Biomedical subjects

E Martinotti

Publications and source records attributed to E Martinotti.

At least 19 recordsLinked to original sources

PubMed2003

Genetic characterization of the three medicinal Echinacea species using RAPD analysis.

The three medicinal species of the Echinacea genus, E. angustifolia DC., E. pallida (Nutt.) Nutt. and E. purpurea (L.) Moench were distinguished using the RAPD (random amplified polymorphic DNA) technique. Species-specific markers were identified from amplicons obtained with four of the twenty 10-mer primers contained in the Operon RAPD kit A. In particular, one marker was identified for E. angustifolia (OPA 20, 1800 pb) and E. pallida (OPA 10, 600 pb) and three markers for E. purpurea (OPA 11 : 1250 pb; OPA 17 : 750, 1800 pb). Genetic distance analysis indicated a high degree of difference among the three species with a relative lower difference between E. angustifolia and E. pallida.

Echinacea↗

PubMed2001

New 5-substituted-1-(2-hydroxybenzoyl)-benzotriazoles, potassium channel activators. IV.

This paper reports the synthesis of a series of new 5-substituted-1-(2-hydroxybenzoyl)-benzotriazoles, which have been tested for their activity as possible activators of potassium channels. In rat aortic rings, the 'opened' derivatives 1a-f, intermediates of synthesis, showed vasorelaxing properties, with appreciable values of potency. However, the most remarkable effects were recorded for the 2-hydroxybenzoylbenzotriazoles 3a-f, which showed full vasorelaxing efficacy and high potency values. The introduction of a 2-hydroxybenzyl substituent in the 1 position of the benzotriazole ring (compound 7) strongly decreased the activity, showing the importance of the electron-acceptor carbonyl function. The best compound, 3b, was further investigated, in order to evaluate the possible mechanism of action involved in the vasodilator activity. In the vascular model, different potassium channel blockers inhibited the effects of the compound, and an increase of the levels of membrane depolarisation induced a significant reduction of the recorded responses. Compound 3b was also tested in a model of isolated rat heart, retroperfused through the aorta and submitted to a global ischemia/reperfusion cycle. In such an experimental condition, 3b showed an interesting cardioprotective activity. All the above observations are in agreement with the hypothesis of a mechanism linked to the activation of potassium channels.

Animals↗

PubMed2001

Some structural changes on triazolyl-benzotriazoles and triazolyl-benzimidazolones as potential potassium channel activators. III.

This paper reports the synthesis and pharmacological evaluation of some compounds, obtained by structural modifications of 1,2,3-triazolyl-benzotriazoles and 1,2,3-triazolyl-benzimidazolones, which had shown activity as potential activators of the big-conductance calcium-activated potassium channels (BK(Ca)). Changes have concerned the introduction of a hinderer substituent in the 5-position of the benzimidazolone (4a, b) and benzotriazole (5a, b) rings, opening of the benzimidazolone ring (7) and substitution of the 1,2,3-triazole ring with a 2-hydroxyphenyl ring (10). Furthermore a series of 3-aryl-benzotriazin-4-one derivatives (13a-e) has been studied, which appears as a modification and/or combination of the benzimidazolone and benzotriazole rings. Only compound 10 shows interesting activity, while the other structural modifications either do not increase (compounds 4 and 5) or reduce (compounds 7 and 13) the pharmacological activity. However, these results provide useful information about structure-activity relationships.

Animals↗

PubMed2001

Adenosine-mediated hypotension in in vivo guinea-pig: receptors involved and role of NO.

1. Adenosine produced a biphasic lowering of the mean BP with a drastic bradycardic effect at the highest doses. The first phase hypotensive response was significantly reduced by the nitric oxide (NO) synthase inhibitor L-NAME. 2. The A(2a)/A(2b) agonist NECA produced hypotensive and bradycardic responses similar to those elicited by adenosine, which were not significantly modified by the A(2b) antagonist enprofylline. 3. The A(2a) agonist CGS 21680 did not significantly influence basal HR while induced a hypotensive response antagonized by the A(2a) selective antagonist ZM 241385, and reduced by both L-NAME and the guanylate cyclase inhibitor methylene blue. 4. The A(1) agonist R-PIA showed a dose-dependent decrease in BP with a drastic decrease in HR at the highest doses. The A(1) selective antagonist DPCPX significantly reduced the bradycardic activity and also the hypotensive responses obtained with the lowest doses while it increased those obtained with the highest ones. 5. The A(1)/A(3) agonist APNEA, in the presence of the xanthinic non-selective antagonist 8-pSPT, maintained a significant hypotensive, but not bradycardic, activity, not abolished by the histamine antagonist diphenhydramine. 6. The selective A(3) agonist IB-MECA revealed a weak hypotensive and bradycardic effect, but only at the highest doses. 7. In conclusion, in the systemic cardiovascular response to adenosine two major components may be relevant: an A(2a)- and NO-mediated hypotension, and a bradycardic effect with a consequent hypotension, via atypical A(1) receptors. Finally, an 8-pSPT-resistant hypotensive response not attributable to A(3) receptor-stimulation or to release of histamine by mastocytes or other immune cells was observed.

Adenosine↗

PubMed2000

Triazolyl-benzimidazolones and triazolyl-benzotriazoles: new potential potassium channel activators. II.

This paper reports the synthesis and pharmacological evaluation of a series of 5-substituted-triazolyl benzotriazoles (2a-f) and the corresponding series of 5-substituted-triazolyl-benzimidazolones (6a-f), as potential activators of the big-conductance calcium-activated potassium channels (BK(Ca)). The synthesis and structure demonstration of the stock compounds of the two series have been described in our previous works, as well as the common starting compounds 4-carboxamido-5-(4-substituted-2amino-anilino)-1,2,3-triazoles (1a-f). The triazolyl-benzotriazoles were obtained by diazotization, while the triazolyl-benzimidazolones were obtained by thermal intramolecular cyclization of ethoxycarbonylamino derivatives or directly with phosgene. Benzimidazolone compounds generally showed little effect whilst the compounds with a benzotriazole ring showed full efficacy, with vasorelaxing properties and potency parameters a little lower than that of the reference compound NS 1619. These effects were significantly reduced by an increased membrane depolarization. This depolarization-sensitive response is in agreement with the pharmacodynamic hypothesis of activation of potassium channels.

Animals↗

PubMed1999

Vascular effects of aqueous crude extracts of Artemisia verlotorum Lamotte (Compositae): in vivo and in vitro pharmacological studies in rats.

Artemisia verlotorum Lamotte (Compositae), growing in almost all the northern hemisphere, is used in folk medicine of some countries of Tuscany, Italy, as a remedy for hypertension. The pharmacological evaluation of the responses evoked by an aqueous dried extract of Artemisia verlotorum on the blood pressure of anaesthetized rats and on in vitro rat isolated aortae showed a marked, but transient, hypotensive activity. This effect was mediated by a strong vasodilator action, closely linked to the release of endothelial nitric oxide and to the nitric oxide-guanosine 3'-5'-cyclic monophosphate (cGMP) pathway, caused by a muscarinic receptor agonism.

Acetylcholine↗

PubMed1999

Experimental and theoretical comparisons between the classical Schild analysis and a new alternative method to evaluate the pA2 of competitive antagonists.

The Schild analysis is undoubtedly the most frequently used powerful diagnostic tool to investigate the nature of an antagonist and, consequently, to evaluate its potency, often expressed as pA2. Nevertheless, different reasons often prevent the experimenter from applying this analysis, leading to use an inhibition curve for the antagonist and to evaluate its potency by means of several approaches, which are generally considered theoretically invalid. In a recent work, a new theoretical approach, mathematically analogous to the Schild one, has been shown. By means of a simplified experimental protocol based on an antagonist inhibition curve (following a control concentration-response curve for the agonist), this method allows a linear regression analysis, giving a slope value absolutely equivalent to the Schild slope and a reliable estimation of the pA2 of a competitive antagonist. In this paper, this new method has been compared with the Schild analysis, to determine the parameters of potency relative to well-known competitive antagonists, on different in vitro isolated preparations. In strips of guinea pig isolated gastric smooth muscle, pirenzepine antagonised the effects of bethanechol. In guinea pig isolated ileum, atropine blocked the contracturant effects of carbamylcholine, while in electrostimulated ileum segments, the inhibitory responses to alpha-methylnoradrenaline were reduced by idazoxan. Finally, in guinea pig isolated spontaneously beating atria, the negative inotropic effects of 5'-N-ethylcarboxamidoadenosine were antagonised by 8-cyclopentyl-1,3-dipropylxanthine. The parameters of potency, relative to all the above competitive antagonists and expressed as pA2, resulted almost equivalent, when calculated by the Schild analysis or by the alternative method. Furthermore, when tested also for the well-known irreversible alkylating agent dibenamine in rat aortic rings stimulated by noradrenaline, the alternative method furnished a profile of clear nonlinearity, unmasking the nature of the antagonism. Finally, the relationships between the results calculated by the alternative analysis or by the Schild analysis and different levels of computer-generated "random noise" (affecting the shape and the position of theoretical curves) were also evaluated, in order to know the robustness of the new method. The two methods proved reliable and almost equivalent in robustness, when applied with different levels of "random noise". These results confirm the Schild analysis as the most accurate tool to study antagonists, since this analysis can furnish the highest number of information and observations on the behaviour of an antagonist. Nevertheless, when limiting conditions prevent a classical Schild analysis and impose the use of an inhibition curve, the new method probably represents the most preferable experimental approach. Indeed, it allows to calculate the antagonist potency, after the evaluation of a slope parameter giving an important information about the possible nature of the antagonism.

Animals↗

PubMed1998

A simplified empirical approach to evaluate the dissociation constant of a full agonist by the irreversible receptor inactivation method.

The estimation of the dissociation constant (Ka) of full agonists represents an essential tool for the classification of drugs and drug receptors, in functional pharmacology. The evaluation of the Ka was a problem until the development of the Furchgott's method (irreversible partial receptor inactivation method), which surely represents the most used analysis for the evaluation of the agonist Ka in experimental protocols on isolated tissues. The Furchgott's method can furnish a reliable estimation of the Ka, but it requires a relatively complicated manipulation of experimental data. In this article, an alternative approach for the evaluation of the Ka is proposed, on the basis of empirical considerations. This method, also based on the partial alkylation of a fraction of receptors, needs only the knowledge of the location parameters of the concentration-response curves and the application of a very simple equation, without any complicated intermediate interpolation of the experimental data.

Algorithms↗

PubMed1998

Intrinsic activity and EC50: the simplest tools for the evaluation of the dissociation constant of a partial agonist.

In functional pharmacology, the Furchgott's analysis and the Waud's analysis are the most widely used among the reliable different methods actually available, for the determination of the dissociation constant of partial agonists. However, they need the application of relatively complicated procedures of manipulation and interpolation of raw data. On the basis of empirical assumptions, this article proposes a new approach, which probably can be considered the simplest method to determine the dissociation constant of a partial agonist, because of the rapid experimental protocol and the easy calculation procedure. Computer-generated concentration-response curves (CRC) for hypothesised partial agonists were analysed by the widely known Waud's analysis and by this new approach. Furthermore, this new analysis was also used to evaluate experimental data from literature, relative to the dissociation constants of alpha-adrenoceptor partial agonists, recorded in rabbit and rat aortae and calculated by the Waud's method. The results obtained by the new approach, both for the computer-generated and for the experimentally studied partial agonists, showed a high level of accuracy, when compared with the classical Waud's analysis.

Adrenergic alpha-Agonists↗

PubMed1998

New 1,2,3-triazole derivatives (ureas, amides, urethanes) with a potential biological interest.

This paper reports the preparation of three series of 1,2,3-triazole derivatives bearing an ureido substituent (compounds 5a-h), a carboxamido substituent (compounds 6a-f) or an urethane substituent (compounds 7a-l). Some compounds were submitted to in vitro functional tests on guinea-pig isolated intestinal preparations and/or to in vitro antitumor and antiviral screening. The tested compounds showed no activity on guinea-pig ileum, except compound 7g, provided of contracturant properties; similarly, no anticancer or antiviral significant activity was found.

Animals↗

PubMed1997

Histaminic bronchospasm potentiated by adenosine: investigation of the mechanisms.

In anaesthetized guinea pigs, adenosine enhances the histamine-induced bronchospasm by means of a mechanism partly involving non-adrenergic-non-cholinergic (NANC) nerves, not related to capsaicin-sensitive neurons (Breschi et al., 1994). In the present paper, we excluded any interference by adenosine with the mediators known to be present in the airway inhibitory NANC system, VIP (vasoactive intestinal polypeptide) and NO (nitric oxide). The use of alpha-chymotrypsin or L-N(G)-nitro-arginine methyl ester (L-NAME) failed to modify the potentiation under study. The effects of adenosine were further investigated by studying whether an increased release of excitatory mediators from non-neural cells, in particular 5-HT (5-hydroxytryptamine, serotonin) and arachidonic products, was involved. In this connection, methysergide did not significantly affect the modulatory action of adenosine, revealing that the release of 5-HT was also not involved. Inhibition was obtained with hydrocortisone and with nordihydroguaiaretic acid, but not with indomethacin or with the mastocyte membrane stabilizer, sodium cromoglycate. This evidence suggests that lipooxygenase products, not derived from mastocytes, probably participate in the potentiating effect of adenosine.

Adenosine↗

PubMed1997

Effects of noise stress on EFS-mediated cholinergic and inhibitory NANC responses in tracheae from normal and sensitized guinea-pigs.

1 The aim of the present research was to study the cholinergic and inhibitory non-adrenergic-non-cholinergic (NANC) responses obtained with electrical field stimulation (EFS) of tracheal tissues from sham- and noise-exposed guinea-pigs. A comparison was also made between normal and ovalbumin (OA)-sensitized animals. 2 In proximal tracheae pretreated with indomethacin (3 microM), propranolol (1 microM), alpha-chymotrypsin (2 U ml-1) and L-NAME (0.1 mM), frequency-dependent responses to EFS (0.1 ms width; 20 V, 0.1-100 Hz, 15 s train duration) were obtained, both contractile and relaxing in nature. The contractile responses were abolished by atropine (1 microM), and did not vary significantly between sham- and noise-exposed guinea-pigs, or between normal and sensitized animals. The NANC relaxing responses, present in spite of the pre-treatment of the tissues with L-NAME and alpha-chymotrypsin, and almost completely abolished by tetrodotoxin (TTX) treatment (10 microM), appeared to be enhanced in noise-exposed guinea-pigs, with respect to sham-exposed animals, but only when the animals were not OA-sensitized. 3 In distal tracheae contracted with histamine (10 microM), the study of the whole inhibitory NANC response (pre-treatment with propranolol, but not with alpha-chymotrypsin and L-NAME), which was mainly TTX-sensitive, revealed a statistically non-significant difference between sham- and noise-exposed guinea-pigs, both normal and OA-sensitized. When distal tracheae were preincubated with alpha-chymotrypsin (2 U ml-1) and L-NAME (0.1 mM), in addition to propranolol, a significant residual inhibitory NANC response to EFS was observed. Surprisingly, in this case, similarly to the evidence obtained in proximal tracheae, a significantly enhanced response was revealed in noise-exposed guinea-pigs with respect to sham-exposed animals. 4 The noise-induced enhancement of the relaxant response disappeared when the tissues were pretreated with the A2 purinergic antagonist 3,7-dimethyl-1-propargylxanthine (DMPX, 1 microM), while it persisted in the presence of the A1 antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, 10 nM). 5 The above data indicate that, while not modifying the cholinergic and the whole inhibitory NANC response to EFS, noise stress selectively influences an inhibitory component of the NANC system in guinea-pig trachea with a mechanism probably involving an enhanced neurally mediated release of adenosine, which relaxes the smooth muscle via A2 receptors. This effect appears to be lacking or masked in sensitized guinea-pigs.

Acetylcholine↗

PubMed1996

A modified aortic multiple-ring preparation for functional studies.

A comparison is made of four rat aortic preparations, that is, single ring, spiral strip, zig-zag strip, and multiple-ring, on the basis of the responses to norepinephrine (NE) and acetylcholine (ACh). The single ring preparation was suitable under isometric, but not under isotonic conditions, because of the small isotonic tension which developed in response to the contractile agonist. The spiral and the zig-zag strips showed a discontinuity in the relaxant activity of the ACh, probably because of the removal, or not, of the endothelium from the preparation. The reproducibility of both contracturant and relaxing responses in the multiple-ring preparation makes this the most suitable of all four for the study of vasoactive drugs.

Acetylcholine↗

PubMed1996

Acetylcholine-induced bronchoconstriction modified by noise exposure in normal but not in sensitized guinea-pigs.

1. The acute (6h) exposure of guinea-pigs to white noise (110 dB) as a stress stimulus, reduced bronchial reactivity to acetylcholine (Ach) (3-1000 micrograms kg-1 i.v.) in anaesthetized animals. 2. The hyporesponsiveness to Ach in stressed animals was not confirmed in vitro on tracheal preparations (Ach 1 x 10(-9)-1 x 10(-4) g ml-1) and disappeared in vivo when the animals were sensitized with ovalbumin (OA, 100 mg kg-1 i.p. + 100 mg kg-1 s.c.). The hyporesponsiveness was also absent in ovalbumin sensitized guinea-pigs exposed to an aerosol of ovalbumin 60 min before testing with Ach. 3. In non-sensitized guinea-pigs, pretreatment with butoxamine (1 mg kg-1 i.v.) or with theophylline (25 mg kg-1 i.v.), completely abolished the effect of noise-exposure. In contrast, pretreatment with L-NG-nitro-arginine methyl ester (L-NAME, 10 mg kg-1 i.v.), alpha-chymotrypsin (2 U kg-1 i.v.) or with enprofylline (10 mg kg-1 i.v.), did not affect it. 4. In conclusion, our experiments reveal inhibitory mechanisms upon Ach-induced bronchoconstriction activated by a stress stimulus and this is absent in sensitized animals. These mechanisms seem to be linked to the adrenergic beta 2-receptors and a role for the purinergic system (via A-receptors) may also be present.

Acetylcholine↗

PubMed1995

Benzodiazepine agonists reverse the effects of noise exposure on central benzodiazepine receptors and cardiac responsiveness.

Rats were submitted to 110 dB white noise exposure for 1, and 6 hours and brain alpha 1, beta 1 and benzodiazepine receptor binding was evaluated with selective ligands. An increase in cerebral benzodiazepine receptor (CBR) concentration, without any significant change in affinity constant, occurred after the 6 h treatment; no change was observed in adrenergic receptor binding at any period of exposure. Both diazepam and clonazepam pre-treatment reversed the effects of noise on CBR binding, confirming a role of these receptors in the response to noise stress. Furthermore, these benzodiazepine agonists influenced the response of cardiac and aortic tissues, which are known to be changed by stress exposure. Diazepam and clonazepam pre-treatment protected cardiac tissue from the effects of 6h noise stress, and a potentiation of aortic responses was detected, although at different times of exposure. The differences between the responses of these peripheral tissues to benzodiazepine treatment suggest that the expression depends on the tissue examined and the period of exposure.

Animals↗

PubMed1995

Central GABA-A receptors exert a tonic inhibitory control on gastric pepsinogen secretion in anaesthetized rats.

1. The purpose of the present study was to analyse the role played by central gamma-aminobutyric acid (GABA-A) receptors in the regulation of gastric basal pepsinogen secretion in anaesthetized rats. 2. The central, but not parenteral, administration of the GABA-A receptor antagonist bicuculline or SR-95531 caused a significant and dose-dependent increase in basal pepsinogen secretion without changes in acid output. The stimulant effect exerted by bicuculline was prevented by atropine or pirenzepine, but not by vagotomy. 3. The central, but not parenteral, administration of the GABA-A receptor agonist muscimol or THIP induced a significant and dose-dependent stimulation of both basal pepsinogen and acid secretion. The excitatory effect exerted by muscimol was prevented by atropine, pirenzepine, or pretreatment with omeprazole, but not by vagotomy. 4. These results suggest that central GABA-A receptors mediate a tonic inhibitory control on gastric pepsinogen secretion, while their phasic activation leads to an excitatory effect on acid output. However, the agonist-induced pepsigogue action appears to be generated peripherally as an indirect consequence of the increase in acid secretion. 5. It is also suggested that central GABA-A receptors affect the gastric secretory functions through non-vagal pathways that are sensitive to the blockade of peripheral cholinergic receptors.

Anesthesia↗

PubMed1994

Adenosine enhances the bronchocontractile response to histamine in anaesthetized and curarized guinea pigs through a mechanism partly blocked by hexamethonium.

The ability of adenosine to potentiate the airway narrowing induced by histamine in anaesthetized and curarized guinea pigs has been investigated in order to establish whether it could be ascribed to a modulatory activity by the nucleoside at the neuronal level. Bilateral vagotomy, atropine (2 mg/kg i.v.), and pretreatment with capsaicin (52 mg/kg s.c. 6 days before the experiment) did not result in any significant protection against the enhancement provoked by the nucleoside of the bronchocontractile effect of histamine. On the contrary, the latter was significantly reduced by the ganglionic blocking agent, hexamethonium (10 mg/kg i.v.). Moreover, the effect of adenosine on airway responsiveness to histamine was not modified in animals treated with propranolol (1 mg/kg i.v.) or guanethidine (20 mg/kg s.c. over a period of 2 days). In conclusion, current data suggest that the purine is able, in our experimental model, to potentiate the bronchospasm induced by histamine by means of a mechanism mediated, at least partly, by non-adrenergic-non-cholinergic nerves not related to capsaicin-sensitive afferent neurons.

Adenosine↗

PubMed1994

Morphofunctional changes in the noradrenergic innervation of the rat cardiovascular system after varying duration of noise stress.

One, six and twelve hs of exposure to acoustic stress showed different influences on the noradrenergic pattern and receptor function of the rat atria and aorta. Moreover, the lipid content of the adrenal cortex and hepatic glycogen were histochemically evaluated in the same animals to correlate these observations with the previous results. The increase in exposure time induced a corresponding increase in sympathetic innervation, which was more evident at cardiac level. The functional results showed that the potency of the agonists on the alpha- and beta-adrenoceptors does not vary, with the exception of 6-h treatment, which affected beta-receptors. By contrast, the M.R.R. of beta-receptors proved to be modified in all treatments, suggesting that noise stress affected mainly postreceptorial mechanisms linked to beta-adrenoceptors rather than their density or affinity; no significant functional modifications were observed when alpha-receptors were considered.

Adrenal Cortex↗