Multiple pathways for telomere tethering: functional implications of subnuclear position for heterochromatin formation

Technical advances in the imaging of GFP derivatives in living cells have improved our ability to determine the position and dynamics of specific chromatin loci. This approach, combined with genetics and functional assays, has shed new light on how nuclear compartments facilitate gene repression in yeast.     

Interaction of poly(L-lysine)-g-poly(ethylene glycol) with supported phospholipid bilayers

Interactions between the graft copolymer poly(L-lysine)-g-poly(ethylene glycol), PLL-g-PEG, and two kinds of surface-supported lipidic systems (supported phospholipid bilayers and supported vesicular layers) were investigated by a combination of microscopic and spectroscopic techniques. It was found that the application of the copolymer to zwitterionic or negatively charged supported bilayers in a buffer of low ionic strength led to their decomposition, with the resulting formation of free copolymer-lipid complexes. The same copolymer had no destructive effect on a supported vesicular layer made up of vesicles of identical composition. A comparison between poly(L-lysine), which did not induce decomposition of supported bilayers, and PLL-g-PEG copolymers with various amounts of PEG side chains per backbone lysine unit, suggested that steric repulsion between the PEG chains that developed upon adsorption of the polymer to the nearly planar surface of a supported phospholipid bilayer (SPB) was one of the factors responsible for the destruction of the SPBs by the copolymer. Other factors included the ionic strength of the buffer used and the quality of the bilayers, pointing toward the important role defects present in the SPBs play in the decomposition process.     

Raman study of poly(alanine-glycine)-based peptides containing tyrosine, valine, and serine as model for the semicrystalline domains of Bombyx mori silk fibroin

For a deeper insight into the structure of Bombyx mori silk fibroin, some model peptides containing tyrosine (Y), valine (V), and serine (S) in the basic (AG)n sequence were synthesized by the solid-phase method and analyzed by Raman spectroscopy in order to clarify their conformation and to evaluate the formation and/or disruption of the ordered structure typical of B. mori silk fibroin upon incorporation of Y, V, and S residues into the basic (AG)n sequence. The Raman results indicated that the silk I structure remains stable only when the Y residue is positioned near the chain terminus; otherwise, a silk I --> silk II conformational transition occurs. The peptides AGVGAGYGAGVGAGYGAGVGAGYG(AG)3 and (AG)3YG(AG)2VGYG(AG)3YG(AG)3 treated with LiBr revealed a prevalent silk II conformation; moreover, the former contained a higher amount of random coil than the latter. This result was explained in relation to the different degrees of interruption of the (AG)n sequence. The Raman analysis of the AGSGAG-containing samples confirmed that the AGSGAG hexapeptide is a good model for the silk II crystalline domain. As the number of AGSGAG repeating units decreased, the random coil content increased. The study of the Y domain (I850/I830 intensity ratio) allowed us to hypothesize that in the packing characteristic of Silk I and Silk II conformations the Y residues experience different environments and hydrogen-bonding arrangements; the packing typical of silk I structure traps the tyrosyl side chains in environments more unfavorable to phenoxyl hydrogen-bonding interactions.     

Automatic generation of accurate subject-specific bone finite element models to be used in clinical studies

Most of the finite element models of bones used in orthopaedic biomechanics research are based on generic anatomies. However, in many cases it would be useful to generate from CT data a separate finite element model for each subject of a study group. In a recent study a hexahedral mesh generator based on a grid projection algorithm was found very effective in terms of accuracy and automation. However, so far the use of this method has been documented only on data collected in vitro and only for long bones. The present study was aimed at verifying if this method represents a procedure for the generation of finite element models of human bones from data collected in vivo, robust, accurate, automatic and general enough to be used in clinical studies. Robustness, automation and numerical accuracy of the proposed method were assessed on five femoral CT data sets of patients affected by various pathologies. The generality of the method was verified by processing a femur, an ileum, a phalanx, a proximal femur reconstruction, and the micro-CT of a small sample of spongy bone. The method was found robust enough to cope with the variability of the five femurs, producing meshes with a numerical accuracy and a computational weight comparable to those found in vitro. Even when the method was used to process the other bones the levels of mesh conditioning remained within acceptable limits. Thus, it may be concluded that the method presents a generality sufficient to cope with almost any orthopaedic application.     

An ectophosphatase activity in Cryptococcus neoformans

There is increasing evidence in the literature showing that fungal pathogens express biologically active ectoenzymes. The expression of surface phosphatases at the cell surface of Cryptococcus neoformans, the etiologic agent of cryptococcosis, was evaluated in the present study. Different isolates of C. neoformans express ectophosphatase activity, which is not influenced by capsule size or serotype. The cryptococcal enzyme is an acid phosphatase, inhibited by classic inhibitors of ectophosphatases, including ammonium molybdate and sodium salts of fluoride and orthovanadate. Only the inhibition of enzyme activity caused by sodium orthovanadate has been shown to be irreversible. The cryptococcal ectoenzyme is also inhibited by Zn2+ and inorganic phosphate, the final product of reactions catalyzed by phosphatases. The ectophosphatase from C. neoformans efficiently releases phosphate groups from different phosphorylated amino acids, giving a higher rate of phosphate removal when phosphothreonine is used as a substrate. Yeast cells with irreversibly inhibited ectophosphatases are less capable of adhering to animal epithelial cells than fungi fully expressing enzyme activity, suggesting that ectoenzyme expression can contribute to the pathogenesis of C. neoformans.     

Synthesis, stability and reactivity of palladium(0) olefin complexes bearing labile or hemi-labile ancillary ligands and electron-poor olefins

An overview of the general features of electron-poor olefin stabilized palladium(0) complexes bearing labile and hemi-labile ancillary ligands is presented. In particular, we have summarized the synthetic methodologies, the ligands commonly used, and the characterization of such complexes. The behavior of these species in solution is also described with particular attention to their fluxional rearrangements and reactivity. Thus, olefin exchange reactions are described and a comprehensive order of coordinative capability of the most widely used electron-poor alkenes is presented. The reactions of the title complexes dealing with olefin isomerization, oxidative addition, and formation of palladacyclopentadiene derivatives are eventually reported together with their main structural characteristics.     

NMR solution structure of the acylphosphatase from Escherichia coli

The solution structure of Escherichia coli acylphosphatase (E. coli AcP), a small enzyme catalyzing the hydrolysis of acylphosphates, was determined by (1)H and (15)N NMR and restrained modelling calculation. In analogy with the other members of AcP family, E. coli AcP shows an alpha/beta sandwich domain composed of four antiparallel and one parallel beta-strand, assembled in a five-stranded beta-sheet facing two antiparallel alpha-helices. The pairwise RMSD values calculated for the backbone atoms of E. coli and Sulfolobus solfataricus AcP, Bovine common type AcP and Horse muscle AcP are 2.18, 5.31 and 5.12 A, respectively. No significant differences are present in the active site region and the catalytic residue side chains are consistently positioned in the structures.     

Which endothelium-derived factors are really important in humans?

The endothelium plays a primary role in the local control of vascular function and structure, mainly by the production and release of NO, a potent vasodilator that also inhibits all the mechanisms involved in the pathogenesis of atherosclerosis, thus protecting the vessel wall against the development of atherosclerosis and thrombosis. Cardiovascular risk factors are associated with endothelial dysfunction, which involves enhanced production of oxygen free radicals that reduce NO availability and the release of contracting factors, including prostanoids and endothelin-1. In humans, endothelium-dependent relaxation can be assessed by tests that explore vascular reactivity. Besides the degree of vasodilation, which represents a crude estimate of endothelial function, the utilization of a complex experimental design, requiring the administration of specific agonists and antagonists, allows detailed exploration of the mediators and mechanisms involved in endothelium-dependent vasodilation. At present, the degree of endothelium-dependent vasodilation (evoked by receptor-operated agonists or the application of mechanical forces) is considered an independent predictor of cardiovascular events. In contrast, scant information is available concerning the clinical relevance of different mediators involved in endothelial function. Further studies are needed in the future to assess the specific impact of different endothelial responses on the clinical outcome in patients with cardiovascular risk factors and disease.     

Are spontaneous fractures possible? An example of clinical application for personalised,multiscale neuro-musculo-skeletal modelling

Elderly frequently present variable degrees of osteopenia, sarcopenia, and neuromotor control degradation. Severely osteoporotic patients sometime fracture their femoral neck when falling. Is it possible that such fractures might occur without any fall, but rather spontaneously while the patient is performing normal movements such as level walking? The aim of this study was to verify if such spontaneous fractures are biomechanically possible, and in such case, which conditions of osteoporosis, sarcopenia, and neuromotor degradation could produce them. To the purpose, a probabilistic multiscale body-organ model validated against controlled experiments was used to predict the risk of spontaneous fractures in a population of 80-years old women, with normal weight and musculoskeletal anatomy, and variable degree of osteopenia, sarcopenia, and neuromotor control degradation. A multi-body inverse dynamics sub-model, coupled to a probabilistic neuromuscular sub-model, and to a femur finite element sub-model, formed the multiscale model, which was run within a Monte Carlo stochastic scheme, where the various parameters were varied randomly according to well defined distributions. The model predicted that neither extreme osteoporosis, nor extreme neuromotor degradation alone are sufficient to predict spontaneous fractures. However, when the two factors are combined an incidence of 0.4% of spontaneous fractures is predicted for the simulated population, which is consistent with clinical reports. When the model represented only severely osteoporotic patients, the incidence of spontaneous fractures increased to 29%. Thus, is biomechanically possible that spontaneous femoral neck fractures occur during level walking, due to a combination of severe osteoporosis and severe neuromotor degradation.     

Feeding ecology and prey preferences of a piscivorous fish in the Lagoa do Peixe National Park, a Biosphere Reserve in Southern Brazil

We investigated the diet, feeding strategy, size-related dietary shifts and prey preferences of South American Hoplias aff. malabaricus in an internationally recognized but poorly investigated Biosphere Reserve in southern Brazil. Fish were caught between April 2008 and March 2009 using a variety of fishing gear. The analysis of 113 individuals revealed a diet essentially composed of fish (16 species), particularly characid species (9). The diet became more diverse and contained larger fish prey with increasing predator size. Feeding strategy analysis revealed a clear specialization towards the consumption of fish. However, individuals did not prey upon particular prey species, instead opportunistically consuming many different fish species, which could be a strategy to avoid intraspecific competition. Characid species were the most important prey, followed by poecillids. A multi-gear sampling of the ichthyofauna revealed that these prey species were the most abundant (Characidae: 61.3%, Poeciliidae 18.8%) of the 14 fish families occurring at the study site, suggesting that the predator exploits the most abundant fish resources available rather than the rarer fish prey. These findings suggest that potential top-down controls exerted by H. aff. malabaricus in this system follow specific food web pathways that seem to be mediated by the abundance of prey resources.