A new level of conotoxin diversity, a non-native disulfide bond connectivity in alpha-conotoxin AuIB reduces structural definition but increases biological activity

alpha-Conotoxin AuIB and a disulfide bond variant of AuIB have been synthesized to determine the role of disulfide bond connectivity on structure and activity. Both of these peptides contain the 15 amino acid sequence GCCSYPPCFATNPDC, with the globular (native) isomer having the disulfide connectivity Cys(2-8 and 3-15) and the ribbon isomer having the disulfide connectivity Cys(2-15 and 3-8). The solution structures of the peptides were determined by NAIR spectroscopy, and their ability to block the nicotinic acetylcholine receptors on dissociated neurons of the rat parasympathetic ganglia was examined. The ribbon disulfide isomer, although having a less well defined structure, is surprisingly found to have approximately 10 times greater potency than the native peptide. To our knowledge this is the first demonstration of a non-native disulfide bond isomer of a conotoxin exhibiting greater biological activity than the native isomer.

Cross-language Perception of Word-final Stops by Native Vietnamese Listeners: Preliminary Results on the Role of Specific, Non-native Phonetic Experience

This study examined the discrimination of word-final stop contrasts (/p/-/t/, /p/-/k/, /t/-/k/) in English and Thai by 12 listeners who speak Vietnamese as their first language (L1). Vietnamese shares specific phonetic realization of stops with Thai, i.e., unreleased final stop and differs from English which allows both released and unreleased final stops. These 12 native Vietnamese (NV) listeners’ discrimination accuracy was compared to that of the two listener groups (Australian English (AE), native Thai (NT)) tested in previous studies. The NV group was less accurate than the native group in discriminating both English and Thai stop contrasts. In particular, for the Thai /t/-/k/ contrast, they were significantly less accurate than the AE listeners. The present findings suggest that experience with specific (i.e., unreleased) and native phonetic realization of sounds may be essential in accurate discrimination of final stop contrasts. The effect of L1 dialect on cross-language speech perception is discussed.

Border Patrol in the Borderless World: Negotiating Intercultural Internet Discussion

Public Internet discussion forums appear to offer limitless opportunities for communication across linguistic, geographical and cultural borders. Closer inspection, however, reveals that cordial intercultural exchanges are far from widespread in this genre. And yet, such forums have a great deal to offer the independent language learner in terms of ease of access, potential for meaningful language practice and feedback, and exposure to different cultural conventions. This paper explores obstacles to the participation of advanced language learners in public forums of this kind through an examination of the speaking positions most readily available to the non-native speaker. A case study of sustained intercultural interaction on a public discussion thread suggests an alternative to these. Here a core of Francophone and Anglophone participants manage to negotiate an intercultural identity in order to pursue their communicative goals. The paper traces the way in which participants shift footing to regulate insider and outsider status on the forum and draws conclusions regarding the conditions for successful intercultural exchange in this genre

Norwegian long-distance reflexives: Lectal variation and speaker attitudes

10 lectal variables were examined with respect to Norwegian speakers' acceptance of long-distance reflexives (LDR), using a questionnaire to elicit grammaticality judgements on 50 potential LDR sentences. A sample of 180 speakers completed the questionnaire. The data was analysed using a general linear model univariate model, and Spearman's correlation. In this sample the results showed that dialect and level of education had significant effects on speakers' acceptance of long-distance reflexives, while sex, age, being a native speaker, having both native-speaker parents, living in the city or the country, and the speaker's attitudes to the two Norwegian writing languages had no influence on speakers' acceptance of long-distance reflexives. It is suggested that the influence of Danish on Norwegian writing and on the southern dialects may be the cause of the observed variation with respect to LDR in Norwegian.

The utility and limitations of chloroplast DNA analysis for identifying native British oak stands and for guiding replanting strategy

We report a method using variation in the chloroplast genome (cpDNA) to test whether oak stands of unknown provenance are of native and/or local origin. As an example, a sample of test oaks, of mostly unknown status in relation to nativeness and localness, were surveyed for cpDNA type. The sample comprised 126 selected trees, derived from 16 British seed stands, and 75 trees, selected for their superior phenotype (201 tree samples in total). To establish whether these two test groups are native and local, their cpDNA type was compared with that of material from known autochthonous origin (results of a previous study which examined variation in 1076 trees from 224 populations distributed across Great Britain). In the previous survey of autochthonous material, four cpDNA types were identified as native; thus if a test sample possessed a new haplotype then it could be classed as non-native. Every one of the 201 test samples possessed one of the four cpDNA types found within the autochthonous sample. Therefore none could be proven to be introduced and, on this basis, was considered likely to be native. The previous study of autochthonous material also found that cpDNA variation was highly structured geographically and, therefore, if the cpDNA type of the test sample did not match that of neighbouring autochthonous trees then it could be considered to be non-local. A high proportion of the seed stand group (44.2 per cent) and the phenotypically superior trees (58.7 per cent) possessed a cpDNA haplotype which matched that of the neighbouring autochthonous trees and, therefore, can be considered as local, or at least cannot be proven to be introduced. The remainder of the test sample could be divided into those which did not grow in an area of overall dominance (18.7 per cent of seed stand trees and 28 per cent of phenotypically superior) and those which failed to match the neighbouring autochthonous haplotype (37.1 per cent and 13.3 per cent, respectively). Most of the non-matching test samples were located within 50 km of an area dominated by a matching autochthonous haplotype (96.0 per cent and 93.5 per cent, respectively), and potentially indicates only local transfer. Whilst such genetic fingerprinting tests have proven useful for assessing the origin of stands of unknown provenance, there are potential limitations to using a marker from the chloroplast genome (mostly adaptively neutral) for classifying seed material into categories which have adaptive implications. These limitations are discussed, particularly within the context of selecting adaptively superior material for restocking native forests.

A corpus-based analysis of transfer effects and connected speech processes in Vietnamese English

This paper presents a corpus-based descriptive analysis of the most prevalent transfer effects and connected speech processes observed in a comparison of 11 Vietnamese English speakers (6 females, 5 males) and 12 Australian English speakers (6 males, 6 females) over 24 grammatical paraphrase items. The phonetic processes are segmentally labelled in terms of IPA diacritic features using the EMU speech database system with the aim of labelling departures from native-speaker pronunciation. An analysis of prosodic features was made using ToBI framework. The results show many phonetic and prosodic processes which make non-native speakers’ speech distinct from native ones. The corpusbased methodology of analysing foreign accent may have implications for the evaluation of non-native accent, accented speech recognition and computer assisted pronunciation- learning.

Modal functions of future tenses in French

The paper disputes two influential claims in the Romance Linguistics literature. The first is that the synthetic future tenses in spoken Western Romance are now rivalled, if not supplanted, as temporal functors by the more recently developed GO futures. The second is that these synthetic futures now have modal rather than temporal meanings in spoken Romance. These claims are seen as reflecting a universal cycle of diachronic change, in which verb forms originally expressing modal (or aspectual) values take on future temporal reference, becoming tenses. The new modal meanings supplant the temporal, which are then taken up by new forms. Challenges to this theory for French are raised on the basis of empirical evidence of two sorts. Positively, future tenses in spoken Romance continue to be used with temporal meaning. Negatively, evidence of modal meaning for these forms is lacking. The evidence comes froma corpora of spoken French, native speaker judgements and verb data from a daily broadsheet. Cumulatively, it points to the reverse of the claims noted above: the synthetic future in spoken French has temporal but little modal meaning.

Structure determination of the three disulfide bond isomers of alpha-conotoxin GI: A model for the role of disulfide bonds in structural stability

The three possible disulfide bonded isomers of alpha-conotoxin GI have been selectively synthesised and their structures determined by H-1 NMR spectroscopy. alpha-Conotoxin GI derives from the venom of Conus geographus and is a useful neuropharmacological tool as it selectively binds to the nicotinic acetylcholine receptor (nAChR), a ligand-gated ion channel involved in nerve signal transmission. The peptide has the sequence ECCNPACGRHYSC-NH2, and the three disulfide bonded isomers are referred to as GI(2-7;3-13), GI(2-13;3-7) and GI(2-3;7-13). The NMR structure for the native isomer GI(2-7;3-13) is of excellent quality, with a backbone pairwise RMSD of 0.16 Angstrom for a family of 35 structures, and comprises primarily a distorted 3(10),, helix between residues 5 to 11. The two non-native isomers exhibit multiple conformers in solution, with the major populated forms being different in structure both from each other and from the native form. Structure-activity relationships for the native GI(2-7;3-13) as well as the role of the disulfide bonds on folding and stability of the three isomers are examined. It is concluded that the disulfide bonds in alpha-conotoxin GI play a crucial part in determining both the structure and stability of the peptide. A trend for increased conformational heterogeneity was observed in the order of GI(2-7;3-13) < GI(2-13;3-7) < GI(2-3;7-13). It was found that the peptide bond joining Cys2 to Cys3 in GI(2-3;7-13) is predominantly trans, rather than cis as theoretically predicted. These structural data are used to interpret the varying nAChR binding of the non-native forms. A model for the binding of native GI(2-7;3-13) to the mammalian nAChR is proposed, with an alpha-subunit binding face made up of Cys2, Asn4, Pro5, Ala6 and Cys7 and a selectivity face, comprised of Arg9 and His10. These two faces orient the molecule between the alpha and delta subunits of the receptor. The structure of the CCNPAC sequence of the native GI(2-7;3-13) is compared to the structure of the identical sequence from the toxic domain of heat-stable enterotoxins, which forms part of the receptor binding region of the enterotoxins, but which has a different disulfide connectivity. (C) 1998 Academic Press Limited.

Cultural aspects of classroom effectiveness: Overseas-educated students in Australian classrooms

Teachers who are new to the country often find themselves as 'the stranger' in their own classroom. Languages education is one area where such overseas-educated teachers are common. The study reported here investigated what cultural factors might influence the classroom performance of such teachers. The early classroom experience of beginning Japanese native speaker teachers and trainees was examined to this end.

Increasing the efficacy of Judas goats by sterilisation and pregnancy termination

The use of Judas goats to locate remnant animals is a potentially powerful tool for enhancing goat-eradication efforts, which are especially important to island conservation. However, current Judas goat methodology falls short of its potential efficacy. Female Judas goats are often pregnant at the time of deployment or become impregnated in the field; pregnant females leave associated goats to give birth, causing downtime of Judas goat operations. Further, male Judas goats may inseminate remnant females. Sterilising Judas goats prior to deployment removes these inefficiencies. Here, we describe two methods (epididymectomy for males and tubal occlusion for females) that sterilise Judas goats while still maintaining sexual motivation and other behaviours associated with intact animals. These surgeries are straightforward, time efficient, and may be conducted in the field by staff with minimal training. Given the widespread and deleterious impacts of non-native herbivores to ecosystems and the importance of Judas operations in detecting animals at low densities, sterilisation and termination of pregnancy should be applied routinely in Judas goat (and possibly other species) programs to increase the efficacy of low-density control operations and eradication campaigns.

Ecology of Leptocoris Hahn (Hemiptera: Rhopalidae) Soapberry Bugs in Australia

Soapberry bugs are worldwide seed predators of plants in the family Sapindaceae. Australian sapinds are diverse and widespread, consisting of about 200 native trees and shrubs. This flora also includes two introduced environmental weeds, plus cultivated lychee (Litchi chinensis Sonn.), longan (Dimocarpus longan Lour.) and rambutan (Nephelium lappaceum L.). Accordingly, Australian soapberry bugs may be significant in ecology, conservation and agriculture. Here we provide the first account of their ecology. We find five species of Leptocoris Hahn in Australia, and list sapinds that do and do not serve as reproductive hosts. From museum and field records we map the continental distributions of the insects and primary hosts. Frequency of occupation varies among host species, and the number of hosts varies among the insects. In addition, differences in body size and beak length are related to host use. For example, the long-beaked Leptocoris tagalicus Burmeister is highly polyphagous in eastern rainforests, where it occurs on at least 10 native and non-native hosts. It aggregates on hosts with immature fruit and commences feeding before fruits dehisce. Most of its continental range, however, matches that of a single dryland tree, Atalaya hemiglauca F. Muell., which has comparatively unprotected seeds. The taxon includes a smaller and shorter-beaked form that is closely associated with Atalaya, and appears to be taxonomically distinct. The other widespread soapberry bug is the endemic Leptocoris mitellatus Bergroth. It too is short-beaked, and colonises hosts phenologically later than L. tagalicus, as seeds become more accessible in open capsules. Continentally its distribution is more southerly and corresponds mainly to that of Alectryon oleifolius Desf. Among all host species, the non-native environmental weeds Cardiospermum L. and Koelreuteria Laxm. are most consistently attacked, principally by L. tagalicus. These recent host shifts have biocontrol implications. In contrast, the sapinds planted as fruit crops appear to be less frequently used at present and mainly by the longer-beaked species.

A modelling approach to estimate the effect of exotic pollinators on exotic weed population dynamics: bumblebees and broom in Australia

The role of mutualisms in contributing to species invasions is rarely considered, inhibiting effective risk analysis and management options. Potential ecological consequences of invasion of non-native pollinators include increased pollination and seed set of invasive plants, with subsequent impacts on population growth rates and rates of spread. We outline a quantitative approach for evaluating the impact of a proposed introduction of an invasive pollinator on existing weed population dynamics and demonstrate the use of this approach on a relatively data-rich case study: the impacts on Cytisus scoparius (Scotch broom) from proposed introduction of Bombus terrestris. Three models have been used to assess population growth (matrix model), spread speed (integrodifference equation), and equilibrium occupancy (lattice model) for C. scoparius. We use available demographic data for an Australian population to parameterize two of these models. Increased seed set due to more efficient pollination resulted in a higher population growth rate in the density-independent matrix model, whereas simulations of enhanced pollination scenarios had a negligible effect on equilibrium weed occupancy in the lattice model. This is attributed to strong microsite limitation of recruitment in invasive C. scoparius populations observed in Australia and incorporated in the lattice model. A lack of information regarding secondary ant dispersal of C. scoparius prevents us from parameterizing the integrodifference equation model for Australia, but studies of invasive populations in California suggest that spread speed will also increase with higher seed set. For microsite-limited C. scoparius populations, increased seed set has minimal effects on equilibrium site occupancy. However, for density-independent rapidly invading populations, increased seed set is likely to lead to higher growth rates and spread speeds. The impacts of introduced pollinators on native flora and fauna and the potential for promoting range expansion in pollinator-limited 'sleeper weeds' also remain substantial risks.

Processing of mutated human proinsulin to mature insulin in the non-endocrine cell line, CHO

Heterologous genes encoding proproteins, including proinsulin, generally produce mature protein when expressed in endocrine cells while unprocessed or partially processed protein is produced in non-endocrine cells. Proproteins, which are normally processed in the regulated pathway restricted to endocrine cells, do not always contain the recognition sequence for cleavage by furin, the endoprotease specific to the constitutive pathway, the principal protein processing pathway in non-endocrine cells. Human proinsulin consists of B-Chain-C-peptide-A-Chain and cleavage at the B/C and C/A junctions is required for processing. The B/C, but not the C/A junction, is recognised and cleaved in the constitutive pathway. We expressed a human proinsulin and a mutated proinsulin gene with an engineered furin recognition sequence at the C/A junction and compared the processing efficiency of the mutant and native proinsulin in Chinese Hamster Ovary cells. The processing efficiency of the mutant proinsulin was 56% relative to 0.7% for native proinsulin. However, despite similar levels of mRNA being expressed in both cell lines, the absolute levels of immunoreactive insulin, normalized against mRNA levels, were 18-fold lower in the mutant proinsulin-expressing cells. As a result, there was only a marginal increase in absolute levels of insulin produced by these cells. This unexpected finding may result from preferential degradation of insulin in non-endocrine cells which lack the protection offered by the secretory granules found in endocrine cells.

Human susceptibility to Schistosoma japonicum in China correlates with antibody isotypes to native antigens

Antibody isotypic responses (IgE, IgA, IgG1, IgG2, IgG3 and IgG4) to Schistosoma japonicum antigens-adult worm (AWA), soluble egg (SEA) and the recombinant proteins TEG (22.6-kDa tegumental antigen, Sj22) and PMY (paramyosin, Sj97)-were measured (in 1998) in a cohort of 179 Chinese subjects 2 years post-treatment. Subjects in the highest intensity re-infection group (> 100 eggs per gram faeces) had significantly higher levels of IgG1 and IgG4 against AWA. Analysis of IgG4/IgE ratios for AWA and SEA linked IgG4 excess to re-infection and IgE excess to non-re-infection. Two years after chemotherapeutic cure, 29 subjects, who were re-infected or never infected but highly water-exposed, were classified as epidemiologically susceptible (n = 15) or epidemiologically insusceptible to infection (n = 14). IgG4 levels against native antigens (AWA and SEA) were higher in susceptibles and IgE levels were higher in insusceptibles but antibody responses to the recombinant proteins (PMY and TEG) showed no clear pattern or difference between susceptibility groups. These and earlier findings provide evidence that immunity develops against schistosomiasis japonica in China and that susceptibility/resistance correlates with antibody isotypes against native schistosome antigens.

Yorta Yorta: The effect of changes in traditional laws and customs in native title determinations

Members of the Yorta Yorta Aboriginal Community v State of Victoria was the first case in which a claim for native title was lodged in a non-remote area of the Australian mainland which was the subject of European settlement at an early stage in Australian history - highlights the difficulties in establishing native title claims in long settled regions of Australia - a failure to recognise the strength of oral tradition in establishing Aboriginal connection with the land.