Cardicola Short, 1953 and Braya n. gen. (Digenea : Sanguinicolidae) from five families of tropical Indo-Pacific fishes

A survey of Pacific coral reef fishes for sanguinicolids revealed that two species of Lutjanidae (Lutjanus argentimaculatus, L. bohar), six species of Siganidae (Siganus corallinus, S. fuscescens, S. lineatus, S. margaritiferus, S. punctatus, S. vulpinus), seven species of Chaetodontidae (Chaetodon aureofasciatus, C. citrinellus, C. flavirostris, C. lineolatus, C. reticulatus, C. ulietensis, C. unimaculatus), three species of Scombridae (Euthynnus affinis, Scomberomorus commerson, S. munroi) and three species of Scaridae (Chlorurus microrhinos, Scarus frenatus, S. ghobban) were infected with morphologically similar sanguinicolids. These flukes have a flat elliptical body, a vestigial oral sucker, a single testis, separate genital pores and a post-ovarian uterus. However, these species clearly belong in two genera based on the position of the testis and genital pores. Sanguinicolids from Lutjanidae, Siganidae, Chaetodontidae and Scombridae belong in Cardicola Short, 1953; the testis originates anteriorly to, or at the anterior end of, the intercaecal field and does not extend posteriorly to it, the male genital pore opens laterally to the sinistral lateral nerve chord and the female pore opens near the level of the ootype ( may be anterior, lateral or posterior to it) antero-dextral to the male pore. Those from Scaridae are placed in a new genus, Braya; the testis originates near the posterior end of the intercaecal field and extends posteriorly to it, the male pore opens medially at the posterior end of the body and the female pore opens posterior to the ootype, antero-sinistral to the male pore. The second internal transcribed spacer (ITS2) of ribosomal DNA from these sanguinicolids and a known species, Cardicola forsteri Cribb, Daintith & Munday, 2000, were sequenced, aligned and analysed to test the distinctness of the putative new species. Results from morphological comparisons and molecular analyses suggest the presence of 18 putative species; 11 are described on the basis of combined morphological and molecular data and seven are not because they are characterised solely by molecular sequences or to few morphological specimens (n= one). There was usually a correlation between levels of morphological and genetic distinction in that pairs of species with the greatest genetic separation were also the least morphologically similar. The exception in this regard was the combination of Cardicola tantabiddii n. sp. from S. fuscescens from Ningaloo Reef ( Western Australia) and Cardicola sp. 2 from the same host from Heron Island ( Great Barrier Reef). These two parasite/ host/location combinations had identical ITS2 sequences but appeared to differ morphologically ( however, this could simply be due to a lack of morphological material for Cardicola sp. 2). Only one putative species ( Cardicola sp. 1) was found in more than one location; most host species harboured distinct species in each geographical location surveyed ( for example, S. corallinus from Heron and Lizard Islands) and some ( for example, S. punctatus, S. fuscescens and Chlorurus microrhinos) harboured two species at a single location. Distance analysis of ITS2 showed that nine species from siganids, three from scombrids and five from scarids formed monophyletic clades to the exclusion of sanguinicolids from the other host families. Cardicola milleri n. sp. and C. chaetodontis Yamaguti, 1970 from lutjanids and chaetodontids, respectively, were the only representatives from those families that were sequenced. Within the clade formed by sanguinicolids from Siganidae there wasa further division of species; species from the morphologically similar S. fuscescens and S. margaritiferus formed a monophyletic group to the exclusion of sanguinicolids from all other siganid species.

Molecular systematics of the Philippine malaria vector Anopheles flavirostris

Allozyme and molecular sequence data from the malaria vector Anopheles flavirostris (Ludlow) (Diptera: Culicidae) were analysed from 34 sites throughout the Philippines, including the type locality, to test the hypothesis that this taxon is a single panmictic species. A finer-scaled allozyme study, of mainly Luzon samples, revealed no fixed genetic differences in sympatric sites and only low levels of variation. We obtained data from partial sequences for the internal transcribed spacer 2 (ITS2) (483 bp), the third domain (D3) (330 bp) of the 28S ribosomal DNA subunit and cytochrome c oxidase subunit I (COI) of mitochondrial DNA (261 bp). No sequence variation was observed for ITS2, only a one base pair difference was observed between Philippine and Indonesian D3 sequences and An. flavirostris sequences were unique, confirming their diagnostic value for this taxon. Sixteen COI haplotypes were identified, giving 25 parsimony informative sites. Neighbour-Joining, Maximum Parsimony, Maximum Likelihood and Bayesian phylogenetic analysis of COI sequences for An. flavirostris and outgroup taxa revealed strong branch support for the monophyly of An. flavirostris, thus confirming that Philippine populations of this taxon comprise a single separate species within the Minimus Subgroup of the Funestus Group. Variation in the behaviour of An. flavirostris is likely to be intraspecific rather than interspecific in origin. © 2006 The Royal Entomological Society.

Phylogeny and evolution of anomalous roots in Daviesia (Fabaceae : Mirbelieae)

The phylogeny of the Australian legume genus Daviesia was estimated using sequences of the internal transcribed spacers of nuclear ribosomal DNA. Partial congruence was found with previous analyses using morphology, including strong support for monophyly of the genus and for a sister group relationship between the clade D. pachyloma and the rest of the genus. A previously unplaced bird-pollinated species, anceps + D. D. epiphyllum, was well supported as sister to the only other bird-pollinated species in the genus, D. speciosa, indicating a single origin of bird pollination in their common ancestor. Other morphological groups within Daviesia were not supported and require reassessment. A strong and previously unreported sister clade of Daviesia consists of the two monotypic genera Erichsenia and Viminaria. These share phyllode-like leaves and indehiscent fruits. The evolutionary history of cord roots, which have anomalous secondary thickening, was explored using parsimony. Cord roots are limited to three separate clades but have a complex history involving a small number of gains (most likely 0-3) and losses (0-5). The anomalous structure of cord roots ( adventitious vascular strands embedded in a parenchymatous matrix) may facilitate nutrient storage, and the roots may be contractile. Both functions may be related to a postfire resprouting adaptation. Alternatively, cord roots may be an adaptation to the low-nutrient lateritic soils of Western Australia. However, tests for association between root type, soil type, and growth habit were equivocal, depending on whether the variables were treated as phylogenetically dependent (insignificant) or independent ( significant).

Systematics and evolution of ticks with a list of valid genus and species names

In recent years there has been much progress in our understanding of the phylogeny and evolution of ticks, in particular the hard ticks (Ixodidae). Indeed, a consensus about the phylogeny of the hard ticks has emerged which is quite different to the working hypothesis of 10 years ago. So that the classification reflects our knowledge of ticks, several changes to the nomenclature of ticks are imminent or have been made. One subfamily, the Hyalomminae, should be sunk, while another, the Bothriocrotoninae, has been created (Klompen, Dobson & Barker, 2002). Bothriocrotoninae, and its sole genus Bothriocroton, have been created to house an early-diverging ('basal') lineage of endemic Australian ticks that used to be in the genus Aponomma. The remaining species of the genus Aponomma have been moved to the genus Amblyomma. Thus, the name Aponomma is no longer a valid genus name. The genus Rhipicephalus is paraphyletic with respect to the genus Boophilus. Thus, the genus Boophilus has become a subgenus of the genus Rhipicephalus (Murrell & Barker, 2003). Knowledge of the phylogenetic relationships of ticks has also provided new insights into the evolution of ornateness and of their life cycles, and has allowed the historical zoogeography of ticks to be studied. Finally, we present a list of the 899 valid genus and species names of ticks as of February 2004.

Supervision practices in allied mental health: Relationships of supervision characteristics to perceived impact and job satisfaction

This study examined whether supervision characteristics impacted on mental health practice and morale, and developed a new Supervision Attitude Scale (SAS). Telephone surveys were conducted with a representative sample of 272 staff from public mental health services across Queensland. Although supervision was widely received and positively rated, it had low average intensity, and assessment and training of skills was rarely incorporated. Perceived impact on practice was associated with acquisition of skills and positive attitudes to supervisors, but extent of supervision was related to impact only if it was from within the profession. Intention to resign was unrelated to extent of supervision, but was associated with positive attitudes to supervisors, accessibility, high impact, and empathy or praise in supervision sessions. The SAS had high internal consistency, and its intercorrelations were consistent with it being a measure of relationship positivity. The study supported the role of supervision in retention and in improving practice. It also highlighted supervision characteristics that might be targeted in training, and provided preliminary data on a new measure.

Evidence of multiple transcription initiation and termination sites within the rDNA intergenic spacer and rRNA readthrough transcription in the urochordate Herdmania curvata

Analysis of the structure of the urochordate Herdmania curvata ribosomal DNA intergenic spacer (IGS) and its role in transcription initiation and termination suggests that rRNA gene regulation in this chordate differs from that in vertebrates. A cloned H, curvata IGS is 1881 bp and composed predominantly of two classes of similar repeat sequences that largely alternate in a tandem array. Southern blot hybridization demonstrates that the IGS length variation within an individual and population is largely the result of changes in internal repeat number. Nuclease S1 mapping and primer extension analyses suggest that there are two transcription initiation sites at the 3' end of the most 3' repetitive element; these sites are 6 nucleotides apart. Unlike mouse, Xenopus, and Drosophila, there is no evidence of transcription starting elsewhere in the IGS. Most sequence differences between the promoter repeat and the other internal repeats are in the vicinity of the putative initiation sites. As in Drosophila, nuclease S1 mapping of transcription termination sites suggest that there is not a definitive stop site and a majority of the pre-rRNAs read through a substantial portion of the IGS. Some transcription appears to proceed completely through the promoter repeat into the adjacent rDNA unit. Analysis of oocyte RNA by reverse transcription-polymerase chain reaction (RT-PCR) confirms that readthrough transcription into the adjacent rDNA unit is occurring in some small IGS length variants; there is no evidence of complete readthrough of IGSs larger than 1.0 kb.

Improved high-performance liquid chromatography determination of methotrexate and its major metabolite in plasma using a poly(styrene-divinylbenzene) column

A sensitive high-performance liquid chromatographic assay has been developed for measuring plasma concentrations of methotrexate and its major metabolite, 7-hydroxymethotrexate. Methotrexate and metabolite were extracted from plasma using solid-phase extraction. An internal standard, aminopterin was used. Chromatographic separation was achieved using a 15-cm poly(styrene-divinylbenzene) (PRP-1(R)) column. This column is more robust than a silica-based stationary phase. Post column, the eluent was irradiated with UV light, producing fluorescent photolytic degradation products of methotrexate and the metabolite. The excitation and emission wavelengths of fluorescence detection were at 350 and 435 nm, respectively. The mobile phase consisted of 0.1 M phosphate buffer (pH 6.5), with 6% N,N-dimethylformamide and 0.2% of 30% hydrogen peroxide. The absolute recoveries for methotrexate and 7-hydroxymethotrexate were greater than 86%. Precision, expressed as a coefficient of variation (n=6), was

Reverse transcription of a naturally occurring nonretroviral RNA produces a precise deletion in the majority of its cDNA products

A precise, reproducible deletion made during in vitro reverse transcription of RNA2 from the icosahedral positive-stranded Helicoverpa armigera stunt virus (Tetraviridae) is described. The deletion, located between two hexamer repeats, is a 50-base sequence that includes one copy of the hexamer repeat. Only the Moloney murine leukemia virus reverse transcriptase and its derivative Superscript I, carrying a deletion of the carboxy-terminal RNase H region, showed this response, indicating a template-switching mechanism different from one proposed that involves a RNase H-dependent strand transfer, Superscript II, however, which carries point mutations to reduce RNase H activity, does not cause a deletion. A possible mechanism involves the enzyme pausing at the 3' side of a stem-loop structure and the 3' end of the nascent DNA strand separating from the template and reannealing to the upstream hexamer repeat.

Epidemiological modelling (including economic modelling) and its role in preventive drug therapy

In contrast to curative therapies, preventive therapies are administered to largely healthy individuals over long periods. The risk-benefit and cost-benefit ratios are more likely to be unfavourable, making treatment decisions difficult. Drug trials provide insufficient information for treatment decisions, as they are conducted on highly selected populations over short durations, estimate only relative benefits of treatment and offer little information on risks and costs. Epidemiological modelling is a method of combining evidence from observational epidemiology and clinical trials to assist in clinical and health policy decision-making. It can estimate absolute benefits, risks and costs of long-term preventive strategies, and thus allow their precise targeting to individuals for whom they are safest and most cost-effective. Epidemiological modelling also allows explicit information about risks and benefits of therapy to be presented to patients, facilitating informed decision-making.

Molecular evidence for definition of genera in the Oxylobium group (Fabaceae : Mirbelieae)

The circumscription of Oxylobium and related genera has been problematic for nearly 200 years. Traditional definitions of genera in the group have relied on morphological features of the leaves, flower, and fruit that overlap extensively between genera. Therefore sequences of cpDNA (trnL-F intron and spacer) and nrDNA (ITS) were used to estimate the phylogeny of the group in an attempt to redefine the genera as monophyletic groups. Oxylobium sens. str. was found to be a well supported clade in both data sets, with the inclusion of Mirbelia oxylobioides. No other genus in the group was supported by these data, except Gastrolobium sens. lat. Some species groups within Chorizema, Mirbelia, and Podolobium were supported but relationships among these, Oxylobium and Gastrolobium differed significantly between the chloroplast and nuclear data sets. No group supported by the molecular data had a morphological synapomorphy, not even Oxylobium or Gastrolobium. Therefore it may be necessary to adopt a much broader generic concept in this group than has been done previously. Incongruence between the two molecular data sets, and very short internal basal branches in both, suggest a rapid early radiation in this group, possibly combined with hybridization and lineage sorting.

How the NHMRC got its fingers burnt

On 20 December 1996, Justice J Finn of the Federal Court of Australia handed down his judgment in the case of the Tobacco Institute of Australia (TIA) Ltd and others v the National Health and Medical Research Council (NHMRC) and others. Justice Finn concluded that, in developing its recommendations for control of environmental tobacco smoke, the NHMRC's Working Party on Passive Smoking erred significantly in regard to the consultative procedures that it employed. As the following discussion shows, the legal decision has profound implications for the NHMRC and the provision of expert advice to Australian governments on matters of health and health policy. The discussion has been prepared by three members of the NHMRC Working Party, but reflects their personal views and not necessarily those of the Working Party as a whole or those of the NHMRC.

A sensitive vorticity gauge using rotated porphyroblasts, and its application to rocks adjacent to the Alpine Fault, New Zealand

Variable aspect ratio porphyroblasts deformed in non-coaxial flow. and internally containing rotated relicts of an external foliation, can be used to characterise plane strain flow regimes. The distribution obtained by plotting the orientation of the long axis of such grains, classified by aspect ratio, against the orientation of the internal foliation is potentially a sensitive gauge of both the bulk shear strain (as previously suggested) and kinematic vorticity number. We illustrate the method using rotated biotite porphyroblasts in the Alpine Schist: a sequence of mid-crustal rocks that have been ramped to the surface along the Alpine Fault. a major transpressional plate boundary. Results indicate that, at distances greater than or equal to similar to1 km from the fault, the rocks have undergone a combination of irrotational fattening and dextral-oblique, normal-sense shear, with a bulk shear strain of similar to0.6 and kinematic vorticity number of similar to0.2. The vorticity analysis is compatible with estimates of strongly oblate bulk strain of similar to 75% maximum shortening. Dextral-reverse transpressional flow characterises higher strain S-tectonite mylonite within similar to1 km of the Alpine Fault. These relationships provide insight into the kinematics of flow and distribution of strain in the hangingwall of the Alpine Fault and place constraints on numerical mechanical models for the exhumation of these mid-crustal rocks. (C) 2001 Elsevier Science Ltd. All rights reserved.

Development of ligase-assisted spacer addition for the measurement of microsatellites

Conventional methods for detecting differences in microsatellite repeat lengths rely on electrophoretic fractionation on long denaturing polyacrylamide gels, a time-consuming and labor-intensive method. Therefore, there is a need for the development of new and rapid approaches to routinely detect such length polymorphisms. The advent of techniques allowing the coupling of DNA molecules to solid surfaces has provided new prospects in the area of mutation. We describe here the development and optimization of the ligase-assisted spacer addition (LASA) method, a novel and rapid procedure based on an ELISA format to measure microsatellite repeat lengths. The LASA assay was successfully applied to a set of 11 bird samples to assess its capability as a genotyping method.

Early Australian experience with infliximab, a chimeric antibody against tumour necrosis factor-alpha, in the treatment of Crohn's disease: is its efficacy augmented by steroid-sparing immunosuppressive therapy?

Background: Tumour necrosis factor-alpha (TNF-alpha) plays an important role in the pathology of Crohn's disease. Infliximab, a chimeric antibody against TNF-alpha, has been shown in controlled clinical trials to be effective in two-thirds of patients with refractory or fistulating Crohn's disease. The factors that determine a clinical response in some patients but not others are unknown. Aims: To document the early Australian experience with infliximab treatment for Crohn's disease and to identify factors that may determine a beneficial clinical response. Methods: Gastroenterologists known to have used infliximab for Crohn's disease according to a compassionate use protocol were asked to complete a spreadsheet that included demographic information, Crohn's disease site, severity, other medical or surgical treatments and a global clinical assessment of Crohn's disease outcome, judged by participating physicians as complete and sustained (remission for the duration of the study), complete but unsustained (remission at 4 weeks but not for the whole study) or partial clinical improvement (sustained or unsustained). Results: Fifty-seven patients were able to be evaluated, with a median follow-up time of 16.4 (4-70) weeks, including 23 patients with fistulae. There were 21 adverse events, including four serious events. Fifty-one patients (89%) had a positive clinical response for a median duration (range) of 11 (2-70) weeks. Thirty patients (52%) had a remission at 4 weeks, 10 of whom had remission for longer than 12 weeks. Forty-two per cent of fistulae closed. Sustained remission (P = 0.065), remission at 4 weeks (P = 0.033) and a positive clinical response of any sort (P = 0.004) were more likely in patients on immunosuppressive therapy, despite there being more smelters in this group. Conclusion: This review of the first Australian experience with infliximab corroborates the reported speed and efficacy of this treatment for Crohn's disease. The excellent response appears enhanced by the concomitant use of conventional steroid-sparing immunosuppressive therapy.