Uncertainty in the evaluation of the Predicted Mean Vote index using Monte Carlo analysis.

Today, evaluation of thermohygrometric indoor conditions is one of the most useful tools for building design and re-design and can be used to determine energy consumption in conditioned buildings. Since the beginning of the Predicted Mean Vote index (PMV), researchers have thoroughly investigated its issues in order to reach more accurate results; however, several shortcomings have yet to be solved. Among them is the uncertainty of environmental and subjective parameters linked to the standard PMV approach of ISO 7730 that classifies the thermal environment. To this end, this paper discusses the known thermal comfort models and the measurement approaches, paying particular attention to measurement uncertainties and their influence on PMV determination. Monte Carlo analysis has been applied on a data series in a "black-box" environment, and each involved parameter has been analysed in the PMV range from -0.9 to 0.9 under different Relative Humidity conditions. Furthermore, a sensitivity analysis has been performed in order to define the role of each variable. The results showed that an uncertainty propagation method could improve PMV model application, especially where it should be very accurate (-0.2 < PMV<0.2 range; winter season with Relative Humidity of 30%).

Evidence of hepatitis E virus (HEV) infection in human and pigs in Sardinia, Italy.

INTRODUCTION: The aim of this study was to determine the seroprevalence of anti-HEV antibodies in humans sera and to study HEV prevalence in swine from different Sardinian farms, testing viral HEV-RNA in bile samples. METHODS: In the first six months of 2008, 532 subjects of whom 402 blood donors and 130 workers at zoonotic risk, were enrolled. Anti-HEV were determined with an enzyme linked immunosorbent assay (ELISA). In positive subjects, RNA was extracted and tested by RT-Nested-PCR. From July 2006 to March 2007, 95 bile samples were collected from randomly selected pigs. RNA was extracted from 250 microl of bile and tested by RT-Nested-PCR. RESULTS: The overall prevalence of anti-HEV antibodies was 4.3%; 5.0% among blood donors and 2.3% among workers at zoonotic risk, with no statistically significant differences between sex, age classes and occupation. The search for HEV-RNA in the subjects positive for antibodies, gave negative results. HEV genome was detected in 6 of the 95 swine bile samples tested. Sequences were clustered within the genotype 3 and are edited on GenBank under accession number: from FJ850960 to FJ850962 and from FJ883000 to FJ883002. DISCUSSION: The overall prevalence of anti-HEV shows that the virus circulates without giving origin to cases of acute hepatitis. The low prevalence value found in workers at zoonotic risk do not apparently support the hypothesis of professional risk. In this study, HEV-RNA was isolated from pigs in Sardinia for the first time confirming the role of swine as HEV reservoir and the possibility of virus transmission to humans.

[The environmental monitoring of the exposure to chemical contamination in operating rooms]. / Monitoraggio ambientale della esposizione a microcontaminanti chimici nelle sale operatorie.

The medical staff which works in an operating room is exposed to danger due to the chemical contamination found in the air. The results of this research depend to hormonal and haematochemical variations. The chemical contamination can be the cause of pathologies of the respiratory organs, the skin, the mucosa and the immune system. After a preventive evaluation of the production processes and the working procedures, some researchers have estimated the environmental risk caused by low concentrations of chemical products. In order to control the levels of the chemical agents, they have used an integrated system set up by a gaschromatography-mass spectrometer that has found some levels of chemical agents peculiar to a low pollution, characterized by the low concentration under the levels of the so-called ACGIH (2006) of hesane, toluene, ethylbenzene, o-xylene and naphthalene. According to the latest studies is very important to develop working methods and scientific knowledges direct to environmental, medical and toxicological problems. They are necessary to guarantee a greater protection of the human health and the safety at work.

[Formation and quality: an integrated pattern of professional growing]. / Formazione e qualità: modello integrato di crescita professionale.

The cultural formation the professional man's knowledges and his way to operate. It represents a resource to elaborate organizational and operative methods compatible with the user's needs. A good formation permeates the professional man's processing technique and it accompanies the definition of strategies, targets and decisions. The formation makes the professional man acquire specialist knowledges and actions to reduce the fragmentation of the human communication and it favours the efficacy of all the interventions for the health and the security of employment. It is necessary to promote many operative methods integrated with a qualitative system in order to favour the managerial abilities of a professional man together with his competences inside our social and productive world.

Structural studies of lysyl-tRNA synthetase: conformational changes induced by substrate binding.

Lysyl-tRNA synthetase is a member of the class II aminoacyl-tRNA synthetases and catalyses the specific aminoacylation of tRNA(Lys). The crystal structure of the constitutive lysyl-tRNA synthetase (LysS) from Escherichia coli has been determined to 2.7 A resolution in the unliganded form and in a complex with the lysine substrate. A comparison between the unliganded and lysine-bound structures reveals major conformational changes upon lysine binding. The lysine substrate is involved in a network of hydrogen bonds. Two of these interactions, one between the alpha-amino group and the carbonyl oxygen of Gly 216 and the other between the carboxylate group and the side chain of Arg 262, trigger a subtle and complicated reorganization of the active site, involving the ordering of two loops (residues 215-217 and 444-455), a change in conformation of residues 393-409, and a rotation of a 4-helix bundle domain (located between motif 2 and 3) by 10 degrees. The result of these changes is a closing up of the active site upon lysine binding.

Active site of lysyl-tRNA synthetase: structural studies of the adenylation reaction.

Aminoacyl-tRNA synthetases play a key role in protein biosynthesis by catalyzing the specific aminoacylation of tRNA. The energy required for the formation of the ester bond between the amino acid carboxylate group and the tRNA acceptor stem is supplied by coupling the reaction to the hydrolysis of ATP. Lysyl-tRNA synthetase from Escherichia coli belongs to the family of class II synthetases and carries out a two-step reaction, in which lysine is activated by being attached to the alpha-phosphate of AMP before being transferred to the cognate tRNA. Crystals of the thermo-inducible E. coli lysyl-tRNA synthetase LysU which diffract to 2.1 A resolution have been used to determine crystal structures of the enzyme in the presence of lysine, the lysyl-adenylate intermediate, and the nonhydrolyzable ATP analogue AMP-PCP. Additional data have been obtained from crystals soaked in a solution containing ATP and Mn(2+). The refined crystal structures give "snapshots" of the active site corresponding to key steps in the aminoacylation reaction and provide the structural framework for understanding the mechanism of lysine activation. The active site of LysU is shaped to position the substrates for the nucleophilic attack of the lysine carboxylate on the ATP alpha-phosphate. No residues are directly involved in catalysis, but a number of highly conserved amino acids and three metal ions coordinate the substrates and stabilize the pentavalent transition state. A loop close to the catalytic pocket, disordered in the lysine-bound structure, becomes ordered upon adenine binding.

Identification and characterization of a DNA primase from the hyperthermophilic archaeon Methanococcus jannaschii.

We report the identification and characterisation of a DNA primase from the thermophilic methanogenic archaeon Methanococcus jannaschii (Mjpri). The analysis of the complete genome sequence of this organism has identified an open reading frame coding for a protein with sequence similarity to the small subunit of the eukaryotic DNA primase (the p50 subunit of the polymerase alpha-primase complex). This protein has been overexpressed in Escherichia coli and purified to near homogeneity. Recombinant Mjpri is able to synthesise oligoribonucleotides on various pyrimidine single-stranded DNA templates [poly(dT) and poly(dC)]. This activity requires divalent cations such Mg(2+), Mn(2+)or Zn(2+), and is additionally stimulated by the monovalent cation K(+). A multiple sequence alignment has revealed that most of the regions that are conserved in eukaryotic p50 subunits are also present in the archaeal primases, including the conserved negatively charged residues, which have been shown to be essential for catalysis in the mouse primase. Of the four cysteine residues that have been postulated to make up a putative Zn-binding motif, two are not present in the archaeal homologue. This is the first report on the biochemical characterisation of an archaeal DNA primase.