The new complexity: new dynamics in clusters and districts

Recently, service clusters have been identified, such as in the case of the logistics and transportation cluster in the Basque Country (e.g. transportation of people and ware, intelligent transport services, etc.) or that of port activities (warehousing services, loading-unloading of goods, shipping service, etc.). Simultaneously, several multi-sector clusters around a specific client are rising. It is the case of the child cluster in France or in Catalunya that includes much more than traditional clusters focused on a final homogeneous product; in fact this includes the production of toys, kids clothes, health products and furniture for children, among others, which belonged to separate sectors and clusters before. In addition to identifying new types of clusters that strengthen the visibility and work of their firms , the great majority of these clusters and districts, e.g. in the Basque Country, have become more ‘complex’ in depth (division and specialization of labor) and extension to a variety of activities that were quite separated from one another, though can now come together with the objective to respond to the new demand of local and global society. For instance, the demand for new renewable sources of energy promotes a change in sectoral/entrepreneurial attitudes, for which such diversified sectors as oil refinery, wind, maritime and solar energy, find representation in the same cluster association and recognize themselves as part of one 'complex' cluster that gains in visibility and scale of operations for global agents such as large industries, large scale construction projects, among others. The identification and the activity of such clusters and districts, sometimes associated in a specific locality, in other occasions to a wider territory, exhibit the growing awareness of the strengths and diversities of the territorial production fabric and the importance businesses and agents attribute to identifying themselves as such in order to develop joint initiatives that can generate important economies of scale and scope that help competing in the new globalized economy, particularly when these local production systems strive to open their way in global production networks and in international markets.

Biofilm as an adaptation strategy to extreme conditions

Biofilm is the most successful and widely distributed form of life on earth, it is not simply structured collections of cells attached to surfaces but is a dynamic complex biological system able to respond to environmental changes. The biofilm characteristics make it unique and central to microbial evolution and adaptation. The ability to establish biofilms is a key trait for microorganisms growing in extreme environments like extreme temperature, high radiation, acidic or alkaline pH values, heavy metal pollution, and high salinity. In this article, we report the main features of biofilm and how these characteristics make biofilms a successful survival strategy in extreme conditions. All aspects examined in this article help to explain why biofilms are a successful survival strategy in extreme conditions and why the ability to establish biofilms is a key trait for microorganisms growing in extreme environments

E-Commerce and Transfer Pricing - Some Selected Issues

The aim of this article is to draft a concise and clear picture concerning the applicability of transfer pricing regime in a typical e-commerce scenario. Provided twolegal and business models (intra-company transaction between head office and branch, and inter-companies transaction between parent company and subsidiary,or between two subsidiaries of the same group), we will assess whether, and to what extent, a website and a server, used by the enterprise or the group to carry on its e-business, can be conceived as a permanent establishment and, then, we will analysewhich methods have to be used in order to apply the arm’s length principle. Our research will predominantly take into account the OECD Model Convention andTransfer Pricing Guidelines

Active human full-length CDKL5 produced in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125

Background: A significant fraction of the human proteome is still inaccessible to in vitro studies since the recombinant production of several proteins failed in conventional cell factories. Eukaryotic protein kinases are difficult-to-express in heterologous hosts due to folding issues both related to their catalytic and regulatory domains. Human CDKL5 belongs to this category. It is a serine/threonine protein kinase whose mutations are involved in CDKL5 Deficiency Disorder (CDD), a severe neurodevelopmental pathology still lacking a therapeutic intervention. The lack of successful CDKL5 manufacture hampered the exploitation of the otherwise highly promising enzyme replacement therapy. As almost two-thirds of the enzyme sequence is predicted to be intrinsically disordered, the recombinant product is either subjected to a massive proteolytic attack by host-encoded proteases or tends to form aggregates. Therefore, the use of an unconventional expression system can constitute a valid alternative to solve these issues. Results: Using a multiparametric approach we managed to optimize the transcription of the CDKL5 gene and the synthesis of the recombinant protein in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 applying a bicistronic expression strategy, whose generalization for recombinant expression in the cold has been here confirmed with the use of a fluorescent reporter. The recombinant protein largely accumulated as a full-length product in the soluble cell lysate. We also demonstrated for the first time that full-length CDKL5 produced in Antarctic bacteria is catalytically active by using two independent assays, making feasible its recovery in native conditions from bacterial lysates as an active product, a result unmet in other bacteria so far. Finally, the setup of an in cellulo kinase assay allowed us to measure the impact of several CDD missense mutations on the kinase activity, providing new information towards a better understanding of CDD pathophysiology. Conclusions: Collectively, our data indicate that P. haloplanktis TAC125 can be a valuable platform for both the preparation of soluble active human CDKL5 and the study of structural–functional relationships in wild type and mutant CDKL5 forms. Furthermore, this paper further confirms the more general potentialities of exploitation of Antarctic bacteria to produce “intractable” proteins, especially those containing large intrinsically disordered regions

Case Report: Anterior Cruciate Ligament Calcification in a Patient With Chondrocalcinosis: Micro-Computed Tomography Presentation

In this case report, an incidental postoperative diagnosis of anterior cruciate ligament (ACL) calcification, associated with calcification of posterior cruciate ligament (PCL) and lateral meniscus insertions, was made using micro-computed tomography (μCT) technology in a knee specimen obtained during a total knee replacement (TKR) surgery due to painful tri-compartmental osteoarthritis (OA) with chondrocalcinosis signs at preoperative X-ray. Anterior cruciate ligament calcification is an uncommon finding, and conventional X-ray and MRI are not so helpful in its identification. μCT scan, in contrast, is of interest because it provides highly spatial three-dimensional information with excellent visualization of bones and calcifications. The μCT technology used in this case report allowed us to perform a detailed analysis and a 3-D reconstruction of the calcium pyrophosphate dihydrate (CPPD) crystal deposition about the knee without the need to section the specimens into slice as performed in previous studies. The 3-D model obtained with μCT scan permits to gain more insight into the shape of the calcification within the fibers of the ligamentous structures of the joint

Development of high-copy number plasmids in Pseudoalteromonas haloplanktis TAC125

Abstract: The Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 (PhTAC125) is considered an interesting alternative host for the recombinant protein production, that can be explored when the conventional bacterial expression systems fail. Indeed, the manufacture of all the difficult-to-express proteins produced so far in this bacterial platform gave back soluble and active products. Despite these promising results, the low yield of recombinant protein production achieved is hampering the wider and industrial exploitation of this psychrophilic cell factory. All the expression plasmids developed so far in PhTAC125 are based on the origin of replication of the endogenous pMtBL plasmid and are maintained at a very low copy number. In this work, we set up an experimental strategy to select mutated OriR sequences endowed with the ability to establish recombinant plasmids at higher multiplicity per cell. The solution to this major production bottleneck was achieved by the construction of a library of psychrophilic vectors, each containing a randomly mutated version of pMtBL OriR, and its screening by fluorescence-activated cell sorting (FACS). The selected clones allowed the identification of mutated OriR sequences effective in enhancing the plasmid copy number of approximately two orders of magnitude, and the production of the recombinant green fluorescent protein was increased up to twenty times approximately. Moreover, the molecular characterization of the different mutant OriR sequences allowed us to suggest some preliminary clues on the pMtBL replication mechanism that deserve to be further investigated in the future. Key points: • Setup of an electroporation procedure for Pseudoalteromonas haloplanktis TAC125. • Two order of magnitude improvement of OriR-derived psychrophilic expression systems. • Almost twenty times enhancement in Green fluorescent protein production

Structural characterization of an all-aminosugar-containing capsular polysaccharide from Colwellia psychrerythraea 34H

Colwellia psychrerythraea strain 34H, a Gram-negative bacterium isolated from Arctic marine sediments, is considered a model to study the adaptation to cold environments. Recently, we demonstrated that C. psychrerythraea 34H produces two different extracellular polysaccharides, a capsular polysaccharide and a medium released polysaccharide, which confer cryoprotection to the bacterium. In this study, we report the structure of an additional capsular polysaccharide produced by Colwellia grown at a different temperature. The structure was determined using chemical methods, and one- and two-dimensional NMR spectroscopy. The results showed a trisaccharide repeating unit made up of only amino-sugar residues: N-acetyl-galactosamine, 2,4-diacetamido-2,4,6-trideoxy-glucose (bacillosamine), and 2-acetamido-2-deoxyglucuronic acid with the following structure: →4)-β-d-GlcpNAcA-(1 →3)-β-d-QuipNAc4NAc-(1 →3)-β-d-GalpNAc-(1 →. The 3D model, generated in accordance with 1H,1H-NOE NMR correlations and consisting of ten repeating units, shows a helical structure. In contrast with the other extracellular polysaccharides produced from Colwellia at 4 °C, this molecule displays only a low ice recrystallization inhibition activity

Business innovation modes and their impact on innovation outputs: Regional variations and the nature of innovation across EU regions

This work contributes to the literature on innovation systems and, in particular, delivers a thorough analysis on business innovation modes across a range of regional contexts. This analysis refers to the strand of literature on STI (Science and Technology-based Innovation) and DUI innovation modes (Innovation based on learning-by-Doing, learning-by-Using, learning-by-Interacting) that have been intensely debated over the past few years. It is a relevant area of research because it discusses the most effective innovation mode adopted by firms and their regions in the context of increasing global competition. In this scientific area, we inquire whether and how the regional context and its specific technological capabilities produce a differentiated impact of STI and DUI innovation modes on innovation outputs, alongside the nature of innovation outputs. In this respect, this study advances the literature on regional innovation systems that have not been analyzed by other scholarly contributions in this strand who have mostly discussed the differentiated impact of innovation modes across individual countries, industries, and business networks. Based on the large heterogeneity of regions across the European geography, we move beyond the set of individual country studies and develop a thorough analysis based on the Community Innovation Survey (CIS 2014) data from the Eurostat office about EU regions. Empirical evidence based on the application of a multiple treatment model suggests that both regional specificities and the nature of innovation matter. In addition, the DUI innovation mode proves to be often more important than expected for most types of innovation output