The development and application of a novel safety-catch linker for BOC-based assembly of libraries of cyclic peptides

Cyclic peptides are appealing targets in the drug-discovery process. Unfortunately, there currently exist no robust solid-phase strategies that allow the synthesis of large arrays of discrete cyclic peptides. Existing strategies are complicated, when synthesizing large libraries, by the extensive workup that is required to extract the cyclic product from the deprotection/cleavage mixture. To overcome this, we have developed a new safety-catch linker. The safety-catch concept described here involves the use of a protected catechol derivative in which one of the hydroxyls is masked with a benzyl group during peptide synthesis, thus making the linker deactivated to aminolysis. This masked derivative of the linker allows BOC solid-phase peptide assembly of the linear precursor. Prior to cyclization, the linker is activated and the linear peptide deprotected using conditions commonly employed (TFMSA), resulting in deprotected peptide attached to the activated form of the linker. Scavengers and deprotection adducts are removed by simple washing and filtration. Upon neutralization of the N-terminal amine, cyclization with concomitant cleavage from the resin yields the cyclic peptide in DMF solution. Workup is simple solvent removal. To exemplify this strategy, several cyclic peptides were synthesized targeted toward the somatostatin and integrin receptors. From this initial study and to show the strength of this method, we were able to synthesize a cyclic-peptide library containing over 400 members. This linker technology provides a new solid-phase avenue to access large arrays of cyclic peptides.

Ecology, shell morphology, anatomy and sperm ultrastructure of the caenogastropod Pyrgula annulata, with a discussion of the relationship between the 'Pyrgulidae' and Caspian and Baikalian rissooideans

The composition of the Pyrgulidae and its relationships to other member families of the caenogastropod superfamily Rissooidea are examined after a consideration of new anatomical (including gross anatomy, sperm ultrastructure), conchological (including protoconch features), ecological, biogeographical and palaeontological data and a re-evaluation of existing literature. Pyrgulidae can be distinguished from hydrobiids unequivocally only with the aid of the radula. Sperm ultrastructural features suggest a very close relationship between the Pyrgulidae, the Hydrobiidae and the Bithyniidae (in fact no family-diagnostic sperm characters can be found to separate these three taxa). Based upon neontological and fossil evidence it is likely that pyrgulids represent a Miocene offshoot from a paratethyal hydrobiid lineage. Pyrgulids may also represent the stock from which the baicaliids arose, in which case the Pyrgulidae must be considered a paraphyletic group. The huge biogeographic gap between the Caspian Sea and Lake Baikal is to some extent bridged by the finding of a Neogene pyrgulid from the Altai Mountains. An alternative scenario for the origin of baicaliids is presented.

Wildlife-based tourism & increased support for nature conservation financially and otherwise: evidence from sea-turtle ecotourism at Mon Repos

The arguments of most conservationists supporting ecotourism have been based on the view that it is environmentally friendly as a resource-use and that receipts from it can counter demands to use the natural resources involved for more extractive economic purposes. But wildlife-based ecotourism can also have positive impacts in itself on the willingness of tourists to pay for wildlife conservation, strengthen the pro-conservation attitudes of tourists, and foster personal actions by them that contribute to wildlife conservation. These aspects are explored in this article on the basis of a survey of tourists visiting Mon Repos Beach near Bundaberg, Queensland, for the purpose of watching marine turtles. The results enable several of the conservation impacts of this experience on tourists to be quantified, and highlight important relationships between specific socio-economic variables and the willingness of tourists to pay for the protection of sea turtles. Furthermore, it is shown that the on-site experiences of ecotourists have positive impacts on the willingness of tourists to pay for the conservation of wildlife, and that willingness to pay is sensitive to whether or not wildlife is seen. It is suggested that in situ ecotourism is likely to be a more powerful force for fostering pro-conservation attitudes and actions among visitors than ex situ wildlife-based tourism in aquaria and zoos.

Delayed emergence of bovine leukemia virus after vaccination with a protective cytotoxic T cell-based vaccine

In a previous study eight MHC class I-matched sheep were vaccinated with a minimal cytotoxic T lymphocyte (CTL) peptide epitope vaccine and were challenged with the retrovirus, bovine leukemia virus (BLV). Half the vaccinated animals remained PCR negative after challenge, whereas the remaining half and the placebo group became PCR positive within 4 weeks postchallenge (Hislop AD, Good MF, Mateo L, Gardner J, Gatei MH, Daniel RCW, Meyers BV, Lavin MF, and Suhrbier A: Nat Med 1998; 4: 1193). Here we show that neither epitope mutations nor processing differences explained why half the peptide-vaccinated animals failed to resist the BLV challenge. However, in these animals the development of BLV-induced lymphosarcomas was significantly delayed compared with the placebo group, suggesting a role for CTLs in preventing retrovirus-induced cancers. Importantly, two of the initially protected animals become PCR positive after similar to1.5 years, indicating extended suppression but not elimination of challenge virus by vaccine-induced CTLs. The late emergence of virus could not be explained by epitope escape mutations or the loss of memory CTL responses. We speculate that high levels of effector CTL may be needed to protect animals from a postchallenge viremia and maintenance of such effector CTLs, rather than memory CTLs, may be required to prevent subsequent emergence of virus from latent pools.

Phylogenetic revision of Agapophytinae subf.n. (Diptera : Therevidae) based on molecular and morphological evidence

Agapophytinae subf.n. is a highly diverse lineage of Australasian Therevidae, comprising eight described and two new genera: Agapophytus Guerin-Meneville, Acupalpa Krober, Acraspisa Krober, Belonalys Krober, Bonjeania Irwin & Lyneborg, Parapsilocephala Krober, Acatopygia Krober, Laxotela Winterton & Irwin, Pipinnipons gen.n. and Patanothrix gen.n. A genus-level cladistic analysis of the subfamily was undertaken using sixty-eight adult morphological characters and c. 1000 base pairs of the elongation factor-1 alpha (EF-1 alpha) protein coding gene. The morphological data partition produced three most parsimonious cladograms, whereas the molecular data partition gave a single most parsimonious cladogram, which did not match any of the cladograms found in the morphological analysis. The level of congruence between the data partitions was determined using the partition homogeneity test (HTF) and Wilcoxon signed ranks rest. Despite being significantly incongruent in at least one of the incongruence tests, the partitions were combined in a simultaneous analysis. The combined data yielded a single cladogram that was better supported than that of the individual partitions analysed separately. The relative contributions of the data partitions to support for individual nodes on the combined cladogram were investigated using Partitioned Bremer Support. The level of support for many nodes on the combined cladogram was non-additive and often greater than the sum of support for the respective nodes on individual partitions. This synergistic interaction between incongruent data partitions indicates a common phylogenetic signal in both partitions. It also suggests that criteria for partition combination based solely on incongruence may be misleading. The phylogenetic relationships of the genera are discussed using the combined data. A key to genera of Agapophytinae is presented, with genera diagnosed and figured. Two new genera are described: Patanothrix with a new species (Pat. skevingtoni) and Pat. wilsoni (Mann) transferred from Parapsilocephala, and Pipinnipons with a new species (Pip. kroeberi). Pipinnipons fascipennis (Krober) is transferred from Squamopygin Krober and Pip. imitans (Mann) is transferred from Agapophytus. Agapophytus bicolor (Krober) is transferred from Parapsilocephala. Agapophytus varipennis Mann is synonymised with Aga, queenslandi Krober and Aga. flavicornis Mann is synonymised with Aga. pallidicornis (Krober).

A phylogeny for the genus Isoodon and a range extension for I-obesulus peninsulae based on mtDNA control region and morphology

Short-nosed bandicoots, Isoodon, have undergone marked range contractions since European colonisation of Australia and are currently divided into many subspecies, the validity of which is debated. Discriminant function analysis of morphology and a phylogeny of Isoodon based on mtDNA control region sequences indicate a clear split between two of the three recognised species, I. macrourus and I. obesulus/auratus. However, while all previously recognised taxa within the I. obesulus/auratus group are morphologically distinct, I. auratus and I. obesulus are not phylogenetically distinct for mtDNA. The genetic divergence between I. obesulus and I. auratus (2.6%) is similar to that found among geographic isolates of the former (I. o. obesulus and I. o. peninsulae: 2.7%). Further, the divergence between geographically close populations of two different species (I. o. obesulus from Western Australia and I. a. barrowensis: 1.2%) is smaller than that among subspecies within I. auratus (I. a. barrowensis and I. auratus from northern Western Australia: 1.7%). A newly discovered population of Isoodon in the Lamb Range, far north Queensland, sympatric with a population of I. m. torosus, is shown to represent a range extension of I. o. peninsulae (350 km). It seems plausible that what is currently considered as two species, I. obesulus and I. auratus, was once one continuous species now represented by isolated populations that have diverged morphologically as a consequence of adaptation to the diverse environments that occur throughout their range. The taxonomy of these populations is discussed in relation to their morphological distinctiveness and genetic similarity.

Developing biodegradable mulch films from starch-based polymers

This paper examines the development of starch-based plastics for use as biodegradable mulch film. A variety of starch-based polymers are blended with high performance biodegradable polyester polymers in order to determine the applicability of films to be processed on a film blowing line and to perform well in mulch film field trials. The process of material formulation, film blowing processing and scale-up and performance properties are highlighted for a successful material. Insights into future developments of starch-derived biodegradable polymers are given.

Advances in application of climate prediction in agriculture

Agricultural ecosystems and their associated business and government systems are diverse and varied. They range from farms, to input supply businesses, to marketing and government policy systems, among others. These systems are dynamic and responsive to fluctuations in climate. Skill in climate prediction offers considerable opportunities to managers via its potential to realise system improvements (i.e. increased food production and profit and/or reduced risks). Realising these opportunities, however, is not straightforward as the forecasting skill is imperfect and approaches to applying the existing skill to management issues have not been developed and tested extensively. While there has been much written about impacts of climate variability, there has been relatively little done in relation to applying knowledge of climate predictions to modify actions ahead of likely impacts. However, a considerable body of effort in various parts of the world is now being focused on this issue of applying climate predictions to improve agricultural systems. In this paper, we outline the basis for climate prediction, with emphasis on the El Nino-Southern Oscillation phenomenon, and catalogue experiences at field, national and global scales in applying climate predictions to agriculture. These diverse experiences are synthesised to derive general lessons about approaches to applying climate prediction in agriculture. The case studies have been selected to represent a diversity of agricultural systems and scales of operation. They also represent the on-going activities of some of the key research and development groups in this field around the world. The case studies include applications at field/farm scale to dryland cropping systems in Australia, Zimbabwe, and Argentina. This spectrum covers resource-rich and resource-poor farming with motivations ranging from profit to food security. At national and global scale we consider possible applications of climate prediction in commodity forecasting (wheat in Australia) and examine implications on global wheat trade and price associated with global consequences of climate prediction. In cataloguing these experiences we note some general lessons. Foremost is the value of an interdisciplinary systems approach in connecting disciplinary Knowledge in a manner most suited to decision-makers. This approach often includes scenario analysis based oil simulation with credible models as a key aspect of the learning process. Interaction among researchers, analysts and decision-makers is vital in the development of effective applications all of the players learn. Issues associated with balance between information demand and supply as well as appreciation of awareness limitations of decision-makers, analysts, and scientists are highlighted. It is argued that understanding and communicating decision risks is one of the keys to successful applications of climate prediction. We consider that advances of the future will be made by better connecting agricultural scientists and practitioners with the science of climate prediction. Professions involved in decision making must take a proactive role in the development of climate forecasts if the design and use of climate predictions are to reach their full potential. (C) 2001 Elsevier Science Ltd. All rights reserved.

A segmentation-based and partial-volume-compensated method for an accurate measurement of lateral ventricular volumes on T-1-weighted magnetic resonance images

Lateral ventricular volumes based on segmented brain MR images can be significantly underestimated if partial volume effects are not considered. This is because a group of voxels in the neighborhood of lateral ventricles is often mis-classified as gray matter voxels due to partial volume effects. This group of voxels is actually a mixture of ventricular cerebro-spinal fluid and the white matter and therefore, a portion of it should be included as part of the lateral ventricular structure. In this note, we describe an automated method for the measurement of lateral ventricular volumes on segmented brain MR images. Image segmentation was carried in combination of intensity correction and thresholding. The method is featured with a procedure for addressing mis-classified voxels in the surrounding of lateral ventricles. A detailed analysis showed that lateral ventricular volumes could be underestimated by 10 to 30% depending upon the size of the lateral ventricular structure, if mis-classified voxels were not included. Validation of the method was done through comparison with the averaged manually traced volumes. Finally, the merit of the method is demonstrated in the evaluation of the rate of lateral ventricular enlargement. (C) 2001 Elsevier Science Inc. All rights reserved.

A risk-based system to penalize and reward line management for occupational safety and health performance

Penalizing line management for the occurrence of lost time injuries has in some cases had unintended negative consequences. These are discussed. An alternative system is suggested that penalizes line management for accidents where the combination of the probability of recurrence and the maximum reasonable consequences such a recurrence may have exceeds an agreed limit. A reward is given for prompt effective control of the risk to below the agreed risk limit. The reward is smaller than the penalty. High-risk accidents require independent investigation by a safety officer using analytical techniques. Two case examples are given to illustrate the system. Continuous safety improvement is driven by a planned reduction in the agreed risk limit over time and reward for proactive risk assessment and control.

Reinfection with Schistosoma haematobium following school-based chemotherapy with praziquantel in four highly endemic villages in Côte d'Ivoire

We present the comparative evaluation of school-based chemotherapy with praziquantel on Schistosoma haematobium reinfection patterns, 6, 12, 18 and 24 months after systematic treatment of schoolchildren in four villages of south-central Côte d'Ivoire. At baseline, very high S. haematobium infection prevalences of 88–94% were found in Taabo Village, located adjacent to a large man-made lake, and in Batera and Bodo, where small dams were constructed. In Assinzé, a village with no man-made environmental alterations, the baseline infection prevalence was significantly lower (67%). The parasitological cure rate, assessed 4 weeks after praziquantel administration in the village with the highest prevalence and intensity of infection, was high (82%), and showed a clear association with infection intensity prior to treatment. Six months after chemotherapy, significant reductions in the prevalence and intensity of infection were observed in all villages. However, infection prevalence was again high in Taabo Village (63%) and in Batera (49%). Different patterns of reinfection occurred in the four villages: rapid reinfection in Taabo Village to reach almost baseline infection prevalence 12 months post-treatment; slow but gradual increase in the prevalence and intensity of infection in Bodo; marked increase in prevalence and intensity of infection during the second year of the follow-up in Assinzé; and prevalence and intensity of infection that remained almost constant between 6 and 24 months post-treatment in Batera. Our study confirms that S. haematobium reinfection patterns largely depend on the local epidemiological setting, which is of central importance to tailoring treatment strategies that are well adapted to these different settings.