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A qualification methodology for advanced rotary shouldered threaded connections is presented. It covers physical tests of connection prototypes and virtual tests with modeling and simulation techniques. The methodology had been applied to qualify a fatigue-resistant threaded connection design that has recently been released for field testing. The qualification process consists of in-lab makeup and breakout tests, on-rig makeup and breakout tests, sealability tests, fatigue tests, torsional yield limit tests, and tensile capacity tests. During the development of the design, modeling and simulation techniques were extensively used to optimize the design prior to physical prototyping and testing. Some of the qualification tests, such as torsional yield limit tests and tensile capacity tests, were carried out by using advanced modeling and simulation techniques. Because fatigue tests under nominal loads take a long time to complete, accelerated fatigue tests under calculated overloads were conducted to assess whether the design would meet the fatigue life requirements with low risk. After positive results were obtained from the accelerated fatigue tests, nominal loads required by the product specifications were then applied to complete the qualification fatigue tests. It was found from the experimental results that the predicted performance of the connections, such as fatigue life, critical sealing pressure, and breakout torque, matched with the physical test results well. Despite the fact that numerous rotary shouldered threaded connection designs have been developed in the oil and gas industry, there has been no formally defined qualification procedure for such designs. The objective of this paper is to introduce, for the first time, a systematic and complete qualification procedure for rotary shouldered threaded connections.

Free standing films of a genetically engineered silk-elastin-like protein (SELP) were prepared using water and formic acid as solvents. Exposure to methanol-saturated air promoted the formation of aggregated β-strands rendering aqueous insolubility and improved the mechanical properties leading to a 10-fold increase in strain-to-failure. The films were optically clear with resistivity values similar to natural rubber and thermally stable up to 180 °C. Addition of glycerol showed to enhance the flexibility of SELP/glycerol films by interacting with SELP molecules through hydrogen bonding, interpenetrating between the polymer chains and granting more conformational freedom. This detailed characterization provides cues for future and unique applications using SELP based biopolymers.

N/A The origins of the modern football codes have attracted considerable attention from historians of sport over the past two decades, resulting in a vigorous debate between the self-described ‘revisionists’, led by Adrian Harvey, and the followers of Eric Dunning, dubbed by their opponents as the ‘orthodox’ view. However, this article argues that both sides commit the same methodological errors: an overestimation of the importance of written rules, an ahistoric view of culture and continuity, and a tendency to view the past through the lens of the present. By re-examining the historical record of early forms of football and presenting a broader contextual perspective for the emergence of the football codes in the 1840 to 1880 period, the article aims to address some of the key historiographical issues that confront historians of sport today.

Cold-active enzymes are produced by organisms, known as psychrophiles, adapted to permanently cold habitats. Low temperatures have an exponential deleterious effct on reaction rates, and thus psychrophilic enzymes have to be adapted to secure appropriate reaction rates in their environment. These enzymes have a high specific activity at low temperatures, in any case higher than that of their mesophilic and thermophilic counterparts, and display a shift of the apparent optimum temperature for activity towards low temperatures as well as a reduced thermal stability and increased flexibility. The increased flexibility may be global, involving the overall edifice, or local, involving only those zones crucial for activity, be they near or distant from the active site. The reduced thermodynamic stability of cold-adapted enzymes is illustrated by a significantly lower stabilisation energy as compared to that of their mesophilic and thermophilic counterparts, yet maximum stability occurs at similar temperatures in all cases. The comparison of their three-dimensional structures with higher temperature-adapted homologues, in conjunction with various mutagenesis studies, has shown that their high activity results from rather discrete molecular changes that tend to decrease the stability of the molecular edifice. Each cold-adapted enzyme however adopts a specific strategy. There is apparently a continuum in the adaptation, with some enzymes showing extremely acute cold adaptation, as illustrated by a severe shift of the activity towards low temperatures, whereas others appear to cover a broader range of temperatures. This probably depends on the specific evolutionary history of the organisms which produce them., Tony Collins is supported by the FCT, the European Social Fund, the Programa Operacional Potencial Humano and the Investigador FCT Programme (IF/01635/ 2014). The FCT is thanked for their funding through EXPL/BBB-BIO/1772/2013-FCOMP-01- 0124-FEDER-041595, the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER- 007569), and the ERDF through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI). All the technical staff at the CBMA is thanked for the skillful technical assistance., info:eu-repo/semantics/publishedVersion

Background: Applying very high gravity (VHG) fermentation conditions to the sugarcane juice (SCJ) bioethanol industry would improve its environmental and economic sustainability without the need for major infrastructure changes or investments. It could enable a decrease in the consumption of biological and natural resources (cane/ land, water and energy) while maintaining acceptable production parameters. The present study attempts to demonstrate and characterise an efective industrially relevant SCJ-VHG fermentation process. Results: An industry-like SCJ-VHG bioethanol production process with 30 and 35 °Bx broth was employed to investigate the efects of both the yeast strain used and nitrogen source supplementation on process yield, process productivity, biomass viability, glycerol concentration and retention-associated gene expression. Process performance was shown to be variably afected by the diferent process conditions investigated. Highest process efciency, with a 17% (w/v) ethanol yield and only 0.2% (w/v) sugar remaining unfermented, was observed with the Saccharomyces cerevisiae industrial strain CAT-1 in 30 °Bx broth with urea supplementation. In addition, efcient retention of glycerol by the yeast strain was identifed as a requisite for better fermentation and was consistent with a higher expression of glycerol permease STL1 and channel FPS1. Urea was shown to promote the deregulation of STL1 expression, overcoming glucose repression. The consistency between Fps1-mediated ethanol secretion and ethanol in the extracellular media reinforces previous suggestions that ethanol might exit the cell through the Fps1 channel. Conclusions: This work brings solid evidence in favour of the utilisation of VHG conditions in SCJ fermentations, bringing it a step closer to industrial application. SCJ concentrated up to 30 °Bx maintains industrially relevant ethanol production yield and productivity, provided the broth is supplemented with a suitable nitrogen source and an appropriate industrial bioethanol-producing yeast strain is used. In addition, the work contributes to a better understanding of the VHG-SCJ process and the variable efects of process parameters on process efciency and yeast strain response. Keywords: Biofuel, Bioethanol, Sugarcane, Saccharomyces cerevisiae, CAT-1, Very high gravity, Process optimisation, Process sustainability, B. Monteiro was supported by the Ph.D. Grant 2011/12185-0 from the Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPESP. P. Ferraz and M. Barroca are supported by the Doctoral Programme in Applied and Environmental Microbiology (DP-AEM) and the FCT by Ph.D. Grants PD/ BD/113814/2015 and PD/BD/113810/2015, respectively. T. Collins thanks the FCT for support through the Investigador FCT Programme (IF/01635/2014). T. Collins and C. Lucas are supported by the strategic programme UID/ BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and the ERDF through COMPETE2020-Programa Operacional Competitividade e Internacionalização (POCI). P. Ferraz, T. Collins and C. Lucas were further funded by the project EcoAgriFood (NORTE-01- 0145-FEDER-000009), supported by the Norte Portugal Regional Operational Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement through the European Regional Development Fund (ERDF)., info:eu-repo/semantics/publishedVersion

Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. Here, custom-designed mutant primers oriented in the inverse direction are used to amplify the entire circular template with incorporation of the required mutation(s). By careful primer design it can be used to perform such diverse modifications as the introduction of point mutations and multiple mutations, the insertion of new sequences, and even sequence deletions. Three primer formats are commonly used; nonoverlapping, partially overlapping and fully overlapping primers, and here we describe the use of nonoverlapping primers for introduction of a point mutation. Use of such a primer setup in the PCR reaction, with one of the primers containing the desired mismatch mutation, results in the amplification of a linear, double-stranded, mutated product. Methylated template DNA is removed from the nonmethylated PCR product by DpnI digestion and the PCR product is then phosphorylated by polynucleotide kinase treatment before being recircularized by ligation, and transformed to E. coli. This relatively simple site-directed mutagenesis procedure is of major importance in biology and biotechnology today where it is commonly employed for the study and engineering of DNA, RNA, and proteins.

Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. Here, custom-designed mutant primers oriented in the inverse direction are used to amplify the entire circular template with incorporation of the required mutation(s). By careful primer design it can be used to perform such diverse modifications as the introduction of point mutations and multiple mutations, the insertion of new sequences, and even sequence deletions. Three primer formats are commonly used; nonoverlapping, partially overlapping and fully overlapping primers, and here we describe the use of nonoverlapping primers for introduction of a point mutation. Use of such a primer setup in the PCR reaction, with one of the primers containing the desired mismatch mutation, results in the amplification of a linear, double-stranded, mutated product. Methylated template DNA is removed from the nonmethylated PCR product by DpnI digestion and the PCR product is then phosphorylated by polynucleotide kinase treatment before being recircularized by ligation, and transformed to E. coli. This relatively simple site-directed mutagenesis procedure is of major importance in biology and biotechnology today where it is commonly employed for the study and engineering of DNA, RNA, and proteins., (undefined), info:eu-repo/semantics/publishedVersion

Today pipeline operators are required to balance operational efficiency, financial performance, and long-term sustainability. Transporting hydrocarbons efficiently and safely is a demanding task, and energy consumption is one of the major operating costs and sources of emissions. New applications based on pipeline simulation can contribute to reducing energy costs and optimizing operations while continuing to achieve the required delivery schedules within the safety and operating margins. This simulation-based approach considers the different energy costs across the pipeline pump/compressor stations, utility contracts, pump/compressor efficiency, and the costs and benefits of using drag-reducing agents (DRAs). It is a valuable tool that allows energy managers and pipeline operators to optimize energy costs. It can be seamlessly integrated into the supervisory control and data acquisition (SCADA) infrastructure, which provides the realtime views and reporting of all energy and process variables, including the cost savings achieved. The SCADA system monitors and controls one or more complete pipelines used for transporting liquid or gas products from one location to another. Each pipeline may be very long and may have several branches.

The first academic global history of the origins and development of rugby football

This article explores the origins and early history of American football in relation to the development of rugby and similar codes in the English-speaking world of the last third of the nineteenth century. It suggests that the traditional narrative description of the emergence of the American game – which is ascribed in large part to the individual initiative of Walter Camp – fails to situate the sport in the context of the wider, transnational dynamics of the development of the various handling codes of football. In particular, it contends that the common assumption that the gridiron game's early development was a sporting expression of American exceptionalism is mistaken and that it only acquired its distinctive national character in the early twentieth century.

Cold-adapted enzymes produced by organisms inhabiting permanently low temperature environments are typically characterized by a high activity at low to moderate temperatures and a poor thermal stability. Such characteristics make these enzymes highly attractive for various applications where they can enable more efficient, cost-effective, and environmentally friendlier processes than higher temperature-adapted enzymes. In this chapter, the biotechnological aspects of coldadapted enzymes and their application in industry are reviewed and discussed with a focus on cleaning/detergents, food and beverages, molecular biology, biomedicine, pharmaceuticals, cosmetics, textiles, biofuels, and materials applications., s T.C. is supported by the Fundac¸~ao para a Cieˆncia e a Tecnologia (FCT), the European Social Fund, the Programa Operacional Potencial Humano and the Investigador FCT Programme (IF/01635/2014). M.B. acknowledges the FCT for grant PD/BD/113810/2015 within the Doctoral Program in Applied and Environmental Microbiology. This work was funded by the European Regional Development Fund (ERDF) through project EcoAgriFood (NORTE-01-0145- FEDER-000009) via the North Portugal Regional Operational Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement. The FCT is thanked for their funding through EngXyl (EXPL/BBB-BIO/1772/2013-FCOMP-01-0124-FEDER-041595) and the strategic program UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569). All the technical staff at the CBMA are thanked for their skillful technical assistance., info:eu-repo/semantics/publishedVersion

Boundary Conditions for Microbial Life at Low Temperatures.- The Climate of Snow and Ice as Boundary Condition for Microbial Life.- Limits for Microbial Life at Subzero Temperatures.- Microbial Diversity in Cold Ecosystems.- Bacteria in Snow and Glacier Ice.- Bacteria in Subglacial Environments.- Bacteria in the Deep Sea: Psychropiezophiles.- Bacteria in Permafrost.- Anaerobic Bacteria and Archaea in Cold Ecosystems.- Cyanobacteria in Cold Ecosystems.- Fungi in Cold Ecosystems.- Cold-Active Viruses.- Molecular Adaptations to Cold Habitats.- Membrane Components and Cold Sensing.- Cold-Shock Proteins.- Fundamentals of Cold-Adapted Enzymes.- Cryoprotectants and Ice-Binding Proteins.- The Role of Exopolymers in Microbial Adaptation to Sea Ice.- Genomic Analysis of Psychrophilic Prokaryotes.- Microalgae in Polar Regions: Linking Functional Genomics and Physiology with Environmental Conditions.- Metagenomics: Microbial Community Genomes Revealed.- Proteomic Studies of Psychrophilic Microorganisms.- Biotechnological Aspects.- Biotechnological Aspects of Cold-Adapted Enzymes.- Heterologous Protein Expression in Psychrophilic Hosts.- Cold-Adapted Fungi as a Source for Valuable Metabolites.- Natural and Stimulated Biodegradation of Petroleum in Cold Marine Environments.- Microbial Adaptation to Boreal Saturated Subsurface: Implications in Bioremediation of Polychlorophenols.- Biological Iron Oxidation and Sulfate Reduction in the Treatment of Acid Mine Drainage at Low Temperatures.

The authors regret that Telma C. Bernardo was inadvertently omitted in the author line-up. The correct authorship order should be as follows: Ana Margarida Pereira, Raul Machado, André da Costa, Artur Ribeiro, Telma C. Bernardo, Tony Collins, Andreia C. Gomes, Isabel B. Leonor, David L. Kaplan, Rui L. Reis, Margarida Casal. Telma C. Bernardo participated in recombinant 6mer+FNII production and purification. The authors regret the error and would like to apologize for any inconvenience caused., (undefined)

Silk-elastin-like proteins (SELPs) are a family of genetically engineered recombinant protein polymers exhibiting mechanical and biological properties suited for a wide range of applications in the biomedicine and materials fields. They are being explored as the next generation of biomaterials but low productivities and use of antibiotics during production undermine their economic viability and safety. We have developed an industrially relevant, scalable, fed-batch process for the high level production of a novel SELP in E. coli in which the commonly used antibiotic selection marker of the expression vector is exchanged for a post segregational suicide system, the separate-component-stabilisation system (SCS). SCS significantly augments SELP productivity but also enhances the product safety profile and reduces process costs by eliminating the use of antibiotics. Plasmid content increased following induction but no significant differences in plasmid levels were discerned when using SCS or the antibiotic selection markers under the controlled fed-batch conditions employed. It is suggested that the absence of competing plasmid-free cells improves host cell viability and enables increased productivity with SCS. With the process developed, 12.8 g L(-1) purified SELP was obtained, this is the highest SELP productivity reported to date and clearly demonstrates the commercial viability of these promising polymers., This work was financed by the European Commission via the 7th Framework Programme Project EcoPlast (FP7-NMP-2009-SME-3), by national funds from the FCT through EXPL/BBB-BIO/1772/2013-FCOMP-010124-FEDER-041595, the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) and a fellowship to SRC (SFRH/BPD/89980/2012), as well as from ERDF through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). T.C. is supported by the FCT, the European Social Fund, the Programa Operacional Potencial Humano and the Investigador FCT Programme (IF/01635/2014). All the technical staff at the CBMA are thanked for their skilful technical assistance., info:eu-repo/semantics/publishedVersion

In June 2013, Brett Gosper, the chief executive of the International Rugby Board (IRB), the governing body of world rugby union, sent out a tweet that read ‘Ever wondered what the most game changing hand ball was in the history of football?’ It was accompanied by a photograph of the famous plaque at Rugby School that commemorates William Webb Ellis picking up the ball and running with it in 1823 (Gosper, 2013).

The objective of this work was to exploit the fibronectin type II (FNII) module from human matrix metalloproteinase-2 as a functional domain for the development of silk-based biopolymer blends that display enhanced cell adhesion properties. The DNA sequence of spider dragline silk protein (6mer) was genetically fused with the FNII coding sequence and expressed in Escherichia coli. The chimeric protein 6mer + FNII was purified by non-chromatographic methods. Films prepared from 6mer + FNII by solvent casting promoted only limited cell adhesion of human skin fibroblasts. However, the performance of the material in terms of cell adhesion was significantly improved when 6mer + FNII was combined with a silk-elastin-like protein in a concentration-dependent behavior. With this work we describe a novel class of biopolymer that promote cell adhesion and potentially useful as biomaterials for tissue engineering and regenerative medicine. Statement of Significance This work reports the development of biocompatible silk-based composites with enhanced cell adhesion properties suitable for biomedical applications in regenerative medicine. The biocomposites were produced by combining a genetically engineered silk-elastin-like protein with a genetically engineered spider-silk-based polypeptide carrying the three domains of the fibronectin type II module from human metalloproteinase-2. These composites were processed into free-standing films by solvent casting and characterized for their biological behavior. To our knowledge this is the first report of the exploitation of all three FNII domains as a functional domain for the development of bioinspired materials with improved biological performance. The present study highlights the potential of using genetically engineered protein-based composites as a platform for the development of new bioinspired biomaterials., This work was supported by Fundação para a Ciência e Tecnologia (FCT – Portugal) Funded Project “Chimera” (PTDC/EBB-EBI/109093/2008), by FCT/MEC through Portuguese funds (PIDDAC) – PEst-OE/BIA/UI4050/2014, by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI). TC is thankful to the FCT, ESF and POPH for its support through the Investigador FCT Programme (IF/01635/2014). ARibeiro thanks FCT for the SFRH\BPD\98388\2013 grant. AMPereira, RMachado and AdaCosta acknowledge FCT for PD/BD/113811/2015, SFRH-BPD/86470/2012 and SFRH/BD/75882/2011 grants, respectively.

The online version of this article (doi:10.1007/s00792-016-0851-9) contains supplementary material, which is available to authorized users., Cuproxidases are a subset of the blue multicopper oxidases that catalyze the oxidation of toxic Cu(I) ions into less harmful Cu(II) in the bacterial periplasm. Cuproxidases from psychrophilic, mesophilic, and thermophilic bacteria display the canonical features of temperature adaptation, such as increases in structural stability and apparent optimal temperature for activity with environmental temperature as well as increases in the binding affinity for catalytic and substrate copper ions. In contrast, the oxidative activities at 25 °C for both the psychrophilic and thermophilic enzymes are similar, suggesting that the nearly temperature-independent electron transfer rate does not require peculiar adjustments. Furthermore, the structural flexibilities of both the psychrophilic and thermophilic enzymes are also similar, indicating that the firm and precise bindings of the four catalytic copper ions are essential for the oxidase function. These results show that the requirements for enzymatic electron transfer, in the absence of the selective pressure of temperature on electron transfer rates, produce a specific adaptive pattern, which is distinct from that observed in enzymes possessing a well-defined active site and relying on conformational changes such as for the induced fit mechanism., This work was supported by the F.R.S-FNRS, Belgium (Fonds de la Recherche Fondamentale et Collective, contract numbers 2.4535.08, 2.4523.11 and U.N009.13 to G.F.) and by the Belgian program of Interuniversity Attraction Poles (iPros P7/44) initiated by the Federal Office for Scientific, Technical and Cultural Affairs. T.C. thanks the FCT and FEDER (POFC-COMPETE) for funding through the Investigador FCT Programme (IF/01635/2014) and project EngXyl (EXPL/BBB-BIO/1772/2013- FCOMP01-0124-FEDER-041595) as well as strategic funding via UID/ BIA/04050/2013. F.R., A.G. and A.C. were FRIA research fellows., info:eu-repo/semantics/publishedVersion

L'importance de la penalite (penalty) dans le rugby a XV trouve son origine dans les classes moyennes anglaises qui concurent ce sport a l'epoque victorienne. Or, ses dirigeants anglais ne souhaitaient pas en general que le rugby se « diffuse » et la crainte de la penalite, a la fois sur le terrain et en dehors, est devenue l'un de ses traits distinctifs. Cet article montre que le processus d'expansion d'un sport ne se reduit pas a une « diffusion » naturelle ou spontanee. Les decisions prises par ses premiers dirigeants concernant son developpement jouent egalement un role essentiel.

The history of professional basketball in the United States has not been well-served by historians of American sport. Whereas baseball overflows with studies of every conceivable era and aspect of ...

This is rich and fascinating book, one that is both a model of ethnography and an important commentary on the social devastation wrought on America's black communities over the past 30 or so years....

Enzymes produced by psychrophilic organisms have successfully overcome the low temperature challenge and evolved to maintain high catalytic rates in their permanently cold environments. As an initial step in our attempt to elucidate the cold-adaptation strategies used by these enzymes we report here the 1H, 15N and 13C assignments for the reduced form of a thiol-disulphide oxidoreductase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125., The NMR spectrometers are part of The National NMR Network (REDE/1517/RMN/2005), supported by ‘‘Programa Operacional Ciência e Inovação (POCTI) 2010’’ and Fundação para a Ciência e a Tecnologia (FCT). This work was funded by FCT, POCTI and FEDER; Projects POCI/BIA-PRO/57263/2004 and PTDC/BIO/70806/2006. TC is holder of a long term EMBO fellowship. MM is thankful to the Fundação para a Ciência e Tecnologia for its support through Programa Ciência 2007., info:eu-repo/semantics/publishedVersion

The increasing demand for oil and gas has incited exploration and production of deeper wells that reach high pressure and high temperature (HPHT) reservoirs. One critical element that is required to this end is rubber seals that can withstand HPHT conditions while meeting the requirements of sealability and structural integrity. Novel nanocomposites that comprise of natural rubber (NR) reinforced by well dispersed, high-concentration carbon nanotubes (CNTs) were recently developed to achieve the desired performance and were experimentally shown to exhibit significantly higher storage modulus than the matrix material. Understanding of the underlying reinforcing mechanism of this class of nanocomposites subjected to large deformation, especially in the real application conditions, has been very limited. In this study, a multiscale modeling method is developed to understand the mechanical behavior of CNT-rubber seals installed in a groove and subjected to high pressure. A micromechanics model is first constructed to evaluate the effective stress-strain responses of a representative volume element under different loading conditions, including uniaxial tension, equal biaxial extension, and planar tension. The effective properties thus established are then inputted into an appropriate hyperelasticity model, which is then used to model a CNT-rubber O-ring installed and pressurized. Sealability and structural integrity are evaluated in terms of contact pressure and strain. The numerical results are compared with the available experimental data. A parametric study is then conducted to assess the effects of CNT concentrations.Copyright © 2015 by ASME

Free standing films of a genetically engineered silk-elastin-like protein (SELP) were prepared using water and formic acid as solvents. Exposure to methanol-saturated air promoted the formation of aggregated β-strands rendering aqueous insolubility and improved the mechanical properties leading to a 10-fold increase in strain-to-failure. The films were optically clear with resistivity values similar to natural rubber and thermally stable up to 180 °C. Addition of glycerol showed to enhance the flexibility of SELP/glycerol films by interacting with SELP molecules through hydrogen bonding, interpenetrating between the polymer chains and granting more conformational freedom. This detailed characterization provides cues for future and unique applications using SELP based biopolymers.

In the present work we explored the ABP-CM4 peptide properties from Bombyx mori for the creation of biopolymers with broad antimicrobial activity. An antimicrobial recombinant protein-based polymer (rPBP) was designed by cloning the DNA sequence coding for ABP-CM4 in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. The new rPBP, named CM4-A200, was purified via a simplified nonchromatographic method, making use of the thermoresponsive behavior of the A200 polymer. ABP-CM4 peptide was also purified through the incorporation of a formic acid cleavage site between the peptide and the A200 sequence. In soluble state the antimicrobial activity of both CM4-A200 polymer and ABP-CM4 peptide was poorly effective. However, when the CM4-A200 polymer was processed into free-standing films high antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts and filamentous fungi was observed. The antimicrobial activity of CM4-A200 was dependent on the physical contact of cells with the film surface. Furthermore, CM4-A200 films did not reveal a cytotoxic effect against both normal human skin fibroblasts and human keratinocytes. Finally, we have developed an optimized ex vivo assay with pig skin demonstrating the antimicrobial properties of the CM4-A200 cast films for skin applications., This work was supported by FEDER through POFC - COMPETE, by Portuguese funds from FCT through Project PEst-OE/BIA/UI4050/2014, and by the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y Leon-JCyL (VA152A12-2 and VA155A12-2), Spain. The authors also thank Matadouro - Central Carnes de Entre Douro e Minho, Lda for their availability to provide pig skin. A.C. and R.M. acknowledge FCT for SFRH/BD/75882/2011 and SFRH-BPD/86470/2012 Grants, respectively. T.C. is thankful to the FCT for its support through Programa Ciencia 2008.

The Antarctic marine environment is characterized by challenging conditions for the survival of native microorganisms. Indeed, next to the temperature effect represented by the Arrhenius law, the viscosity of the medium, which is also significantly enhanced by low temperatures, contributes to slow down reaction rates. This review analyses the different challenges and focuses on a key element of life at low temperatures: cold-adapted enzymes. The molecular characteristics of these enzymes are discussed as well as the adaptation strategies which can be inferred from the comparison of their properties and three-dimensional structures with those of their mesophilic counterparts. As these enzymes display a high specific activity at low and moderate temperatures associated with a relatively high thermosensitivity, the interest in these properties is discussed with regard to their current and possible applications in biotechnology.

This paper examines the history of English rugby union football in the period between the 1895 split in the sport and the end of the Edwardian era. It argues that during this period, the RFU's definition of amateurism was constantly being refined and adjusted to take into account the changing fortunes of the game. However, underlying these shifts was a constant desire to create a sport that was fashioned around the needs of young middle-class men to create a social and recreational environment which they controlled, unthreatened by the prospect of working-class domination or commercial exigencies.

The ability of psychrophiles to survive and proliferate at low temperatures implies that they have overcome key barriers inherent to permanently cold environments. These challenges include: reduced enzyme activity; decreased membrane fluidity; altered transport of nutrients and waste products; decreased rates of transcription, translation and cell division; protein cold-denaturation; inappropriate protein folding; and intracellular ice formation. Cold-adapted organisms have successfully evolved features, genotypic and/or phenotypic, to surmount the negative effects of low temperatures and to enable growth in these extreme environments. In this review, we discuss the current knowledge of these adaptations as gained from extensive biochemical and biophysical studies and also from genomics and proteomics.

Xylanases have long been used in the baking industry for improving dough stability and flexibility and for increasing bread volume and crumb structure. Only xylanases from glycoside hydrolase families 10 and 11 appear to have been tested in this application and only those from the latter family have as yet found application. Interestingly, enzymes with a putative xylanase activity are also found in glycoside hydrolase families 5, 7, 8 and 43, but apparently these have not, as yet, been tested in baking. Baking trials were used to determine the effectiveness of a psychrophilic and a mesophilic family 8 xylanolytic enzyme as well as a psychrophilic family 10 xylanase and a currently used family 11 commercial mesophilic xylanase. The potential of family 8 xylanases as technological aids in baking was clearly demonstrated as both the psychrophilic enzyme from Pseudoalteromonas haloplanktis TAH3a and the mesophilic enzyme from Bacillus halodurans C-125 had a positive effect on loaf volume. In contrast, the psychrophilic family 10 enzyme from Cryptococcus adeliae TAE85 was found to be ineffective.

In 1989 the historian Christopher Hill wondered why the North of England, with all its apparent cultural, social and geographical distinctiveness, had never achieved a separate state or even seen the formation of a 'northern separatist movement' .1 Although the broader questions of the historical development of the North of England are beyond our immediate scope, this paper does seek to shed light on the question of Northern English identity through the medium of the quintessentially Northern English cultural experience of the annual Rugby League Challenge Cup Final atWembley stadium where between 1929 to 1999, tens of thousands of people from Cumberland, Lancashire and Yorkshire travelled down to London to watch the final of the Rugby League Challenge Cup, traditionally on the first Saturday of May. It was a unique event in British sport.

Xylanases are hydrolytic enzymes which randomly cleave the beta 1,4 backbone of the complex plant cell wall polysaccharide xylan. Diverse forms of these enzymes exist, displaying varying folds, mechanisms of action, substrate specificities, hydrolytic activities (yields, rates and products) and physicochemical characteristics. Research has mainly focused on only two of the xylanase containing glycoside hydrolase families, namely families 10 and 11, yet enzymes with xylanase activity belonging to families 5, 7, 8 and 43 have also been identified and studied, albeit to a lesser extent. Driven by industrial demands for enzymes that can operate under process conditions, a number of extremophilic xylanases have been isolated, in particular those from thermophiles, alkaliphiles and acidiphiles, while little attention has been paid to cold-adapted xylanases. Here, the diverse physicochemical and functional characteristics, as well as the folds and mechanisms of action of all six xylanase containing families will be discussed. The adaptation strategies of the extremophilic xylanases isolated to date and the potential industrial applications of these enzymes will also be presented.