652 research outputs found

    Transposon Tn7 Preferentially Inserts into GAA‱TTC Triplet Repeats under Conditions Conducive to Y‱R‱Y Triplex Formation

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    BACKGROUND: Expansion of an unstable GAA*TTC repeat in the first intron of the FXN gene causes Friedreich ataxia by reducing frataxin expression. Structure formation by the repeat has been implicated in both frataxin repression and GAA*TTC instability. The GAA*TTC sequence is capable of adopting multiple non-B DNA structures including Y*R*Y and R*R*Y triplexes. Lower pH promotes the formation of Y*R*Y triplexes by GAA*TTC. Here we used the bacterial transposon Tn7 as an in vitro tool to probe whether GAA*TTC repeats can attract a well-characterized recombinase. METHODOLOGY/PRINCIPAL FINDINGS: Tn7 showed a pH-dependent preference for insertion into uninterrupted regions of a Friedreich ataxia patient-derived repeat, inserting 48, 39 and 14 percent of the time at pH 7, pH 8 and pH 9, respectively. Moreover, Tn7 also showed orientation and region specific insertion within the repeat at pH 7 and pH 8, but not at pH 9. In contrast, transposon Tn5 showed no strong preference for or against the repeat during in vitro transposition at any pH tested. Y*R*Y triplex formation was reduced in predictable ways by transposon interruption of the GAA*TTC repeat. However, transposon interruptions in the GAA*TTC repeats did not increase the in vitro transcription efficiency of the templates. CONCLUSIONS/SIGNIFICANCE: We have demonstrated that transposon Tn7 will recognize structures that form spontaneously in GAA*TTC repeats and insert in a specific orientation within the repeat. The conditions used for in vitro transposition span the physiologically relevant range suggesting that long GAA*TTC repeats can form triplex structures in vivo, attracting enzymes involved in DNA repair, recombination and chromatin modification

    Georeferenced cartography dataset of the La Fossa crater fumarolic field at Vulcano Island (Aeolian Archipelago, Italy): conversion and comparison of data from local to global positioning methods

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    The present study illustrates the procedures applied for the coordinate system conversion of the historical fumarole positions at La Fossa crater, to allow their comparison with newly acquired global positioning system (GPS) data. Due to the absence of ground control points in the field and on both the old Gauss Boaga and the new UTM WGS 1984 maps, we had to model the transformation errors between the two systems using differential GPS techniques. Once corrected, the maps show a residual Easting shifting, due to erroneous georeferencing of the original base maps; this is corrected by morphological comparative methods. The good correspondence between the corrected positions of the historical data and the results of the new GPS survey that was carried out in 2009 highlights the good quality of the old surveys, although they were carried out without the use of accurate topographical instruments

    Large-scale numerical modeling of hydro-acoustic waves generated by tsunamigenic earthquakes

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    Abstract. Tsunamigenic fast movements of the seabed generate pressure waves in weakly compressible seawater, namely hydro-acoustic waves, which travel at the sound celerity in water (about 1500 m s−1). These waves travel much faster than the counterpart long free-surface gravity waves and contain significant information on the source. Measurement of hydro-acoustic waves can therefore anticipate the tsunami arrival and significantly improve the capability of tsunami early warning systems. In this paper a novel numerical model for reproduction of hydro-acoustic waves is applied to analyze the generation and propagation in real bathymetry of these pressure perturbations for two historical catastrophic earthquake scenarios in Mediterranean Sea. The model is based on the solution of a depth-integrated equation, and therefore results are computationally efficient in reconstructing the hydro-acoustic waves propagation scenarios

    A persistent RNA·DNA hybrid formed by transcription of the Friedreich ataxia triplet repeat in live bacteria, and by T7 RNAP in vitro

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    Expansion of an unstable GAA·TTC repeat in the first intron of the FXN gene causes Friedreich ataxia by reducing frataxin expression. Deficiency of frataxin, an essential mitochondrial protein, leads to progressive neurodegeneration and cardiomyopathy. The degree of frataxin reduction correlates with GAA·TTC tract length, but the mechanism of reduction remains controversial. Here we show that transcription causes extensive RNA·DNA hybrid formation on GAA·TTC templates in bacteria as well as in defined transcription reactions using T7 RNA polymerase in vitro. RNA·DNA hybrids can also form to a lesser extent on smaller, so-called ‘pre-mutation’ size GAA·TTC repeats, that do not cause disease, but are prone to expansion. During in vitro transcription of longer repeats, T7 RNA polymerase arrests in the promoter distal end of the GAA·TTC tract and an extensive RNA·DNA hybrid is tightly linked to this arrest. RNA·DNA hybrid formation appears to be an intrinsic property of transcription through long GAA·TTC tracts. RNA·DNA hybrids have a potential role in GAA·TTC tract instability and in the mechanism underlying reduced frataxin mRNA levels in Friedreich Ataxia

    Grazing and control of coral reef community structure by Diadema antillarum Philippi (Echinodermata: Echinoidea): A preliminary study

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    The grazing activities of the tropical echinoid Diadema antillarum Philippi can markedly influence the structure of the shallow-water epibenthic coral reef community. The elimination of an entire Diadema population from a patch reef in St. Croix, U.S. Virgin Islands resulted in a great increase in macro-algal biomass, alterations in species composition, shifts in dominance, an increase in species numbers, and a decrease in equitability...

    Identification of D179H, a novel missense GJB2 mutation in a Western Sicily family

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    The main purpose of this study was to describe a novel missense mutation (p.D179H) found in a Western Sicily family and to examine the genetic and audiologic profiles of all family members by performing a GJB2 and GJB6 mutations analysis and a complete audiologic assessment. The proband was a 3-month-old infant with a congenital profound sensorineural hearing loss; direct sequencing of the GJB2 revealed the presence of a c.35delG mutation in the heterozygous state and a heterozygous G[C transition at nucleotide 535 in trans; this novel mutation, called p.D179H, resulted in an aspartic acid to histidine change at codon 179. It was also evidenced in the heterozygous state in two members of this family, both with normal hearing. No GJB6 mutations were evidenced in all subjects studied. Considering the genotypic and phenotypic analysis of all family members, we suggest, differently from the p.D179 N mutation previously reported, a recessive mode of inheritance. Functional studies on p.D179H have to be performed to confirm our hypothesis

    New insights into North Sea deep crustal structure and extension from transdimensional ambient noise tomography

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    SUMMARY The deep crustal structure beneath the North Sea is poorly understood since it is constrained by only a few seismic reflection and refraction profiles. However, it is widely acknowledged that the mid to lower crust plays important roles in rift initiation and evolution, particularly when large-scale sutures and/or terrane boundaries are present, since these inherited features can focus strain or act as inhibitors to extensional deformation. Ancient tectonic features are known to exist beneath the iconic failed rift system of the North Sea, making it an ideal location to investigate the complex interplay between pre-existing regional heterogeneity and rifting. To this end, we produce a 3-D shear wave velocity model from transdimensional ambient seismic noise tomography to constrain crustal properties to ∌30 km depth beneath the North Sea and its surrounding landmasses. Major North Sea sedimentary basins appear as low shear wave velocity zones that are a good match to published sediment thickness maps. We constrain relatively thin crust (13–18 km) beneath the Central Graben depocentres that contrasts with crust elsewhere at least 25–30 km thick. Significant variations in crustal structure and rift symmetry are identified along the failed rift system that appears to be related to the locations of Laurentia–Avalonia–Baltica palaeoplate boundaries. We constrain first-order differences in structure between palaeoplates; with strong lateral gradients in crustal velocity related to Laurentia–Avalonia–Baltica plate juxtaposition and reduced lower crustal velocities in the vicinity of the Thor suture, possibly representing the remnants of a Caledonian accretionary complex. Our results provide fresh insight into the pivotal roles that ancient terranes can play in the formation and failure of continental rifts and may help explain the characteristics of other similar continental rifts globally.</jats:p

    Extended mild-slope equations for compressible fluids

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    In this paper we derive new forms of the mildslope equation (MSE) for water waves in a weakly compressible fluid on a slowly varying bathymetry, with surface and bottom disturbances. The MSE is a powerful tool to model the refraction-diffraction dynamics of water waves propagating on a variable bathymetry [1]. Traditionally, mild-slope models are derived by assuming that the wave steepness is small, the fluid is inviscid and incompressible and the motion is irrotational. Furthermore, no disturbances are normally considered both on the free surface and at the bottom of the fluid domain [2]. In this paper we shall find new expressions of the MSE by relaxing the incompressibility hypothesis and considering both surface and bottom disturbances. We shall name the set of new formulae as the extended acoustic-gravity mild-slope equations (EAG-MSE). Such a system of equations can be implemented in numerical models for the early detection of coastal flooding based on the hydro-acoustic precursors of surface gravity waves (see [3]–[5])

    Drug utilization, safety, and effectiveness of exenatide, sitagliptin, and vildagliptin for type 2 diabetes in the real world: Data from the Italian AIFA Anti-diabetics Monitoring Registry

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    AbstractBackground and aimsIn Italy, the reimbursed use of incretin mimetics and incretin enhancers was subject to enrollment of patients into a web-based system recording the general demographic and clinical data of patients. We report the utilization data of glucagon-like peptide 1 (GLP1) receptor agonists and dipeptidylpeptidase-4 (DPP4) inhibitors in clinical practice as recorded by the Italian Medicines Agency (AIFA) Monitoring Registry.Methods and resultsFrom February 2008 to August 2010, 75,283 patients with type 2 diabetes were entered into the registry and treated with exenatide, sitagliptin, or vildagliptin. The treatment was administered to patients in a wide range of ages (≄75 years, n = 6125 cases), body mass index (BMI) (≄35 kg/m2, n = 22,015), and metabolic control (HbA1c ≄ 11% ((96 mmol/mol), n = 3151). Overall, 1116 suspected adverse drug reactions were registered, including 12 cases of acute pancreatitis (six on exenatide). Hypoglycemic episodes mainly occurred in combination with sulfonylureas. Treatment discontinuation for the three drugs (logistic regression analysis) was negatively associated with the male gender and positively with baseline HbA1c, diabetes duration, and, limitedly to DPP-4 inhibitors, with BMI. Treatment discontinuation (including loss to follow-up, accounting for 21–26%) was frequent. Discontinuation for treatment failure occurred in 7.7% of cases (exenatide), 3.8% (sitagliptin), and 4.1% (vildagliptin), respectively, corresponding to 27–40% of all discontinuations, after excluding lost to follow-up. HbA1c decreased on average by 0.9–1.0% (9 mmol/mol). Body weight decreased by 3.5% with exenatide and by 1.0–1.5% with DPP-4 inhibitors.ConclusionsIn the real world of Italian diabetes centers, prescriptions of incretins have been made in many cases outside the regulatory limits. Nevertheless, when appropriately utilized, incretins may grant results at least in line with pivotal trials
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