ANCIENT LAW – UM CLÁSSICO REVISITADO 150 ANOS DEPOIS

Este artigo pretende realçar a importância desempenhada pela obra de Henry James Sumner Maine na formação da antropologia jurídica e da sociologia do direito. Mediante a recuperação da tese central de sua obra Ancient Law, procura ressaltar o papel por ela desempenhado no delineamento de uma nova forma de abordagem da relação entre direito e sociedade. Para tanto, recupera traços gerais das análises de Norbert Rouland e de Niklas Luhmann acerca da importância das ideias do autor.

Secondary metabolites from two species of ·Tolpis and their biological activities

[EN] Phytochemical research of two Tolpis species, T. proustii and T. lagopoda, led to the isolation of three new compounds: 30-chloro-3beta-acetoxy-22alfa-hydroxyl-20(21)- taraxastene (1), 3beta,22alfa-diacetoxy-30-ethoxy-20(21)-taraxastene (2) and 3beta,28-dihydroxy- 11alfa-hydroperoxy-12-ursene (3). The structures of the new compounds were elucidated by means of extensive IR, NMR, and MS data and by comparison of data reported in the literature. The in vitro antioxidant activities of the extracts were assessed by the DPPH and ABTS scavenging methods. The cytotoxicity of several known compounds and its derivatives was also assessed against human myeloid leukemia K-562 and K-562/ADR cell lines.

Processo biotecnologico a cellule immobilizzate per la produzione di acidi grassi volatili da acque di vegetazione

Le acque di vegetazione (AV) costituiscono un serio problema di carattere ambientale, sia a causa della loro elevata produzione sia per l’ elevato contenuto di COD che oscilla fra 50 e 150 g/l. Le AV sono considerate un refluo a tasso inquinante fra i più elevati nell’ambito dell’industria agroalimentare e la loro tossicità è determinata in massima parte dalla componente fenolica. Il presente lavoro si propone di studiare e ottimizzare un processo non solo di smaltimento di tale refluo ma anche di una sua valorizzazione, utlizzandolo come materia prima per la produzione di acidi grassi e quindi di PHA, polimeri biodegradabili utilizzabili in varie applicazioni. A tale scopo sono stati utilizzati due bioreattori anaerobici a biomassa adesa, di identica configurazione, con cui si sono condotti due esperimenti in continuo a diverse temperature e carichi organici al fine di studiare l’influenza di tali parametri sul processo. Il primo esperimento è stato condotto a 35°C e carico organico pari a 12,39 g/Ld, il secondo a 25°C e carico organico pari a 8,40 g/Ld. Si è scelto di allestire e mettere in opera un processo a cellule immobilizzate in quanto questa tecnologia si è rivelata vantaggiosa nel trattamento continuo di reflui ad alto contenuto di COD e carichi variabili. Inoltre si è scelto di lavorare in continuo poiché tale condizione, per debiti tempi di ritenzione idraulica, consente di minimizzare la metanogenesi, mediata da microrganismi con basse velocità specifiche di crescita. Per costituire il letto fisso dei due reattori si sono utilizzati due diversi tipi di supporto, in modo da poter studiare anche l’influenza di tale parametro, in particolare si è fatto uso di carbone attivo granulare (GAC) e filtri ceramici Vukopor S10 (VS). Confrontando i risultati si è visto che la massima quantità di VFA prodotta nell’ambito del presente studio si ha nel VS mantenuto a 25°C: in tale condizione si arriva infatti ad un valore di VFA prodotti pari a 524,668 mgCOD/L. Inoltre l’effluente in uscita risulta più concentrato in termini di VFA rispetto a quello in entrata: nell’alimentazione la percentuale di materiale organico presente sottoforma di acidi grassi volatili era del 54 % e tale percentuale, in uscita dai reattori, ha raggiunto il 59 %. Il VS25 rappresenta anche la condizione in cui il COD degradato si è trasformato in percentuale minore a metano (2,35 %) e questo a prova del fatto che l’acidogenesi ha prevalso sulla metanogenesi. Anche nella condizione più favorevole alla produzione di VFA però, si è riusciti ad ottenere una loro concentrazione in uscita (3,43 g/L) inferiore rispetto a quella di tentativo (8,5 g/L di VFA) per il processo di produzione di PHA, sviluppato da un gruppo di ricerca dell’università “La Sapienza” di Roma, relativa ad un medium sintetico. Si può constatare che la modesta produzione di VFA non è dovuta all’eccessiva degradazione del COD, essendo questa nel VS25 appena pari al 6,23%, ma piuttosto è dovuta a una scarsa concentrazione di VFA in uscita. Questo è di buon auspicio nell’ottica di ottimizzare il processo migliorandone le prestazioni, poiché è possibile aumentare tale concentrazione aumentando la conversione di COD in VFA che nel VS25 è pari a solo 5,87%. Per aumentare tale valore si può agire su vari parametri, quali la temperatura e il carico organico. Si è visto che il processo di acidogenesi è favorito, per il VS, per basse temperature e alti carichi organici. Per quanto riguarda il reattore impaccato con carbone attivo la produzione di VFA è molto ridotta per tutti i valori di temperatura e carichi organici utilizzati. Si può quindi pensare a un’applicazione diversa di tale tipo di reattore, ad esempio per la produzione di metano e quindi di energia.

Eocene nappe tectonics and late Alpine extension in the Central Anatolide belt, western Turkey

Strukturgeologische Untersuchungen belegen, daß die Anatoliden der Westtürkei im Eozän durch die Plazierung der Kykladischen Blauschiefereinheit entlang einer durchbrechenden Überschiebung auf die Menderes-Decken unter grünschieferfaziellen Metamorphosebedingungen entstanden.Die kykladischen Blauschiefer in der Westtürkei enthalten Relikte eines prograden alpinen Gefüges (DA1), welches hochruckmetamorph von Disthen und Chloritoid poikiloblastisch überwachsen wurde. Dieses Mineralstadium dauerte noch während des Beginns des nachfolgenden Deformationsereignisses (DA2) an, welches durch NE-gerichtete Scherung und Dekompression charakterisiert ist. Die nachfolgende Deformation (DA3) war das erste Ereignis, das beide Einheiten, sowohl die kykladische Blauschifereinheit als auch die Menderes-Decken, gemeinsam erfaßte. Der Überschiebungskontakt zwischen der kykladischen Blauschiefereinheit und den Menderes-Decken ist eine DA3-Scherzone: die ‘Cycladic-Menderes Thrust’ (CMT). Entlang der CMT-Überschiebungsbahn wurden die kykladischen Blauschiefer gegen veschiedene Einheiten der MN plaziert. Die CMT steigt nach S zum strukturell Hangenden hin an und kann daher als eine durchbrechende Überschiebung entlang einer nach S ansteigenden Rampe betrachtet werden. In den kykladischen Blauschiefern überprägen DA3-Strukturen, die im Zusammenhang mit der CMT stehen, hochdruckmentamorphe Gefüge.In den Menderes-Decken, dem Liegenden der CMT, wird DA3 durch regional vebreitete Gefügeelemente dokumentiert, die im Zusammenhang mit S-gerichteten Schersinnindikatoren stehen. DA3-Gefüge haben die Decken intern deformiert und bilden jene Scherzonen, welche die Decken untereinander abgrenzen. In der Çine-Decke können granitische Gesteine in Orthogneise und Metagranite unterteilt werden. Die Deformationsgeschichte dieser Gesteine dokumentiert zwei Ereignisse. Ein frühes amphibolitfazielles Ereignis erfaßte nur die Orthogneise, in denen vorwiegend NE-SW orientierte Lineare und NE-gerichtete Schersinnindikatoren entstanden. Die jüngeren Metagranite wurden sowohl durch vereinzelte DA3-Scherzonen, als auch in einer großmaßstäblichen DA3-Scherzone am Südrand des Çine-Massivs deformiert. In DA3-Scherzonen sind die Lineare N-S orientiert und die zugehörigen Schersinnindikatoren zeigen S-gerichtete Scherung unter grünschieferfaziellen Bedingungen an. Diese grünschieferfaziellen Scherzonen überprägen die amphibolitfaziellen Gefüge in den Orthogneisen. Magmatische Zirkone aus einem Metagranit, der einen Orthogneiss mit Top-NE Gefügen durchschlägt, ergaben ein 207Pb/206Pb-Alter von 547,2±1,0 Ma. Dies deutet darauf hin, daß DPA proterozoischen Alters ist. Dies wird auch durch die Tatsache gestützt, daß triassische Granite in der Çine- und der Bozdag-Decke keine DPA-Gefüge zeigen. Die jüngeren Top-S-Gefüge sind wahrscheinlich zur gleichen Zeit entstanden wie die ältesten Gefüge der Bayindir-Decke.Das Fehlen von Hochdruck-Gefügen im Liegenden der CMT impliziert eine Exhumierung der kykladischen Blauschiefer von mehr ca. 35 km, bevor diese im Eozän auf die Menderes-Decken aufgeschoben wurden. Die substantiellen Unterschiede bezüglich in der tektonometamorphen Geschichte der kykladischen Blauschiefer und der Menderes-Decken widersprechen der Modellvorstellung eines lateral kontinuierlichen Orogengürtels, nach der die Menderes-Decken als östliche Fortsezung der kykladischen Blauschiefer angesehen werden.Die Analyse spröder spätalpiner Deformationsstrukturen und das regionale Muster mit Hilfe von Spaltspurdatierung modellierter Abkühlalter deuten darauf hin, daß die Struktur des Eozänen Deckenstapels durch miozäne bis rezente Kernkomplex-Bildung stark modifiziert wurde. Eine großmaßstäbliche Muldenstruktur im zentralen Teil der Anatoliden hat sich als Folge zweier symmetrisch angeordneter Detachment-Systeme von initial steilen zu heute flachen Orientierungen im Einflußbreich von ’Rolling Hinges’ gebildet. Die Detachment-Störungen begrenzen den ‘Central Menderes metamorphic core complex’ (CMCC). Das Muster der Apatit-Spaltspuralter belegt, daß die Bildung des CMCC im Miozän begann. Durch die Rück-Deformierung von parallel zur Foliation konstruierten Linien gleicher Abkühlalter kann gezeigt werden, daß die Aufwölbung im Liegenden der Detachments zur Entstehung der Muldenstruktur führte. Das hohe topographische Relief im Bereich des CMCC ist eine Folge der Detachment-Störungen, was darauf hindeutet daß der obere Mantel in den Prozeß mit einbezogen gewesen ist.

Reverse genetic studies of Benyvirus - Polymyxa betae molecular interaction: Role of the RNA4-encoded protein in virus transmission

Beet necrotic yellow vein virus (BNYVV), the leading infectious agent that affects sugar beet, is included within viruses transmitted through the soil from plasmodiophorid as Polymyxa betae. BNYVV is the causal agent of Rhizomania, which induces abnormal rootlet proliferation and is widespread in the sugar beet growing areas in Europe, Asia and America; for review see (Peltier et al., 2008). In this latter continent, Beet soil-borne mosaic virus (BSBMV) has been identified (Lee et al., 2001) and belongs to the benyvirus genus together with BNYVV, both vectored by P. betae. BSBMV is widely distributed only in the United States and it has not been reported yet in others countries. It was first identified in Texas as a sugar beet virus morphologically similar but serologically distinct to BNYVV. Subsequent sequence analysis of BSBMV RNAs evidenced similar genomic organization to that of BNYVV but sufficient molecular differences to distinct BSBMV and BNYVV in two different species (Rush et al., 2003). Benyviruses field isolates usually consist of four RNA species but some BNYVV isolates contain a fifth RNA. RNAs -1 contains a single long ORF encoding polypeptide that shares amino acid homology with known viral RNA-dependent RNA polymerases (RdRp) and helicases. RNAs -2 contains six ORFs: capsid protein (CP), one readthrough protein, triple gene block proteins (TGB) that are required for cell-to-cell virus movement and the sixth 14 kDa ORF is a post-translation gene silencing suppressor. RNAs -3 is involved on disease symptoms and is essential for virus systemic movement. BSBMV RNA-3 can be trans-replicated, trans-encapsidated by the BNYVV helper strain (RNA-1 and -2) (Ratti et al., 2009). BNYVV RNA-4 encoded one 31 kDa protein and is essential for vector interactions and virus transmission by P. betae (Rahim et al., 2007). BNYVV RNA-5 encoded 26 kDa protein that improve virus infections and accumulation in the hosts. We are interest on BSBMV effect on Rhizomania studies using powerful tools as full-length infectious cDNA clones. B-type full-length infectious cDNA clones are available (Quillet et al., 1989) as well as A/P-type RNA-3, -4 and -5 from BNYVV (unpublished). A-type BNYVV full-length clones are also available, but RNA-1 cDNA clone still need to be modified. During the PhD program, we start production of BSBMV full-length cDNA clones and we investigate molecular interactions between plant and Benyviruses exploiting biological, epidemiological and molecular similarities/divergences between BSBMV and BNYVV. During my PhD researchrs we obtained full length infectious cDNA clones of BSBMV RNA-1 and -2 and we demonstrate that they transcripts are replicated and packaged in planta and able to substitute BNYVV RNA-1 or RNA-2 in a chimeric viral progeny (BSBMV RNA-1 + BNYVV RNA-2 or BNYVV RNA-1 + BSBMV RNA-2). During BSBMV full-length cDNA clones production, unexpected 1,730 nts long form of BSBMV RNA-4 has been detected from sugar beet roots grown on BSBMV infected soil. Sequence analysis of the new BSBMV RNA-4 form revealed high identity (~100%) with published version of BSBMV RNA-4 sequence (NC_003508) between nucleotides 1-608 and 1,138-1,730, however the new form shows 528 additionally nucleotides between positions 608-1,138 (FJ424610). Two putative ORFs has been identified, the first one (nucleotides 383 to 1,234), encode a protein with predicted mass of 32 kDa (p32) and the second one (nucleotides 885 to 1,244) express an expected product of 13 kDa (p13). As for BSBMV RNA-3 (Ratti et al., 2009), full-length BSBMV RNA-4 cDNA clone permitted to obtain infectious transcripts that BNYVV viral machinery (Stras12) is able to replicate and to encapsidate in planta. Moreover, we demonstrated that BSBMV RNA-4 can substitute BNYVV RNA-4 for an efficient transmission through the vector P. betae in Beta vulgaris plants, demonstrating a very high correlation between BNYVV and BSBMV. At the same time, using BNYVV helper strain, we studied BSBMV RNA-4’s protein expression in planta. We associated a local necrotic lesions phenotype to the p32 protein expression onto mechanically inoculated C. quinoa. Flag or GFP-tagged sequences of p32 and p13 have been expressed in viral context, using Rep3 replicons, based on BNYVV RNA-3. Western blot analyses of local lesions contents, using FLAG-specific antibody, revealed a high molecular weight protein, which suggest either a strong interaction of BSBMV RNA4’s protein with host protein(s) or post translational modifications. GFP-fusion sequences permitted the subcellular localization of BSBMV RNA4’s proteins. Moreover we demonstrated the absence of self-activation domains on p32 by yeast two hybrid system approaches. We also confirmed that p32 protein is essential for virus transmission by P. betae using BNYVV helper strain and BNYVV RNA-3 and we investigated its role by the use of different deleted forms of p32 protein. Serial mechanical inoculation of wild-type BSBMV on C. quinoa plants were performed every 7 days. Deleted form of BSBMV RNA-4 (1298 bp) appeared after 14 passages and its sequence analysis shows deletion of 433 nucleotides between positions 611 and 1044 of RNA-4 new form. We demonstrated that this deleted form can’t support transmission by P. betae using BNYVV helper strain and BNYVV RNA-3, moreover we confirmed our hypothesis that BSBMV RNA-4 described by Lee et al. (2001) is a deleted form. Interesting after 21 passages we identifed one chimeric form of BSBMV RNA-4 and BSBMV RNA-3 (1146 bp). Two putative ORFs has been identified on its sequence, the first one (nucleotides 383 to 562), encode a protein with predicted mass of 7 kDa (p7), corresponding to the N-terminal of p32 protein encoded by BSBMV RNA-4; the second one (nucleotides 562 to 789) express an expected product of 9 kDa (p9) corresponding to the C-terminal of p29 encoded by BSBMV RNA-3. Results obtained by our research in this topic opened new research lines that our laboratories will develop in a closely future. In particular BSBMV p32 and its mutated forms will be used to identify factors, as host or vector protein(s), involved in the virus transmission through P. betae. The new results could allow selection or production of sugar beet plants able to prevent virus transmission then able to reduce viral inoculum in the soil.

Influence of genetic variants and post transcriptional factors on imatinib transporters as determinants of the pharmacological response

Introduction – Although imatinib (IM) is a recognized gold standard in chronic myeloid leukemia (CML) therapy, resistance has emerged in a significant proportion of patients. Aim – The aim of this study was: (1) to investigate the role of genetic variants in genes encoding for IM transporters, as candidate of IM responsiveness and (2) to test the influence of miRNAs on IM response, focusing on efflux transporters. Methods – As a first step, a panel of polymorphisms (SNPs) was genotyped in a subgroup population of 189 patients enrolled in the Tyrosine Kinase Inhibitor Optimization and Selectivity (TOPS) trial. The association with cytogenetic response and molecular response (MR) was assessed for each SNP. As a second step, an in vitro IM-resistant model (K-562 CML cell line) was established. miRNAs profiles were analyzed using Taqman arrays and in silico search was performed for miRNAs deregulated after IM treatment. mRNA and protein expression were quantified using TaqMan realtime PCR and Western blotting, respectively. Results – (1) Among Caucasian patients, ABCB1 rs60023214 significantly correlated with complete MR (P = 0.005). Concerning SNPs combination in IM uptake transporters, the associations with treatment outcomes were statistically significant for both major and complete MR (P = 0.005 and P = 0.01, respectively). (2) ABCB1 protein was not expressed under any conditions of treatment, differently from ABCG2. Two deregulated miRNAs, namely miR-212 and miR-328, were identified to be inversely correlated with ABCG2 (r2= 0.57; p=0.03 and r2=0.47; p=0.06, respectively). Experiments of loss and gain of function confirmed the functional influence of these miRNAs on ABCG2. Conclusion – The multiple candidate gene approach identified single and combination of SNPs that can be proposed as predictor of IM response. The in vitro study suggested that IM resistance could be mediated by miRNA-dependent mechanism. Further studies are needed to validate these preliminary findings.

Nocturnal periodic breathing during acclimatization at very high altitude at Mount Muztagh Ata (7,546 m)

Quantitative data on ventilation during acclimatization at very high altitude are scant. Therefore, we monitored nocturnal ventilation and oxygen saturation in mountaineers ascending Mt. Muztagh Ata (7,546 m).

A randomised determination of the effect of fluvastatin and atorvastatin on top of dual antiplatelet treatment on platelet aggregation after implantation of coronary drug-eluting stents. The EFA-Trial

Drug-drug interaction between statins metabolised by cytochrome P450 3A4 and clopidogrel have been claimed to attenuate the inhibitory effect of clopidogrel. However, published data regarding this drug-drug interaction are controversial. We aimed to determine the effect of fluvastatin and atorvastatin on the inhibitory effect of dual antiplatelet therapy with acetylsalicylic acid (ASA) and clopidogrel. One hundred one patients with symptomatic stable coronary artery disease undergoing percutaneous coronary intervention and drug-eluting stent implantation were enrolled in this prospective randomised study. After an interval of two weeks under dual antiplatelet therapy with ASA and clopidogrel, without any lipid-lowering drug, 87 patients were randomised to receive a treatment with either fluvastatin 80 mg daily or atorvastatin 40 mg daily in addition to the dual antiplatelet therapy for one month. Platelet aggregation was assessed using light transmission aggregometry and whole blood impedance platelet aggregometry prior to randomisation and after one month of receiving assigned statin and dual antiplatelet treatment. Platelet function assessment after one month of statin and dual antiplatelet therapy did not show a significant change in platelet aggregation from 1st to 2nd assessment for either statin group. There was also no difference between atorvastatin and fluvastatin treatment arms. In conclusion, neither atorvastatin 40 mg daily nor fluvastatin 80 mg daily administered in combination with standard dual antiplatelet therapy following coronary drug-eluting stent implantation significantly interfere with the antiaggregatory effect of ASA and clopidogrel.