Prevalence of hepatitis E virus in Italian pig herds. Preliminary results

Hepatitis E virus (HEV) is the causative agent of hepatitis E, and is an unenveloped positive sense single-stranded RNA wus. Swine HEV strains are genetically closely related to human strains from the same area, suggesting the occurrence of zoonotic transmission. Recently, human cases of hepatitis E have been linked to the consumption of raw or undercooked meat or organs from deer, wild boars or p1gs. The disease is now considered an emerging food-borne transmitted zoonosis. During 2006, a pilot investigation was performed to determine the prevalence of HEV in pig farms located in Northern Italy. 274 faecal samples were collected from healthy fattening animals (3-4 and 8-9 months of age) and from healthy breeding animals (gilts and sows) from 6 different farms, and analyzed using a Nested-RT-PCR targeting the open reading frame 2 (ORF2) region. Stool samples were suspended in water, and viral RNA extraction was performed using a commercial kit. Extracted viral RNA was subjected to RT-PCR amplification using degenerate primers conA 1-conS1 for the first amplification, and degenerate primers conA2-conS2 for the nested PCR, yielding a final fragment of 145 bp. HEV RNA was detected in sixty-nine of the 274 (25.2%) examined samples. None of the six farms resulted negative and the prevalence within the farms ranged between 2% and 60.5% For the characterization of the strains, randomly selected positive samples were subjected to nucleotide sequencing, and aligned with those present in the NCBI Data Bank Sequence analysis showed that all stra1ns were Swine Hepatitis E belonging to Genotype 3. These preliminary results confirm that swine HEV is widespread in Italian swine farms

Mutation of tyrosine 492/493 in the kinase domain of ZAP-70 affects multiple T-cell receptor signaling pathways.

The protein-tyrosine kinase ZAP-70 is implicated, together with the Src kinase p56lck, in controlling the early steps of the T-cell antigen receptor (TCR) signaling cascade. To help elucidate further the mechanism by which ZAP-70 regulates these initial events, we used a dominant-negative mutant approach. We overexpressed in the Jurkat T-cell line ZAP-70 mutated on Tyr-492 and Tyr-493 in the putative regulatory loop of its kinase domain. This mutant inhibited TCR-induced activation of nuclear factor of activated T cells by interfering with both intracellular calcium increase and Ras-regulated activation of extracellular signal-regulated kinases. Moreover, TCR-induced phosphorylation of pp36-38, thought to play a role upstream of these pathways, was found to be reduced. In contrast, overexpression of wild-type ZAP-70 induced constitutive activation of nuclear factor of activated T cells. The ZAP-70 mutant studied here could be phosphorylated on tyrosine when associated to the TCR ζ chain and was able to bind p56lck. This result demonstrates that Tyr-492 and Tyr-493 are not responsible for the Src homology domain 2-mediated association of p56lck with ZAP-70. Our data are most consistent with a model in which recruitment to the TCR allows ZAP-70 autophosphorylation and binding to p56lck, which in turn phosphorylates Tyr-492 and/or Tyr-493 with consequent up-regulation of the ZAP-70 kinase activity. ZAP-70 will then be able to effectively control phosphorylation of its substrates and lead to gene activation

Detection of serum antibodies to hepatitis E virus in domestic pigs in Italy using a recombinant swine HEV capsid protein

Background: The hepatitis E virus (HEV) has been detected in both humans and animals, particularly pigs, worldwide. Several evidences, including human infection following consumption of raw contaminated meat, suggest a zoonotic transmission of HEV. In Italy, large circulation of genotype 3 HEV has been reported in swine, and recent studies have confirmed the involvement of this genotype in autochthonous human cases. Result: In this study 111 sera collected from healthy pigs in two Italian regions were tested for anti-HEV IgG antibodies. For specific HEV antibody detection in swine, we developed ELISA and Western blotting methods, using a truncated capsid (ORF2) protein lacking the first 111 amino acids of a swine HEV genotype 3 strain. The ORF2-based ELISA revealed anti-HEV antibodies in 104 out of 111 pigs compared with 102 detected with a commercial ELISA kit. A lower number of sera reacted with the recombinant ORF2 protein in a Western blotting format (81/111). Using a Latent class analysis (LCA), the estimated sensitivities for ELISA-ORF2 and ELISA-kit tests were 0.961 and 0.936, respectively, whereas specificities were 0.599 and 0.475. The estimated sensitivity of Western blotting was 0.775, and the specificity was 0.944. Conclusions: The overall results confirm the high prevalence of HEV seropositive healthy pigs in Italy. Through comparisons with a commercial ELISA test, the swine genotype 3 HEV antigen produced in this study was proven suitable to detect anti-HEV antibodies in pig sera by both ELISA and Western Blotting

Correlation between micro and macrostructural biaxial behavior of ascending thoracic aneurysm: a novel experimental technique

Mechanical properties and microstructural modifications of vessel tissues are strongly linked, as established in the state of the art of cardiovascular diseases. Techniques to obtain both mechanical and structural information are reported, but the possibility to obtain real-time microstructural and macrostructural data correlated is still lacking. An experimental approach to characterize the aortic tissue is presented. A setup integrating biaxial traction and Small Angle Light Scattering (SALS) analysis is described. The system was adopted to test ex-vivo aorta specimens from healthy and aneusymatic (aTAA) cases. A significant variation of the fiber dispersion with respect to the unloaded state was encountered during the material traction. The corresponding microstructural and mechanical data were successfully used to fit a given anisotropic constitutive model, with satisfactory R2 values (0.97±0.11 and 0.96±0.17, for aTAA and healthy population, respectively) and fiber dispersion parameters variations between the aTAA and healthy populations (0.39±0.23 and 0.15±0.10). The method integrating the biaxial/SALS technique was validated, allowing for real-time synchronization between mechanical and microstructural analysis of anisotropic biological tissues

Complete Reversible Refolding of a G-Protein Coupled Receptor on a Solid Support

The factors defining the correct folding and stability of integral membrane proteins are poorly understood. Folding of only a few select membrane proteins has been scrutinised, leaving considerable deficiencies in knowledge for large protein families, such as G protein coupled receptors (GPCRs). Complete reversible folding, which is problematic for any membrane protein, has eluded this dominant receptor family. Moreover, attempts to recover receptors from denatured states are inefficient, yielding at best 40-70% functional protein. We present a method for the reversible unfolding of an archetypal family member, the β1-adrenergic receptor, and attain 100% recovery of the folded, functional state, in terms of ligand binding, compared to receptor which has not been subject to any unfolding and retains its original, folded structure. We exploit refolding on a solid support, which could avoid unwanted interactions and aggregation that occur in bulk solution. We determine the changes in structure and function upon unfolding and refolding. Additionally, we employ a method that is relatively new to membrane protein folding; pulse proteolysis. Complete refolding of β1-adrenergic receptor occurs in n-decyl-β-D-maltoside (DM) micelles from a urea-denatured state, as shown by regain of its original helical structure, ligand binding and protein fluorescence. The successful refolding strategy on a solid support offers a defined method for the controlled refolding and recovery of functional GPCRs and other membrane proteins that suffer from instability and irreversible denaturation once isolated from their native membranes

Comparison between off-resonance and electron Bernstein waves heating regime in a microwave discharge ion sourcea)

A microwave discharge ion source (MDIS) operating at the Laboratori Nazionali del Sud of INFN, Catania has been used to compare the traditional electron cyclotron resonance (ECR) heating with an innovative mechanisms of plasma ignition based on the electrostatic Bernstein waves (EBW). EBW are obtained via the inner plasma electromagnetic-to-electrostatic wave conversion and they are absorbed by the plasma at cyclotron resonance harmonics. The heating of plasma by means of EBW at particular frequencies enabled us to reach densities much larger than the cutoff ones. Evidences of EBW generation and absorption together with X-ray emissions due to high energy electrons will be shown. A characterization of the discharge heating process in MDISs as a generalization of the ECR heating mechanism by means of ray tracing will be shown in order to highlight the fundamental physical differences between ECR and EBW heating

A generalized Hamiltonian Constraint Operator in Loop Quantum Gravity and its simplest Euclidean Matrix Elements

We study a generalized version of the Hamiltonian constraint operator in nonperturbative loop quantum gravity. The generalization is based on admitting arbitrary irreducible SU(2) representations in the regularization of the operator, in contrast to the original definition where only the fundamental representation is taken. This leads to a quantization ambiguity and to a family of operators with the same classical limit. We calculate the action of the Euclidean part of the generalized Hamiltonian constraint on trivalent states, using the graphical notation of Temperley-Lieb recoupling theory. We discuss the relation between this generalization of the Hamiltonian constraint and crossing symmetry.Comment: 35 pp, 20 eps figures; minor corrections, references added; version to appear in Class. Quant. Gra

broadband visible light emission from nominally undoped and hbox cr 3 doped garnet nanopowders

Synthetic garnet nanopowders of Y 3 Al 5 O 12 (YAG) and Gd 3 Ga 5 O 12 (GGG) were produced, and the occurrence of a broadband bright visible emission by nominally undoped YAG and GGG and Cr 3+ doped GGG, depending on the environment pressure, as well as exciting on the pumping power, was demonstrated. The results indicate that high-intensity infrared laser irradiation in samples not only leads to heating (melting effects) but also produces visible broadband emission. Low pressure of the powders' environment favors the white light emission by lowering the threshold pumping power. A hypothesis on the nature of the emission is presented

Integration of optical telecommunications and radio access networks to assure quality of service

n/

Prediction of responsiveness of gait variables to rehabilitation training in Parkinson's disease

Background: Gait disorders represent one of the most disabling features of Parkinson's disease, which may benefit from rehabilitation. No consistent evidence exists about which gait biomechanical factors can be modified by rehabilitation and which clinical characteristic can predict rehabilitation-induced improvements. Objectives: The aims of the study were as follows: (i) to recognize the gait parameters modifiable by a short-term rehabilitation program; (ii) to evaluate the gait parameters that can normalize after rehabilitation; and (iii) to identify clinical variables predicting improvements in gait function after rehabilitation. Methods: Thirty-six patients affected by idiopathic Parkinson's disease in Hoehn-Yahr stage 1-3 and 22 healthy controls were included in the study. Both clinical and instrumental (gait analysis) evaluations were performed before and after a 10-weeks rehabilitation treatment. Time-distance parameters, lower limb joint, and trunk kinematics were measured. Results: At baseline evaluation with matched speed, almost all gait parameters were significantly different between patients and healthy controls. After the 10-weeks rehabilitation, most gait parameters improved, and spatial asymmetry and trunk rotation normalized. Multiple linear regression of gender combined with Unified Parkinson's Disease Rating Scale-III predicted both ΔSpeed and ΔStep length of both sides; gender combined with Unified Parkinson's Disease Rating Scale-II predicted ΔCadence; age combined with Hoehn-Yahr score and disease duration predicted 1trunk rotation range of motion. Conclusions: Impaired gait parameters are susceptible to improvement by rehabilitation, and younger men with Parkinson's disease who are less severely affected and at early disease stage are more susceptible to improvements in gait function after a 10-weeks rehabilitation program