Tactically ambiguous performance: the 1:1 model of Mies’s Krefeld Golf Clubhouse Project by Robbrecht en Daem architecten

A temporary and fragmentary 1:1 model of Mies van der Rohe’s unexecuted golf clubhouse project for Krefeld was designed by Robbrecht en Daem architecten and built largely in plywood near to the project's original site in the summer of 2013. This article analyses the project against the background of the debate on contemporary facsimile monument reconstructions in Germany and beyond. It argues that the design of this 1:1 model, which is not a replica nor a reconstruction, employs particular design strategies to enact the absent building in an ambiguous and self-critical way. Depending on their position in and around the pavilion, visitors alternately experienced this structure as an immersive evocation of Miesian architecture or as a self-exposing scenographic device. The theatricality of this alternation is compared to the spatio-temporal experience of tratteggio restoration techniques and to conceptual approaches to 1:1 (re)materialisations in architecture and art exhibitions. This theatrical alternation is valued as a critical complexification on the image-materiality axis of reconstructions, challenging the polarised positions in the debate about the (conditions of) legitimacy of new-old-monuments.     

Structured inspections of code

Cleanroom programming and code inspections independently provide evidence that it is more efficient to postpone the testing of code to a later stage than is usually done. This paper argues that an additional gain in quality and efficiency of development can be obtained by structuring inspections by means of an inspection protocol. The written part of such a protocol is prepared by the programmer before the inspection. It is modelled on Floyd's method for the verification of flowcharts. However, the protocol differs from Floyd's method in being applicable in practice. Structured inspections gain this advantage by not attempting to be a proof; they are no more than an articulation of existing forms of inspection. With the usual method of structured programming it may be difficult to prepare the inspection protol. On the other hand, ‘assertion-driven programming’ (of which an example is included in this paper) not only facilitates protocol preparation, but also the coding itself.     

The Role of the Lymphatic System in the Pathogenesis and Treatment of Inflammatory Bowel Disease

Although the number of therapeutic options for the treatment of inflammatory bowel disease (IBD) has increased in recent years, patients suffer from decreased quality of life due to non-response or loss of response to the currently available treatments. An increased understanding of the disease’s etiology could provide novel insights for treatment strategies in IBD. Lymphatic system components are generally linked to immune responses and presumably related to inflammatory diseases pathophysiology. This review aims to summarize findings on immune-mediated mechanisms in lymphoid tissues linked with IBD pathogenesis and (potential) novel treatments. Enhanced innate and adaptive immune responses were observed in mesenteric lymph nodes (MLNs) and other lymphoid structures, such as Peyer’s patches, in patients with IBD and in animal models. Furthermore, the phenomenon of lymphatic obstruction in the form of granulomas in MLNs and lymphatic vessels correlates with disease activity. There is also evidence that abnormalities in the lymphatic stromal components and lymph node microbiome are common in IBD and could be exploited therapeutically. Finally, novel agents targeting lymphocyte trafficking have been added to the treatment armamentarium in the field of IBD. Overall, gut-associated lymphoid tissue plays a key role in IBD immunopathogenesis, which could offer novel therapeutic targets.     

Modulation Excitation X-Ray Absorption Spectroscopy to Probe Surface Species on Heterogeneous Catalysts

The advantages and open questions of the combination of modulation excitation spectroscopy and phase sensitive detection (PSD) with X-ray absorption spectroscopy (XAS) for the analysis of heterogeneous catalysts at work are reviewed. The characteristic spectral signatures of two different edges (Pd K and Pt L₃) are described in relation to the red-ox chemistry of Pd/Al₂O₃ and Pt/Al₂O₃ with respect to NO reduction by CO and CO oxidation, respectively. Both examples demonstrate that PSD makes XAS sensitive to potentially active species for the catalytic reaction.     

Structuring and financing international BOO/BOT desalination projects

There has been a recent increase in desalination projects in an effort to meet the growing demand for clean water. Desalination plants, like other large public service infrastructure, are capital intensive with long asset lives. This makes them prime vehicles to introduce private sector investment both equity and debt. Key to attracting the private sector, particularly International investors, is a well-developed commercial framework for the project following principles widely seen and understood by private investors. Desalination plants can easily fit into well-tested structures, which optimize the risk profile for each stakeholder, e.g. the consumer, the public sector authority, the equity investor/developer, the lenders; and reward each of them appropriately. This paper seeks to introduce a typical project financing structure and compare desalination to other infrastructure projects, which are also competing in the International arena for private investment. Given the success of recent private sector investment in desalination, and the growing demand for clean drinking water, it could be expected that this industry should continue to attract its fair share of the International investor community.     

Pyrethrins Protect Pyrethrum Leaves Against Attack by Western Flower Thrips, <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis>

Pyrethrins are active ingredients extracted from pyrethrum flowers (Tanacetum cinerariifolium), and are the most widely used botanical insecticide. However, several thrips species are commonly found on pyrethrum flowers in the field, and are the dominant insects found inside the flowers. Up to 80 % of western flower thrips (WFT, Frankliniella occidentalis) adults died within 3 days of initiating feeding on leaves of pyrethrum, leading us to evaluate the role of pyrethrins in the defense of pyrethrum leaves against WFT. The effects of pyrethrins on WFT survival, feeding behavior, and reproduction were measured both in vitro and in planta (infiltrated leaves). The lethal concentration value (LC50) for pyrethrins against WFT adults was 12.9 mg/ml, and pyrethrins at 0.1 % (w/v) and 1 % (w/v) had significantly negative effects on feeding, embryo development, and oviposition. About 20-70 % of WFT were killed within 2 days when they were fed chrysanthemum leaves containing 0.01-1 % pyrethrins. Chrysanthemum leaves containing 0.1 % or 1 % pyrethrins were significantly deterrent to WFT. In a no-choice assay, the reproduction of WFT was reduced significantly when the insects were fed leaves containing 0.1 % pyrethrins, and no eggs were found in leaves containing 1 % pyrethrins. Our results suggest that the natural concentrations of pyrethrins in the leaves may be responsible for the observed high mortality of WFT on pyrethrum.     

Optical Control of Antibody Activity by Using Photocleavable Bivalent Peptide–DNA Locks

Antibody-based molecular recognition plays a central role in today's life sciences, ranging from immunoassays to molecular imaging and antibody-based therapeutics. Control over antibody activity by using external triggers such as light could further increase the specificity of antibody-based targeting. Here we present bivalent peptide–DNA ligands containing photocleavable linkers as a noncovalent approach by which to allow photoactivation of antibody activity. Light-triggered cleavage of the 3-amino-3-(2-nitrophenyl)propionic acid peptide linker converted the high-affinity bivalent peptide–DNA lock into weakly binding monovalent ligands, effectively restoring antibody targeting of cell-surface receptors. In this work, a proof of principle was provided with an anti-hemagglutinin antibody, but the molecular design of the lock is generic and applicable to any monoclonal antibody for which an epitope or mimotope of sufficient affinity is available.     

Directed evolution: From a staphylococcal lipase to a phospholipase

The lipase of Staphylococcus aureus (SAL) is able to degrade lipids and p‐nitrophenylesters but is not active on phospholipid substrates. Interestingly, the homologous lipase from Staphylococcus hyicus is highly active on phospholipids. In order to investigate the molecular basis for this difference in substrate specificity, phospholipase activity was introduced into SAL by directed evolution strategy. In this approach, sequential rounds of error‐prone PCR were performed in combination with a screening of the resulting mutant libraries. The screening was based on a high‐throughput plate assay and a subsequent chromogenic assay in 96‐well plate format to accurately determine the enzymatic activities in cell lysates of a selected number of clones. After 4 rounds of error‐prone PCR, two products were obtained, displaying a 7.8‐ and 9.2‐fold increase in absolute phospholipase activity and a 5.9‐ and 6.9‐fold increase in phospholipase/ lipase activity ratio. A final round of DNA shuffling with these two products and wildtype (WT)‐SAL was performed to combine beneficial mutations and to eliminate neutral or deleterious mutations. This procedure yielded a best variant containing 6 amino acid mutations displaying a 11.6‐fold increase in absolute phospholipase activity and a 11.5‐fold increase in phospholipase/lipase ratio as compared to the starting point. The character of the mutations and their possible effects on substrate specificity are discussed.     

Synthesis and Biological Characterization of (3R,4R)‐4‐(2‐(Benzhydryloxy)ethyl)‐1‐((R)‐2‐hydroxy‐2‐phenylethyl)‐piperidin‐3‐ol and Its Stereoisomers for Activity toward Monoamine Transporters

A novel series of optically active molecules based on a 4‐(2‐(benzhydryloxy)ethyl)‐1‐((R)‐2‐hydroxy‐2‐phenylethyl)‐piperidin‐3‐ol template were developed. Depending on stereochemistry, the compounds exhibit various degrees of affinity for three dopamine, serotonin, and norepinephrine transporters. These molecules have the potential for treating several neurological disorders such as drug abuse, depression, and attention deficit hyperactivity disorder.

     

His-tags as Zn(II) binding motifs in a protein-based fluorescent sensor

Fluorescent indicators that allow real-time imaging of Zn(II) in living cells are invaluable tools for understanding Zn(II) homeostasis. Genetically encoded sensors based on fluorescence resonance energy transfer between fluorescent protein domains have important advantages over synthetic probes. We discovered that hexahistidine tags have a strong tendency to dimerize upon binding of Zn(II) in solution and we used this principle to develop a new protein-based sensor for Zn(II). Enhanced cyan and yellow fluorescent proteins were connected by long flexible peptide linkers and His-tags were incorporated at both termini of this fusion protein. The resulting sensor CLY9-2His allows the ratiometric fluorescent detection of Zn(II) in the nanomolar range. In addition, CLY9-2His is selective over the physiologically relevant metal ions Fe(II), Mn(II), Ca(II) and Mg(II). Our approach demonstrates the potential of using small peptides as metal-binding ligands in chelating fluorescent protein chimeras.