Molecular characterization of Italian rice cultivars

Random amplified polymorphic DNA (RAPD) is an efficient tool to detect DNA polymorphisms and to distinguish different clones and cultivars. We have applied the RAPD technique to distinguish, starting from a single rice seed, among some Italian rice cultivars from market and from National Institute for research on food and nutrition, Italy. We also estimated the genomic similarity among the rice cultivars object of this study taking advantage of the amplification patterns. Despite the low genomic variability among Italian rice cultivars, RAPD technique allowed us to distinguish among them. This indicates that RAPD can be used routinely to verify the identity of Italian rice cultivars in order to prevent fraudulent commercial activity.     

Evidence that mitochondrial respiration is a source of potentially toxic oxygen free radicals in intact rabbit hearts subjected to ischemia and reflow

Previous in vitro studies have shown that isolated mitochondria can generate oxygen radicals. However, whether a similar phenomenon can also occur in intact organs is unknown. In the present study, we tested the hypothesis that resumption of mitochondrial respiration upon reperfusion might be a mechanism of oxygen radical formation in postischemic hearts, and that treatment with inhibitors of mitochondrial respiration might prevent this phenomenon. Three groups of Langendorff-perfused rabbit hearts were subjected to 30 min of global ischemia at 37 degrees C, followed by reflow. Throughout ischemia and early reperfusion the hearts received, respectively: (a) 5 mM KCl (controls), (b) 5 mM sodium amobarbital (Amytal, which blocks mitochondrial respiration at Site I, at the level of NADH dehydrogenase), and (c) 5 mM potassium cyanide (to block mitochondrial respiration distally, at the level of cytochrome c oxidase). The hearts were then processed to directly evaluate oxygen radical generation by electron paramagnetic resonance spectroscopy, or to measure oxygen radical-induced membrane lipid peroxidation by malonyl dialdehyde (MDA) content of subcellular fractions. Severity of ischemia, as assessed by 31P-nuclear magnetic resonance measurements of cardiac ATP, phosphocreatine, and pH, was similar in all groups. Oxygen-centered free radical concentration averaged 3.84 +/- 0.54 microM in reperfused control hearts, and it was significantly reduced by Amytal treatment (1.98 +/- 0.26; p < 0.05), but not by KCN (2.58 +/- 0.96 microM; p = not significant (NS)), consistent with oxygen radicals being formed in the mitochondrial respiratory chain at Site I. Membrane lipid peroxidation of reperfused hearts was also reduced by treatment with Amytal, but not with KCN. MDA content of the mitochondrial fraction averaged 0.75 +/- 0.06 nM/mg protein in controls, 0.72 +/- 0.06 in KCN-treated hearts, and 0.54 +/- 0.05 in Amytal-treated hearts (p < 0.05 versus both groups). Similarly, MDA content of lysosomal membrane fraction was 0.64 +/- 0.09 nM/mg protein in controls, 0.79 +/- 0.15 in KCN-treated hearts, and 0.43 +/- 0.06 in Amytal-treated hearts (p < 0.05 versus both groups). Since the effects of Amytal are known to be reversible, in a second series of experiments we investigated whether transient mitochondrial inhibition during the initial 10 min of reperfusion was also associated with beneficial effects on subsequent recovery of cardiac function after wash-out of the drug. At the end of the experiment, recovery of left ventricular end-diastolic and of developed pressure was significantly greater in those hearts that had been treated with Amytal during ischemia and early reflow, as compared to untreated hearts.(ABSTRACT TRUNCATED AT 400 WORDS)     

A dirichlet process elaboration diagnostic for binomial goodness of fit

Useful model checking tools can be constructed by measuring the distance between a prior distribution that concentrates most of its mass around a model of interest, and the resulting posterior distribution. In this paper we use this approach to construct a diagnostic measure for detecting lack of fit in discrete data, with special focus on binomial data. We begin by constructing a suitable probability model “around” the model of interest, via a Dirichlet Process elaboration. We derive the resulting diagnostic and show that, approximately, it is the sum of two terms: the first is the logarithm of the Bayes factor and the second is proportional to the Pearson chi-square statistics. We give details of a simulation algorithm for computing the diagnostic and illustrate its use in an application to biomedical data.     

SRAM-Based FPGA Systems for Safety-Critical Applications：A Survey on Design Standards and Proposed Methodologies

As the ASIC design cost becomes affordable only for very large-scale productions, the FPGA technology is currently becoming the leading technology for those applications that require a small-scale prod...     

Studies of the hydrogen held by solids: XXII. The surface chemistry of reduced molybdenaalumina catalysts

The surface hydroxyl concentrations of a fresh molybdenaalumina catalyst (8% Mo) and of the alumina from which it was made have been determined as a function of the temperature of pretreatment. Similar data were obtained for catalysts reduced with H₂ or with CO. In all cases, the hydroxyl concentrations decreased with increasing pretreatment temperatures. The difference between the curves for the parent alumina and the molybdenaalumina preparation made from it provided a measure of the number of hydroxyl groups eliminated as the epitaxial monolayer of molybdena was grown onto the surface. The values obtained (1.7 ± 0.6 OH/Mo) showed that the surface hydroxyl groups of alumina are replaced by molybdate anions. When the catalyst was reduced with CO to about $\text{e}\text{Mo}\text{= 1.5}$ (average valence, Mo^+4.5), the curve obtained was almost identical with that for the unreduced catalyst, but when the catalyst was reduced with H₂, values for the retained hydrogen were higher than for the oxidized catalyst and approached those of the parent alumina as its evacuation temperature was increased to 550 °C. This increase in hydroxyl concentration was in agreement with earlier deductions.The hydroxyl region of the infrared spectra of similar preparations was recorded. Four distinct bands could be characterized for the parent alumina at 3780, 3740, 3705, and 3650 cm^?1 and a shoulder at 3795 cm^?1. The same bands were present on the oxidized catalyst, but with lower intensities and with altered intensity ratios; i.e., some bands were affected more than others as hydroxyl groups were replaced by molybdena species. Spectra from catalysts reduced with CO were indistinguishable in the OH region from those for the unreduced catalyst. No new bands appeared when the catalysts were reduced with H₂, but the intensities of bands attributable to alumina OH increased with the 3795 cm^?1 band strengthening noticeably more than the others. Thus, the new hydroxyl groups introduced on reduction are probably alumina OH rather than MoOH as previously supposed. A form of hydrogen which is chemisorbed but which can be removed from the catalyst as H₂ on evacuation at the reduction temperature also appeared in the OH region, mainly as a continuous contribution to the low frequency edge. From absorption coefficients derived from the present data, it was deduced that about twice as many hydrogen atoms were present in the H₂ formed than were supplied by these OH groups; i.e., the chemisorption appears heterolytic with half the atoms unseen by ir. A search was made for a band attributable to MoH, but without success. A brief study was made of this adsorption process, which was found to be slow but reversible, and to have a positive pressure dependence. When the catalyst was reduced with CO, rather than with H₂, a portion of the CO remained irreversibly chemisorbed in electronically comparable amounts. Infrared spectra of such samples contained a band at about 1585 cm^?1 attributable to a carbonate species. Data for the two reducing gases differed in that no reversibly chemisorbed CO was observed. At room temperature, CO was also chemisorbed as a linear species with the stretching frequency (2190 cm^?1) higher than that of the gaseous molecule (2143 cm^?1).     

Muscle Proteomic Profile before and after Enzyme Replacement Therapy in Late-Onset Pompe Disease

Mutations in the acidic alpha-glucosidase (GAA) coding gene cause Pompe disease. Late-onset Pompe disease (LOPD) is characterized by progressive proximal and axial muscle weakness and atrophy, causing respiratory failure. Enzyme replacement therapy (ERT), based on recombinant human GAA infusions, is the only available treatment; however, the efficacy of ERT is variable. Here we address the question whether proteins at variance in LOPD muscle of patients before and after 1 year of ERT, compared withhealthy age-matched subjects (CTR), reveal a specific signature. Proteins extracted from skeletal muscle of LOPD patients and CTR were analyzed by combining gel based (two-dimensional difference gel electrophoresis) and label-free (liquid chromatography-mass spectrometry) proteomic approaches, and ingenuity pathway analysis. Upstream regulators targeting autophagy and lysosomal tethering were assessed by immunoblotting. 178 proteins were changed in abundance in LOPD patients, 47 of them recovered normal level after ERT. Defects in oxidative metabolism, muscle contractile protein regulation, cytoskeletal rearrangement, and membrane reorganization persisted. Metabolic changes, ER stress and UPR (unfolded protein response) contribute to muscle proteostasis dysregulation with active membrane remodeling (high levels of LC3BII/LC3BI) and accumulation of p62, suggesting imbalance in the autophagic process. Active lysosome biogenesis characterizes both LOPD PRE and POST, unparalleled by molecules involved in lysosome tethering (VAMP8, SNAP29, STX17, and GORASP2) and BNIP3. In conclusion this study reveals a specific signature that suggests ERT prolongation and molecular targets to ameliorate patient’s outcome.     

Thermo‐responsive albumin hydrogels with LCST near the physiological temperature

This paper deals with the synthesis of thermoresponsive microspheres with proteic structure exhibiting a transition temperature close to the body temperature. The hydrogels were synthesized by free radical polymerization of methacrylate Bovine Serum Albumin (BSA‐MA) as crosslinker, and 2‐hydroxyethyl methacrylate (HEMA) and/or N‐isopropylacrylamide (NIPAAm), as hydrophilic and thermoresponsive monomers, respectively. The modification of the hydrophilic/hydrophobic balance in the polymerization feed allows to modulate the volume phase transition temperature of the macromolecular network. The hydrogels were characterized by infrared spectroscopy and thermal analyses, which showed negative thermoresponsive behavior for all compositions and, by increasing the content of the hydrophilic moieties in the network, the transition temperature was ranged from 34.2 to 36.8°C. To test the preformed materials as drug carriers, diclofenac diethyl ammonium salt was chosen and drug entrapment percent was determined. Drug release profiles, in media at different temperature, depend on the crosslinking degree and on the composition of the hydrogels. By using semiempirical equations, the release mechanism was extensively studied and the diffusional contribute evaluated. The physic‐chemical characteristics of thermoresponsive materials confirm the applicability of the microspheres as drug delivery device. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Rate‐dependent self‐healing behavior of an ethylene‐co‐methacrylic acid ionomer under high‐energy impact conditions

In this work, the mechanical and the self‐healing behaviors of an ethylene‐co‐methacrylic acid ionomer were investigated in different testing conditions. The self‐healing capability was explored by ballistic impact tests at low‐velocity, midvelocity, and hypervelocity bullet speed; different experimental conditions such as sample thickness and bullet diameter were examined; in all impact tests, spherical projectiles were used. These experiments, in particular those at low and midspeed, allowed to define a critical ratio between sample thickness and bullet diameter below which full repair was not observed. After ballistic damage, the healing efficiency was evaluated by applying a pressure gradient through tested samples. Subsequently, morphology analysis of the affected areas was made observing all tested samples by scanning electron microscope. This analysis revealed different characteristic features of the damaged zones affected at different projectile speed. Stress–strain curves in uniaxial tension performed at different temperatures and strain rates revealed yield strength and postyield behavior significantly affected by these two parameters. A rise of temperature during high strain rate tests in the viscoplastic deformation region was also detected. This behavior has a strong influence on the self‐repairing mechanism exhibited by the studied material during high‐energy impact tests. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 1949–1958, 2013     

Development of novel peptidomimetics containing a vinyl sulfone moiety as proteasome inhibitors

Proteasome inhibition is a topic of great interest in anticancer research. The proteolytic activity of this multicatalytic complex relies on three subunits, β1, β2 and β5, containing a caspase-like, a trypsin-like and a chymotrypsin-like active site, respectively. Several studies have demonstrated that, of the three activities, the chymotrypsin-like activity was the most necessary for cell viability and protein processing. Thus, most efforts towards the development of proteasome inhibitors have focused on the selective inhibition of the β5 subunit active site. Herein, we report the design and synthesis of a series of conformationally constrained tripeptidyl vinyl sulfones were determined to be good inhibitors of the chymotrypsin-like activity of proteasome, with K(I) values in the sub-micromolar to micromolar range. These compounds were also tested against bovine pancreatic α-chymotrypsin and human cathepsin B and L, revealing a good selectivity for the target enzyme over these related enzymes.