Identifying design criteria for urban system last-mile solutions -A multi-stakeholder perspective

This is the author accepted manuscript. The final version is available from Taylor & Francis via http://dx.doi.org/10.1080/09537287.2016.1147099This study presents a novel approach to design and evaluate ‘last-mile’ solutions – encompassing the social and economic perspectives of key stakeholders. While urban system initiatives have been implemented in practice, theoretical gaps remain at the operational design level. A theoretical framework is developed, based on design criteria identified from a critical synthesis of supply chain and operations management literature, and ‘operationalised’ using an in-depth case study demonstrating implementation of a Consumer Choice Portal-Package Consolidation Centre solution, within a densely populated urban geography. Findings suggest that there is a need to re-define the role of institutional actors beyond that of the traditional governance task, to one of being able to facilitate performance outcomes. Similarly, industrial efficiency dimensions need to be reorientated to include consumer participation, social considerations and multi-stakeholder service outcomes. Finally, implications for operations theory and practising managers in city logistics are highlighted, with suggested directions for future research.TS

Precursors of Cytochrome Oxidase in Cytochrome-Oxidase-Deficient Cells of Neurospora crassa

Three different cell types of Neurospora crassa deficient in cytochrome oxidase were studied: the nuclear mutant cni-1, the cytoplasmic mutant mi-1 and copper-depleted wild-type cells. * 1. The enzyme-deficient cells have retained a functioning mitochondrial protein synthesis. It accounted for 12–16% of the total protein synthesis of the cell. However, the analysis of mitochondrial translation products by gel electrophoresis revealed that different amounts of individual membrane proteins were synthesized. Especially mutant cni-1 produced large amounts of a small molecular weight translation product, which is barely detectable in wild-type. * 2. Mitochondrial preparations of cytochrome-oxidase-deficient cells were examined for precursors of cytochrome oxidase. The presence of polypeptide components of cytochrome oxidase in the mitochondria was established with specific antibodies. On the other hand, no significant amounts of heme a could be extracted. * 3. Radioactively labelled components of cytochrome oxidase were isolated by immunoprecipitation and analysed by gel electrophoresis. All three cell types contained the enzyme components 4–7, which are translated on cytoplasmic ribosomes. The mitochondrially synthesized components 1–3 were present in mi-1 mutant and in copper-depleted wild-type cells. In contrast, components 2 and 3 were not detectable in the nuclear mutant cni-1. Both relative and absolute amounts of these polypeptides in the enzyme-deficient cells were quite different from those in wild-type cells. * 4. The components of cytochrome oxidase found in the enzyme-deficient cells were tightly associated with the mitochondrial membranes. * 5. Processes, which affect and may control the production of enzyme precursors or their assembly to a functional cytochrome oxidase are discussed

Living in a cloud: intraspecific facilitation by allelochemical mediated grazing protection 1 2

Abstract 23 Dinoflagellates are a major cause of harmful algal blooms, with consequences for coastal marine 24 ecosystem functioning and services. Alexandrium tamarense is one of the most abundant and 25 widespread toxigenic species in the temperate northern and southern hemisphere, and produces 26 paralytic shellfish poisoning toxins as well as lytic allelochemical substances. These bioactive 27 compounds may support the success of A. tamarense and its ability to form blooms. Here we 28 investigate grazing of highly (Alex2) and moderately (Alex4) allelochemical active A. tamarense 29 strains and a non-allelochemical active conspecific (Alex5) by the heterotrophic dinoflagellate 30 Polykrikos kofoidii in monoclonal and mixed experimental set-ups. While Alex4 and particularly 31 Alex5 were strongly grazed by P. kofoidii in monoclonal set-ups, both strains did grow well in 32 the mixed assemblages (Alex4+Alex5 and Alex2+Alex5). Hence, the allelochemical active 33 strains facilitate the non-active strain by protecting the population against grazing. This 34 allelochemical mediated facilitation resembles associational resistance observed in various 35 macroalgal and plant communities. Occurring intraspecifically it may partly explain the high 36 genotypic and phenotypic diversity of Alexandrium populations. It is furthermore conceivable 37 that these diverse populations comprise multiple cooperative traits that may support mutual 38 intraspecific facilitation, which in turn will promote the success of this notorious harmful algal 39 bloom species. 40 4

Calcium and Vitamin D increase mRNA levels for the growth control hIK1 channel in human epidermal keratinocytes but functional channels are not observed

BACKGROUND: Intermediate-conductance, calcium-activated potassium channels (IKs) modulate proliferation and differentiation in mesodermal cells by enhancing calcium influx, and they contribute to the physiology of fluid movement in certain epithelia. Previous reports suggest that IK channels stimulate proliferative growth in a keratinocyte cell line; however, because these channels indirectly promote calcium influx, a critically unique component of the keratinocyte differentiation program, an alternative hypothesis is that they would be anti-proliferative and pro-differentiating. This study addresses these hypotheses. METHODS: Real-time PCR, patch clamp electrophysiology, and proliferation assays were used to determine if human IK1 (hIK1) expression and function are correlated with either proliferation or differentiation in cultured human skin epidermal keratinocytes, and skin biopsies grown in explant culture. RESULTS: hIK1 mRNA expression in human keratinocytes and skin was increased in response to anti-proliferative/pro-differentiating stimuli (elevated calcium and Vitamin D). Correspondingly, the hIK1 agonist 1-EBIO inhibited keratinocyte proliferation suggesting that the channel could be anti-proliferative and pro-differentiating. However, this proliferative inhibition by 1-EBIO was not reversed by a panel of hIK1 blockers, calling into question the mechanism of 1-EBIO action. Subsequent patch clamp electrophysiological analysis failed to detect hIK1 channel currents in keratinocytes, even those expressing substantial hIK1 mRNA in response to calcium and Vitamin D induced differentiation. Identical electrophysiological recording conditions were then used to observe robust IK1 currents in fibroblasts which express IK1 mRNA levels comparable to those of keratinocytes. Thus, the absence of observable hIK1 currents in keratinocytes was not a function of the electrophysiological techniques. CONCLUSION: Human keratinocyte differentiation is stimulated by calcium mobilization and influx, and differentiation stimuli coordinately upregulate mRNA levels of the calcium-activated hIK1 channel. This upregulation is paradoxical in that functional hIK1 channels are not observed in cultured keratinocytes. It appears, therefore, that hIK1 does not contribute to the functional electrophysiology of primary human keratinocytes, nor intact human skin. Further, the results indicate caution is required when interpreting experiments utilizing pharmacological hIK1 modulators in human keratinocytes

Minimally Invasive Surgical Approaches and Traditional Total Hip Arthroplasty: A Meta-Analysis of Radiological and Complications Outcomes

BACKGROUND: Minimally invasive total hip arthroplasty (MITHA) remains considerably controversial. Limited visibility and prosthesis malposition increase the risk of post-surgical complications compared to those of the traditional method. METHODS: A meta-analysis was undertaken of all published databases up to May 2011. The studies were divided into four subgroups according to the surgical approach taken. The radiological outcomes and complications of minimally invasive surgery were compared to traditional total hip arthroplasty (TTHA) using risk ratio, mean difference, and standardized mean difference statistics. RESULTS: In five studies involving the posterolateral approach, no significant differences were found between the MITHA groups and the TTHA groups in the acetabular cup abduction angle (p = 0.41), acetabular anteversion (p = 0.96), and femoral prosthesis position (p = 0.83). However, the femoral offset was significantly increased (WMD = 3.00; 95% CI, 0.40-5.60; p = 0.02). Additionally, there were no significant differences among the complications in both the groups (dislocations, nerve injury, infection, deep vein thrombosis, proximal femoral fracture) and revision rate (p>0.05). In three studies involving the posterior approach, there were no significant differences in radiological outcomes or all other complications between MITHA or TTHA groups (p>0.05). Three studies involved anterolateral approach, while 2 studies used the lateral approach. However, the information from imaging and complications was not adequate for statistical analysis. CONCLUSIONS: Posterior MITHA seems to be a safe surgical procedure, without the increased risk of post-operative complication rates and component malposition rates. The posterolateral approach THA may lead to increased femoral offset. The current data are not enough to reach a positive conclusion that lateral and anterolateral approaches will result in increased risks of adverse effects and complications at the prosthesis site

Stability of domain structures in multi-domain proteins

Multi-domain proteins have many advantages with respect to stability and folding inside cells. Here we attempt to understand the intricate relationship between the domain-domain interactions and the stability of domains in isolation. We provide quantitative treatment and proof for prevailing intuitive ideas on the strategies employed by nature to stabilize otherwise unstable domains. We find that domains incapable of independent stability are stabilized by favourable interactions with tethered domains in the multi-domain context. Stability of such folds to exist independently is optimized by evolution. Specific residue mutations in the sites equivalent to inter-domain interface enhance the overall solvation, thereby stabilizing these domain folds independently. A few naturally occurring variants at these sites alter communication between domains and affect stability leading to disease manifestation. Our analysis provides safe guidelines for mutagenesis which have attractive applications in obtaining stable fragments and domain constructs essential for structural studies by crystallography and NMR

Genetic Rearrangements Can Modify Chromatin Features at Epialleles

Analogous to genetically distinct alleles, epialleles represent heritable states of different gene expression from sequence-identical genes. Alleles and epialleles both contribute to phenotypic heterogeneity. While alleles originate from mutation and recombination, the source of epialleles is less well understood. We analyze active and inactive epialleles that were found at a transgenic insert with a selectable marker gene in Arabidopsis. Both converse expression states are stably transmitted to progeny. The silent epiallele was previously shown to change its state upon loss-of-function of trans-acting regulators and drug treatments. We analyzed the composition of the epialleles, their chromatin features, their nuclear localization, transcripts, and homologous small RNA. After mutagenesis by T-DNA transformation of plants carrying the silent epiallele, we found new active alleles. These switches were associated with different, larger or smaller, and non-overlapping deletions or rearrangements in the 3′ regions of the epiallele. These cis-mutations caused different degrees of gene expression stability depending on the nature of the sequence alteration, the consequences for transcription and transcripts, and the resulting chromatin organization upstream. This illustrates a tight dependence of epigenetic regulation on local structures and indicates that sequence alterations can cause epigenetic changes at some distance in regions not directly affected by the mutation. Similar effects may also be involved in gene expression and chromatin changes in the vicinity of transposon insertions or excisions, recombination events, or DNA repair processes and could contribute to the origin of new epialleles

A Molecular and Co-Evolutionary Context for Grazer Induced Toxin Production in Alexandrium tamarense

Marine dinoflagellates of the genus Alexandrium are the proximal source of neurotoxins associated with Paralytic Shellfish Poisoning. The production of these toxins, the toxin biosynthesis and, thus, the cellular toxicity can be influenced by abiotic and biotic factors. There is, however, a lack of substantial evidence concerning the toxins' ecological function such as grazing defense. Waterborne cues from copepods have been previously found to induce a species-specific increase in toxin content in Alexandrium minutum. However, it remains speculative in which context these species-specific responses evolved and if it occurs in other Alexandrium species as well. In this study we exposed Alexandrium tamarense to three copepod species (Calanus helgolandicus, Acartia clausii, and Oithona similis) and their corresponding cues. We show that the species-specific response towards copepod-cues is not restricted to one Alexandrium species and that co-evolutionary processes might be involved in these responses, thus giving additional evidence for the defensive role of phycotoxins. Through a functional genomic approach we gained insights into the underlying molecular processes which could trigger the different outcomes of these species-specific responses and consequently lead to increased toxin content in Alexandrium tamarense. We propose that the regulation of serine/threonine kinase signaling pathways has a major influence in directing the external stimuli i.e. copepod-cues, into different intracellular cascades and networks in A. tamarense. Our results show that A. tamarense can sense potential predating copepods and respond to the received information by increasing its toxin production. Furthermore, we demonstrate how a functional genomic approach can be used to investigate species interactions within the plankton community

Synthetic Mimic of Antimicrobial Peptide with Nonmembrane-Disrupting Antibacterial Properties

Proteolysis in dairy lactic acid bacteria has been studied in great detail by genetic, biochemical and ultrastructural methods. From these studies the picture emerges that the proteolytic systems of lactococci and lactobacilli are remarkably similar in their components and mode of action. The proteolytic system consists of an extracellularly located serine-proteinase, transport systems specific for di-tripeptides and oligopeptides (> 3 residues), and a multitude of intracellular peptidases. This review describes the properties and regulation of individual components as well as studies that have led to identification of their cellular localization. Targeted mutational techniques developed in recent years have made it possible to investigate the role of individual and combinations of enzymes in vivo. Based on these results as well as in vitro studies of the enzymes and transporters, a model for the proteolytic pathway is proposed. The main features are: (i) proteinases have a broad specificity and are capable of releasing a large number of different oligopeptides, of which a large fraction falls in the range of 4 to 8 amino acid residues; (ii) oligopeptide transport is the main route for nitrogen entry into the cell; (iii) all peptidases are located intracellularly and concerted action of peptidases is required for complete degradation of accumulated peptides.