A novel method for selection of chymotrypsin inhibitors from a phage peptide library

A novel screening strategy has been developed for the identification of alpha-chymotrypsin inhibitors from a phage peptide library. In this strategy, the standard affinity selection protocol was modified by adding a proteolytic cleavage period to avoid recovery of alpha-chymotrypsin substrates. After four cycles of selection and further activity assay, a group of related peptides were identified by DNA sequencing. These peptides share a consensus sequence motif as (S/T)RVPR(R/H). Then, a corresponding short peptide (Ac-ASRVPRRG-NH2) was synthesized chemically and proved to be an inhibitor of alpha-chymotrypsin. The present work provides a useful way for searching proteinase inhibitors without detailed knowledge of the molecular structure.

Selection of a strain of Aspergillus for the production of citric acid from pineapple waste in solid-state fermentation

Aspergillus foetidus ACR I 3996 (=FRR 3558) and three strains of Aspergillus niger ACM 4992 (=ATCC 9142), ACM 4993 (=ATCC 10577), ACM 4994 (=ATCC 12846) were compared for the production of citric acid from pineapple peel in solid-state fermentation. A. niger ACM 4992 produced the highest amount of citric acid, with a yield of 19.4 g of citric acid per 100 g of dry fermented pineapple waste under optimum conditions, representing a yield of 0.74 g citric acid/g sugar consumed. Optimal conditions were 65% (w/w) initial moisture content, 3% (v/w) methanol, 30 degrees C, an unadjusted initial pH of 3.4, a particle size of 2 mm and 5 ppm Fe2+. Citric acid production was best in flasks, with lower yields being obtained in tray and rotating drum bioreactors.

Anthropometric-based selection and sprint kayak training in children

A 12 week kayak training programme was evaluated in children who either had or did not have the anthropometric characteristics identified as being unique to senior elite sprint kayakers. Altogether, 234 male and female school children were screened to select 10 children with and 10 children without the identified key anthropometric characteristics. Before and after training, the children completed an all-out 2 min kayak ergometer simulation test; measures of oxygen consumption, plasma lactate and total work accomplished were recorded. In addition, a 500 m time trial was performed at weeks 3 and 12. The coaches were unaware which 20 children possessed those anthropometric characteristics deemed to favour development of kayak ability. All children improved in both the 2 min ergometer simulation test and 500 m time trial. However, boys who were selected according to favourable anthropometric characteristics showed greater improvement than those without such characteristics in the 2 min ergometer test only. In summary, in a small group of children selected according to anthropometric data unique to elite adult kayakers, 12 weeks of intensive kayak training did not influence the rate of improvement of on-water sprint kayak performance.

Effects of egg load on the host-selection behaviour of Helicoverpa armigera (Hubner) (Lepidoptera : Noctuidae)

We examined the effect of age-specific fecundity, mated status, and egg load on host-plant selection, by Helicoverpa armigera under laboratory conditions. The physiological state of a female moth (number of mature eggs produced) greatly influences her host-plant specificity and propensity to oviposit (oviposition motivation). Female moths were less discriminating against cowpea (a low-ranked host) relative to maize (a high-ranked host) as egg load increased. Similarly, increased egg load led to a greater propensity to oviposit on both cowpea and maize. Distribution of oviposition with age of mated females peaked shortly after mating and declined steadily thereafter until death. Most mated females (88%) carried only a single spermatophore, a few females (12%) contained two. The significance of these findings in relation to host-plant selection by H. armigera, and its management, are discussed.

Effects of data characteristics on the results of reserve selection algorithms

We tested the effects of four data characteristics on the results of reserve selection algorithms. The data characteristics were nestedness of features (land types in this case), rarity of features, size variation of sites (potential reserves) and size of data sets (numbers of sites and features). We manipulated data sets to produce three levels, with replication, of each of these data characteristics while holding the other three characteristics constant. We then used an optimizing algorithm and three heuristic algorithms to select sites to solve several reservation problems. We measured efficiency as the number or total area of selected sites, indicating the relative cost of a reserve system. Higher nestedness increased the efficiency of all algorithms (reduced the total cost of new reserves). Higher rarity reduced the efficiency of all algorithms (increased the total cost of new reserves). More variation in site size increased the efficiency of all algorithms expressed in terms of total area of selected sites. We measured the suboptimality of heuristic algorithms as the percentage increase of their results over optimal (minimum possible) results. Suboptimality is a measure of the reliability of heuristics as indicative costing analyses. Higher rarity reduced the suboptimality of heuristics (increased their reliability) and there is some evidence that more size variation did the same for the total area of selected sites. We discuss the implications of these results for the use of reserve selection algorithms as indicative and real-world planning tools.

A rapid selection/amplification procedure for high-level expression of recombinant protein in a metal-amplifiable mammalian expression system

We describe a strategy for the selection and amplification of foreign gene expression in Chinese hamster ovary (CHO) cells employing a metallothionein gene-containing expression vector. This report describes an amplification procedure that results in an enrichment of clones exhibiting high levels of recombinant protein production and reduces the labour required for screening recombinant cell lines.

Life-history consequences of divergent selection on egg size in Drosophila melanogaster

Life histories are generally assumed to evolve via antagonistic pleiotropy (negative genetic correlations) among traits, and trade-offs between life-history traits are typically studied using either phenotypic manipulations or selection experiments. We investigated the trade-off between egg size and fecundity in Drosophila melanogaster by examining both the phenotypic and genetic relationships between these traits after artificial selection for large and small eggs, relative to female body size. Egg size responded strongly to selection in both directions, increasing in the large-egg selected lines and decreasing in the small-egg selected lines. Phenotypic correlations between egg size and fecundity in the large-egg selected lines were negative, but no relationship between these traits occurred in either the control or small-egg selected lines. There was no negative genetic correlation between egg size and fecundity. Total reproductive allocation decreased in the small-egg selected lines but did not increase in the large-egg lines. Our results have three implications. First, our selection procedure may have forced females selected for large eggs into a physiological trade-off not reflected in a negative genetic correlation between these traits. Second, the lack of a negative genetic correlation between egg size and number suggests that the phenotypic trade-off frequently observed between egg size and number in other organisms may not evolve over the short term via a direct genetic trade-off whereby increases in egg size are automatically accompanied by decreased fecundity. Finally, total reproductive allocation may not evolve independently of egg size as commonly assumed.

Role of chemical cues from cotton in mediating host selection and oviposition behaviour in Helicoverpa armigera (Hubner) (Lepidoptera : Noctuidae)

We investigated the role of chemoreception in the host selection and oviposition behaviour of Helicoverpa armigera in the laboratory using five cotton genotypes and synthetic volatile terpenes. Female moths oviposited on substrates treated with methanol, ethanol, acetone and pentane extracts of leaves, squares and flowers of the cotton genotypes. Phytochemicals soluble in pentane were the most efficient in eliciting oviposition behaviour. In a two-way bioassay, pentane extracts of leaves or squares of a Multiple Host-plant Resistance genotype (MHR11), Deltapine commercial (DP90), and Smith Red Leaf (SRL) received significantly more eggs than solvent-treated controls. Extracts of squares of the native genotype Gossypium nelsonii did not receive more eggs. Females preferred DP90 and MHR11 to SRL and G. nelsonii. Female moths also laid more eggs on pentane extracts of MHR11 flowers than MHR11 leaves from preflowering, early flowering and peak-flowering plants. In a flight chamber, female moths used olfactory cues at short range to mediate oviposition and discrimination between host plants. Egg-laying, mated females were attracted at a distance (1.5 m) to volatile compounds released by whole plants and odours emanating from filter papers treated with synthetic volatile terpenes. Individually, the terpenes did not stimulate any significant oviposition response. However, there was a significant oviposition response to a mixture of equal volumes of the terpenes (trans-beta-caryophyllene, alpha-pinene, beta-pinene, myrcene, beta-bisabolol, and alpha-humulene). Conversely, antennectomised (moths with transected antennae), egg-laying, mated females did not stimulate any significant oviposition response. The significance of these findings in relation to H. armigera hostplant selection are discussed.

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.

An object-oriented neural network approach to short-term traffic forecasting

This paper discusses an object-oriented neural network model that was developed for predicting short-term traffic conditions on a section of the Pacific Highway between Brisbane and the Gold Coast in Queensland, Australia. The feasibility of this approach is demonstrated through a time-lag recurrent network (TLRN) which was developed for predicting speed data up to 15 minutes into the future. The results obtained indicate that the TLRN is capable of predicting speed up to 5 minutes into the future with a high degree of accuracy (90-94%). Similar models, which were developed for predicting freeway travel times on the same facility, were successful in predicting travel times up to 15 minutes into the future with a similar degree of accuracy (93-95%). These results represent substantial improvements on conventional model performance and clearly demonstrate the feasibility of using the object-oriented approach for short-term traffic prediction. (C) 2001 Elsevier Science B.V. All rights reserved.

Quasars from a complete spectroscopic survey

With the advent of multi-fibre spectrographs such as the 'Two-Degree Field' (2dF) instrument at the Angle-Australian Telescope, quasar surveys that are free of any preselection of candidates and any biases this implies have become possible for the first time. The first of these is that which is being undertaken as part of the Fornax Spectroscopic Survey, a survey of the area around the Fornax Cluster of galaxies, and aims to obtain the spectra of all objects in the magnitude range 16.5 < b(j) < 19.7. To date, 3679 objects in the central pi -deg(2) area have been successfully identified from their spectral characteristics. Of these, 71 are found to be quasars, 61 with redshifts 0.3 < z < 2.2 and 10 with redshifts z > 2.2. Using this complete quasar sample, a new determination of quasar number counts is made, enabling an independent check of existing quasars surveys. Cumulative counts per square degree at a magnitude limit of b(j) < 19.5 are found to be 11.5 +/- 2.2 for 0.3 < z < 2.2, 2.22 +/- 0.93 for z > 2.2 and 13.7 +/- 3.1 for z > 0.3. Given the likely detection of extra quasars in the Fornax survey, we make a more detailed examination of existing quasar selection techniques. First, looking at the use of a stellar criterion, four of the 71 quasars are 'non-stellar' on the basis of the automated plate measuring facility (APM) b(j) classification, however inspection shows all are consistent with stellar, but misclassified due to image confusion. Examining the ultraviolet excess and multicolour selection techniques, for the selection criteria investigated, ultraviolet excess would find 69 +/- 6 per cent of our 0.3 < z < 2.2 quasars and only 50(-18)(+14), per cent of our z > 2.2 quasars, while the completeness level for multicolour selection is found to be 90(-4)(+3) per cent for 0.3 < z < 2.2 quasars and 80(-12)(+14) per cent for z > 2.2 quasars. The extra quasars detected by our all-object survey thus have unusually red star-like colours, and this appears to be a result of the continuum shape rather than any emission features. An intrinsic dust extinction model may, at least partly, account for the red colours.

Structuring real-time Object-Z specifications

This paper presents a means of structuring specifications in real-time Object-Z: an integration of Object-Z with the timed refinement calculus. Incremental modification of classes using inheritance and composition of classes to form multi-component systems are examined. Two approaches to the latter are considered: using Object-Z's notion of object instantiation and introducing a parallel composition operator similar to those found in process algebras. The parallel composition operator approach is both more concise and allows more general modelling of concurrency. Its incorporation into the existing semantics of real-time Object-Z is presented.

The Fornax Cluster Spectroscopic Survey: a sample of confirmed cluster dwarfs

The Fornax Cluster Spectroscopic Survey (FCSS) project utilizes the Two-degree Field (2dF) multi-object spectrograph on the Anglo-Australian Telescope (AAT). Its aim is to obtain spectra for a complete sample of all 14 000 objects with 16 5 less than or equal to b(j) less than or equal to 19 7 irrespective of their morphology in a 12 deg(2) area centred on the Fornax cluster. A sample of 24 Fornax cluster members has been identified from the first 2dF field (3.1 deg(2) in area) to be completed. This is the first complete sample of cluster objects of known distance with well-defined selection limits. Nineteen of the galaxies (with -15.8 < M-B < 12.7) appear to be conventional dwarf elliptical (dE) or dwarf S0 (dS0) galaxies. The other five objects (with -13.6 < M-B < 11.3) are those galaxies which were described recently by Drinkwater et al. and labelled 'ultracompact dwarfs' (UCDs). A major result is that the conventional dwarfs all have scale sizes alpha greater than or similar to 3 arcsec (similar or equal to300 pc). This apparent minimum scale size implies an equivalent minimum luminosity for a dwarf of a given surface brightness. This produces a limit on their distribution in the magnitude-surface brightness plane, such that we do not observe dEs with high surface brightnesses but faint absolute magnitudes. Above this observed minimum scale size of 3 arcsec, the dEs and dS0s fill the whole area of the magnitude-surface brightness plane sampled by our selection limits. The observed correlation between magnitude and surface brightness noted by several recent studies of brighter galaxies is not seen with our fainter cluster sample. A comparison of our results with the Fornax Cluster Catalog (FCC) of Ferguson illustrates that attempts to determine cluster membership solely on the basis of observed morphology can produce significant errors. The FCC identified 17 of the 24 FCSS sample (i.e. 71 per cent) as being 'cluster' members, in particular missing all five of the UCDs. The FCC also suffers from significant contamination: within the FCSS's field and selection limits, 23 per cent of those objects described as cluster members by the FCC are shown by the FCSS to be background objects.

Response selection deficits in melancholic but not nonmelancholic unipolar major depression

One consistent functional imaging finding from patients with major depression has been abnormality of the anterior cingulate cortex (ACC). Hypoperfusion has been most commonly reported, but some studies suggest relative hyperperfusion is associated with response to somatic treatments. Despite these indications of the possible importance of the ACC in depression there have been relatively few cognitive studies ACC function in patients with major depression. The present study employed a series of reaction time (RT) tasks involving selection with melancholic and nonmelancholic depressed patients, as well as age-matched controls. Fifteen patients with unipolar major depression (7 melancholic, 8 nonmelancholic) and 8 healthy age-matched controls performed a series of response selection tasks (choice RT, spatial Stroop, spatial stimulus-response compatibility (SRC), and a combined Stroop + SRC condition). Reaction time and error data were collected. Melancholic patients were significantly slower than controls on all tasks but were slower than nonmelancholic patients only on the Stroop and Stroop + SRC conditions. Nonmelancholic patients did not differ from the control group on any task. The Stroop task seems crucial in differentiating the two depressive groups, they did not differ on the choice RT or SRC tasks. This may reflect differential task demands, the SRC involved symbolic manipulation that might engage the dorsal ACC and dorsolateral prefrontal cortex (DLPFC) to a greater extent than the, primarily inhibitory, Stroop task which may engage the ventral ACC and orbitofrontal cortex (OFC). This might suggest the melancholic group showed a greater ventral ACC-OFC deficit than the nonmelancholic group, while both groups showed similar dorsal ACC-DLPFC deficit.