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Ricchelli F.,CNR Institute of Biomedical Technologies | Sileikyte J.,University of Padua | Bernardi P.,University of Padua
Biochimica et Biophysica Acta - Bioenergetics | Year: 2011

The mitochondrial permeability transition is an increase of permeability of the inner mitochondrial membrane to ions and solutes with an exclusion size of about 1500 Da. It is generally accepted that the permeability transition is due to opening of a high-conductance channel, the permeability transition pore. Although the molecular nature of the permeability transition pore remains undefined, a great deal is known about its regulation and role in pathophysiology. This review specifically covers the characterization of the permeability transition pore by chemical modification of specific residues through photoirradiation of mitochondria after treatment with porphyrins. The review also illustrates the basic principles of the photodynamic effect and the mechanisms of phototoxicity and discusses the unique properties of singlet oxygen generated by specific porphyrins in discrete mitochondrial domains. These experiments provided remarkable information on the role, interactions and topology of His and Cys residues in permeability transition pore modulation and defined an important role for the outer membrane 18 kDa translocator protein (formerly known as the peripheral benzodiazepine receptor) in regulation of the permeability transition. © 2011 Elsevier B.V.

Navarro B.,CNR Institute of Plant virology | Gisel A.,CNR Institute of Biomedical Technologies | Rodio M.E.,CNR Institute of Plant virology | Delgado S.,Polytechnic University of Valencia | And 2 more authors.
Plant Journal | Year: 2012

How viroids, tiny non-protein-coding RNAs (∼250-400 nt), incite disease is unclear. One hypothesis is that viroid-derived small RNAs (vd-sRNAs; 21-24 nt) resulting from the host defensive response, via RNA silencing, may target for cleavage cell mRNAs and trigger a signal cascade, eventually leading to symptoms. Peach latent mosaic viroid (PLMVd), a chloroplast-replicating viroid, is particularly appropriate to tackle this question because it induces an albinism (peach calico, PC) strictly associated with variants containing a specific 12-14-nt hairpin insertion. By dissecting albino and green leaf sectors of Prunus persica (peach) seedlings inoculated with PLMVd natural and artificial variants, and cloning their progeny, we have established that the hairpin insertion sequence is involved in PC. Furthermore, using deep sequencing, semi-quantitative RT-PCR and RNA ligase-mediated rapid amplification of cDNA ends (RACE), we have determined that two PLMVd-sRNAs containing the PC-associated insertion (PC-sRNA8a and PC-sRNA8b) target for cleavage the mRNA encoding the chloroplastic heat-shock protein 90 (cHSP90), thus implicating RNA silencing in the modulation of host gene expression by a viroid. Chloroplast malformations previously reported in PC-expressing tissues are consistent with the downregulation of cHSP90, which participates in chloroplast biogenesis and plastid-to-nucleus signal transduction in Arabidopsis. Besides PC-sRNA8a and PC-sRNA8b, both deriving from the less-abundant PLMVd (-) strand, we have identified other PLMVd-sRNAs potentially targeting peach mRNAs. These results also suggest that sRNAs derived from other PLMVd regions may downregulate additional peach genes, ultimately resulting in other symptoms or in a more favorable host environment for viroid infection. © 2012 Blackwell Publishing Ltd.

Correa Leite M.L.,CNR Institute of Biomedical Technologies
Clinical and Applied Thrombosis/Hemostasis | Year: 2011

Aim: To examine the relationship between some blood parameters and mild kidney dysfunction. Participants and Methods: A total of 719 Italian men aged 42 to 74 years from a population-based survey carried out in the town of Bollate (Milan). General linear models were used to examine the variations in plasma fibrinogen, hematocrit, platelet counts, mean platelet volume, and uric acid across levels of kidney function (estimated on the basis of glomerular filtration rate [GFR]), adjusting for age, education, smoking, alcohol consumption, physical activity (evaluated as TV watching, engaging in sport practice, and walking/cycling), waist circumference, arm muscle area, high-density lipoprotein (HDL)-cholesterol, triglycerides, hypertension, diabetes, cardiovascular disease history, and nonsteroid anti-inflammatory, diuretic, and antihypertensive drug use. Results: Plasma fibrinogen and hematocrit levels increased, and platelet counts and mean platelet volume significantly decreased as GFR fell to <80 or <70 mL/min per 1.73 m 2; stratified analysis revealed an association with serum uric acid levels. Alterations compatible with an increased cardiovascular risk were particularly evident among the participants with higher uric acid levels, whereas those indicative of platelet dysfunction were found among participants with lower levels. Conclusions: Parameters affecting hemostasis and blood viscosity are altered when kidney function is only slightly reduced, and the patterns of these relationships seem to be influenced by the levels of serum uric acid, whose easy and inexpensive measurement could have prognostic value. © The Author(s) 2011.

Rizzi E.,CNR Institute of Biomedical Technologies
Methods in Molecular Biology | Year: 2015

The pyrosequencing methodology was applied in 2005 by 454 Lifesciences to the emerging field of next generation sequencing (NGS), revolutionizing the way of DNA sequencing. In the last years the same strategy grew up and was technologically updated, reaching a high throughput in terms of amount of generated sequences (reads) per run and in terms of length of sequence up to values of 800–1,000 bases. These features of pyrosequencing perfectly fit to bacterial genome sequencing for the de novo assemblies and resequencing as well. The approaches of shotgun and paired ends sequencing allow the bacterial genome finishing providing a high-quality data in few days with unprecedented results. © Springer Science+Business Media New York 2015.

Castellana S.,University of Bari | Vicario S.,CNR Institute of Biomedical Technologies | Saccone C.,CNR Institute of Biomedical Technologies | Saccone C.,University of Bari
Genome Biology and Evolution | Year: 2011

The mitochondrial genome is a fundamental component of the eukaryotic domain of life, encoding for several important subunits of the respiratory chain, the main energy production system in cells. The processes by means of which mitochondrial DNA (mtDNA) replicates, expresses itself and evolves have been explored over the years, although various aspects are still debated. In this review, we present several key points in modern research on the role of evolutionary forces in affecting mitochondrial genomes in Metazoa. In particular, we assemble the main data on their evolution, describing the contributions of mutational pressure, purifying, and adaptive selection, and how they are related. We also provide data on the evolutionary fate of the mitochondrial synonymous variation, related to the nonsynonymous variation, in comparison with the pattern detected in the nucleus. Elevated mutational pressure characterizes the evolution of the mitochondrial synonymous variation, whereas purging selection, physiologically due to phenomena such as cell atresia and intracellular mtDNA selection, guarantees coding sequence functionality. This enables mitochondrial adaptive mutations to emerge and fix in the population, promoting mitonuclear coevolution. © The Author(s) 2010.

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