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Gli ultimi terremoti severi avvenuti in Italia hanno mostrato l’estrema vulnerabilità delle costruzioni storiche. In particolare una delle principali criticità osservate ha riguardato gli archi e le volte. Risulta evidente che una conoscenza approfondita del comportamento dinamico di questi elementi strutturali rappresenta un passo fondamentale per la salvaguardia del patrimonio storico. Normalmente le volte vengono studiate con metodi semplificati o, in alternativa, con metodi FEM più avanzati. Ad oggi, sperimentazioni sul comportamento dinamico di questi elementi strutturali rappresentano un vuoto nell’ingegneria strutturale che va necessariamente colmato. Nell’ambito del progetto di ricerca PROVACI, (Tecnologie per la PROtezione e la VAlorizzazione di Complessi di Interesse culturale) rientrante nel Programma Operativo Nazionale per la Ricerca e la Competitività PON-REC 2007-20013 (PON 01_02324), è stata svolta una campagna di prove per approfondire il comportamento dinamico di volte in muratura per effetto di azioni sismiche. Allo scopo è stato appositamente realizzato e, successivamente, testato, presso il dipartimento di strutture per l’ingegneria e l’architettura della facoltà di Napoli Federico II2, mediante prove dinamiche su tavola vibrante, un campione di volta in muratura in scala reale. La presente memoria descrive approfonditamente tutta la campagna di prove svolta, che ha riguardato molteplici test eseguiti sul campione e, successivamente, il rinforzo dello stesso, a valle del quale è stata svolta un’ulteriore campagna di prove. Viene quindi confrontata la risposta strutturale del campione prima e dopo il rinforzo potendone quantificare l’incremento prestazionale raggiunto.

DVT is common among critically ill patients. A rapid and accurate diagnosis is essential for patient care. We assessed the accuracy and timeliness of intensivist-performed compression ultrasonography studies (IP-CUS) for proximal lower extremity DVT (PLEDVT) by comparing results with the formal vascular study (FVS) performed by ultrasonography technicians and interpreted by radiologists.We conducted a multicenter, retrospective review of IP-CUS examinations performed in an ICU by pulmonary and critical care fellows and attending physicians. Patients suspected of having DVT underwent IP-CUS, using a standard two-dimensional compression ultrasonography protocol for the diagnosis of PLEDVT. The IP-CUS data were collected prospectively as part of a quality-improvement initiative. The IP-CUS interpretation was recorded and timed at the end of the examination on a standardized report form. An FVS was then ordered, and the FVS result was used as the criterion standard for calculating sensitivity and specificity. Time delays between the IP-CUS and FVS were recorded.A total of 128 IP-CUS were compared with an FVS. Eighty-one percent of the IP-CUS were performed by fellows with2 years of clinical ultrasonography experience. Prevalence of DVT was 20%. IP-CUS studies yielded a sensitivity of 86% and a specificity of 96% with a diagnostic accuracy of 95%. Median time delay between the ordering of FVS and the FVS result was 13.8 h.Rapid and accurate diagnosis of proximal lower extremity DVT can be achieved by intensivists performing compression ultrasonography at the bedside.

Negli ultimi anni le richieste da parte degli utenti finali di veicoli sempre più performanti in termini di comfort acustico e le stringenti normative europee, hanno spinto i progettisti a rivolgere la propria attenzione verso la qualità del suono percepita a bordo, guardando a questa come un importante obiettivo da raggiungere già durante le prime fasi di pre-design. In quest'ottica si inserisce l'NVH-CAE, ovvero quel settore che studia le problematiche NVH (Noise Vibration and Harshness) utilizzando gli strumenti CAE (Computer Aided Engineering). Il presente articolo ha come fine la stima del rumore percepito dagli occupanti di un veicolo commerciale, causato dalle vibrazioni superficiali del power-train; sebbene le fonti di rumore di un veicolo sono molteplici, quella proveniente dal propulsore, quando il veicolo è fermo, risulta essere la preponderante alle basse frequenze. Una volta completato il modello CAD e realizzato quello ad elementi finiti, si è proceduto al calcolo dei modi strutturali ed acustici propedeutici alla interazione fluido-struttura (FSI); le analisi, modali e della risposta accoppiata, sono state effettuate con il software VPS di ESI-Group. Le fonti di rumore introdotte nel modello numerico analizzato sono state ricavate sperimentalmente mediante l'utilizzo di una sonda Microflown in campo vicino con il motore a 1500 e poi a 2000 rpm. È stato misurato, inoltre, tramite l'utilizzo di microfoni, il rumore percepito ad entrambe le orecchie dal driver, in maniera tale da effettuare poi una validazione numerico-sperimentale del modello creato. A valle dell'introduzione del materiale poroelastico (PEM) in tutte quelle zone dell'abitacolo in cui è classica la sua applicazione, è stato possibile osservare la diminuzione del rumore percepito. Si è proceduto infine al confronto con i risultati sperimentali, sia in termini di spettro in frequenza del rumore misurato sia in termini di overall level. In particolare, gli scarti percentuali tra il rumore globale percepito dai microfoni virtuali e quelli sperimentali, sia a 1500 che a 2000 rpm, sono risultati essere molto contenuti.

The aim of this study was to assess the genetic diversity of four autochthonous cattle breeds of Tuscany and their relationships in comparison with Italian Friesian and Italian Brown, using amplified fragment length polymorphism markers. A total of 212 individuals were genotyped with three primer combinations generating 102 polymorphic markers. Average expected heterozygosity ranged from 0.23 in Mucca Pisana to 0.26 in Chianina, Italian Friesian, Italian Brown and Maremmana. The differences resulted not significant (Kruskall-Wallis test, p = 0.416). Gst-B index revealed that 86% of the total genetic variance is retained within population and only 14% is accounted by the between populations component. Multivariate analysis at individual and population level indicated that: (i) Calvana and Chianina are quite separate from the other breeds as an effect of the bottleneck experienced or as a signature of different origin; (ii) Podolian, Maremmana and Italian Brown clustered with the double purpose Mucca Pisana, revealing their contribution to its admixed genetic make up; (iii) Italian Friesian behaved always as out group. The 'analysis of molecular variance' recovered a significant subdivision clustering the six populations into three groups: Italian Friesian and Italian Brown versus Maremmana and Mucca Pisana versus Chianina and Calvana (6% of the total variance).

The BH3-interacting domain death agonist (Bid) is a pro-apoptotic member of the B-cell lymphoma-2 (Bcl-2) protein family. Previous studies have shown that stress reduces levels of Bcl-2 in brain regions implicated in the pathophysiology of mood disorders, whereas antidepressants and mood stabilizers increase Bcl-2 levels. The Bcl-2 protein family has an essential role in cellular resilience as well as synaptic and neuronal plasticity and may influence mood and affective behaviors. This study inhibited Bid in mice using two pharmacological antagonists (BI-11A7 and BI-2A7); the selective serotonin reuptake inhibitor citalopram was used as a positive control. These agents were studied in several well-known rodent models of depression—the forced swim test (FST), the tail suspension test (TST), and the learned helplessness (LH) paradigm—as well as in the female urine sniffing test (FUST), a measure of sex-related reward-seeking behavior. Citalopram and BI-11A7 both significantly reduced immobility time in the FST and TST and attenuated escape latencies in mice that underwent the LH paradigm. In the FUST, both agents significantly improved duration of female urine sniffing in mice that had developed helplessness. LH induction increased the activation of apoptosis-inducing factor (AIF), a caspase-independent cell death constituent activated by Bid, and mitochondrial AIF expression was attenuated by chronic BI-11A7 infusion. Taken together, the results suggest that functional perturbation of apoptotic proteins such as Bid and, alternatively, enhancement of Bcl-2 function, is a putative strategy for developing novel therapeutics for mood disorders.

The activity of a set of anilide inhibitors of the Hill reaction was modeled using the traditional Hansch approach and Comparative Molecular Field Analysis (CoMFA). In the “best” Hansch model the most relevant parameters were the hydrophobic constants associated to substituents at the 3- and 4-positions of the benzene ring (π3 and π4) and the B 1 Verloop's parameter describing the “minimum width” of the substituent attached to the carbonyl. Successively, a combined “Hansch–CoMFA” analysis was performed using as descriptors the steric field of the acyl substituents in conjunction with the π3 and π4 constants multiplied by proper weighting factors. The results of this latter type of analysis were significantly better than those obtained through the traditional Hansch approach. The predictive ability of the “Hansch–CoMFA” model was further tested by predicting the activity of a large number of anilides not included in the training set. The residuals of such predictions indicated that the Hansch–CoMF...

The mucosa around implants and the gingiva around teeth respond to plaque formation with the development of an inflammatory lesion which has similar magnitude and histological features. Different cell types in inflamed and healthy periodontal and peri-implant tissues are capable of producing a variety of important pro-inflammatory and anti-inflammatory cytokines and growth factors which mediate the host response. The aim of this study is to compare the expression levels of anti-inflammatory cytokines detectable in the peri-implant soft tissue of two single-implant crowns supported either by zirconia or titanium abutments. Two frozen samples of peri-implant soft tissue of two single-implant crowns supported either by zirconia or titanium abutments were treated to obtain mRNA. The mRNA extracted from these specimens was converted in cDNA and analyzed with “SuperArray GEArray Q Series Human Inflammatory Cytokine/Receptor Gene Array kit”, planned for studying 96 genes involved in inflammatory response. Data showed that gene expression levels of anti-inflammatory cytokines were higher in specimens sampled from the zirconia abutment compared with those from the titanium abutment. It was considered important to detect the mRNA levels of the anti-inflammatory mediators in healthy peri-implant tissues to verify the biological tolerability of zirconia compared with titanium abutments. The difference detected in cytokine expression could be due to the intrinsic biological tolerability of zirconia ceramics or to a lesser bacterial accumulation.

The verification of the breed origin of animal products is relevant for food safety and authenticity. We assessed the suitability of AFLP molecular markers in the assignment of cattle individuals to their breed of origin. Three hundred and ninety-six animals belonging to 16 cattle breeds genotyped with 141 AFLP markers were used as reference data set. Assignment was performed with likelihood (aflpop) and Bayesian (structure) methods. The Bayesian approach was superior to the likelihood algorithm with respect to (i) the correct assignment of simulated individuals to their breed of origin (93% vs. 81% respectively), (ii) the correct assignment of 44 sampled Romagnola animals (91% vs. 45% respectively) and (iii) the correct classification of animals belonging to a breed that was not included within the reference dataset. Thus, AFLP profiling in combination with the Bayesian approach seems a useful tool for breed assignment.

The implant/abutment interface represents a crucial element to maintain the structural stability of the implant restoration and to prevent mechanical problems that can compromise the long-term success of the implant treatment.In this study deformation of the external hexagonal and internal octagonal extension of 2 implants after applying different values of insertion torque has been tested. The implants were secured in a customized resin baseplate, with the implant neck exposed, then different values of insertion torque were applied by an electronic torque controller.Five internal octagonal implants were tested for 60 seconds to different torque magnitudes (12 Ncm, 21 Ncm, 27 Ncm, 36 Ncm, 42 Ncm). Two external hexagonal implants were tested for 60 seconds to a torque value of 27 Ncm and of 36 Ncm.After torquing tests, each implant was examined by a scanning electron microscope (SEM) medium and high magnification to evaluate the deformation of the 2 different antirotational systems. The SEM analysis of the internal octagonal and external hexagonal antirotational systems of the TBR implant showed a deformation proportional to the magnitude of the applied torque during implant insertion. The geometric deformation of the antirotational extension is capable of compromising its function after a torque magnitude of 36 Ncm or superior to this value. Moreover it was demonstrated a more evident alteration of the external hexagonal extension in comparison with the octagonal system after applying a torque value of 27 Ncm.

Implant impression and transfer of the implant position to a working cast for rehabilitative purposes not always represent a predictable reconstructive step. The variable results depend on the transfer technique utilized and on the accuracy of the impression materials. The aim of this study was to compare the accuracy of the direct and indirect impression techniques and to analyze the distortion of different impression materials. An edentulous acrylic resin mandible was used as reference model: in the cuspid position 2 laboratory analogs were inserted perpendicularly to the occlusal plane and fastened with resin. Two impressions for each indirect and direct recording techniques with 4 different impression materials were made. Polyvinyl siloxanes were used for the repositioning implant level method. Polyethers were used for the ''pick up'' impression technique. Analysis of cast distortion was measured computing the inter-implant position on the developed models compared to that of the master model. Measurements were recorded by a digital caliper. The results of this in vitro study demonstrate that the direct impression technique is preferable to the indirect one and that the Impregum polyether impression material is the most precise when used in association with a direct technique.

Ureaplasma parvum and Ureaplasma urealyticum, also known as biovar parvum and biovar T960, respectively, could be associated with several disorders in men, women, and mainly, in newborn children with under weight. Several methods have been developed in order to identify the species or biovars of ureaplasmas. We developed a Multiplex-PCR method using the UPS-UPSA and UUS2-UUA2 primers, specific for U. parvum and U. urealyticum, respectively. This Multiplex-PCR method was used to identify cultures of clinical positive samples to Ureaplasma spp. by the "MYCOFAST Evolution-2" Kit. Of 56 positive cultures to Ureaplasma spp. from newborn children, 70% were U. parvum and 30% U. urealyticum; in 76 positive samples in women, 83% corresponded to U. parvum and 17% to U. urealyticum, while in 63 positive samples of men, 76% identified U. parvum and 24% U. urealyticum. The PCR-multiplex method showed specificity for the identification of the biovars or species of ureaplasmas of clinical interest.

Biphosphonates are a large class of drugs which inhibits the resorbing activity of osteoclasts. The aim of this study was to evaluate the influence of topical application of the biphosphonate alendronate on the integration process of endosseus implants placed in a partially edentulous patient. After incision and elevation of a mucoperiosteal flap and carrying out the osteotomy, 2 SLA implants 10 mm long and 3.5 mm wide were inserted in the area of # 4.5 and 4.6. Before placement of # 4.5, a paste made of grinded tablets of alendronate and Rifocin was applied in the osteotomy. Fifteen days after stage I, an evaluation of the osteoblastic peri-implant activity was carried out using single photon emission computerized tomography. Healing was within normal limits. Four months after stage I, the 2 implants were uncovered and considered clinically and radiographically integrated. The osteoblastic activity was judged similar bet-ween the 2 implants in terms of quality of biological response. Osseous scintigraphy and the achievement of implant osseointegration demonstrate the lack of toxicity of the topical application of the biphosphonate alendronate during the initial osseous healing. However, new experimental studies and clinical trials are necessary to confirm the results obtained.

Oral implantology is considered an acceptable treatment modality to replace missing teeth when a recipient site with a sufficient horizontal and vertical bone volume exists. Guided bone regeneration is based on the principle of selective exclusion of non osteogenic cells from the bony defect by a barrier membrane. Several supporting systems of the barrier membrane have been successfully used to treat non space-mantaining de-fects. A 35-year-old female affected by a partial edentulism in the molar area of the right mandible showed 8 mm of bone height of the residual ridge above the alveolar canal. A vertical ridge augmentation was performed using a Gore-Tex membrane in conjunction with a microplate and an irradiated freeze-dried cortico-cancellous allograft. Six months after the surgical procedure a 11.5 mm by 5 mm threaded implant was placed into the regenerated bone. After three months of healing, the implant was uncovered and considered integrated. Before the implant placement, a biopsy of the regenerated bone was arried out by a triphine. The histological examination of the bony core showed the attainment of biological goals of guided bone regeneration. The surgical technique followes in this case report has been effective in regenerating a vertical bone volume sufficient for implant treatment.

The field of NMR spectroscopy has grown beyond expectations from the first historic observation of nuclear magnetic resonance in 1946 by Bloch and Purcell, to the first generation of protein structures 20 years ago, to the present where NMR structures now represent 15 to 20% of those submitted to the Protein Data Bank (PDB). However, the applications of NMR go far beyond structure determination. Since NMR is a non-invasive technique that provides a wealth of information about structure and dynamics, it is an ideal analytical method that is now being used in every step of the drug discovery and development process. Indeed, NMR is undergoing a renaissance in the pharmaceutical industry as a plethora of new applications have been developed, ranging from in vitro screening methods in early stage discovery to in vivo magnetic resonance imaging techniques used to assess drug efficacy in clinical trials. In this review, we will present an overview of the role of NMR in these areas, with a special focus on how NMR is poised to revolutionize the high-throughput structural characterization of protein-ligand interactions.

How substrate affinity is modulated by nucleotide binding remains a fundamental, unanswered question in the study of 70 kDa heat shock protein (Hsp70) molecular chaperones. We find here that the Escherichia coli Hsp70, DnaK, lacking the entire alpha-helical domain, DnaK(1-507), retains the ability to support lambda phage replication in vivo and to pass information from the nucleotide binding domain to the substrate binding domain, and vice versa, in vitro. We determined the NMR solution structure of the corresponding substrate binding domain, DnaK(393-507), without substrate, and assessed the impact of substrate binding. Without bound substrate, loop L3,4 and strand beta3 are in significantly different conformations than observed in previous structures of the bound DnaK substrate binding domain, leading to occlusion of the substrate binding site. Upon substrate binding, the beta-domain shifts towards the structure seen in earlier X-ray and NMR structures. Taken together, our results suggest that conformational changes in the beta-domain itself contribute to the mechanism by which nucleotide binding modulates substrate binding affinity.

Residual heteronuclear dipolar couplings obtained from partially oriented protein samples can provide unique NMR constraints for protein structure determination. However, partial orientation of protein samples also causes severe 1H line broadening resulting from residual 1H-1H dipolar couplings. In this communication we show that band-selective 1H homonuclear decoupling during data acquisition is an efficient way to suppress residual 1H-1H dipolar couplings, resulting in spectra that are still amenable to solution NMR analysis, even with high degrees of alignment. As an example, we present a novel experiment with improved sensitivity for the measurement of one-bond 1HN-15N residual dipolar couplings in a protein sample dissolved in magnetically aligned liquid crystalline bicelles.

The 23 kDa two-domain periplasmic chaperone FimC from Escherichia coli is required for the assembly of type-1 pili, which are filamentous, highly oligomeric protein complexes anchored to the outer bacterial membrane that mediate adhesion of pathogenic E. coli strains to host cell surfaces. Here we identified the contact sites on the surface of the NMR structure of FimC that are responsible for the binding of the 28 kDa mannose-binding type-1 pilus subunit FimH by 15N and 1H NMR chemical shift mapping, using transverse relaxation-optimized spectroscopy (TROSY). The FimH-binding surface of FimC is formed nearly entirely by the N-terminal domain, and its extent and shape indicate that FimC binds a folded form of the pilus subunits.

Magnetic resonance angiography (MRA) and transoesophageal echocardiography together are very useful imaging techniques in the evaluation of thoracic aortic pseudoaneurysms. A 63 year old hypertensive man was admitted to a community hospital by his primary care physician with three weeks history of night sweats, chills, fever, …

Summary The Gram-negative α -Proteobacterium Wolbachia pipientis has been described as an obligate endosymbiont in many arthropod species, where it induces a variety of reproductive alterations, including parthenogenesis. Recently, this microorganism has also been detected in the parthenogenetic collembolan Folsomia candida . Here, we confirm the occurrence of the endosymbiont also in two Italian parthenogenetic populations of F. candida using ultrastructural (electron microscopy) and molecular (PCR screening on two bacterial genes) evidence. The strain isolated in the Italian populations has almost-identical gene sequences compared to that previously isolated in other populations of F. candida . In addition, we discovered a population of Folsomia cf. candida , which showed the presence of both males and females. This population is not infected by Wolbachia . A screening of two mitochondrial genes ( COI and COII ) showed that the bisexual population has high levels of genetic divergence in comparison with the parthenogenetic ones, even suggesting the possibility that it belongs to a different species. Furthermore, the remarkably high levels of genetic divergence between the two parthenogenetic populations suggests a possible influence of Wolbachia on inducing such differentiation, and, in the long term, speciation.

We have recently identified a series of compounds which efficiently inhibit Anthrax lethal factor (LF) metallo-protease. Here we present further structure activity relationship and CoMFA (Comparative Molecular Field Analysis) studies on newly derived inhibitors. The obtained 3D QSAR model was subsequently compared with the X-ray structure of the complex between LF and a representative compound. Our studies form the basis for the rational design of additional compounds with improved activity and selectivity.

In this paper, we report a simple structure-based iterative optimizations (SUBITO) strategy to identify and optimize new protein ligands and inhibitors. The approach is based on a combination of NMR-based screening and computational docking methods and enabled the identification of novel chemical leads among hundreds of thousands of commercially available compounds by screening only a few hundred compounds from a scaffold library followed by iterative screening steps where only few dozen compounds are tested. As an application, we report on the discovery of a novel class of non-peptide reversible caspase inhibitors, with IC50 values in the low micromolar range. © 2005 American Chemical Society.

The authors examined some in silico approaches to identify Akt (protein kinase B) inhibitors. Experimental validation of selected compounds was achieved using a fluorescence-based enzymic assay and a substrate phosphorylation assay involving the protein GSK-3. The authors report on success and failure obtained by using several strategies including FlexX, GOLD, and CSCORE, where the 100-200 top-scoring compds. from a 50,000-compd. library were exptl. tested. This study led to the identification of low micromolar Akt1 inhibitors.

In this review, the use of general NMR spectroscopy techniques to detect ligand binding and to monitor enzyme kinetics and inhibition, which appear particularly useful in hit identification and validation, is reiterated. Furthermore, the use of NMR-based strategies for lead optimisations that are based on either iterative derivatisations of an initial core structure or on linking fragments that occupy adjacent pockets in the target's binding site will also be described. Several recent examples will be reported and the use of these techniques in cases when the three dimensional structure of the target protein is known will be discussed. © 2004 Ashley Publications Ltd.