ddRAD-seq reveals the genetic structure and detects signals of selection in Italian brown trout.

BACKGROUND: Brown trout is one of the most widespread fresh-water fish species in Europe. The evolutionary history of and phylogenetic relationships between brown trout populations are complex, and this is especially true for Italian populations, which are heavily influenced in different ways by stocking practices. The characterization of the genetic structure of Italian brown trout populations may give information on the risk of losing endemic Italian populations due to lack of genetic diversity or to admixture with stocking populations. The identification of signatures of selection, and the information deriving from dense genotyping data will help genotype-informed breeding programs. We used a ddRAD-seq approach to obtain more than 100,000 single nucleotide polymorphisms (SNPs), and to characterize the population structure and signatures of selection in 90 brown trout samples. RESULTS: Italian brown trout populations are genetically differentiated, although the stocking practices have introduced strong admixture in endemic Italian trout, especially with the Atlantic lineage. Most of the analysed populations showed high levels of kinship and inbreeding. We detected putative signatures of selection using different approaches, and investigated if the regions were enriched for functional categories. Several regions putatively under selection and characterized by a reduction in heterozygosity across all the studied populations are enriched for genes involved in the response to viral infections. CONCLUSIONS: Our results, which show evidence of admixture with the Atlantic lineage (commonly used for stocking), confirm the need for controlling stocking practices, in order to avoid the erosion of the endemic gene pool; given the apparently high levels of kinship and inbreeding in local populations, our results also show the need to take action for increasing gene diversity. In addition, we used the genetically-distinct lineages to detect signatures of selection and we identified putative signatures of selection in several regions associated with resistance to infectious diseases. These constitute candidate regions for the study of resistance to infections in wild and farmed trout.

Evidence of Phytotoxicity and Genotoxicity in Hordeum vulgare L. Exposed to CeO2 and TiO2 Nanoparticles.

Engineered nanoscale materials (ENMs) are considered emerging contaminants since they are perceived as a potential threat to the environment and the human health. The reactions of living organisms when exposed to metal nanoparticles (NPs) or NPs of different size are not well known. Very few studies on NPs-plant interactions have been published, so far. For this reason there is also great concern regarding the potential NPs impact to food safety. Early genotoxic and phytotoxic effects of cerium oxide NPs (nCeO2) and titanium dioxide NPs (nTiO2) were investigated in seedlings of Hordeum vulgare L. Caryopses were exposed to an aqueous dispersion of nCeO2 and nTiO2 at, respectively 0, 500, 1000, and 2000 mg l(-1) for 7 days. Genotoxicity was studied by Randomly Amplified Polymorphism DNA (RAPDs) and mitotic index on root tip cells. Differences between treated and control plants were observed in RAPD banding patterns as well as at the chromosomal level with a reduction of cell divisions. At cellular level we monitored the oxidative stress of treated plants in terms of reactive oxygen species (ROS) generation and ATP content. Again nCeO2 influenced clearly these two physiological parameters, while nTiO2 were ineffective. In particular, the dose 500 mg l(-1) showed the highest increase regarding both ROS generation and ATP content; the phenomenon were detectable, at different extent, both at root and shoot level. Total Ce and Ti concentration in seedlings was detected by ICP-OES. TEM EDSX microanalysis demonstrated the presence of aggregates of nCeO2 and nTiO2 within root cells of barley. nCeO2 induced modifications in the chromatin aggregation mode in the nuclei of both root and shoot cells.

Variation in Heavy Metal Accumulation and Genetic Diversity at a Regional Scale Among Metallicolous and Non-Metallicolous Populations of the Facultative Metallophyte Biscutella laevigata subsp. laevigata.

Biscutella laevigata is a facultative metallophyte, with populations on non-metalliferous and metalliferous soils. Some of its metallicolous populations have been shown to hyperaccumulate thallium or lead in nature. Only Tl hyperaccumulation has been experimentally confirmed. We aimed to compare the patterns of metal (hyper)accumulation and genetic diversity among populations of B. laevigata subsp. laevigata in NE Italy. None of the populations exhibited foliar hyperaccumulation of Cu, Zn, or Pb. The root-to-shoot accumulation rates for these metals were unchanged or decreased rather than enhanced in the metallicolous populations, in comparison with the non-metallicolous ones. Hyperaccumulation of Tl was confined to the population of the Cave del Predil mine. This population was genetically very distinct from the others, as demonstrated by AFLP-based cluster analysis. The two other mine populations did not surpass the threshold for Tl hyperaccumulation, but showed enhanced foliar Tl concentrations and root-to-shoot translocation rates, in comparison with the non-metallicolous populations. Genetic analysis suggested that adaptation to metalliferous soil must have been independently evolved in the metallicolous populations.

A randomized trial comparing eptifibatide vs. placebo in patients with diffuse coronary artery disease undergoing drug-eluting stent implantation: design of the INtegrilin plus STenting to Avoid myocardial Necrosis Trial.

BACKGROUND: Despite the availability of several potent antithrombotic agents, the optimal antiplatelet regimen in elective patients undergoing complex percutaneous coronary interventions is still debated. Aim of the INtegrilin plus STenting to Avoid myocardial Necrosis Trial will be to assess the safety and efficacy of routine usage of the glycoprotein IIb/IIIa inhibitor eptifibatide in patients already treated with aspirin and clopidogrel and undergoing implantation of at least two drug-eluting stents in the same lesion, thus identifying a clinically stable but anatomically complex patient subset. DESIGN: This will be a single-blind, placebo-controlled multicenter randomized trial. METHODS: Patients with stable coronary artery disease, who are undergoing percutaneous coronary intervention by means of implantation of greater than 33 mm of drug-eluting stents (e.g. with two 23-mm drug-eluting stents or one 32-mm and one 12-mm drug-eluting stent), will be randomized, after administration of aspirin and clopidogrel (600 mg loading dose recommended), to eptifibatide and unfractioned heparin according to the ESPRIT protocol or placebo and unfractioned heparin. Blood draws for creatine kinase-MB mass, total creatine kinase, and cardiac troponin levels will be taken at baseline, 6 and 12 h postprocedurally. Patients will be followed for clinical events by direct visit or phone contact up to 6 months. The primary endpoint of the study will be the rate of abnormal values of creatine kinase-MB mass after percutaneous coronary intervention. Secondary endpoints will be the composite of cardiac death, nonfatal myocardial infarction, urgent target vessel revascularization, and thrombotic bailout glycoprotein IIb/IIIa inhibitor therapy within 180 days, and in-hospital, 1-month and 6-month major adverse cardiovascular events, defined as the composite of cardiac death, nonfatal myocardial infarction or urgent target vessel revascularization. IMPLICATIONS: The INtegrilin plus STenting to Avoid myocardial Necrosis Trial study will test for the first time the beneficial impact of routine glycoprotein IIb/IIIa inhibition on top of dual oral antiplatelet treatment in clinically stable yet anatomically complex patients undergoing drug-eluting stents implantation. Results of this single-blind randomized trial will provide important insights to improve the management strategy of patients and outcomes in the current drug-eluting stents era.

Long-term follow-up of Tako-Tsubo-like syndrome: a retrospective study of 22 cases.

OBJECTIVES: To assess the late outcome of the Tako-Tsubo like syndrome in a community hospital in northern Italy. METHODS AND RESULTS: We reviewed 2233 patients who were admitted from 2001 to 2006 with diagnosis of acute coronary syndrome. Twenty-two patients (1%) presenting clinical and instrumental characteristics of Tako-Tsubo like syndrome were included in the study and prospectively underwent clinical and echocardiographic follow-up. All patients were women; aged 76 +/- 7 years; 82% experienced a stress before the acute episode; 50% reported chest pain and dyspnoea also days before. Mean troponin peak value was 3.6 +/- 3.3 microg/l. Mean acute echocardiographic ejection fraction was 40 +/- 7%. Eighteen percent of them presented major in-hospital complications. At a mean follow-up time of 27 +/- 19 months, 2 patients (9%) died because of ischemic stroke and renal failure, respectively, 14 (63%) were asymptomatic, 1 (5%) declared a paroxysmal episode of atrial fibrillation, and 5 (23%) still lamented dyspnoea or atypical chest pain. In all patients, typical apical ballooning disappeared and an increase in mean ejection fraction to 60 +/- 4% was observed. CONCLUSION: After complications are promptly recognized and treated in the acute phase, prognosis of Tako-Tsubo like syndrome appears to be good at long-term follow-up, with a complete recovery of normal left ventricular function.

A rare case of idiopathic spontaneous coronary artery dissection causing acute myocardial infarction.

Spontaneous coronary artery dissection is a rare clinical event and often a fatal cause of ischaemic heart disease occurring predominantly in young or middle-aged otherwise healthy patients. We present the case of a 59-year-old female patient who was admitted to our coronary care unit with acute anterior myocardial infarction. Coronary angiography showed a long dissection involving the left coronary artery. Emergency coronary artery bypass grafting was performed unsuccessfully.

Transcranial Doppler monitoring during stenting of the carotid bifurcation: evaluation of two different distal protection devices in preventing embolization.

PURPOSE: To compare the efficacy of 2 emboli protection devices in preventing embolization during carotid artery stenting (CAS). METHODS: The GuardWire distal occlusion system (n=19) and the distal FilterWire EX (n=12) were compared in 31 consecutive patients (24 men; mean age 71+/-10 years) monitored with transcranial Doppler for microembolic signals before, during, and after CAS. The choice of the protection device was based on availability and on the patency of the contralateral carotid artery. RESULTS: The baseline characteristics were similar in the patients treated under protection from either device. Placement and retrieval of the protection device, stenting, and postdilation were technically successful in all patients. Two patients suffered a transient ischemic attack shortly after the procedure; no other adverse cardiovascular events occurred at 30 days. Compared to the GuardWire, the use of the FilterWire was associated with more microembolic signals during stent deployment (77.4+/-33.5 versus 1.07+/-1.94, p<0.0001), postdilation (63.9+/-21.0 versus 2.06+/-2.58, p<0.0001), and retrieval of the protection device (21.4+/-15.4 versus 10.9+/-8.3, p=0.051). Consequently, the total amount of microembolic signals during the procedure was higher when the filter device was employed (183.0+/-42.1 versus 31.7+/-12.0, p<0.0001). CONCLUSION: The distal occlusion device appears to be more effective than the filter in reducing distal embolization detected by transcranial Doppler monitoring.

Modifications of minimal luminal diameter during the 24 hours following percutaneous treatment of in-stent restenosis.

The early renarrowing after percutaneous coronary interventions (PCI) for in-stent restenosis (ISR) is a phenomenon that has received little investigation. Therefore, we assessed the angiographic minimal luminal diameter (MLD) before, immediately after, and 24 hours after the procedure in 30 ISR's treated with balloon angioplasty (BA) or rotational atherectomy (RA). The MLD was 0.68+/-0.48 mm at baseline, 2.42+/-0.38 mm after the procedure, and 2.25+/-0.39 mm at 24 hours (p < 0.0001 for all comparisons). Therefore, a small but significant decrease in MLD between post-procedure and 24 hours, corresponding to an early loss of 0.18+/-0.16 mm, was evident. The early loss was not significantly different in ISR's treated with BA or RA (0.15+/-0.16 mm and 0.20+/-0.16 mm, respectively). No significant correlation between early loss and lesion length, diameter stenosis, reference diameter, and acute gain was found. However, no increase of the 24-hour diameter stenosis to more than 50% was observed. We conclude that the magnitude of early luminal loss after PCI of ISR is small and therefore does not generally constitute a clinical problem.

Distal protection with a filter device during coronary stenting in patients with stable and unstable angina.

BACKGROUND: Filter protection after percutaneous coronary intervention (PCI) is now available to prevent distal embolization. The aims of this study were (1) to evaluate the microembolization phenomenon during procedures of stent implantation in native coronary arteries of patients with stable and unstable angina, (2) to assess the amount and characteristics of the debris captured by the Angioguard, and (3) to investigate the relation between clinical and angiographic variables and pathological data. METHODS AND RESULTS: Elective coronary stenting with the use of a protective filter was attempted in 39 consecutive coronary artery lesions with >60% stenosis (mean, 67.6+/-8.79%). Debris was present in 75.6% of the filters. Particle size ranged from 47.16 to 2503.48 microm (mean, 518.83+/-319.61 microm) in the major axis. Particles >300 microm were found in 24 of 28 filters with debris (85.7%), and particles >1000 microm were present in 10 of 28 filters (35.7%). Patients with unstable angina had greater particles (mean maximum longitudinal diameter, 1098.33+/-714.3 microm) than those with stable angina (412.91+/-453 microm; P<0.001). The presence of unstable angina (OR, 65; CI, 1.2 to 3420; P=0.03) and age >67 years (OR, 42; CI, 1 to 1698; P=0.04) were found to be the only independent predictors of embolic particle size. CONCLUSIONS: By limiting embolization, protective devices may prevent a number of potentially unfavorable events, thereby improving outcome. Our data support the use of these devices, especially in lesions with higher embolic potential, such as those occurring in older patients and in those with unstable angina.