Organization of aerobactin, hemolysin, and antibacterial resistance genes in lactose negative Escherichia coli strains of serotype O4 isolated from children with dhiarrea

Epidemiologically related, non-lactose-fermenting (NLF) Escherichia coli strains of serotype O4 have been isolated at a high frequency from children with diarrhea in Somalia (M. Nicoletti, F. Superti, C. Conti, A. Calconi, and C. Zagaglia, J. Clin. Microbiol. 26:524-529, 1988). In order to define the virulence potential of these strains, we characterized the replication properties of their high-molecular-weight plasmids and studied the genetic locations and organization of the aerobactin (aer) and hemolysin (hly) determinants encoded by 23 NLF O4 E. coli strains. Southern blot hybridizations, mobilization assays of nonconjugative plasmids, and incompatibility-exclusion experiments conducted with a conjugative incompatibility group FI (IncFI) plasmid showed that (i) 20 out of the 23 strains examined harbor a 160- to 180-kb IncFI plasmid that shares homology with the basic replicons RepFIA, RepFIB, and (except for the plasmid of one strain) RepFIC, and 22 strains also contain a 40- to 140-kb IncFII plasmid sharing homology with the RepFIIA replicon; (ii) the IncFI plasmid is nonconjugative and carries antibiotic resistance genes; (iii) the aer system is located on the IncFI plasmids and/or the chromosomes in the three strains not harboring IncFI, and it is found in an inverted orientation; (iv) the hly determinants are located on the chromosome, and their genetic organization is well conserved and closely resembles that of the reference hemolytic plasmid pHly152; and (v) Hly- mutants obtained by transposon insertion mutagenesis are not cytotoxic to HeLa cell monolayers, indicating that hemolysin is responsible for the high cytotoxic activity we have previously reported for these strains. The structural organization of the plasmid-encoded aer operon, together with the finding that those plasmids also carry antibiotic resistance genes, indicates that the IncFI plasmid of the NLF O4 E. coli strains studied more closely resembles aer-encoding virulence IncFI Salmonella R plasmids than E. coli ColV plasmids. The data presented here cannot rule out whether the strains examined are potentially intestinal or extraintestinal pathogens. Nevertheless, the genetic organization of the virulence genes, together with the epidemiological behavior and the wide spectrum of antibiotic resistance of the NLF O4 E. coli strains, indicates that these strains are structured as typical E. coli pathogenic isolates of human origin

Preface: Introduction to special issue

Peer ReviewedPostprint (author’s final draft

A Service Station Location Model to Explore Prospects and Policies for Alternative Transport Fuels: A Case of CNG Distribution in Italy

CNG is an example of alternative gaseous fuel whose market development requires supply infrastructure (pipelines), refuelling stations and alternative vehicles to exist at the same time, which is known as the \u2018\u2018chicken and egg dilemma\u2019\u2019. In this chapter, a case study of limited or locally nonexistent market development for CNG in an Italian frontier region is analyzed and a mixed integer non linear programming model is introduced to evaluate the effect of incentive measures envisaged by the regional government to foster refuelling station development. It is found that, taking an entrepreneurs\u2019 perspective of maximizing profits, even with substantial capital grants investors are more likely to choose higher demand areas, in spite of fiercer competition, rather than areas without stations. Subsidies should be more specifically targeted to critical areas to be efficient

Mapping knowledge management research: a bibliometric overview

In recent years, knowledge management (KM) has consistently attained considerably growing research attention. Consequently, several literature reviews have been performed addressing different topic areas of KM. This paper seeks to present a comprehensive bibliometric and network analysis on KM to understand its development from the perspective of academic communities. Subsequently, it seeks to identify the structure of associations between prior and current themes, predict emerging trends and offer a longitudinal perspective on KM research. This study used web of science database and the initial sample was trimmed down by considering only the articles contributing to KM literature, and further 8,721 KM papers published in the last 30 years were systematically evaluated. The descriptive statistics and science mapping methods employing co-citation analysis were performed with VOSviewer software. In the descriptive analysis, we have analysed publication trends over time, geographical localization of the contributing institutions, journals, most prolific authors, top-performing institutions and most cited articles. Science mapping analysis is based on co-word analysis and co-citations analysis, namely articles’ co-citations and authors’ co-citations. The main findings of this paper will help researchers and academicians to develop knowledge in a specific sub-field by analysing the research outcomes of the papers included in the body of literature. First published online 28 December 202

Scanning Electron Microscopy, X-Ray Microanalysis and Immunohistochemistry on Worn Soft Contact Lenses

The deposits accumulated on the surfaces of soft contact lenses are a cause of problems for the wearer of these lenses, as the deposits are never completely removed by the available washing solutions. Therefore it appears of interest to investigate the composition of these deposits. In this paper we review the major findings in the literature and, in addition, present our personal experience. We have studied new, continuously and daily worn soft contact lenses by scanning electron microscopy (SEM), X-ray microanalysis and immunohistochemistry. We have carefully evaluated preparative methods, and we can conclude that SEM and X-ray microanalysis are best carried out on unfixed, air-dried lenses. The deposits present consist mainly of mucus, especially on the tarsal side of the lenses. Chloride and potassium, coming from the tear fluid, as well as sulfur, derived from proteins, were found. Calcium was very rarely detected. IgG, IgA, IgE and C3c complement fractions were found only on the outer surfaces and not within the lens. We believe that the best characterization of the deposits is achieved by means of correlative techniques on the same lens. In fact, this approach integrates morphology and composition

Coupled Natural Fusion Enzymes in a Novel Biocatalytic Cascade Convert Fatty Acids to Amines

[Image: see text] Tambjamine YP1 is a pyrrole-containing natural product. Analysis of the enzymes encoded in the Pseudoalteromonas tunicata “tam” biosynthetic gene cluster (BGC) identified a unique di-domain biocatalyst (PtTamH). Sequence and bioinformatic analysis predicts that PtTamH comprises an N-terminal, pyridoxal 5′-phosphate (PLP)-dependent transaminase (TA) domain fused to a NADH-dependent C-terminal thioester reductase (TR) domain. Spectroscopic and chemical analysis revealed that the TA domain binds PLP, utilizes l-Glu as an amine donor, accepts a range of fatty aldehydes (C(7)–C(14) with a preference for C(12)), and produces the corresponding amines. The previously characterized PtTamA from the “tam” BGC is an ATP-dependent, di-domain enzyme comprising a class I adenylation domain fused to an acyl carrier protein (ACP). Since recombinant PtTamA catalyzes the activation and thioesterification of C(12) acid to the holo-ACP domain, we hypothesized that C(12) ACP is the natural substrate for PtTamH. PtTamA and PtTamH were successfully coupled together in a biocatalytic cascade that converts fatty acids (FAs) to amines in one pot. Moreover, a structural model of PtTamH provides insights into how the TA and TR domains are organized. This work not only characterizes the formation of the tambjamine YP1 tail but also suggests that PtTamA and PtTamH could be useful biocatalysts for FA to amine functional group conversion

Evolution in prostheses for sprinters with lower-limb amputation

Depuis une quinzaine d'années, les progrès techniques en appareillage ont été le facteur déterminant de la progression des performances des sportifs amputés de membre inférieur. Pour l'amputé tibial, la prothèse de course comprend un manchon gel et une emboîture solidarisés par un accrochage distal ou un vide virtuel. Par ses qualités dynamiques, le pied en fibre de carbone, garni de pointes, assure des performances remarquables. Pour l'amputé fémoral, équipé des mêmes pieds prothétiques, le genou est à biellettes et à contrôle des phases d'appui et pendulaire. Par rapport au coureur valide, le temps d'appui sur le membre appareillé est raccourci tandis que celui sur le membre sain est allongé. L'asymétrie du sprint de l'amputé tibial est discrète. C'est le travail des extenseurs de hanche qui est la principale compensation au déficit de propulsion dû à l'amputation. Chez l'amputé fémoral, l'absence de genou aggrave l'asymétrie. L'extension totale du genou prothétique, précoce en fin de phase oscillante et persistant pendant toute la phase d'appui, impose une compensation par une augmentation d'extension de la hanche controlatérale. Les transferts de charge de travail entre côté amputé et sain, par l'intermédiaire d'une hyperlordose lombaire, mettent en jeu le bassin, le tronc et les épaules. Les amputés sportifs font progresser la recherche en appareillage. Leurs orthoprothésistes acquièrent avec eux un savoir-faire dont bénéficient leurs patients non-sportifs.For about 15 years, technical advances in prosthetic treatment have been the main factor in the increased performance of athletes with lower-limb amputation. For trans-tibial amputation, the prosthesis for sprinting is composed of a gel liner and a socket joined by a locking or virtual vacuum liner. Because of these dynamic properties, the carbon prosthetic foot equipped with tacks ensures outstanding performance. For trans-femoral amputation, a hydraulic swing and a stance control unit are added to the same prosthesis. In comparison with the able-bodied runner, athletes with amputation have smaller loading times in the prosthetic limb and larger ones in the sound limb. The length of the energy-storing prosthetic foot is determined by the “up-on-the-toes” running gait. The sprinting gait with trans-tibial amputation is almost symmetrical. The hip extensor effort is the main compensation of propulsion reduction with lower-limb amputation. With trans-femoral amputation, the lack of knee increases the asymmetry. The total prosthetic knee extension (early in late-swing phase and lasting during total stance phase) compensates with extension of both hips, especially the opposite one. The amputation and sound limb load transfer with lumbar hyperlordosis concern the pelvis, trunk and shoulders. Because of athletes with amputation, research in prosthetic treatment has progressed. The development of orthotics and prostheses for such athletes has benefited non-athletes with amputation