Need for integrative thinking to fight against emerging infectious diseases. Proceedings of the 5th seminar on emerging infectious diseases, March 22, 2016 - current trends and proposals.

We present here the proceedings of the 5th seminar on emerging infectious diseases, held in Paris on March 22nd, 2016, with seven priority proposals that can be outlined as follows: encourage research on the prediction, screening and early detection of new risks of infection; develop research and surveillance concerning transmission of pathogens between animals and humans, with their reinforcement in particular in intertropical areas ("hot-spots") via public support; pursue aid development and support in these areas of prevention and training for local health personnel, and foster risk awareness in the population; ensure adapted patient care in order to promote adherence to treatment and to epidemic propagation reduction measures; develop greater awareness and better education among politicians and healthcare providers, in order to ensure more adapted response to new types of crises; modify the logic of governance, drawing from all available modes of communication and incorporating new information-sharing tools; develop economic research on the fight against emerging infectious diseases, taking into account specific driving factors in order to create a balance between preventive and curative approaches.

Low-loss reciprocal optical terminals for two-way time-frequency transfer.

We present the design and performance of a low-cost, reciprocal, compact free-space terminal employing tip/tilt pointing compensation that enables optical two-way time-frequency transfer over free-space links across the turbulent atmosphere. The insertion loss of the terminals is ∼1.5 dB with total link losses of 15 dB, 24 dB, and 50 dB across horizontal, turbulent 2-km, 4-km, and 12-km links, respectively. The effects of turbulence on pointing control and aperture size, and their influence on the terminal design, are discussed.

Gas temperature dependent sticking of hydrogen on cold amorphous water ice surfaces of interstellar interest.

Using the King and Wells method, we present experimental data on the dependence of the sticking of molecular hydrogen and deuterium on the beam temperature onto nonporous amorphous solid water ice surfaces of interstellar interest. A statistical model that explains the isotopic effect and the beam temperature behavior of our data is proposed. This model gives an understanding of the discrepancy between all known experimental results on the sticking of molecular hydrogen. Moreover, it is able to fit the theoretical results of Buch et al. [Astrophys. J. 379, 647 (1991)] on atomic hydrogen and deuterium. For astrophysical applications, an analytical formula for the sticking coefficients of H, D, H(2), D(2), and HD in the case of a gas phase at thermal equilibrium is also provided at the end of the article.

OH formation from O and H atoms physisorbed on a graphitic surface through the Langmuir-Hinshelwood mechanism: a quasi-classical approach.

We study the quasi-classical dynamics of OH formation on a graphitic surface through the Langmuir-Hinshelwood (LH) mechanism when both O and H ground-state atoms are initially physisorbed on the surface. The model proceeds from previous theoretical work on the LH formation of the H 2 molecule on graphite [Morisset, S.; Aguillon, F.; Sizun, M.; Sidis, V. J. Chem. Phys. 2004, 121, 6493; ibid 2005, 122, 194704]. The H-graphite system is first revisited with a view to get a tractable DFT-GGA computational prescription for the determination of atom physisorption onto graphitic surfaces. The DZP-RPBE combination is found to perform well; it is thereafter used along with MP2 calculations to determine the physisorption characteristics of atomic oxygen on graphitic surfaces. We also deal with chemisorption. In accordance with previous work, we find that O chemisorbs on graphite in a singlet spin state epoxy-like conformation. In the triplet state we find only "metastable" chemisorption with an activation barrier of 0.2 eV. The physisorption results are then used in the LH dynamics calculation. We show that in the [0.15 meV, 12 meV] relative collision energy range of the reacting O and H atoms on the surface, the OH molecule is produced with a large amount of internal energy ( approximately = 4eV) and a significant translation energy (>or=100 meV) relative to the surface.

Diagnostic specificity of the African swine fever virus antibody detection enzyme-linked immunosorbent assay in feral and domestic pigs in the United States.

African swine fever (ASF) is a highly contagious haemorrhagic disease of pigs that has the potential to cause mortality nearing 100% in naïve animals. While an outbreak of ASF in the United States' pig population (domestic and feral) has never been reported, an introduction of the disease has the potential to cause devastation to the pork industry and food security. During the recovery phase of an outbreak, an antibody detection diagnostic assay would be required to prove freedom of disease within the previously infected zone and eventually nationwide. Animals surviving an ASF infection would be considered carriers and could be identified through the persistence of ASF viral antibodies. These antibodies would demonstrate exposure to the disease and not vaccination, as there is no ASF vaccine available. A well-established commercial enzyme-linked immunosorbent assay (ELISA) detects antibodies against ASF virus (ASFV), but the diagnostic specificity of the assay had not been determined using serum samples from the pig population of the United States. This study describes an evaluation of the World Organization for Animal Health (OIE)-recommended Ingezim PPA COMPAC ELISA using a comprehensive cohort (n = 1791) of samples collected in the United States. The diagnostic specificity of the assay was determined to be 99.4% (95% confidence interval (CI): [98.9, 99.7]). The result of this study fills a gap in understanding the performance of the Ingezim PPA COMPAC ELISA in the ASF naïve pig population of the United States.

Sequence and expression of the bpdC1C2BADE genes involved in the initial steps of biphenyl/chlorobiphenyl degradation by Rhodococcus sp. M5.

The nucleotide (nt) sequence of the bpdC1C2BADE genes which encode the first three enzymes in the biphenyl (BP) degradation pathway of Gram+ Rhodococcus sp. M5 (formerly Arthrobacter M5) was determined. Except for the ferredoxin component (BpdB) of the initial BP dioxygenase, the predicted amino acid (aa) sequences of the remaining proteins are found to be more closely related to the counterpart proteins (TodC1C2BADE) present in the toluene-degrader, Pseudomonas putida F1, than those of three BP-degrading pseudomonads. The cloned bpd genes were verified by their expression in the Escherichia coli T7 RNA polymerase/promoter system. In E. coli, BpdA was able to complement TodC1C2B in indigo biosynthesis, although the M5 native or cloned BP dioxygenase does not carry out this reaction.

Cloning, sequence and expression of a linear plasmid-based and a chromosomal homolog of chloroacetaldehyde dehydrogenase-encoding genes in Xanthobacter autotrophicus GJ10.

The degradation of 1,2-dichloroethane (DCE) by Xanthobacter autotrophicus GJ10 proceeds via chloroacetaldehyde (CAA), a toxic intermediate in the cells if it is not metabolized further by the NAD(+)-dependent CAA dehydrogenases. Here, we describe the cloning, sequence and expression in Escherichia coli of aldA, a plasmid-located CAA dehydrogenase-encoding gene of GJ10 as well as a chromosomal homolog, designated aldB. The DNA-predicted amino acid (aa) sequences of the two proteins (505 aa in AldA and 506 aa in AldB) are 84% identical. The cloned aldA and aldB genes were verified by their expression in the E. coli T7 polymerase/promoter and the pUC lac promoter systems. The expression level of AldA and its enzymatic activity towards CAA were both higher than those of AldB. In a hybrid construct, the 3'end of aldB was able to complement, although not completely, the corresponding portion of aldA to produce a functional gene. Both AldA and AldB proteins of GJ10 share the highest degree of sequence identity with an acetaldehyde dehydrogenase (ALDH) encoded by acoD of Alcaligenes eutrophus (77.3-78% identity). Together with at least three other ALDHs of prokaryotic origin, these proteins apparently form a special class of ALDHs whose expressions are dependent on RpoN factors. By pulsed-field gel electrophoresis the 225-kb pXAU1 plasmid encoding aldA was shown to be linear.

Sequence and expression of the todGIH genes involved in the last three steps of toluene degradation by Pseudomonas putida F1.

The todFC1C2BADE gene cluster in Pseudomonas putida F1 encodes enzymes for the first four steps of toluene degradation, leading to the formation of 2-hydroxypenta-2,4-dienoate (HPD). Here, we report the nucleotide (nt) sequence and expression of the remaining three genes of the tod pathway, downstream from todE and arranged in the order, todGIH. The deduced amino acid (aa) sequences of TodG [HPD hydratase (268 aa)], TodH [4-hydroxy-2-oxovalerate (HO) aldolase (352 aa)] and TodI [acylating aldehyde (AA) dehydrogenase (316 aa)] are compared with the isofunctional proteins present in the meta-cleavage pathways of other bacteria. New sequence motifs are identified. The highly conserved TodH and TodI sequences are potentially useful DNA probes for biomonitoring purposes.

A fusion plasmid for the synthesis of lipopeptide-antigen chimeras in Escherichia coli.

Lipopeptides are potential vaccine candidates with a built-in adjuvant property. To circumvent the present chemical route of synthesis for lipopeptide-antigen conjugates, the lipoprotein property of the pColE2-P9-encoded lysis protein, CelB, was used to create the bacterial fusion plasmid, pKLY3, to produce lipopeptide-antigen chimeras in Escherichia coli. Plasmid pKLY3 is a derivative of pKK233-2 with the origin of replication of the single-stranded DNA phage, fl. Under control of the promoter, ptrc, is the 5' end of the celB gene coding for a lipoprotein signal peptide and the first five amino acids (aa) (CQANY) of the mature lysis protein. As model systems for the synthesis of small and large lipopeptide-antigens, DNA sequences coding for the P2 peptide and E. coli alkaline phosphatase (PhoA) were fused in frame to the region of celB coding for a lipoprotein signal peptide and CQANY. P2 is a 12-aa peptide including a tyrosine phosphorylation site of the epidermal growth factor receptor (EGF-R). Inducible expression of stable lipohexapeptide CQANYV, lipo-CQANY-P2, and lipo-CQANYA-PhoA, was demonstrated. Similar expression was obtained for lipo-CIEGR-P2 and lipo-CIEGRA-PhoA in which IEGR is a cleavage recognition site for the blood coagulation factor, Xa. Like QANY, IEGR is predicted to form a beta-turn structure. The presence of a lipid moiety on the products was confirmed by demonstrating the incorporation of radioactive palmitic acid and inhibition of processing by globomycin. The lipid-modified peptides were also identified by incorporation of radioactive tyrosine, and the nature of the P2 peptide was verified immunologically.(ABSTRACT TRUNCATED AT 250 WORDS)

TTX blocks baclofen-induced phase shifts of the mammalian circadian pacemaker in vitro.

The mammalian circadian pacemaker, located in the suprachiasmatic nucleus (SCN), expresses 24-h rhythms when isolated in vitro. The GABA(A) agonist, muscimol, induces phase advances during the mid-subjective day, while the GABA(B) agonist, baclofen, induces both daytime phase advances and nighttime phase delays. Here, we present evidence that tetrodotoxin (TTX) completely blocks baclofen-induced phase shifts in vitro, but does not block in vitro phase advances induced by muscimol. These results suggest that GABA(A), but not GABA(B), receptors are located on SCN pacemaker cells.

Aircraft automation: the problem of the pilot interface.

Aircraft operations, particularly in the IFR environment, are rapidly becoming very complex. Studies have shown that this complexity can frequently lead to accidents and incidents. Results of studies performed at NASA and elsewhere are presented to show that one of the major themes evident in both the accidents and incidents and in the research performed to solve the problems associated with them is that of human error. Examples of various incidents and blunders, recorded in several studies, illustrate and emphasize the hypothesis: "As systems become more and more automated and complex, the more they become prone to human error. The problem can be eliminated or reduced only if good human factor principles are incorporated in the implementation of the systems, to guarantee a good man/machine interface". Aircraft systems technology, however, (e.g.: electronics, avionics, automation) is evolving and developing at a very high rate. Examples of research are presented showing where this emerging technology has been employed to reduce the complexity and enhance the safety and utility of the aircraft operations.

Social consequences of long term impairments and disabilities: conceptual approach and assessment of handicap.

OBJECTIVES: To present a conceptual model of disablement adapted from the WHO model and to conduct a pilot study with a measurement tool (LIFE-H) of the concepts of life habits and handicap situations. DESIGN: Content validity and test-retest reliability study. SETTING: General community. PARTICIPANTS: A panel of 12 experts of rehabilitation for the process of content validity and 49 individuals with spinal cord disorders (adults and children) for the reliability study. OUTCOMES MEASURES: a person's life habits (activities of daily living and social roles). RESULTS: The LIFE-H questionnaire was designed to assess the handicap situations observed in daily life of individuals with disability. The experts concluded that the LIFE-H items covered most of a person's life habits (ADL and social roles) and that it could be used to determine the appearance of handicap situations. The LIFE-H total score showed a good level of reliability for the children and the adult samples (ICC = 0.73 and 0.74, respectively). Taken individually, a majority of life habit categories have shown a moderate to high reliability level (ICC > or = 0.50) while a few life habit categories such as the interpersonal relationship or nutrition showed a lower reliability level. CONCLUSION: The development of LIFE-H allows fulfillment of the need to determine the disruptions in life habits of persons with disabilities.

[Assaults in psychiatric wards : Experience and perceptions of mental health workers at the Pavillon Roland-Saucier du Complexe hospitalier de la Sagamie.]. / Les agressions en milieu psychiatrique. Vécu et perceptions des intervenants du Pavillon Roland-Saucier du Complexe hospitalier de la Sagamie.

This article presents results of a study conducted at the Pavillon Roland-Saucier, the psychiatric ward of the Complexe hospitalier de la Sagamie. The objective was to explore with a phenomenological approach the issue of assaults by patients of mental health workers and in particular the impact of theses assaults on their professional life. Thirty workers of various professional categories, selected by a stratified random procedure, have accepted to be interviewed. The analysis of theses interviews has allowed to draw important elements on the basis which various recommendations have been suggested in order to improve the situation.

Ultrastructural study of nucleolar changes in rat embryos during diapause and reactivation.

Changes in nucleolar ultrastructure were studied in preimplantation rat embryos before diapause (d 5), during diapause (d 6, 7 and 8) and after reactivation brought about either by in vitro culture for 24 h (d 7 and 8) or by estradiol-17 beta treatment of the mothers (d 11). Before diapause, fully developed nucleoli contained several low-density fibrillar centers surrounded by a dense fibrillar component and an abundant granular component. This type of nucleolus indicates a high activity of ribosomal RNA synthesis. During diapause, nucleoli revealed a disorganization of the fibrillar and granular elements typical of a diminution in transcriptional activity. During reactivation, nucleoli progressively returned to a reticulo-fibrillar configuration characteristic of the onset of intense transcriptional activity. It is concluded that the structure of the nucleolus in rat preimplantation embryos corresponds to the level of transcriptional activity and is a reliable model for studying structure-function relationships during early development.

Characterization of a new solvent-responsive gene locus in Pseudomonas putida F1 and its functionalization as a versatile biosensor.

A new gene cluster, designated sepABC and a divergently transcribed sepR, was found downstream of the two-component todST phosphorelay system that regulates toluene degradation (the tod pathway) in Pseudomonas putida F1 (PpF1). The deduced amino acid sequences encoded by sepABC show a high homology to bacterial proteins known to be involved in solvent efflux or multidrug pumps. SepA, SepB and SepC are referred to be periplasmic, inner membrane and outer membrane efflux proteins respectively. Effects on growth of various PpF1 mutants compared to that of the wild type in the presence of toluene indicated a possible protective role of the solvent efflux system in a solvent-stressed environment. Growth tests with the complemented mutants confirmed the involvement of the Sep proteins in conferring solvent tolerance. The sepR gene encodes a 260-residue polypeptide that is a member of the E. coli IclR repressor protein family. The repressor role of SepR was established by conducting tests with a sep-lacZ transcriptional fusion in Escherichia coli and PpF1, expression of SepR as a maltose-binding fusion protein in a DNA binding assay, and mRNA analysis. Southern hybridization experiments and analysis of the P. putida KT2440 genome sequence indicated that sepR is a relatively rare commodity compared to homologues of the sepABC genes. We developed a whole-cell bioluminescent biosensor, PpF1G4, which contains a chromosomally based sep-lux transcriptional fusion. The biosensor showed significant induction of the sepABC genes by a wide variety of aromatic molecules, including benzene, toluene, ethylbenzene, and all three isomers of xylene (BTEX), naphthalene, and complex mixtures of aliphatic and aromatic hydrocarbons. PpF1G4 represents a second-generation biosensor that is not based on a catabolic promoter but is nonetheless inducible by aromatic pollutants and moreover functional under nutrient-rich conditions.

Influence of sex steroids on the production of prostaglandins F2 alpha and E2 and response to oxytocin in cultured epithelial and stromal cells of the bovine endometrium.

The uterus is a primary target for sex steroid action in vivo during the estrous cycle and pregnancy. Cell cultures have been used to determine the specific function of the different cell types forming the uterus. We used endometrial cell cultures previously characterized in our laboratory to study the effect of estradiol (E) and progesterone (P4) on prostaglandin (PG) production and on regulation of the response of the cells to oxytocin (OT). The studies were performed on confluent cultures of epithelial cells grown as a monolayer either on plastic or on filter inserts to allow basal-apical polarization. As described previously, prostaglandin F2 alpha (PGF 2 alpha) production was greater (3.7-fold, p < 0.0001) than prostaglandin E2 (PGE2) production in epithelial cells, and the opposite was true in stromal cells (PGE2 9.9-fold > PGF2 alpha, p < 0.0001). In epithelial cells, the basal production of PGE2 (-61.6%, p < 0.0001) and PGF2 alpha (-51.7%, p < 0.0001) was reduced significantly by E and increased significantly by P4 (PGE2, + 30.0% [p < 0.002]; PGF2 alpha, + 22.2% [p < 0.006]). No significant effect of sex steroids on the basal production of PGs was detected in stromal cells. OT stimulated the production of PGF2 alpha (6.7-fold, p < 0.0001) and PGE2 (9.1-fold, p < 0.0001) in epithelial but not stromal cells. Treatment of the cells with E significantly (p < 0.001) increased OT-stimulated PGF2 alpha production in both the epithelial and stromal cells and that of PGE2 in epithelial cells only. The effect of steroids and OT was similar in polarized (filter) and nonpolarized (plastic) epithelial cells. Analysis of the vectorial secretion of PGs in epithelial cells grown on filter inserts revealed that PGF2 alpha is preferentially secreted in the basal (p < 0.001) compared to the apical compartment. The direction of secretion was not influenced by steroid or OT treatments. The results suggest that epithelial cells of the endometrium are a preferred target for the regulation of PG synthesis by sex steroids and OT.

The NlaIV restriction and modification genes of Neisseria lactamica are flanked by leucine biosynthesis genes.

The genes encoding the Neisseria lactamica restriction endonuclease IV (R.NlaIV) and its cognate DNA methyltransferase (M.NlaIV), both of which recognize the sequence GGNNCC, have been cloned in Escherichia coli and overexpressed using the T7 polymerase/promoter system. Analysis of a sequenced 3.58 kb fragment established the gene order, leuD-M.NlaIV-R.NlaIV-leuB. The predicted primary sequence of M.NlaIV (423 amino acids) shows the highest degree of identity to a pair of cytosine-specific methyltransferases, M.BanI (44.9%) and M.HgiCI (44.3%), which recognize the sequence GGYRCC (Y, pyrimidines; R, purines). In contrast, the R.NlaIV protein sequence (243 amino acids) is unique in the existing data-base, a situation that holds for most endonucleases. Flanking the NlaIV modification and restriction genes are homologues of the leuD and leuB genes of enteric bacteria, which code for enzymes in the leucine biosynthesis pathway. This gene context implies a possible new mode of gene regulation for the RM.NlaIV system, which would involve a mechanism similar to the recently discovered leucine/Lrp regulon in E. coli.

Microbial contamination of embryos and semen during long term banking in liquid nitrogen.

We report on microbial contamination of embryos and semen cryopreserved in sealed plastic straws and stored for 6-35 years in liquid nitrogen. There were 32 bacterial and 1 fungal species identified from randomly drawn liquid nitrogen, frozen semen, and embryos samples stored in 8 commercial and 8 research facility liquid nitrogen (LN) tanks. The identified bacteria represented commensal or environmental microorganisms and some, such as Escherichia coli, were potential or opportunistic pathogens for humans and animals. Stenotrophomonas maltophilia was the most common contaminant identified from the samples and was further shown to significantly suppress fertilization and embryonic development in vitro. Analysis of the strains by pulsed field gel electrophoresis revealed restriction patterns with no relatedness indicating that there was no apparent cross-contamination of S. maltophilia strains between the germplasm and liquid nitrogen samples. In addition, no transmission of bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) from infected semen and embryos straws to clean germplasm stored in the same LN tanks or LN was detected.

Identification of a transcriptional activator (ChnR) and a 6-oxohexanoate dehydrogenase (ChnE) in the cyclohexanol catabolic pathway in Acinetobacter sp. Strain NCIMB 9871 and localization of the genes that encode them.

We identified chnR, a gene encoding an AraC-XylS type of transcriptional activator that regulates the expression of chnB, the structural gene for cyclohexanone monooxygenase (CHMO) in Acinetobacter sp. strain NCIMB 9871. The gene sequence of chnE, which encodes an NADP(+)-linked 6-oxohexanoate dehydrogenase, the enzyme catalyzing the fifth step of cyclohexanol degradation, was also determined. The gene arrangement is chnB-chnE-chnR. The predicted molecular masses of the three polypeptides were verified by radiolabeling by using the T7 expression system. Inducible expression of cloned chnB in Escherichia coli depended upon the presence of chnR. A transcriptional chnB::lacZ fusion experiment revealed that cyclohexanone induces chnB expression in E. coli, in which a 22-fold increase in activity was observed.