17 research outputs found
Dowodzenie udzielenia dyspensy od przeszkody pokrewieństwa w wyrokach Roty Rzymskiej c. Turnaturi z dnia 25 stycznia 2001 r. i c. De Angelis z dnia 14 lutego 2003 r.
No full textThe author of the presented article focused his attention on the issue of the process of granting a dispensation from the impediment of consanguinity in two verdicts of the Roman Rota – c. Turnaturi of 25th January 2001 and c. De Angelis of 14th February 2003. He proved that the contrary verdicts in the same matter resulted from two different methods of argumentation used by the panel of judges. In the verdict of c. Turnaturi what was taken into consideration was the collected documentation but also other circumstances of the case while in the verdict of c. De Angelis the documents were the main focus of attention. The author believes that in the last sentence the method of decision-making was of positivist nature. 
The Process of Granting a Dispensation from the Impediment of Consanguinity in the Verdicts of the Roman Rota: c. Turnaturi of 25th January 2001 and c. De Angelis of 14th February 2003
No full textThe author of the presented article focused his attention on the issue of the process of granting a dispensation from the impediment of consanguinity in two verdicts of the Roman Rota – c. Turnaturi of 25th January 2001 and c. De Angelis of 14th February 2003. He proved that the contrary verdicts in the same matter resulted from two different methods of argumentation used by the panel of judges. In the verdict of c. Turnaturi what was taken into consideration was the collected documentation but also other circumstances of the case while in the verdict of c. De Angelis the documents were the main focus of attention. The author believes that in the last sentence the method of decision-making was of positivist nature.
Synthesis and Structure-Activity Relationships of (−)-cis-N-Normetazocine-Based LP1 Derivatives
No full text(−)-cis-N-Normetazocine represents a rigid scaffold able to mimic the tyramine moiety of endogenous opioid peptides, and the introduction of different N-substituents influences affinity and efficacy of respective ligands at MOR (mu opioid receptor), DOR (delta opioid receptor), and KOR (kappa opioid receptor). We have previously identified LP1, a MOR/DOR multitarget opioid ligand, with an N-phenylpropanamido substituent linked to (−)-cis-N-Normetazocine scaffold. Herein, we report the synthesis, competition binding and calcium mobilization assays of new compounds 10−16 that differ from LP1 by the nature of the N-substituent. In radioligand binding experiments, the compounds 10−13, featured by an electron-withdrawing or electron-donating group in the para position of phenyl ring, displayed improved affinity for KOR (Ki = 0.85−4.80 μM) in comparison to LP1 (7.5 μM). On the contrary, their MOR and DOR affinities were worse (Ki = 0.18−0.28 μM and Ki = 0.38−1.10 μM, respectively) with respect to LP1 values (Ki = 0.049 and 0.033 μM). Analogous trends was recorded for the compounds 14−16, featured by indoline, tetrahydroquinoline, and diphenylamine functionalities in the N-substituent. In calcium mobilization assays, the compound 10 with a p-fluorophenyl in the N-substituent shared the functional profile of LP1 (pEC50MOR = 7.01), although it was less active. Moreover, the p-methyl- (11) and p-cyano- (12) substituted compounds resulted in MOR partial agonists and DOR/KOR antagonists. By contrast, the derivatives 13−15 resulted as MOR antagonists, and the derivative 16 as a MOR/KOR antagonist (pKBMOR = 6.12 and pKBKOR = 6.11). Collectively, these data corroborated the critical role of the N-substituent in (−)-cis-N-Normetazocine scaffold. Thus, the new synthesized compounds could represent a template to achieve a specific agonist, antagonist, or mixed agonist/antagonist functional profile
Confronto dell'Antibioticoresistenza di Pseudomonas aeruginosa verso tutti gli altri batteri Gram negativi non fermentanti il glucosio: risultati di uno studio osservazionale condotto su isolati da matrici ambientali di un policlinico universitario romano.
No full textOBIETTIVI: In Italia il fenomeno dell’antibioticoresistenza ha assunto dimensioni preoccupanti; i dati epidemiologici rilevati a livello europeo dimostrano, infatti, come l’Italia sia tra i Paesi con frequenze piu elevate di microrganismi resistenti agli antibiotici ed anche tra quelli con i consumi più elevati. Uno studio osservazionale, della durata di 6 anni (Settembre 2004-Settembre 2010), e stato condotto per compararel’antibioticoresistenza di Pseudomonas aeruginosacon quella di gli altri batteri gram negativi non fermentantiil glucosio (BGNnF), tutti isolati da differentimatrici acquose provenienti da reparti a medio ed alto rischio di un Policlinico Universitario di Roma.
METODI: E’ stata testata la suscettibilita degli isolati a 19 antibiotici singolarmente ed a due, tre e quattro antibiotici contemporaneamente. I ceppi sono stati sottoposti a studio della resistenza con sistemi automatici miniaturizzati (ATB PSE 5, Bio- Merieux). Il test del chi-quadro (chi2) e stato utilizzato per valutare l’associazione tra l’antibioticoresistenza di P.aeruginosa e quella dei BGNnF; la significativita statistica e stata fissata ad un valore di p <0.05.
RISULTATI: Sono stati prelevati ed analizzati 2.016 campioni di cui 116 positivi: 40 (34.5%) per presenza di P. aeruginosa e 76 (65.5%) di BGNnF. E’ stata rilevata un’associazione statisticamente significativa delle resistenze (p < 0,001) di P. aeruginosa rispetto ai BGNnF ai seguenti antibiotici: Amikacina (chi2 = 18,56), Ampicillina-sulbactam (chi2 = 25,37), Colistina (chi2 = 40,65), Cotrimoxazolo (chi2 = 54,01), Gentamicina (chi2 = 25,72) e Tobramicina (chi2 = 23,17); non si e, invece, evidenziataun’associazione statisticamente significativa nei confronti di Imipenem (chi2 = 0,37; p = 0,539), Meropenem(chi2= 1,64; p = 0,199), Ceftazidima (chi2= 1,04; p= 0,30) e Cefepima (chi2 = 0,02; p = 0,877), considerati antibiotici di ultima generazione.
CONCLUSIONI: I risultati sopra riportati suggeriscono che i ceppi di P. aeruginosa isolati sono ancora suscettibili agli antibiotici di ultima generazione; ciò conferma l’importanza di un uso prudente e “giudizioso” di questi ultimi. La comprensione del ruolo dell’ecologia microbica ambientale e dei profili di antibioticoresistenza potrebbe aiutare a prevedere e controllare lo sviluppo e l’evoluzione di future resistenze anche nei ceppi clinici. Pertanto, sarebbe opportuno continuare a monitorare costantemente, con cadenze temporali prestabilite, l’evoluzione di questo fenomeno, integrando le migliori evidenze scientifiche relative ai ceppi ambientali con quelle inerenti i ceppi clinici, per tutelare l’igiene dell’ambiente e, al tempo stesso, la sicurezza dei pazienti
Beyond peptidase activity of cyanobacterial phytochelatin synthases: the case of Nostoc sp., Gleiterinema sp., Gloeobacter violaceous
No full textThe enzyme phytochelatin synthase (PCS) is a γ-glutamylcysteine dipeptidyl (trans)peptidase (EC 2.3.2.15), belonging to the clan CA of the papain-like cysteine proteases. The PCS catalyzes the prompt formation of some peculiar thiol-peptide compounds, the so-called “phytochelatins”, starting from the reduced form of glutathione (GSH) via a transpeptidation reaction. Phytochelatins (PCs) are thiol-peptide compounds whose general structure is (γ-glutamate–cysteine)n–glycine, with n usually ranging from 2 to 5 (Grill et al., 1985). Due to the thiol group of the cysteine residues, PCs can bind cadmium (Cd) and other thiophilic heavy metals and prevent them from circulating in the cytosol, thus dramatically reducing their toxicity. It is now well known that land plants, as well as several marine and freshwater algae (e.g. some members of Chlorophyta, Chrysophyceae, Phaeophyceae, Rhodophyta), some fungi, lichens and even some animal species do actually produce PCs in response to heavy metal stress, in particular Cd. PCS is indeed of particular interest from an evolutionary prospect, due to its constitutive expression and its widespread presence in nature. Recently, some PCS-like enzymes, sharing significant sequence homologies with land plant PCSs, were identified in cyanobacteria and in some gamma- and beta- proteobacteria. Previous evidences suggested that predicted product of PCS gene of Nostoc sp. (alr0975) contains the conserved N-terminal domain, but not the variable C-terminal domain found in eukaryotic PCSs (Tsuji et al., 2004; Vivares et al., 2005). Proteins encoded by the cyanobacterial genes seem to be primitive forms of PCS and to represent an early stage in the evolution of the enzyme in photoautotrophic organisms, since up to now only peptidase and almost absent transpeptidase activities have been reported for cyanobacteria (Tsuji et al., 2005). However, studies on functional characterization of prokaryotic PCS are scant, despite their importance as fundamental landmarks in evolution. Thus, the aim of our study is to investigate possible functional and molecular differences between phototrophic eukaryotic and prokaryotic PCS, by studying the enzymes in three cyanobacterial strains (Nostoc sp. PCC 7120, Geitlerinema sp. PCC 7407 and Gloeobacter violaceous PCC 7421). Preliminary HPLC-mass-spectrophotometry results evidenced a noteworthy PC synthesis in all the strains upon Cd-exposure, thus demonstrating that cyanobacterial PCS-like enzymes do possess transpeptidase activity, likewise eukaryotic PCSs. This evidence highlights a remarkable evolutionary conservation of PCS functionality between cyanobacteria, algae and land plants
Novel N-normetazocine Derivatives with Opioid Agonist/Sigma-1 Receptor Antagonist Profile as Potential Analgesics in Inflammatory Pain
No full textAlthough opioids and nonsteroidal anti-inflammatory drugs (NSAIDs) are the most common drugs used in persistent pain treatment; they have shown many side effects. The development of new analgesics endowed with mu opioid receptor/delta opioid receptor (MOR/DOR) activity represents a promising alternative to MOR-selective compounds. Moreover, new mechanisms, such as sigma-1 receptor (σ(1)R) antagonism, could be an opioid adjuvant strategy. The in vitro σ(1)R and σ(2)R profiles of previous synthesized MOR/DOR agonists (−)-2R/S-LP2 (1), (−)-2R-LP2 (2), and (−)-2S-LP2 (3) were assayed. To investigate the pivotal role of N-normetazocine stereochemistry, we also synthesized the (+)-2R/S-LP2 (7), (+)-2R-LP2 (8), and (+)-2S-LP2 (9) compounds. (−)-2R/S-LP2 (1), (−)-2R-LP2 (2), and (−)-2S-LP2 (3) compounds have Ki values for σ1R ranging between 112.72 and 182.81 nM, showing a multitarget opioid/σ1R profile. Instead, (+)-2R/S-LP2 (7), (+)-2R-LP2 (8), and (+)-2S-LP2 (9) isomers displayed a nanomolar affinity for σ1R, with significative selectivity vs. σ2R and opioid receptors. All isomers were evaluated using an in vivo formalin test. (−)-2S-LP2, at 0.7 mg/kg i.p., showed a significative and naloxone-reversed analgesic effect. The σ1R selective compound (+)-2R/S-LP2 (7), at 5.0 mg/kg i.p., decreased the second phase of the formalin test, showing an antagonist σ1R profile. The multitarget or single target profile of assayed N-normetazocine derivatives could represent a promising pharmacological strategy to enhance opioid potency and/or increase the safety margin
New Insights into the Opioid Analgesic Profile of <i>cis</i>-(−)-<i>N</i>-Normetazocine-derived Ligands
No full textIn this work, we report on the in vitro and in vivo pharmacological properties of LP1 analogs to complete the series of structural modifications aimed to generate compounds with improved analgesia. To do that, the phenyl ring in the N-substituent of our lead compound LP1 was replaced by an electron-rich or electron-deficient ring and linked through a propanamide or butyramide spacer at the basic nitrogen of the (−)-cis-N-normetazocine skeleton. In radioligand binding assays, compounds 3 and 7 were found to display nanomolar binding affinity for the μ opioid receptor (MOR) (Ki = 5.96 ± 0.08 nM and 1.49 ± 0.24 nM, respectively). In the mouse vas deferens (MVD) assay, compound 3 showed an antagonist effect against DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin), a highly selective MOR prototype agonist, whereas compound 7 produced naloxone reversible effect at MOR. Moreover, compound 7, as potent as LP1 and DAMGO at MOR, was able to reduce thermal and inflammatory pain assessed by the mouse tail-flick test and rat paw pressure thresholds (PPTs) measured by a Randall–Selitto test
(2S)-N-2-methoxy-2-phenylethyl-6,7-benzomorphan compound (2S-LP2): Discovery of a biased mu/delta opioid receptor agonist
No full textThe pivotal role of the stereocenter at the N-substituent of the 6,7-benzomorphan scaffold was investigated combining synthetic and pharmacological approaches. 2R- and 2S-diastereoisomers of the multitarget MOR/DOR antinociceptive ligand LP2 (1) were synthesized and their pharmacological profile was evaluated in in vitro and vivo assays. From our results, 2S-LP2 (5) showed an improved pharmacological profile in comparison to LP2 (1) and 2R-LP2 (4). 2S-LP2 (5) elicited an antinociceptive effect with a 1.5- and 3-times higher potency than LP2 (1) and R-antipode (4), respectively. In vivo effect of 2S-LP2 (5) was consistent with the improved MOR/DOR efficacy profile assessed by radioligand binding assay, to evaluate the opioid receptor affinity, and BRET assay, to evaluate the capability to promote receptor/G-protein and receptor/β-arrestin 2 interaction. 2S-LP2 (5) was able to activate, with different efficacy, G-protein pathway over β-arrestin 2, behaving as biased agonist at MOR and mainly at DOR. Considering the therapeutic potential of both multitarget MOR/DOR agonism and functional selectivity over G-protein, the 2S-LP2 (5) biased multitarget MOR/DOR agonist could provide a safer treatment opportunity
Exploiting the power of stereochemistry in drug action: 3-[(2S,6S,11S)-8-Hydroxy-6,11-dimethyl-1,4,5,6-tetrahydro-2,6-methano-3-benzazocin-3(2H)-yl]-N-phenylpropanamide as potent Sigma-1 receptor antagonist
No full text(+)-(2S,6S,11S)- and (-)-(2R,6R,11R)-Benzomorphan derivatives have a different binding affinity for sigma-1 (σ1R) and opioid receptors, respectively. In this study, we describe the synthesis of the (+)-enantiomer [(+)-LP1] of the benzomorphan MOR agonist/DOR antagonist LP1 [(-)-LP1]. The binding affinity of both (+)-LP1 and (-)-LP1 for σ1R and sigma-2 receptor (σ2R) was tested. Moreover, (+)-LP1 opioid receptor binding affinity was also investigated. Finally, (+)-LP1 was tested in a mouse model of inflammatory pain. Our results showed a nanomolar σ1R and binding affinity for (+)-LP1. Both (+)-LP1 and (-)-LP1 elicited a significant analgesic effect in a formalin test. Differently from (-)-LP1, the analgesic effect of (+)-LP1 was not reversed by naloxone, suggesting a σ1R antagonist profile. Furthermore, σ1R agonist PRE-084 was able to unmask the σ1R antagonistic component of the benzomorphan compound. (+)-LP1 could constitute an useful lead compound to develop new analgesics based on mechanisms of action alternative to opioid receptor activation
