Conformational selection of inhibitors and substrates by proteolytic enzymes: Implications for drug design and polypeptide processing

Inhibitors of proteolytic enzymes (proteases) are emerging as prospective treatments for diseases such as AIDS and viral infections, cancers, inflammatory disorders, and Alzheimer's disease. Generic approaches to the design of protease inhibitors are limited by the unpredictability of interactions between, and structural changes to, inhibitor and protease during binding. A computer analysis of superimposed crystal structures for 266 small molecule inhibitors bound to 48 proteases (16 aspartic, 17 serine, 8 cysteine, and 7 metallo) provides the first conclusive proof that inhibitors, including substrate analogues, commonly bind in an extended beta-strand conformation at the active sites of all these proteases. Representative superimposed structures are shown for (a) multiple inhibitors bound to a protease of each class, (b) single inhibitors each bound to multiple proteases, and (c) conformationally constrained inhibitors bound to proteases. Thus inhibitor/substrate conformation, rather than sequence/composition alone, influences protease recognition, and this has profound implications for inhibitor design. This conclusion is supported by NMR, CD, and binding studies for HIV-1 protease inhibitors/ substrates which, when preorganized in an extended conformation, have significantly higher protease affinity. Recognition is dependent upon conformational equilibria since helical and turn peptide conformations are not processed by proteases. Conformational selection explains the resistance of folded/structured regions of proteins to proteolytic degradation, the susceptibility of denatured proteins to processing, and the higher affinity of conformationally constrained 'extended' inhibitors/substrates for proteases. Other approaches to extended inhibitor conformations should similarly lead to high-affinity binding to a protease.

Using siting algorithms in the design of marine reserve networks

Using benthic habitat data from the Florida Keys (USA), we demonstrate how siting algorithms can help identify potential networks of marine reserves that comprehensively represent target habitat types. We applied a flexible optimization tool-simulated annealing-to represent a fixed proportion of different marine habitat types within a geographic area. We investigated the relative influence of spatial information, planning-unit size, detail of habitat classification, and magnitude of the overall conservation goal on the resulting network scenarios. With this method, we were able to identify many adequate reserve systems that met the conservation goals, e.g., representing at least 20% of each conservation target (i.e., habitat type) while fulfilling the overall aim of minimizing the system area and perimeter. One of the most useful types of information provided by this siting algorithm comes from an irreplaceability analysis, which is a count of the number of, times unique planning units were included in reserve system scenarios. This analysis indicated that many different combinations of sites produced networks that met the conservation goals. While individual 1-km(2) areas were fairly interchangeable, the irreplaceability analysis highlighted larger areas within the planning region that were chosen consistently to meet the goals incorporated into the algorithm. Additionally, we found that reserve systems designed with a high degree of spatial clustering tended to have considerably less perimeter and larger overall areas in reserve-a configuration that may be preferable particularly for sociopolitical reasons. This exercise illustrates the value of using the simulated annealing algorithm to help site marine reserves: the approach makes efficient use of;available resources, can be used interactively by conservation decision makers, and offers biologically suitable alternative networks from which an effective system of marine reserves can be crafted.

Efficiency, costs and trade-offs in marine reserve system design

With marine biodiversity conservation the primary goal for reserve planning initiatives, a site's conservation potential is typically evaluated on the basis of the biological and physical features it contains. By comparison, socio-economic information is seldom a formal consideration of the reserve system design problem and generally limited to an assessment of threats, vulnerability or compatibility with surrounding uses. This is perhaps surprising given broad recognition that the success of reserve establishment is highly dependent on widespread stakeholder and community support. Using information on the spatial distribution and intensity of commercial rock lobster catch in South Australia, we demonstrate the capacity of mathematical reserve selection procedures to integrate socio-economic and biophysical information for marine reserve system design. Analyses of trade-offs highlight the opportunities to design representative, efficient and practical marine reserve systems that minimise potential loss to commercial users. We found that the objective of minimising the areal extent of the reserve system was barely compromised by incorporating economic design constraints. With a small increase in area (< 3%) and boundary length (< 10%), the economic impact of marine reserves on the commercial rock lobster fishery was reduced by more than a third. We considered also how a reserve planner might prioritise conservation areas using information on a planning units selection frequency. We found that selection frequencies alone were not a reliable guide for the selection of marine reserve systems, but could be used with approaches such as summed irreplaceability to direct conservation effort for efficient marine reserve design.

Establishing representative no-take areas in the Great Barrier Reef : Large scale implementation of theory on marine protected areas

The Great Barrier Reef Marine Park, an area almost the size , of Japan, has a new network of no-take areas that significantly improves the protection of biodiversity. The new marine park zoning implements, in a quantitative manner, many of the theoretical design principles discussed in the literature. For example, the new network of no-take areas has at least 20% protection per bioregion, minimum levels of protection for all known habitats and special or unique features, and minimum sizes for no-take areas of at least 10 or 20 kat across at the smallest diameter Overall, more than 33% of the Great Barrier Reef Marine Park is now in no-take areas (previously 4.5%). The steps taken leading to this outcome were to clarify to the interested public why the existing level of protection wets inadequate; detail the conservation objectives of establishing new no-take areas; work with relevant and independent experts to define, and contribute to, the best scientific process to deliver on the objectives; describe the biodiversity (e.g., map bioregions); define operational principles needed to achieve the objectives; invite community input on all of The above; gather and layer the data gathered in round-table discussions; report the degree of achievement of principles for various options of no-take areas; and determine how to address negative impacts. Some of the key success factors in this case have global relevance and include focusing initial communication on the problem to be addressed; applying the precautionary principle; using independent experts; facilitating input to decision making; conducting extensive and participatory consultation; having an existing marine park that encompassed much of the ecosystem; having legislative power under federal law; developing high-level support; ensuring agency Priority and ownership; and being able to address the issue of displaced fishers.

Sketch design drawing, Hill House, Chapel Hill, Brisbane

Sketch floor plan and diagrams.

Sketch design drawing, Hill House, Chapel Hill, Brisbane

Three-dimensional projection sketch showing roof forms and wall finishes.

Hydraulic Design of Stepped Spillways and Downstream Energy Dissipators for Embankment Dams

In recent years, the design flows of many dams were re-evaluated, often resulting in discharges larger than the original design. In many cases, the occurrence of the revised flows could result in dam overtopping because of insufficient storage and spillway capacity. An experimental study was conducted herein to gain a better understanding of the flow properties in stepped chutes with slopes typical of embankment dams. The work was based upon a Froude similitude in large-size experimental facilities. A total of 10 configurations were tested including smooth steps, steps equipped with devices to enhance energy dissipation and rough steps. The present results yield a new design procedure. The design method includes some key issues not foreseen in prior studies : e.g., gradually varied flow, type of flow regime, flow resistance. It is believed that the outcomes are valid for a wide range of chute geometry and flow conditions typical of embankment chutes.

Design, Science and Conceptual Analysis

Philosophers expend considerable effort on the analysis of concepts, but the value of such work is not widely appreciated. This paper principally analyses some arguments, beliefs, and presuppositions about the nature of design and the relations between design and science common in the literature to illustrate this point, and to contribute to the foundations of design theory.

Research, knowledge and design

The discussion about relations between research and design has a number of strands, and presumably motivations. Putting aside the question whether or not design or “creative endeavour” should be counted as research, for reasons to do with institutional recognition or reward, the question remains how, if at all, is design research? This question is unlikely to have attracted much interest but for matters external to Architecture within the modern university. But Architecture as a discipline now needs to understand research much better than in the past when ‘research’ was whatever went on in building science, history or people/environment studies. In this paper, I begin with some common assumptions about design, considered in relation to research, and suggest how the former can constitute or be a mode of the latter. Central to this consideration is an understanding of research as the production of publicly available knowledge. The method is that of conceptual analysis which is much more fruitful than is usually appreciated. This work is part of a larger project in philosophy of design, in roughly the analytical tradition.