Public Space in the New Urban Agenda: Research into Implementation

The New Urban Agenda is a landmark international framework for urbanisation for the next two decades, adopted by acclamation by all 193 countries of the United Nations. Nonetheless, implementation remains an enormous challenge, as does the related need for research evidence to inform practice. This thematic issue brings together research from a number of participants of the Future of Places conference series, contributin new research to inform the development and implementation of the New Urban Agenda, and with a focus on the fundamental topic of public space creation and improvement

Potential of light and temperature exploitation for Accelerated Shelf Life Studies (ASLT) for sauces

The advantage of being “First to Market” is tremendous, but keeping a high and consistent level of quality is determinant as well for the success of a food company. Low quality products cause damage to the brand and monetary loses in the long run. Quality is strongly influenced by a correct estimation of shelf life, but full length shelf life tests demands large inputs of time and money. The need for a more efficient method of estimating shelf life is therefore required, and the use of accelerated storage tests has gained in popularity in recent years. “Accelerated Shelf Life Tests” is by definition a method that allows the estimation of shelf life through short term storage tests. This is done by converting the accelerated storage results mathematically to represent normal storage conditions, often using different kinetic models. This project intends to evaluate the potential of using accelerated shelf life as a method for shelf life estimation for wet sauces. Two sauces was evaluated in this study; Pizza topping and Taco sauce with previously known shelf life of nine months and 18 months respectively. The shelf life was estimated by exposing the samples to high temperatures and light during a time period of eight weeks in Climate Chambers (Sanyo Gallenkamp Prime Incubator, INC-000- MA1.9). The light source was a LED lamp that emitted light around 680 to 770 lux and the samples was stored at 22° C, 30° C and 40° C. The samples were then evaluated by sensory analysis and by measuring pigment degradation. The result showed some inconsistencies with the theoretical aspects of the study. The Pizza topping was estimated to have a shelf life of eight months, and the Taco sauce was predicted to maintain quality for about 17 months. These values correspond well to the current estimated shelf life used. However, the results yielded different estimations depending on how the results were calculated. Q10 – modelling, a method that deduces a conversion factor that allows for direct translation of accelerated storage test results into normal storage condition yielded considerably shorter estimated shelf life values while the use of the Arrhenius equation seemed to results in more realistic values. In addition, the colour analysis resulted in different results when compared to the sensory analysis. The recommendation is that accelerated shelf life tests have the potential to be a valuable tool when predicting product shelf life in a fast-paced innovation environment. However, due to the inconsistencies of the results it is recommended to perform further investigations before adopting accelerated shelf life tests as a standard method for shelf life estimation

Main street plot scale in urban design for inclusive economies : Stockholm case studies

This paper explores evidence that entrepreneurial opportunities for migrants and other lower income populations can be expanded in part through increasing the presence of fine grained scales of plots and plates along main streets, as part of a systematic urban design strategy. It describes that systematic strategy herein. The paper encompasses the study of three main streets with varying plot sizes in the inner city of Stockholm, Sweden, and examines the outcomes for different types and scales of businesses. After presenting the findings, analysis and conclusions, larger questions of urban design for more inclusive economies are discussed

Structure of the poly-C9 component of the complement membrane attack complex

The membrane attack complex (MAC)/perforin-like protein complement component 9 (C9) is the major component of the MAC, a multi-protein complex that forms pores in the membrane of target pathogens. In contrast to homologous proteins such as perforin and the cholesterol-dependent cytolysins (CDCs), all of which require the membrane for oligomerisation, C9 assembles directly onto the nascent MAC from solution. However, the molecular mechanism of MAC assembly remains to be understood. Here we present the 8 Å cryo-EM structure of a soluble form of the poly-C9 component of the MAC. These data reveal a 22-fold symmetrical arrangement of C9 molecules that yield an 88-strand pore-forming β-barrel. The N-terminal thrombospondin-1 (TSP1) domain forms an unexpectedly extensive part of the oligomerisation interface, thus likely facilitating solution-based assembly. These TSP1 interactions may also explain how additional C9 subunits can be recruited to the growing MAC subsequent to membrane insertion

Cryo electron microscopy to determine the structure of macromolecular complexes

Cryo-electron microscopy (cryo-EM) is a structural molecular and cellular biology technique that has experienced major advances in recent years. Technological developments in image recording as well as in processing software make it possible to obtain three-dimensional reconstructions of macromolecular assemblies at near-atomic resolution that were formerly obtained only by X-ray crystallography or NMR spectroscopy. In parallel, cryo-electron tomography has also benefitted from these technological advances, so that visualization of irregular complexes, organelles or whole cells with their molecular machines in situ has reached subnanometre resolution. Cryo-EM can therefore address a broad range of biological questions. The aim of this review is to provide a brief overview of the principles and current state of the cryo-EM field

Preparation and topology of the Mediator middle module

Mediator is the central coactivor complex required for regulated transcription by RNA polymerase (Pol) II. Mediator consists of 25 subunits arranged in the head, middle, tail and kinase modules. Structural and functional studies of Mediator are limited by the availability of protocols for the preparation of recombinant modules. Here, we describe protocols for obtaining pure endogenous and recombinant complete Mediator middle module from Saccharomyces cerevisiae that consists of seven subunits: Med1, 4, 7, 9, 10, 21 and 31. Native mass spectrometry reveals that all subunits are present in equimolar stoichiometry. Ion-mobility mass spectrometry, limited proteolysis, light scattering and small-angle X-ray scattering all indicate a high degree of intrinsic flexibility and an elongated shape of the middle module. Protein–protein interaction assays combined with previously published data suggest that the Med7 and Med4 subunits serve as a binding platform to form the three heterodimeric subcomplexes, Med7N/21, Med7C/31 and Med4/9. The subunits, Med1 and Med10, which bridge to the Mediator tail module, bind to both Med7 and Med4

Two conserved modules of Schizosaccharomyces pombe Mediator regulate distinct cellular pathways

Mediator is an evolutionary conserved coregulator complex required for transcription of almost all RNA polymerase II-dependent genes. The Schizosaccharomyces pombe Mediator consists of two dissociable components—a core complex organized into a head and middle domain as well as the Cdk8 regulatory subcomplex. In this work we describe a functional characterization of the S. pombe Mediator. We report the identification of the S. pombe Med20 head subunit and the isolation of ts alleles of the core head subunit encoding med17+. Biochemical analysis of med8ts, med17ts, Δmed18, Δmed20 and Δmed27 alleles revealed a stepwise head domain molecular architecture. Phenotypical analysis of Cdk8 and head module alleles including expression profiling classified the Mediator mutant alleles into one of two groups. Cdk8 module mutants flocculate due to overexpression of adhesive cell-surface proteins. Head domain-associated mutants display a hyphal growth phenotype due to defective expression of factors required for cell separation regulated by transcription factor Ace2. Comparison with Saccharomyces cerevisiae Mediator expression data reveals that these functionally distinct modules are conserved between S. pombe and S. cerevisiae

Cellular glycosylation affects Herceptin binding and sensitivity of breast cancer cells to doxorubicin and growth factors

Alterations in protein glycosylation are a key feature of oncogenesis and have been shown to affect cancer cell behaviour perturbing cell adhesion, favouring cell migration and metastasis. This study investigated the effect of N-linked glycosylation on the binding of Herceptin to HER2 protein in breast cancer and on the sensitivity of cancer cells to the chemotherapeutic agent doxorubicin (DXR) and growth factors (EGF and IGF-1). The interaction between Herceptin and recombinant HER2 protein and cancer cell surfaces (on-rate/off-rate) was assessed using a quartz crystal microbalance biosensor revealing an increase in the accessibility of HER2 to Herceptin following deglycosylation of cell membrane proteins (deglycosylated cells Bmax: 6.83 Hz; glycosylated cells Bmax: 7.35 Hz). The sensitivity of cells to DXR and to growth factors was evaluated using an MTT assay. Maintenance of SKBR-3 cells in tunicamycin (an inhibitor of N-linked glycosylation) resulted in an increase in sensitivity to DXR (0.1 µM DXR P<0.001) and a decrease in sensitivity to IGF-1 alone and to IGF-1 supplemented with EGF (P<0.001). This report illustrates the importance of N-linked glycosylation in modulating the response of cancer cells to chemotherapeutic and biological treatments and highlights the potential of glycosylation inhibitors as future combination treatments for breast cancer