NMR fingerprinting) and bioactivities of South-African Pelargonium capitatum (L.) L’Herit. (Geraniaceae) essential oil [Articolo su rivista]
A., Guerrini; D., Rossi; G., Paganetto; M., Tognolini; M., Muzzoli; Romagnoli, Carlo; F., Antonioni; S., Vertuani; A., Medici; A., Bruni; C., Useli; E., Tamburini; R., Bruni; G., Sacchetti

Chemical fingerprinting of commercial Pelargonium capitatum (Geraniaceae) essential oil samples of south African origin was performed by GC, GC/MS, and 13 C- and 1H-NMR. Thirty-seven compounds were identified, among which citronellol (32.71%) and geraniol (19.58%) were the most abundant. NMR Spectra of characteristic chemicals were provided. Broad-spectrum bioactivity properties of the oil were evaluated and compared with those of commercial Thymus vulgaris essential oil with the aim to obtain a functional profile in terms of efficacy and safety. P. capitatum essential oil provides a good performance as antimicrobial, with particular efficacy against Candida albicans strains. Antifungal activity performed against dermatophyte and phytopathogen strains revealed the latter as more sensitive, while antibacterial activity was not remarkable against both Gram-positive and Gram-negative bacteria. P. capitatum oil provided a lower antioxidant activity (IC 50 ) than that expressed by thyme essential oil, both in the 1,1- diphenyl-2-picrylhydrazyl (DPPH) and b-carotene bleaching tests. Results in photochemiluminescence (PCL) assay were negligible. To test the safety aspects of P. capitatum essential oil, mutagenic and toxicity properties were assayed by Ames test, with and without metabolic activation. Possible efficacy of P. capitatum essential oil as mutagenic protective agent against NaN 3 , 2-nitrofluorene, and 2-aminoanthracene was also assayed, providing interesting and significant antigenotoxic properties

2011 - Pyrazolo[3,4-c]isothiazole and isothiazolo[4,3-d]isoxazolederivatives as antifungal agents [Articolo su rivista]
C. B., Vicentini; Romagnoli, Carlo; S., Manfredini; D., Rossi; D., Mares

Context: The diseases of plants and humans due to pathogenic fungi are increasing. Among the substances used tocombat fungi, the azoles are of primary interest, both in agricultural field both in health. To avoid fungal resistancephenomena, the synthesis and tests of new derivatives are necessary.Objective: This article discusses the synthesis and the antifungal activity of pyrazolo[3,4-c]isothiazole andisothiazolo[4,3-d]isoxazole derivatives against three fungi that are pathogenic only for plants and two fungi that areopportunistic in humans and plants.Materials and Methods: The compounds were prepared starting from 2-cyano-3-ethoxy-2-butenethioamide. Theantifungal activity of the compounds was determined by measuring the inhibition of growth of the fungi tested at20, 50, and 100 μg/mL in comparison with the controls.Results: Results demonstrated that several compounds were able to control the mycelial growth of the tested fungi,even if they showed different sensitivity to the different azole-derivatives. In general Magnaporthe grisea (T.T. Hebert)Yaegashi & Udagawa was the most sensitive fungus, being blocked almost entirely by 4-chloro derivative even at 20μg/mL, a concentration at which the reference commercial compound tricyclazole was nearly ineffective.Discussion and conclusion: These findings demonstrate that the pyrazolo[3,4-c]isothiazole derivatives have a widespectrum of activity on phytopathogenic and opportunistic fungi. In particular the 4-chloro derivative seems to havea great potential as new product to combat M. grisea in the agricultural field.

2010 - Antifungal activity of essential oil from fruits of Indian Cuminum cyminum [Articolo su rivista]
Romagnoli, Carlo; Andreotti, Elisa; S., Maietti; Mares, R. M. a. h. e. n. d. r. a. D.

The essential oil of fruits of Cuminum cyminum L. (Apiaceae), from India, was analyzed by GC and GC-MS, and its antifungal activity was tested on dermatophytes and phytopathogens, fungi, yeasts and some new Aspergilli. The most abundant components were cumin aldehyde, pinenes, and p-cymene, and a fraction of oxygenate compounds such as alcohol and epoxides. Because of the large amount of the highly volatilecomponents in the cumin extract, we used a modified recent technique to evaluate the antifungal activity only of the volatile parts at doses from 5 to 20 μL of pure essential oil. Antifungal testing showed that Cuminum cyminum is active in general on all fungi but in particular on the dermatophytes, where Trichophyton rubrum was the most inhibited fungus also at the lowest dose of 5 μL. Less sensitive to treatmentwere the phytopathogens.

2010 - La scienza delle piante officinali: passato, presente e futuro [Altro]
Benvenuti, Stefania; P., Paltrinieri; Pellati, Federica; Romagnoli, Carlo; Rossi, Tiziana; Zanoli, Paola

Il Congresso “La scienza delle piante officinali: passato, presente e futuro” si è svolto presso il Centro Servizi Didattici della Facoltà di Medicina e Chirurgia e si è proposto come un’opportunità di incontro fra la realtà del settore produttivo e quello della formazione, della ricerca e dell’innovazione nello studio delle piante officinali e dei principi biologicamente attivi in esse contenuti. Non sono mancati i riferimenti storici e culturali dal momento che l’Archivio di Stato e la Biblioteca Estense di Modena e hanno aderito a tale iniziativa e hanno ospitato una sezione di questo congresso, in cui saranno presentate opere e documenti relativi al mondo delle piante officinali.La Facoltà di Farmacia, che ha da anni istituito un corso di laurea di primo livello in Tecniche Erboristiche, è attivamente impegnata nella formazione di esperti del settore delle piante officinali e ha laureato numerosi giovani che hanno apportato in questo ambito un notevole contributo culturale e professionale. L’attività didattica è sempre stata accompagnata da una continua e approfondita attività di ricerca dei Docenti, che hanno lavorato in stretta collaborazione con aziende del territorio, nazionali ed internazionali. Ne sono testimonianza le numerose pubblicazioni scientifiche prodotte dai Ricercatori del nostro Ateneo in questo campo di indagine.Patrocinato dalla Società Botanica Italiana, dalla Società di Farmacognosia, da CO.N.P.T.ER e da Comune e Provincia di Modena, oltre che dai Dipartimenti Universitari di Scienze Farmaceutiche e di Scienze Biomediche, che hanno ricevuto il sostegno della Fondazione Cassa di Risparmio di Modena e di aziende e società del settore, il convegno ha riunito docenti ed esperti provenienti da tutta Italia per discutere di aspetti legati alla coltivazione delle piante officinali, al loro utilizzo nella formulazione di prodotti erboristici, fitoterapici e fitocosmetici, nonché dell’importanza del controllo di qualità e della fitovigilanza.

2008 - Le Legnose dell'Orto Botanico: II. Juglans L. [Articolo su rivista]
Bosi, Giovanna; Dallai, Daniele; Maffettone, Luigi; Rinaldi, Rossella; Romagnoli, Carlo; Mazzanti, Marta

Secondo contributo alla conoscenza di alcune entità legnose interessanti per aspetto, portamento e maestosità dell’Orto Botanico dell’Università degli Studi di Modena e Reggio Emilia. In questo articolo sono presentate le specie presenti in Orto afferenti al genere Juglans, con particolare riguardo a Juglans regia, meritevole di attenzione sia dal punto di vista botanico che per i suoi molteplici rapporti, passati e attuali, con l’uomo.

2008 - Three New Species of Aspergillus from Amazonian Forest Soil (Ecuador). [Articolo su rivista]
D., Mares; Andreotti, Elisa; M. E., Maldonado; P., Pedrini; C., Colalongo; Romagnoli, Carlo

From an undisturbed natural forest soil inEcuador, three fungal strains of the genus Aspergillus wereisolated. Based on molecular and morphological featuresthey are described as three new species, named A. quitensis,A. amazonicus, and A. ecuadorensis.

2007 - Synthetic Pyrazole derivatives as growth inhibitors of some phytopathogenic fungi [Articolo su rivista]
C. B., Vicentini; Romagnoli, Carlo; Andreotti, Elisa; D., Mares

The present study was carried out to investigate the antifungal activity of pyrazole/isoxazole-3-carboxamido-4-carboxylic acids, 4-oxo-5-substituted pyrazolo[3,4-d]pyrimidine-6-thiones, and N-alkyl/aryl-N′-(4-carbethoxy-3-pyrazolyl)thioureas against Pythium ultimum, Botrytis cinerea, and Magnaporthegrisea. The results on growth inhibition showed differences in the sensitivity of the threefungi to the tested substances, and in general P. ultimum was shown to be the most sensitive. On allphytopathogens the best results within the pyrazole/isoxazolecarboxamide series are given bythe compounds with the carboxamide and carboxylic groups in positions 3 and 4; the presence ofthese groups seems to be critical for biological activity in this series of compounds. Among thepyrazolopyrimidines the derivative supplied with the benzylic group was the most active on the threefungi and in particular against P. ultimum. Several compounds belonging to the thiourea series areable to inhibit selectively M. grisea at 50 and 10 μg mL-1, doses at which the reference commercialcompound tricyclazole had low or no effect.

2006 - Antimutagenic, antioxidant and antimicrobial properties of Maytenus krukovii bark [Articolo su rivista]
R., Bruni; D., Rossi; M., Muzzoli; Romagnoli, Carlo; G., Paganetto; E., Besco; F., Choquecillo; K., Peralta; W. S., Lora; G., Sacchetti

The hydroalcoholic extract of Maytenus krukovii bark was investigated for its in vitro mutageno-protective activities by means of the Ames Salmonella/microsome assay. The extract showed an inhibitory effect in both T98 and T100 strains against the mutagenic activity of promutagen 2-aminoanthracene but was not protective against directly acting mutagens sodium azide and 2-nitrofluorene. When tested as a radical scavenger and antioxidant it produced a dose-dependent inhibition. The extract did not show significant antibacterial properties, and was weakly active against dermatophyte and phytopathogenic fungi, but inhibited the growth of phytopathogen Pithyum ultimum. (c) 2006 Elsevier B.V. All rights reserved.

2006 - Emerging antifungal azoles and effects on Magnaporthe grisea [Articolo su rivista]
D., Mares; Romagnoli, Carlo; Andreotti, Elisa; G., Forlani; S., Guccione; C. B., Vicentini

Derivatives of pyrazolo[1,5-a][1, 3, 5]triazine-2,4-dione,pyrazolo[1,5-c][1, 3, 5] thiadiazine-2-one, pyrazolo[3,4-d][1, 3]thiazine-4-one, and pyrazolo[3,4-d][1, 3]thiazine-4-thione were screened for antifungal activity against the causal agent of rice blast disease, Magnaporthe grisea. The compounds were tested at doses ranging from 10 to 200 mu g ml(-1), using the commercial fungicide tricyclazole as reference compound. All triazine derivatives inhibited the growth and pigmentation of the mycelia less effectively than tricyclazole. The thiadiazine derivatives proved to be more effective than their triazine counterparts, but only 4-(butylimino)-7-methylpyrazolo [1, 5-c] [1, 3,5] thiadiazine-2-one (2h) and 4-(cyclohexylimino) -7- methylpyrazolo [1, 5-c] [1, 3,5] thiadiazine-2-one (2j) were more effective than tricyclazole. Pyrazolo[3,4-d][1,3]thiazine-4-one derivatives were active only at the highest doses, whereas members of the pyrazolo[3,4-d][1,3]thiazine-4-thione series inhibited fungal growth at the lowest concentrations used, at which tricyclazole had no effect. A dose-dependent mechanism might be responsible for this effect, with lipophilicity as the governing factor. Within a given set, the presence of a cyclohexyl or an n-butyl group generally increased antifungal activity, with respect to both growth inhibition and cell de-pigmentation of the mycelium, suggesting that a higher lipophilicity might improve transport inside the cells. SEM and TEM of M. grisea hyphae showed that treatment with the most active substance (2h) caused significant ultrastructural effects, particularly on the endomembrane system, suggesting a mechanism of action similar to that of most azole fungicides. Dissimilarities were also observed, with no alterations of the cell wall evident. In conclusion, several compounds showed greater inhibition than tricyclazole, and therefore provide useful new chemistry for control of M. grisea infections.

2006 - Reperti carpologici in due boccali (Convento benedettino di S. Antonio in Polesine - Ferrara, fine XV-XVI sec. d.C.): possibili documenti di antiche preparazioni officinali [Relazione in Atti di Convegno]
Romagnoli, Carlo; Bosi, Giovanna; Mazzanti, Marta

During the archaeological excavations in the Benedictine Convent of S. Antonio in Polesine- FE, were collected six vessels dated back to the end of XV and the begin of XVI century. Results of archaeocarpological analyses carried out in their fills (and in the sediments round the vessels), were reported. Only two vessels (Pot n° 1 – US 46 and Pot n° 5 – US 420) contained seeds/fruits and the aim of this work was to identify the plant species and the most probably use of the content of each vessel. The seeds/fruits (sf), in mediocre conservation state, belonged to few and different taxa. All the taxa were common medicinal plants. This fact, together with various other clues, indicated that their presence in the vessels was not casual, but that they were introduced with a precise aim; we hypothesized that the vessel probably contained preparations for therapeutical use. In particular we think that the plants contained in the Pot n° 1 (Ficus carica, Galium cf. aparine, Sambucus ebulus, Umbelliferae) probably could be used as a topic remedy for skin diseases. The Pot n° 5 contained, instead, abundant sf of five plants (Chenopodium/Atriplex, Euphorbia peplus, Malva sp., Mercurialis annua, Portulaca oleracea); for all these species it was possible to found a synergic action, on the gastro enteric apparatus. So our hypothesis was that this preparation possessed strong cathartic action due to Euphorbia peplus, and Mercurialis annua, alleviated by the antispasmolytic action of the other plants. In conclusion, even if we are conscious that may appear hazardous our attempt to understand the possible therapeutic use of these mixtures prepared long time ago with different knowledges and way of life, we retain that our hypothesis, based on the reference to books dated back to that age, could be enough realistic and not in contrast with the current knowledgements on these plants.

2006 - Semi e frutti in boccali del monastero benedettino di S. Antonio in Polesine: indizi di antiche preparazioni officinali? [Capitolo/Saggio]
Mazzanti, Marta; Bosi, Giovanna; Romagnoli, Carlo

La presenza di semi e frutti all'interno di boccali rinvenuti in indagini archeologiche presso il monastero benedettino di S. Antonio a Ferrara, ha portato ad ipotesi di antiche preparazioni officinali

2005 - Caratterizzazione chimica e funzionale di fonti vegetali di interesse officinale. [Articolo su rivista]
Mares, D; Medici, A; Pedrini, P; Tosi, B; Sacchetti, G; Romagnoli, Carlo; VICENTINI C., B; Guerrini, A; Maietti, S; Andreotti, Elisa; Rambaldi, A; Muzzoli, M.

La ricerca è impostata su criteri multidisciplinari con finalità applicative. Circa la caratterizzazione chimica della fonte vegetale vengono applicate strategie estrattive ed analitiche al fine di avere una completa impronta biochimica. Le finalità applicative spaziano da ricerche di enzimi per biotrasformazioni alla determinazione dell'eventuale attività biologica( antifungina, antibatterica, antiossidante)dei fitocomplessi e/o delle classi chimiche isolate

2005 - Chemical characterization and antifungal activity of essential oil of capitula from wild Indian Tagetes patula L. [Articolo su rivista]
Romagnoli, Carlo; R., Bruni; Andreotti, Elisa; M. K., Rai; C. B., Vicentini; D., Mares

The essential oil extracted by steam distillation from the capitula of Indian Tagetes patula, Asteraceae, was evaluated for its antifungal properties and analyzed by gas chromatography and gas chromatography-mass spectrometry. Thirty compounds were identified, representing 89.1% of the total detected. The main components were piperitone (24.74%), piperitenone (22.93%), terpinolene (7.8%), dihydro tagetone (4.91%), cis-tagetone (4.62%), limonene (4.52%), and allo-ocimene (3.66%). The oil exerted a good antifungal activity against two phytopathogenic fungi, Botrytis cinerea and Penicillium digitatum, providing complete growth inhibition at 10 μ l/ml and 1.25 μ l/ml, respectively. The contribution of the two main compounds, piperitone and piperitenone, to the antifungal efficacy was also evaluated and ultrastructural modifications in mycelia were observed via electron microscopy, evidencing large alterations in hyphal morphology and a multisite mechanism of action.

2005 - Chicory extracts from Cichorium intybus L. as potential antifungals [Articolo su rivista]
D., Mares; Romagnoli, Carlo; B., Tosi; Andreotti, Elisa; G., Chillemi; F., Poli

Abstract In this work extracts from roots of the common vegetable Cichorium intybus L., highly appreciated for its bitter taste, were studied to investigate their possible biological activity on fungi from a variety of ecological environments: some are parasites on plants (phytopathogens) or of animals and humans (zoophilic and anthropophilic dermatophytes), others live on the soil and only seldom parasitize animals (geophilic dermatophytes). The extracts were ineffective on geophilic species and on tested phytopathogens, with the exception of Pythium ultimum, whereas they inhibited the growth of zoophilic and anthropophilic dermatophytes, in particular Trichophyton tonsurans var. sulfureum, whose treatment caused morphological anomalies, here observed by scanning electron microscopy. This behaviour is discussed on the basis of the presence in the chicory extract of the two main sesquiterpene lactones, 8-deoxylactucin and 11β,13-dihydrolactucin.

2005 - Le legnose dell'Orto Botanico di Modena. I. Taxus L. e Taxus baccata L. [Articolo su rivista]
Torri, Paola; Dallai, Daniele; Bosi, Giovanna; Maffettone, Luigi; Romagnoli, Carlo; Mazzanti, Marta

Primo contributo alla conoscenza di alcune entità legnose interessanti per aspetto, portamento e maestosità dell'Orto Botanico dell'Università degli Studi di Modena e Reggio Emilia. In questo articolo è presentato il tasso (Taxus baccata L.) meritevole di attenzione sia dal punto di vista botanico che per i suoi molteplici rapporti, passati ed attuali, con l'uomo

2004 - Antifungal activity of Tagetes patula extracts on some phytopathogenic fungi: ultrastructural evidence on Pythium ultimum [Articolo su rivista]
D., Mares; B., Tosi; F., Poli; Andreotti, Elisa; Romagnoli, Carlo

Methanol extract, obtained from Tagetes patula plant, was assayed against three phytopathogenic fungi: Botrytis cinerea, Fusarium moniliforme and Pythium ultimum. The antifungal activity was tested both in the dark and in the light, using two different Lighting systems. The data showed that the extract proved to have a dose-dependent activity on all the fungi with a marked difference between treatments in the tight than in the dark. Good growth inhibition was observed in fungi only when these were treated with the highest dose of the extract and irradiated, whereas the same dose gave only a modest inhibition when the experiment was conducted in the dark. At 5 and 10 mug/ml in the dark, growth increased. The results indicated that the presence of a luminous source enhances the antifungal activity, with small differences between UV-A and solar spectrum Light. SEM and TEM observations on Pythium ultimum revealed that the Tagetes patulo extract induced alterations on cell fungal membranes with a photoactivation mechanism possibly involving the production of free radicals and Leading to a premature aging of the mycelium.

2004 - Chemical composition and biological activities of Ishpingo essential oil, a traditional Ecuadorian spice from Ocotea quixos (lam.) Kosterm. (Laureaceae) flower calices. [Articolo su rivista]
R., Bruni; A., Medici; Andreotti, Elisa; C., Fantin; M., Muzzoli; M., Dehesa; Romagnoli, Carlo; G., Sacchetti

The essential oil of Ishpingo (Ocotea quixos, Lauraceae) fruit calices was analysed by GC (gas chromatography) and GC-MS (gas chromatography mass spectrometry). Fourty-four compounds were identified. The main components detected were transcinnamaldehyde (27.9%), methylcinnamate (21.6%), 1,8-cincole (8.0%), benzaldehyde (3.6%), and beta-selinene (2.1%). In vitro antioxidant properties of the essential oil, obtained by DPPH (1,1-diphenyl-2-picrylhydrazyl) and beta-carotene bleaching assays, were also evaluated. The oil exerted a relatively good capacity to act as a non-specific donor of hydrogen atoms or electrons when checked by the diphenylpicrylhydrazyl assay, quenching 52% of the radical. On the other hand, it showed weak effects in inhibiting oxidation of linoleic acid when assayed by the beta-carotene bleaching test. Antibacterial activity of the essential oil was also checked against gram positive (Enterococcus foecalis, Staphylococcus aureus) and gram negative strains (Escherichia coli, Pseudomonas aeruginosa). The oil also showed a dose-dependent antifungal activity against Candida albicans, Saccharomyces cerevisiae, phytopathogen Pythium ultimum and dermatophyte Trichophyton mentagrophytes.

2004 - Synthesis and antifungal action of new tricyclazole analogues [Articolo su rivista]
D., Mares; Romagnoli, Carlo; Andreotti, Elisa; M., Manfrini; Cb, Vicentini

Melanins are very important pigments for the survival and longevity of fungi, so their biosynthesis inhibition is a new biochemical target aiming at the discovery of selective fungicides. In this work is described the synthesis of new pyrazolo-thiazolo-triazole compounds, analogues of tricyclazole (a commercial antifungal product that acts by inhibiting melanin synthesis), and their biological activity was studied on some dermatophytes and phytopathogens. The compounds poorly inhibited the growth and pigmentation of fungi tested and were less efficient than tricyclazole. Electron microscopy on Botrytis cinerea showed that treatment with the most active compound caused abnormally thickened and stratified walls in fungi, whose ultrastructure was, in contrast, generally normal. The fungus treated with tricyclazole, on the other hand, appeared to be drastically altered, so as to become completely disorganized. These results suggest that the new azole compounds employ an action mechanism similar to that of other azoles, but dissimilar to that of tricyclazole.

2003 - A new tecnique for the evaluation of antifungal activity of an alcohol extract of Eugenia caryophyllata Thunberg on Pennicillium digitatum [Capitolo/Saggio]
Romagnoli, Carlo; Sacchetti, G.

The purpose of the present work was to test the activity of an extract of cloves E. caryophyllata Thunberg using the phytopathogen Penicillium digitatum as test fungus and to identify which components are most likely responsible for its antifungal activity. This plant is well known from the phytochemical point of view and for its antibacterial, antifungine activity and other biological properties. To obtain results with this fungus that shows a very fast growth a new, more appropriate technique, counting the number of colonies formed within 48 hours, had to be set up. The extract and the pure standard eugenol were highly active, totally preventing fungal growth, even at the lowest doses, karyophyllene did not shown any effect.

2002 - Antifungal activity of 5 new synthetic compounds vs. Trichophyton rubrum and Epidermophyton floccosum [Articolo su rivista]
Romagnoli, Carlo; Mares, D; Bruni, A; Andreotti, Elisa; Manfrini, M; Vicentini, Cb

The antifungal activity of five new synthetic compounds was evaluated on two dermatophytes: Epidermophyton floccosum and Trichophyton rubrum. The data showed that the imidazo-pyrazole and pyrazolo-thiazoles were not particularly effective, while the two pyrazole-thiocyanates proved highly active on both fungi. The most active 5-amino-3-methyl-1-phenylpyrazolo-4-thiocyanate was chosen to perform SEM and TEM morphological studies on both fungi. Both SEM and TEM observations revealed interesting alterations on the two dermatophytes, particularly involving the endomembrane system.

2002 - Antitungal activity of Tagetes patula extracts [Articolo su rivista]
D., Mares; B., Tosi; Romagnoli, Carlo; F., Poli

In the present paper the methanol extracts obtained from 10 cultivars of Tagetes patula were assayed on two phytopathogenic fungi: Botrytis cinerea and Fusarium moniliforme, B. cinerea showed a high dose-dependent inhibition, with a marked difference between light and dark treatment. F. moniliforme seems to be a more resistant test that does not appear to be affected by the different treatment conditions (light-dark) even at the highest dose. However, it can be asserted that Tagetes patula is a possible source of antifungal substances and that thiophene activity is, in general, strongly increased by UV-A irradiation. During the experiment the method of chromatographic plates was used to evaluate plant extracts bioactivity. The obtained data indicate that it is a rapid method than can be used as an alternative to Petri dish tests.

2002 - Development of new fungicides against Magnaporthe grisea: synthesis and Biological activity of pyrazolo[3,4-d]thiazine, pyrazolo[3,4-d]pyrimidine derivatives [Articolo su rivista]
Cb, Vicentini; G., Forlani; M., Manfrini; Romagnoli, Carlo; D., Mares

Some pyrazolo[3,4-d]pyrimidin-4(5H)-thione, pyrazolo[3,.4-d][1,3]thiazin-4-one/thione, and pyrazolo[1,5-c][1,3,5]thiadiazine-4-one/thione derivatives were synthesized and screened for antifungal activity,against the causal agent of rice blast disease, Magnaporthe grisea. In all cases a remarkable inhibition of fungal growth was found in the range from 10 to 200 mug mL(-1). Several compounds were able to control mycelium growth at a rate of 10 mug mL(-1), a concentration at which the reference compound tricyclazole was completely ineffective. At least in the case of the most active substance, at the same dose the growth of seedlings or cultured cells of rice was substantially unaffected. Results allowed definition of structural requirements either to maintain or to enhance mycotoxic activity.

2002 - Mannan changes induced by 3-methyl-5-aminoisoxazole-4-thiocyanate, a new azole derivative, on Epidermophyton floccosum [Articolo su rivista]
D., Mares; Romagnoli, Carlo; B., Tosi; R., Benvegnu; A., Bruni; C. B., Vicentini

The antifungal activity of 3-methyl-5-aminoisoxazole-4-thiocyanate, a new azole derivative, was studied on the dermatophyte Epidermophyton floccosum. The compound strongly inhibited the in vitro growth of two different strains of the fungus and even induced profound morphogenetic anomalies. Optical and electron microscopy showed that such treatment targets the endomembrane system, particularly the plasmalemma, causing abnormal extrusion of the wall mannans. This results in improper arrangement of the different parietal materials; the walls are thus weak and subject to subapical rupture which terminates cell growth and elongation of the hypha. The morphological results and the preliminary biochemical data on fungal sterols suggest that this compound employs an action mechanism similar to that of other azoles used in therapy.

2002 - Rapid techniques for extraction of vitamin E isomers from Amaranthus caudatus seeds: ultrasonic and supercritical fluid extraction [Articolo su rivista]
R., Bruni; A., Guerrini; S., Scalia; Romagnoli, Carlo; G., Sacchetti

Supercritical fluid extraction (SFE) of seeds of Amaranthus caudatus (Amaranthaceae) and the use of ultrasound as a co-adjuvant in the extraction process were compared with methods traditionally used in the extraction of tocopherols and fatty acids. The use of readily available ultrasound equipment as an adjunct to the classical methods employed for the extraction of tocols provided qualitatively acceptable results more rapidly and more economically. SFE gave quantitatively better yields in shorter times, with solvent-free extracts obtained under conditions that minimised the degradation of thermolabile components. No significant variations were observed in the profile of the fatty acids extracted from amaranth oil by SFE or other methods, thus confirming the qualitative comparability of the faster supercritical extraction with the more time-consuming classical techniques even when processed with the aid of ultrasound. Copyright

2002 - Tocopherol, fatty acids and sterol distributions in wild Ecuadorian Theobroma subincanum (Sterculiaceae) seeds [Articolo su rivista]
R., Bruni; A., Medici; A., Guerrini; S., Scalia; F., Poli; Romagnoli, Carlo; M., Muzzoli; G., Sacchetti

Ecuadorian Theobroma subincanum (Sterculiaceae) seed parts were analyzed to determine quali-quantitative tocopherol distribution. Fatty acids and sterols in the embryos, teguments and endosperm were also evaluated with an aim to better-characterize the plant matrix for its potential use as a source of phytochemicals, for the cocoa butter processing industry and/or as a new source of low cost natural products in the cosmetic, drug and alimentary/nutraceutical industries. HPLC for tocopherols and gas-chromatography (GC), GC-mass spectrometry (MS) analyses for fatty acids profile (FAP) and phytosterols were performed. Tocopherols were particularly abundant in the embryo, with quali-quantitative data similar to wheat germ oil whereas, in the teguments and endosperm, the concentrations of tocopherols were lower. The fatty acid profile and phytosterol characterization of the seed parts showed qualitative homogeneous data. In the endosperm, 80% of the entire FAP consisted of oleic and stearic acid while, among sterols, cycloartenol was more abundant in endosperm than ill embryos and teguments. Accordingly, T. subincanum seeds can be proposed as possible substitutes in the cocoa processing industry and as a potential source of vitamin E isomers. (C) 2002 Elsevier Science Ltd. All rights reserved.

2001 - Secretory tissue ultrastructure in Tagetes patula L. (Asteraceae) and thiophene localization through X-ray microanalysis [Articolo su rivista]
Sacchetti, G; Romagnoli, Carlo; Bruni, A. AND POLI F.

Transmission electron microscopy has been used to determine the ultrastructure of secretory tissues in seedlings (10 days old plants) and in flowering plants (50 days old plants) of Tagetes patula. All the secretory structures in seedling and in flowering plants showed a schizogenous origin. In particular, in T. patula seedlings canals were examined in roots and hypocotyl while secretory cavities were found in cotyledons. In flowering T patula plants, canals in epicotyl and in flower corolla, as well as secretory cavities in leaves and flower bracts were examined. Plastids related to the synthesis and accumulation of lipid substances involved in the secretory process were then identified in some glandular structures. X-ray microanalysis showed the presence of sulfur in the secretory tissue cells and, above all, in the secretion itself. This supports the hypothesis of direct involvement of the T patula glandular structures in the production of thiophene compounds

2001 - Thiophene extracts from cultivated Tagetes patula plants [Articolo su rivista]
Mares, D; Tosi, B; Romagnoli, Carlo; Sacchetti, G. AND POLI F.

Tagetes patula plants have been cultivated in field in large quantity to produce raw material as source of extracts containing thiophenes. The thiophene contents of seeds and plants both fresh and dried from 10 cultivar were analyzed. The adult plants derived from seeds with a total thiophene content lower than 100 g g-1 showed a similar low total thiophene content. The seeds with a high total thiophene content gave plants with the same qualitative and quantitative phytochemical results. The total thiophene content of the dried plants is nearly twice the yield from fresh material. However, considering the commercial needs which could require large amounts of vegetal raw, the extraction from dried material might be more useful. Above all, this work suggests also that, in order to obtain plants with high thiophene contents, an accurate selection of high thiophene containing seeds is necessary

2000 - Effectiveness of Four New Pyrazole-Pyrimidines on Phytopathogens: Ultrastructural Evidences on Pythium ultimum [Articolo su rivista]
Mares, D; Romagnoli, Carlo; Sacchetti, G; Bruni, A; AND VICENTINI, C. B.

Four newly synthesized molecules derived from pyrazole-pyrimidine were assayed on Botrytis cinerea (Micheli), Fusarium moniliforme (Sheld) and Pythium ultimum (Trow). All proved effective in inhibiting the growth of the phytopathogens at all test concentrations (10, 20, 50, 100 µg/ml). The most effective compound was 1-(3)nitrophenyl-6-trifluoromethylpyrazolo3,4-dpyrimidine-4(5H)-thione (CF33). Ultrastructural studies on P. ultimum treated with CF33 revealed alterations in the normal hyphal shape and at high concentration, plasmolysis and damage to the wall texture was observed. At 20 µg/ml different vesicles were seen in the cytoplasm: some appeared quite dense, and specific cytochemical reactions indicated that they were most likely peroxysomes; other vesicles seem to be vacuoles of varying content. In some cases there was disintegration of the nuclear envelope. Effects on membrane lipids and interference in protein synthesis are hypothesized as possible mechanism of action of the molecule

1999 - Alterations in spore productions in Trichophyton rubrum treated “in vitro” with 1-amino-6-methyl-4-phenylpyrazolo-[3,4-d]-1,2,3,triazole [Articolo su rivista]
Mares, D; Romagnoli, Carlo; Sacchetti, G; Vicentini, C. B. AND BRUNI A.

The newly synthesized triazole, 1-amino-6-methyl-4-phenylpyrazolo [3,4-d]-1,2,3-triazole (V5), was tested on the dermatophyte Trichophyton rubrum. Optical and electron microscopy showed that the treatment suppressed the various forms of saprophytic conidia, induced the formation of resistant spores (chlamidospores) and accelerated the formation of parasitic conidia (arthroconidia). In conclusion, this new triazole did not reveal any fungistatic or fungicidal activity and could not be considered as an antifungal substance. On the contrary, it stimulated the development of resistant and parasitic forms of spores.

1999 - Glandular trichomes of Calceolaria adscendens Lidl. (Scrophulariaceae): histochemistry, development and ultrastructure [Articolo su rivista]
Sacchetti, G; Romagnoli, Carlo; Nicoletti, M; DI FABIO, A; Bruni, A. AND POLI F.

This paper reports the results of a study of the morphology and development of glandular trichomes in leaves ofCalceolaria adscendensLidl. using light and electron microscopy. Secretory trichomes started as outgrowths of epidermal cells; subsequent divisions gave rise to trichomes made up of a basal epidermal cell, a stalk cell and a two-celled secretory head. Ultrastructural characteristics of trichome cells were typical of terpene-producing structures. Previous phytochemical studies had revealed thatC. adscendensproduces diterpenes. Comparison withC. volckmanni,which produces triterpenes, and has trichomes with eight-celled secretory heads, suggests that there could be a relationship between the type of glandular trichome and the class of terpene produced. Further work is needed to test the hypothesis and to develop trichome characters as taxonomic tools

1999 - Laticifer tissue distribution and alkaloid location in Vinca sardoa (Stearn) Pign. (Apocynaceae), an endemic plant of Sardinia (Italy). [Articolo su rivista]
Sacchetti, G.; Ballero, M.; Serafini, M.; Romagnoli, Carlo; Bruni, A.; Poli, F.

Vinca sardoa (Apocynaceae) is an endemic plant of Sardinia (Italy) used in local folk medicine. And yet it has never been described in detail, particularly in regard to its secretory structures. The present research uses optical and electron microscopy to identify the laticifer cells in all plant organs, including the roots, where such cells had never previously been reported. Moreover, as opposed to what has been described in similar studies, these laticifer cells are articulated. In addition, histochemical tests were performed to identify the alkaloids in the latex and to quickly identify the same laticifer tissues in fresh samples. These tests always proved positive, indicating a close relationship between laticifer cells and the presence of alkaloids in the plant. TEM also showed that the laticifer cells have well-preserved cytoplasm and organelles which lead one to conclude that metabolic activity is still intact

1998 - Antifungal activity of six newly synthesized triazoles [Articolo su rivista]
Romagnoli, Carlo; Sacchetti, G; Vicentini, C. B. AND MARES D.

Six newly synthesized triazoles were tested on the pathogen Nannizzia cajetani in order to evaluate their antifungal activity. At the doses used (10, 50 e 100 µg/ml) none of the six substances achieved MIC although they all gave a dose-dependent response. The highest degree of inhibition was achieved with 1-amino-6-methyl-4-phenylpyrazolo[3,4-d]-1,2,3-triazole at the highest dose tested. Scanning Electron Microscopy of the samples treated with this triazole showed the total disappearance of the macroconids. The data are discussed on the basis of the structure-activity relationship.

1998 - Morphologycal study of Trichophyton rubrum; ultrastructural findings after treatment with 4-amino-3-methyl-1-phenylpyrazolo-(3,4-c)isothiazole [Articolo su rivista]
Mares, D; Romagnoli, Carlo; Sacchetti, G; Vicentini, C. B. AND BRUNI A.

The antifungal activity of 4-amino-3-methyl-1-phenylpyrazolo-(3,4-c)isothiazole was studied on Trichophyton rubrum. The compound, at concentrations between 20 and 100 g ml-1, induces a remarkable reduction in the growth and causes deep morphogenetic anomalies. The ultrastructural modifications have demonstrated that the compound targets the cell membrane of the fungus, breaking down not only the endomembrane system, but also the “outer” membrane, with consequent extrusion of materials in the medium. The results suggests a mechanism of action similar to other azoles clinically utilized

1998 - The photodynamic effect of 5-(4-hydroxy-1-butinyl)-2,2’-bithienyl on dermatophytes [Articolo su rivista]
Romagnoli, Carlo; Mares, D; Sacchetti, G. AND BRUNI A.

The thiophene 5-(4-hydroxy-1-butinyl)2,2[prime prime or minute]-bithienyl (BBTOH) strongly inhibited in vitro eight different dermatophytes. Epidermophyton floccosum proved most sensitive to all doses of BBTOH when applied in conjunction with uv-A irradiation. BBTOH also proved quite active against Nannizia cajetani, the only dermatophyte which was also strongly inhibited when treated (50 [mu]g ml[minus sign]1) and kept in the dark. For this reason, N. cajetani was chosen as the test organism for TEM and SEM aimed at determining what treatment-induced ultrastructural and morphological modifications had occurred. TEM revealed that the photoactive mechanism of BBTOH was similar to that of 2,2[prime prime or minute][ratio]5[prime prime or minute],2[double prime or second]-terthienyl ([alpha]-T). SEM, on the other hand, showed that early culture aging resulted from treatment.

1997 - Internal secretory structures and preliminary investigation and flavonoid and coumarin content in Santolina insularis (Asteraceae) [Articolo su rivista]
Sacchetti, G; Romagnoli, Carlo; Ballero, M; Tosi, B. AND POLI F.

UV light investigation has shown that vegetative and flowering plants of Santolina insularis have internal secretory structures similar to those of many other Asteraceae. In the root two arrays of ducts arise abutting the endodermis, while in the stem single or unconnected pairs of ducts are localized between the vascular bundles. The leaves and bracts present ducts vein-associated. A preliminary phytochemical investigation employing HPTLC evidenced the presence of some fluorescent flavonoids and coumarins. The fluorescence of the secretion and the presence of fluorescent metabolites in the plant organs, lead to assume that the secretory system is involved in producing and/or storing the secondary metabolites which justify the use of this species in Sardinian traditional medicine.

1997 - Phytochemical screening of coumarins and flavonoids in crude drugs of Asteraceae [Articolo su rivista]
Tosi, B; Donini, A; Romagnoli, Carlo; Sacchetti, G. AND DALL'OLIO G.

Alcoholic and watery extracts prepared from crude drugs of Helichrysum italicum (Roth) Don and Helichrysum stoechas (L.) Moench (Asteraceae), obtained from the Italian herbal market, were analyzed to evaluate the scopoletin content. The results indicate the constant presence of scopoletin, but with remarkable differences among the three suppliers considered. In any case H. stoechas preparations showed the highest concentration of the compound. The possible synergic role of scopoletin to the medicinal properties of H. italicum and H. stoechas preparations is argued.

1997 - Presence of scopoletin in commercial crude drugs of Helichrysum italicum and Helichrysum stoechas (Asteraceae) [Articolo su rivista]
B. TOSI, DONINI A; Tirillini, B; PIETROGRANDE M., C; Sacchetti, G; Romagnoli, Carlo; Dall'Olio, G.

A phytochemical investigation was performed on the concentration of several coumarins and flavonoids present in dried plant materials of thirty two species of Asteraceae, which represent the most sold crude drugs in the Italian herbal market. Qualitative and quantitative evaluation of coumarins and flavonoids content was performed by High Performance Thin Layer Chromatography (HPTLC) and UV detection. Because of their pattern of assumption (pharmaceutical preparations and health beverages), both the alcoholic and watery extracts of the drugs were analyzed. The results obtained from watery and alcoholic extracts were very similar; therefore, in this paper we refer only the data of the alcoholic extracts. The qualitative and quantitative distribution of flavonoids was more homogeneous than the one of coumarins. Since both coumarins and flavonoids are bioactive compounds with numerous pharmacological activities, it is important to ascertain their content in herbal preparations of broad use to avoid toxicity in humans.

1997 - Protoanemonin-induced cytotoxic effects in Euglena gracilis [Articolo su rivista]

The changes that protoanemonin, an antimicrobial agent of natural origin, brought about in the alga Euglena gracilis were studied with light and electron microscopy and with flow cytometry. The compound proved lethal for the alga at a dose of 5x10(-5)m. At the sublethal dose of 3.5x10(-5)m it caused: marked inhibition of growth; increase in the average cell volume; inhibition of cytokinesis and induction of coenocytic organisms; loss of the flagellum and stigma. Most nuclei were arrested in the G2/M phase of the cellular cycle. The chloroplasts were still well organized, but there was a conspicuous decrease in carotenoids and chlorophylls a and bwith a corresponding increase in pheophytins. The multifarious alterations seen in Euglena are discussed on the basis of a possible interaction between this lactone and several enzymatic systems.

1996 - Antifungal properties of the 3-methyl-5-aminoisoxazole-4-thiocyanate on four yeasts. An Unusual dose-dependent effect on Rhodotorula glutinis Harrison [Articolo su rivista]
D., Mares; Romagnoli, Carlo; A., Donini; A., Bruni

Deux activités antifongiques du 3-méthyl-5-aminoisoxazole-4-thiocyanate ont été étudiées chez les espèces de levures : Candida albicans, Saccharomyces cerevisiae, Schizosaccharomyces pombe, et Rhodotorula glutinis. Une inhibition significative de la croissance de chacune des levures a été évidente. Des modifications ultrastructurales ont révélé que le composé, à demi-dose de la concentration inhibitrice minimale (MIC) (1,25 μg.mL-1), causait une transformation de la levure vers la forme mycéllale par formation d'un tube germinal chez le R. glutinis. L'accumulation de quantités anormales de substances pariétales après traitement avec la MIC est discutée en relation avec la production de chitine et d'ergostérol.Two antifungal activities of 3-methyl-5-aminoisoxazole-4-thiocyanate have been studied in yeast species: Candida albicans, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Rhodotorula glutinis. A significant inhibition of growth of each yeast was evident. Ultrastructural changes showed that the compound, half-dose of the minimum inhibitory concentration (MIC) (1.25 μg.mL-1), causing a transformation of the yeast to form mycéllale by formation of a germ tube at the R. glutinis. The accumulation of abnormal amounts of substances parietal after treatment with the MIC is discussed in relation to the production of chitin and ergosterol.

1996 - Antifungal screening of seven new azole derivatives [Articolo su rivista]
D., Mares; Romagnoli, Carlo; C. B., Vicentini; G. SACCHETTI AND A., Bruni

Two strains of the dermatophyte Epidermophyton floccosum were treated with seven new synthetic azole derivatives to determine their possible antifungal activity. Among these, two diazopyrazoles (1-phenyl-3-methyl-4-diazo-5-methanesulphonamid and 1,3-dimethyl-4-diazo-5-methanesulphonamid) and the triazole (4-methyl-6-t-butylpyrazolo[3,4-d][1,2,3]triazole) were inactive at the highest dose tested (100 μg/ml). The 4-methyl-6-phenyl-pyrazolo[3,4-d][1,2,3]triazole produced only a partial inhibition of growth, whereas the most active compounds were the two thiadiazines(6H-3-phenyl-5-methyl-7-(3,4-dichlorophenyl)pyrazolo[3,4-c] [1,2,5]thiadiazine-2,2-dioxide and 7-methyl-2-phenyl-4H-pyrazolo[1,5-c][1,3,5]thiadiazine-4-one), and an aminoisoxazole (3-methyl-5-aminoisoxazole-4-thiocyanate), which induced a remarkable and sometimes complete reduction in the growth rate of the fungus. The structure-activity relationships are discussed.

1996 - Antimicrobial activity of some commercial extracts of propolis prepared with different solvents [Articolo su rivista]
B., Tosi; A., Donini; Romagnoli, Carlo; Bruni, A.

Some commercial extracts of propolis obtained with different solvents were tested to evaluate their antibacterial and antifungal activity. All propolis preparations exhibited antimicrobial activity, particularly against Gram-positive bacteria, yeasts and dermatophytes with zones of inhibition ranging from 3 to 30 mm. Against yeasts and dermatophytes, oil, ethanol and propylene glycol solutions showed an inhibition for more 2 weeks, while the glycerine solution maintained inhibition only for some days. The results indicate that the solvent employed for the extraction may enhance the potency of the antimicrobial activity of propolis. Consistency in the properties and characteristics of propolis were related to the formulation of extraction procedures.

1995 - Antibacterial effects of the essential oil of Santolina insularis (Genn. ex Fiori) Arrig. (Asteraceae) [Articolo su rivista]
G., Sacchetti; Romagnoli, Carlo; D., Mares; L., Bonsignore; F., Poli

The antimicrobial activity of the essential oil of Santolina insularis (Genn. ex Fiori) Arrig. (Asteraceae), an aromatic plant endemic on the island of Sardinia (Italy), eas evaluated. The results reveald significant antibacterial activity against Gram-positive and Gram-negative bacteria.

1995 - Antifungal effects of the 3-methyl-5-aminoisoxazole-4-thiocyanate on some dermatophytes [Articolo su rivista]
Romagnoli, Carlo; C. B. VICENTINI AND D., Mares

This preliminary research demonstratesthe dose dependent antimycotic effect of 3-methyl-5-aminoisoxazole-4-thiocyanate against anumber of animal and human dermatophytes. The in vitro results suggest potential medicalapplications.

1995 - Characterization of coumarins in Chamomile preparations sold in Italy [Articolo su rivista]
B., Tosi; Romagnoli, Carlo; E. MENZIANI ANDREOLI AND A., Bruni

The umbelliferone/herniarin ratio in herbal preparations of chamomile was determined by high-performance thin-layer chromatographic analysis (HPTLC). Plant material was extracted in 150 ml of warm H2O for 10 min, then the residual plant material was extracted with 150 ml of MeOH for 1 h. The herbal preparations containing ligulate florets showed higher content of coumarin if compared to other parts of the flower-heads. The ratio umbelliferonel hemiarin was lower than 1:5 in technological preparations.

1994 - Antifungal activity of some 2,2':5',2" terthiophene derivatives [Articolo su rivista]
D., Mares; Romagnoli, Carlo; R., Rossi; A., Carpita; M. CIOFALO AND A., Bruni

The dermatophyte Microsporum cookei Ajello was treated with nine new natural and synthetic 2,2':5',2"-terthiophenes to determine their possible antifungal activity. In the dark the thiophenes were inactive, while when photoactivated with UV-A they induced a remarkable reduction in the growth rate of the fungus. The only exception was (E)-N-(2-methylpropyl)-3-(2,2':5',2"-terthien-5-yl)-propenamide , which was not fungistatic even at the highest dose tested (24 microM). The more active compounds were 3'-methoxy-2,2':5',2"-terthiophene and 3'-methylthio-2,2':5',2"-terthiophene, whose activity seems to be related to the presence of a substituent in the 3' position of the central ring of thiophenes. Transmission electron microscopic observations demonstrated the photoactive nature of the synthetic molecules to be similar to that of alpha-terthienyl, a natural thiophene present in some Asteraceae. The dark treatment caused only the accumulation of the compound in vacuoles, without other evident alterations. After UV-A irradiation the activated thiophene causes severe modifications to the endomembrane system, probably via oxygen-dependent mechanism.

1994 - Antifungal effects of α-terthienyl from Tagetes patula on five dermatophytes [Articolo su rivista]
Romagnoli, Carlo; D., Mares; M. P. FASULO AND A., Bruni

Alpha-terthienyl (αT), a thiophene compound isolated from Tagetes patula (Asteraceae), exhibits antifungal activity towards five strains of dermatophytes (Trichophyton mentagrophytes, T. rubrum, T. violaceum, Epidermophyton floccosum, Microsporum cookei). αT plus UVA irradiation for 90 min concentrations between 6 μm and 24 μm. Between 1 and 10 days after irradiation, the fungal growth was reduced or arrested with marked responses for T. mentagrophytes, T. rubrum and M. cookei. The ultrastructural events that occur in Trichophylus mentagrophytes after treatment were studied by electron microscopy. In the dark, αT caused no important ultrastructural modifications within the hyphae. After UVA irradiation the photoactive compound caused damage to membranes of the nucleus, mitochondria and endoplasmic reticulum. Plamolytic and autolytic changes resulted in plasma membranes breakage and in cell wall aberrations. UVA activated αT would appear to target protein-rich membranes of the dermatophytes.

1994 - Effect of ω-conotoxin and verapamil on antinociceptive, behavioural and thermoregulatory responses to opioids in the rats [Articolo su rivista]
S., Spampinato; E., Speroni; P., Govoni; E., Pistacchio; Romagnoli, Carlo; G., Murari; S., Ferri

This study with the rat evaluated the contribution of omega-conotoxin GVIA-(omega-CgTx) and verapamil-sensitive Ca2+ channels in behavioural, antinociceptive and thermoregulatory responses to intracerebroventricular (i.c.v.) injection of [D-Ala2,NMePhe4,Gly-ol5]enkephalin (DAMGO), [D-Pen2,D-Pen5]enkephalin (DPDPE) and dynorphin A-(1-17), which are selective agonists for putative mu, delta and kappa-opioid receptors, respectively. The rats treated with omega-CgTx (8-32 pmol i.c.v.) showed transient, dose-dependent shaking behaviour, hyperalgesia and hypothermia which gradually disappeared within 4 h. The behaviour of the rats was normal by 24 h. Histological examination of brain sections showed morphological alterations of neurons in the hippocampus, medial-basal hypothalamus and pyriform cortex. antinociception, catalepsy and thermoregulatory responses elicited by DAMGO (0.4 and 2.0 nmol) were significantly prolonged and potentiated by verapamil (20 pmol i.c.v. 15 min before) or omega-CgTx (8 pmol 24 h before). Antinociception and hypothermia induced by DPDPE were antagonized by verapamil and omega-CgTx, whereas only omega-CgTx prevented the behavioural arousal observed after DPDPE. Similarly, hypothermia induced by dynorphin A-(1-17) (5.0 nmol) and by the kappa-opioid receptor agonist U50,488H (215 nmol) was antagonized by the two Ca2+ channel blockers but only omega-CgTx prevented the barrel rolling and bizarre postures caused by the opioid peptide.

1993 - Antidermatophytic activity of herniarin in Chamomile preparations [Articolo su rivista]
Mares, D; Romagnoli, Carlo; Bruni, A.

Fungal cultures of Microsporum cookei were incubated with aqueous extract of german chamomile [Chamomilla recutita (L.) Rauschert], herniarin (100 and 23 μg/ml) and umbelliferone (100 and 5.5 μg/ml). At the end of the incubation period, the mycelia were irradiated with UVA for 90 min. The fungal morphology was compared with that of control cultures by transmission and scanning electron microscopy. No fungistatic and ultrastructural anomalies were observed for umbelliferone after UVA irradiation. On the contrary herniarin and chamomile extracts show a clear antifungal activity.The main changes were due to inhibition of hyphal outgrowth, apical furcation, alteration of nuclear morphology, deposition of minute electron-dense vesicles in the cytoplasm and thickening of the cell wall. Abnormalities were also involving the mitochondrial feature. These results suggest that the antidermatophytic activity of german chamomile preparations is due to combined effect of herniarin and UVA irradiation.

1993 - Cytological characterization of a giant strain of Euglena gracilis obtained from dark-starved cultures [Articolo su rivista]
D., Mares; Romagnoli, Carlo; M. RUBINI AND M. P., Fasulo

Euglena gracilis green cells were dark-starved for four months. After this period almost the entire population died, while a few giant, viable cells appeared in the culture. The giantism was maintained after repeated subcultures in growth medium in light or dark conditions. However, the phenomenon was not permanent, and the morphological characteristics of the wild-type Euglena were gradually restored.In giants cells nuclei enlarged greatly, DNA content increased and the Golgi apparatus greatly proliferated. Chloroplasts and mitochondria increased in number and size often presented structural modifications when compared with normal Euglena. Importntly, in the giants cells that were maintained in darkness in resting or growth conditions chloroplasts persisted as structured organelles which appeared red-fluorescent under UV ilumination.Wheter giantism is a phenotypic or a genotypic change is still debated. In our case, the evolution of this phenomenon, chiefly the enhanced DNA content, suggests that teratism is a multiploid mutation with the possibility of a return to the normoploid condition. Cnstitutive chloroplasts are present in most algae, except for a few species, among which is Euglena gracilis. The persistence of differentiated plastids in darkness in giant Euglena is considered to be return to an ancestral condition and may, be phylogenetically important.

1993 - Effects of spermine and spermi¬dine on cell division and wall morphogenesis in Saccharomyces cerevisiae [Articolo su rivista]
F., Poli; Romagnoli, Carlo; G. DALL'OLIO AND M. P., Fasulo

The effect of polyamines (PA) on the synthesis and deposition of wall constituents in Saccharomyces cerevisiae was investigated. Spermidine (Spd) in various doses was ineffective whereas spermine (Sp) in the same concentrations caused a marked inhibition (25-60%) of cell proliferation accompanied by evident morphogenetic malformations. Sp-treated cells were elongated, grouped in small clusters, and showed malformed septa and aberrant wall thickenings. The PATAg technique revealed that the aberrations consisted of an abnormal accumulation of both reactive materials, like 1,3-beta-glucans, and unstained chitin components. Since 1,3-beta-glucan synthases and chitin synthases are inserted in the plasma membrane, whose anionic sites interacted with the cation groups of Sp, it is assumed that the molecule determines a condition resulting in an unregulated activation of the two enzymes. The fact that Spd, which contains three cationic groups instead of the four contained in Sp, is without effect suggests that a compound must have at least four cation sites in order to affect the cell division and wall morphogenesis of S. cerevisiae.

1993 - Non-peptide ligands for opioid receptors. Design of k-specific agonists [Articolo su rivista]
G., Ronsisvalle; L., Paquinucci; M. S., Pappalardo; F., Vittorio; G., Fronza; Romagnoli, Carlo; E., Pistacchio; S. SPAMPINATO AND S., Ferri

A series of phenyl carboxy esters 5a-d derived from N-(cyclopropylmethy1)normetazocine wassynthesized and evaluated for its selectivity at p, K, and 6 opioid receptors. Compound 5a, although43 times less potent than the reference compound U50488, was specific for K receptors, having nodetectable affinity for either p or 6 receptors. Greater binding affinity was seen with thediastereoisomer having the 1'R,2'S stereochemistry in the cyclopropyl ring of the nitrogensubstituent, which was only 12 times less active than U50488. Antinociceptive activity in themouse tail flick was only slightly lower than that of U50488 (ED60 = 7.66 us 4.52 mg/kg). Naloxonefully prevented antinociception induced by (1'R,2'S)-5a at the doses of 2.0 mg/kg. Compound(1'R,2'S)-5a is one of the most K-selective non-peptide compounds reported to date. The implicationsof these results in terms of requirements for K ligands are discussed.

1993 - Ultrastructural localization of an intracellular pigment in 3-chlorobenzoate co-metabolizing Pseudomonas fluorescens [Articolo su rivista]
F., Fava; D., DI GIOIA; Romagnoli, Carlo; L. MARCHETTI AND D., Mares

A strain of Pseudomonas fluorescens was capable of co-metabolizing 3-chlorobenzoic acid with the production of a chlorinated catechol black pigment. A peroxidase and another enzymatic activity referred to as a polyphenol oxidase were found to be involved in the oxidation of 4-chlorocatechol to 4-chloro-1,2-benzoquinone, i.e. in the production of highly reactive substrates for pigment formation. Therefore, P. fluorescens cells were seen to take an active part not only in 3-chlorobenzoate mineralization but also in overall pigment production. pH was found to be a key parameter in the regulation of the activity of P. fluorescens oxidoreductive enzymes. Ultrastructural investigations showed that electron dense granules of pigment were distributed throughout the cytoplasm of Pseudomonas fluorescens cells grown in presence of 3-chlorobenzoate, as confirmed also by Thiéry cytochemical investigations.In these cells, an extensive contraction of the cytoplasm as well as a significant damage to the cell wall after two days of incubation, suggested that pigment production caused a premature death of the cells accompanied by the leakage of the cell content. Pigment production seemed to occur mostly in the cytoplasmic context where the electron dense material accumulates until it is released in the medium after the cell lysis.