Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

The cell envelope of mycobacteria is a complex multilaminar structure that protects the cell from stresses encountered in the environment, and plays an important role against the bactericidal activity of immune system cells. The outermost layer of the mycobacterial envelope typically contains species-specific glycolipids. Depending on the mycobacterial species, the major glycolipid localized at the surface can be either a phenolglycolipid or a peptidoglycolipid (GPL). Currently, the mechanism of how these glycolipids are addressed to the cell surface is not understood. In this study, by using a transposon library of Mycobacterium smegmatis and a simple dye assay, six genes involved in GPLs synthesis have been characterized. All of these genes are clustered in a single genomic region of approximately 60 kb. We show by biochemical analyses that two non-ribosomal peptide synthetases, a polyketide synthase, a methyltransferase and a member of the MmpL family are required for the biosynthesis of the GPLs backbone. Furthermore, we demonstrate that a small integral membrane protein of 272 amino acids named Gap (gap: GPL addressing protein) is specifically required for the transport of the GPLs to the cell surface. This protein is predicted to contain six transmembrane segments and possesses homologues across the mycobacterial genus, thus delineating a new protein family. This Gap family represents a new paradigm for the transport of small molecules across the mycobacterial envelope, a critical determinant of mycobacterial virulence

Assessing the role of EO in biodiversity monitoring: options for integrating in-situ observations with EO within the context of the EBONE concept

The European Biodiversity Observation Network (EBONE) is a European contribution on terrestrial monitoring to GEO BON, the Group on Earth Observations Biodiversity Observation Network. EBONE’s aims are to develop a system of biodiversity observation at regional, national and European levels by assessing existing approaches in terms of their validity and applicability starting in Europe, then expanding to regions in Africa. The objective of EBONE is to deliver: 1. A sound scientific basis for the production of statistical estimates of stock and change of key indicators; 2. The development of a system for estimating past changes and forecasting and testing policy options and management strategies for threatened ecosystems and species; 3. A proposal for a cost-effective biodiversity monitoring system. There is a consensus that Earth Observation (EO) has a role to play in monitoring biodiversity. With its capacity to observe detailed spatial patterns and variability across large areas at regular intervals, our instinct suggests that EO could deliver the type of spatial and temporal coverage that is beyond reach with in-situ efforts. Furthermore, when considering the emerging networks of in-situ observations, the prospect of enhancing the quality of the information whilst reducing cost through integration is compelling. This report gives a realistic assessment of the role of EO in biodiversity monitoring and the options for integrating in-situ observations with EO within the context of the EBONE concept (cfr. EBONE-ID1.4). The assessment is mainly based on a set of targeted pilot studies. Building on this assessment, the report then presents a series of recommendations on the best options for using EO in an effective, consistent and sustainable biodiversity monitoring scheme. The issues that we faced were many: 1. Integration can be interpreted in different ways. One possible interpretation is: the combined use of independent data sets to deliver a different but improved data set; another is: the use of one data set to complement another dataset. 2. The targeted improvement will vary with stakeholder group: some will seek for more efficiency, others for more reliable estimates (accuracy and/or precision); others for more detail in space and/or time or more of everything. 3. Integration requires a link between the datasets (EO and in-situ). The strength of the link between reflected electromagnetic radiation and the habitats and their biodiversity observed in-situ is function of many variables, for example: the spatial scale of the observations; timing of the observations; the adopted nomenclature for classification; the complexity of the landscape in terms of composition, spatial structure and the physical environment; the habitat and land cover types under consideration. 4. The type of the EO data available varies (function of e.g. budget, size and location of region, cloudiness, national and/or international investment in airborne campaigns or space technology) which determines its capability to deliver the required output. EO and in-situ could be combined in different ways, depending on the type of integration we wanted to achieve and the targeted improvement. We aimed for an improvement in accuracy (i.e. the reduction in error of our indicator estimate calculated for an environmental zone). Furthermore, EO would also provide the spatial patterns for correlated in-situ data. EBONE in its initial development, focused on three main indicators covering: (i) the extent and change of habitats of European interest in the context of a general habitat assessment; (ii) abundance and distribution of selected species (birds, butterflies and plants); and (iii) fragmentation of natural and semi-natural areas. For habitat extent, we decided that it did not matter how in-situ was integrated with EO as long as we could demonstrate that acceptable accuracies could be achieved and the precision could consistently be improved. The nomenclature used to map habitats in-situ was the General Habitat Classification. We considered the following options where the EO and in-situ play different roles: using in-situ samples to re-calibrate a habitat map independently derived from EO; improving the accuracy of in-situ sampled habitat statistics, by post-stratification with correlated EO data; and using in-situ samples to train the classification of EO data into habitat types where the EO data delivers full coverage or a larger number of samples. For some of the above cases we also considered the impact that the sampling strategy employed to deliver the samples would have on the accuracy and precision achieved. Restricted access to European wide species data prevented work on the indicator ‘abundance and distribution of species’. With respect to the indicator ‘fragmentation’, we investigated ways of delivering EO derived measures of habitat patterns that are meaningful to sampled in-situ observations

Efficacy of Anakinra for Various Types of Crystal-Induced Arthritis in Complex Hospitalized Patients: A Case Series and Review of the Literature

International audienceBackground. There are few data on anakinra use after failure of conventional medications for crystal-induced peripheral arthritis and/or crowned dens syndrome among complex hospitalized patients. Methods. We retrospectively analyzed the outcome of six patients affected with subacute crystal-induced arthritis who had received anakinra in second or third line therapy, including three patients with crowned dens syndrome and three others with gouty arthritis. Patients' comorbidities, reasons for anakinra use and associated drugs, and outcomes were recorded. Results. All patients presented with elevated inflammatory syndrome, systemic symptoms with poly/oligoarthritis. Except for absolute contraindications, all patients were previously treated with full or decreased dose of NSAID, colchicine, and/or glucocorticoids, with unsatisfactory response. All three gouty patients exhibited complete responses in all acute involvements under anakinra within 3 to 5 days, including one of them who needed the reintroduction of colchicine treatment that was previously unsuccessful. Crowned dens syndrome patients, including two with pseudogout and one with subacute hydroxyapatite deposition disease, needed 9 to 11 days to achieve complete response. Tolerance to anakinra was good. Conclusion. In case series of complex hospitalized patients, anakinra showed good activity in crowned dens syndrome and associated crystal-induced peripheral arthritis, with longer treatment duration than in gouty arthritis

Tomato protoplast DNA transformation: physical linkage and recombination of exogenous DNA sequences

Tomato protoplasts have been transformed with plasmid DNA's, containing a chimeric kanamycin resistance gene and putative tomato origins of replication. A calcium phosphate-DNA mediated transformation procedure was employed in combination with either polyethylene glycol or polyvinyl alcohol. There were no indications that the tomato DNA inserts conferred autonomous replication on the plasmids. Instead, Southern blot hybridization analysis of seven kanamycin resistant calli revealed the presence of at least one kanamycin resistance locus per transformant integrated in the tomato nuclear DNA. Generally one to three truncated plasmid copies were found integrated into the tomato nuclear DNA, often physically linked to each other. For one transformant we have been able to use the bacterial ampicillin resistance marker of the vector plasmid pUC9 to 'rescue' a recombinant plasmid from the tomato genome. Analysis of the foreign sequences included in the rescued plasmid showed that integration had occurred in a non-repetitive DNA region. Calf-thymus DNA, used as a carrier in transformation procedure, was found to be covalently linked to plasmid DNA sequences in the genomic DNA of one transformant. A model is presented describing the fate of exogenously added DNA during the transformation of a plant cell. The results are discussed in reference to the possibility of isolating DNA sequences responsible for autonomous replication in tomato.

Robust Optimization for Smart Machining Systems: An Enabler for Agile Manufacturing

ABSTRACT This paper reports our efforts towards developing a mathematical and information framework for optimization of machining processes within a Smart Machining System (SMS). An SMS uses diverse integrated technologies that enable an enterprise to: (1) produce the first and every product correct; (2) improve the response of the production system to changes in demand (just in time); (3) realize rapid and agile manufacturing; and (4) provide data to the rest of the enterprise as needed. Optimization of machining processes is an important component of an SMS and contributes to realizing these capabilities. Based on a prototype, we demonstrate the concepts for robust optimization within an SMS and develop requirements and challenges for robust optimization in an SMS

The development of a New Process for Phosphate Thickening

In phosphate beneficiation process, thickening is used to concentrate the slurry or sludge in order to increase its solid content and recover the maximum of water. The addition of flocculants helps to form larger particles that can thicken out quickly. However, the high consumption of flocculant in the thickeners increases the cost of the operation. The purpose of this research is to study the effect of flocculant on sedimentation velocities of different particles size in order to develop a new thickening process which ensure the maximum thickening rate and a minimum flocculant consumption.To achieve this goal, initially the impact of flocculant on different size fraction was studied. The objective of this step is to define the cut-size from which the flocculant has no considerable effect on thickening. This granulometric slice can decant by a simple free sedimentation without needing the flocculant. After that, a hydrocyclone was dimensioned and modeled in order to eliminate this granulometric slice which will undergo free sedimentation. This allowed to design a new thickening technology that targets only fine particles overflow of the hydro-cyclone and that require the addition of flocculant. This technology will significantly reduce the flocculant consumption and ensure a better water recovery in the thickening process

Is M. ulcerans able to colonize neuronal cells?

Buruli ulcer, or Mycobacterium ulcerans infection, is an emerging disease, principally diagnosed in humid tropical countries and inducing large skin ulcers. These lesions are painless, a distinct feature that suggests that the mycolactone toxin and/or M. ulcerans impedes the signal transmission by the nervous system. In this context, the aim of this work was to study the interaction between M. ulcerans and neuronal cells by using in vitro and in vivo models. We showed that a virulent strain of M. ulcerans is able to enter into neurons cultivated from neonatal rat hippocampus. On the contrary, this phenomenon was less observed with a mycolactone-deficient strain. To support these data, we analysed nerve fibres from mouse-infected tissues and few bacilli were found in close contact with nerve fibres. The invasion process established by M. ulcerans to colonize the nervous system remains uncharacterised, but we hypothesise that this ability could be involved in the painless of the M. ulcerans infection

Efficacy of Continuous Interleukin 1 Blockade in Mevalonate Kinase Deficiency: A Multicenter Retrospective Study in 13 Adult Patients and Literature Review

OBJECTIVE: To report efficacy and tolerance of interleukin 1 blockade in adult patients with mevalonate kinase deficiency (MKD). METHODS: We retrospectively collected data on 13 patients with MKD who had received anakinra (n = 10) and canakinumab (n = 7). RESULTS: Anakinra resulted in complete or partial remission in 3/10 and 5/10 patients, respectively, and no efficacy in 2/10, but a switch to canakinumab led to partial remission. Canakinumab resulted in complete or partial remission in 3/7 and 4/7 patients, respectively. CONCLUSION: These data support frequent partial responses, showing a better response with canakinumab. The genotype and therapeutic outcomes correlation should help in the personalization of treatment

Knowledge representation for product and processes development planning in collaborative environments

Efficiency in the management of integrated product and processes development is a basic requisite to guarantee competitiveness and success for manufacturing companies. This means that operational management of activities and human and material resources is extremely important, especially in virtual OKP (one-of-a-kind production) systems, and must cover related aspects of their capabilities and social character as well as assignment criteria. In this context, and to facilitate collaborative resources management, an ontology focused on resources and capabilities is proposed in this work. This ontology supports the necessary knowledge for generic and collaborative process planning, providing a shared common semantic for all the members of the virtual company. This work differs from other proposed ontologies in the area of process planning where the resources considered are all those elements that participate in the execution of the different activity types involved in this wide and complex process. The ontology directly covers the shared, social nature of the resources, the agentive behaviour of many of them and a characterisation of their capabilities, thus providing specific solutions to the needs of the collaborative integrated development of products, processes and resources (CIDP2R) process.This work has been possible thanks to funding received from the Ministry of Science and Education through the COAPP Research Project - reference DPI2007-66871-C02-01/02.Solano García, L.; Rosado Castellano, P.; Romero Subirón, F. (2014). Knowledge representation for product and processes development planning in collaborative environments. International Journal of Computer Integrated Manufacturing. 27(8):787-801. doi:10.1080/0951192X.2013.834480S78780127