Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.

Communicating with the community about grieving: A description and review of the foundations of a broken leg analogy of grieving

With loss permeating the lives of all people throughout the entire life span and its potential for serious long-term deleterious effects, providing effective care for those experiencing situations of loss would be an important element of any preventive community-based mental health promotion approach. The low use of mental health services by those confronted with loss, and the lack of such services in many areas, makes it imperative to enhance the competency of the broader community to provide appropriate care during times of loss. Contributory to such care is a sound knowledge of the literature concerning loss and grieving. The broken leg analogy of grief presented in this article integrates the traditional and emerging grief theories and empirical evidence into an analogy of grieving that can be easily communicated to the general community, is relevant to many losses. and vet upholds the differing positions concerning grieving held by various schools of psychiatric thought.

Analysis of the mouse transcriptome based on functional annotation of 60,770 full-length cDNAs

Only a small proportion of the mouse genome is transcribed into mature messenger RNA transcripts. There is an international collaborative effort to identify all full-length mRNA transcripts from the mouse, and to ensure that each is represented in a physical collection of clones. Here we report the manual annotation of 60,770 full-length mouse complementary DNA sequences. These are clustered into 33,409 'transcriptional units', contributing 90.1% of a newly established mouse transcriptome database. Of these transcriptional units, 4,258 are new protein-coding and 11,665 are new non-coding messages, indicating that non-coding RNA is a major component of the transcriptome. 41% of all transcriptional units showed evidence of alternative splicing. In protein-coding transcripts, 79% of splice variations altered the protein product. Whole-transcriptome analyses resulted in the identification of 2,431 sense-antisense pairs. The present work, completely supported by physical clones, provides the most comprehensive survey of a mammalian transcriptome so far, and is a valuable resource for functional genomics.

DNA vaccine coding for the full-length infectious Kunjin virus RNA protects mice against the New York strain of West Nile virus

A plasmid DNA directing transcription of the infectious full-length RNA genome of Kunjin (KUN) virus in vivo from a mammalian expression promoter was used to vaccinate mice intramuscularly. The KUN viral cDNA encoded in the plasmid contained the mutation in the NS1 protein (Pro-250 to Leu) previously shown to attenuate KUN virus in weanling mice. KUN virus was isolated from the blood of immunized mice 3-4 days after DNA inoculation, demonstrating that infectious RNA was being transcribed in vivo; however, no symptoms of virus-induced disease were observed. By 19 days postimmunization, neutralizing antibody was detected in the serum of immunized animals. On challenge with lethal doses of the virulent New York strain of West Nile (WN) or wild-type KUN virus intracerebrally or intraperitoneally, mice immunized with as little as 0.1-1 mug of KUN plasmid DNA were solidly protected against disease. This finding correlated with neutralization data in vitro showing that serum from KUN DNA-immunized mice neutralized KUN and WN,viruses with similar efficiencies. The results demonstrate that delivery of an attenuated but replicating KUN virus via a plasmid DNA vector may provide an effective vaccination strategy against virulent strains of WN virus.

Rock fracture mean trace length estimation and confidence interval calculation using maximum likelihood methods

There has been a resurgence of interest in the mean trace length estimator of Pahl for window sampling of traces. The estimator has been dealt with by Mauldon and Zhang and Einstein in recent publications. The estimator is a very useful one in that it is non-parametric. However, despite some discussion regarding the statistical distribution of the estimator, none of the recent works or the original work by Pahl provide a rigorous basis for the determination a confidence interval for the estimator or a confidence region for the estimator and the corresponding estimator of trace spatial intensity in the sampling window. This paper shows, by consideration of a simplified version of the problem but without loss of generality, that the estimator is in fact the maximum likelihood estimator (MLE) and that it can be considered essentially unbiased. As the MLE, it possesses the least variance of all estimators and confidence intervals or regions should therefore be available through application of classical ML theory. It is shown that valid confidence intervals can in fact be determined. The results of the work and the calculations of the confidence intervals are illustrated by example. (C) 2003 Elsevier Science Ltd. All rights reserved.

The use of 16s rDNA restriction fragment length polymorphism analysis for the identification of non-fermenting gram negative bacilli in a cystic fibrosis microbiology referral service

The utility of 16s rDNA restriction fragment length polymorphism (RFLP) analysis for the partial genomovar differentiation of Burkholderia cepacia complex bacterium is well documented. We compared the 16s rDNA RFLP signatures for a number of non-fermenting gram negative bacilli (NF GNB) LMG control strains and clinical isolates pertaining to the genera Burkholderia, Pseudomonas, Achromobacter (Alcaligenes), Ralstonia, Stenotrophomonas and Pandoraea. A collection of 24 control strain (LMG) and 25 clinical isolates were included in the study. Using conventional PCR, a 1.2 kbp 16s rDNA fragment was generated for each organism. Following restriction digestion and electrophoresis, each clinical isolate RFLP signature was compared to those of the control strain panel. Nineteen different RFLP signatures were detected from the 28 control strains included in the study. TwentyoneyTwenty- five of the clinical isolates could be classified by RFLP analysis into a single genus and species when compared to the patterns produced by the control strain panel. Four clinical B. pseudomallei isolates produced RFLP signatures which were indistinguishable from B. cepacia genomovars I, III and VIII. The identity of these four isolates were confirmed using B. pseudomallei specific PCR. 16s rDNA RFLP analysis can be a useful identification strategy when applied to NF GNB, particularly for those which exhibit colistin sulfate resistance. The use of this molecular based methodology has proved very useful in the setting of a CF referral laboratory particularly when utilised in conjunction with B. cepacia complex and genomovar specific PCR techniques. Species specific PCR or sequence analysis should be considered for selected isolates; especially where discrepancies between epidemiology, phenotypic and genotypic characteristics occur.

Influence of wearing an unstable shoe on thigh and leg muscle activity and venous response in upright standing

Purpose: To quantify the effect of unstable shoe wearing on muscle activity and haemodynamic response during standing. Methods: Thirty volunteers were divided into 2 groups: the experimental group wore an unstable shoe for 8 weeks, while the control group used a conventional shoe for the same period. Muscle activity of the medial gastrocnemius, tibialis anterior, rectus femoris and biceps femoris and venous circulation were assessed in quiet standing with the unstable shoe and barefoot. Results: In the first measurement there was an increase in medial gastrocnemius activity in all volunteers while wearing the unstable shoe. On the other hand, after wearing the unstable shoe for eight weeks these differences were not verified. Venous return increased in subjects wearing the unstable shoe before and after training. Conclusions: The unstable shoe produced changes in electromyographic characteristics which were advantageous for venous circulation even after training accommodation by the neuromuscular system.

Effects of the length and timing of nighttime naps on task performance and physiological function

OBJECTIVE: To examine the effects of the length and timing of nighttime naps on performance and physiological functions, an experimental study was carried out under simulated night shift schedules. METHODS: Six students were recruited for this study that was composed of 5 experiments. Each experiment involved 3 consecutive days with one night shift (22:00-8:00) followed by daytime sleep and night sleep. The experiments had 5 conditions in which the length and timing of naps were manipulated: 0:00-1:00 (E60), 0:00-2:00 (E120), 4:00-5:00 (L60), 4:00-6:00 (L120), and no nap (No-nap). During the night shifts, participants underwent performance tests. A questionnaire on subjective fatigue and a critical flicker fusion frequency test were administered after the performance tests. Heart rate variability and rectal temperature were recorded continuously during the experiments. Polysomnography was also recorded during the nap. RESULTS: Sleep latency was shorter and sleep efficiency was higher in the nap in L60 and L120 than that in E60 and E120. Slow wave sleep in the naps in E120 and L120 was longer than that in E60 and L60. The mean reaction time in L60 became longer after the nap, and faster in E60 and E120. Earlier naps serve to counteract the decrement in performance and physiological functions during night shifts. Performance was somewhat improved by taking a 2-hour nap later in the shift, but deteriorated after a one-hour nap. CONCLUSIONS: Naps in the latter half of the night shift were superior to earlier naps in terms of sleep quality. However performance declined after a 1-hour nap taken later in the night shift due to sleep inertia. This study suggests that appropriate timing of a short nap must be carefully considered, such as a 60-min nap during the night shift.

Relação entre a unidade de biofeedback de pressão, deslocamento do centro de pressão e atividade eletromiográfica durante o active straight leg raising em indivíduos com e sem dor lombo-pélvica.

Introdução: A unidade de biofeedback de pressão, em indivíduos com dor lombo-pélvica, é utilizada durante exercícios de estabilização segmentar, no entanto ainda carece de evidência científica. Objetivo: Determinar a relação entre a unidade de biofeedback de pressão (UBFP), o deslocamento do centro de pressão no sentido médio-lateral (COPml) e a atividade eletromiográfica abdominal durante o active straight leg raising (ASLR) em indivíduos com e sem dor lombo-pélvica, bem como identificar diferenças entre os grupos. Metodologia: Estudo transversal analítico em 18 estudantes universitários voluntários com dor lombo-pélvica crónica inespecífica (GCD) e em 20 sem dor (GSD). Durante o ASLR (desafio postural dinâmico) foram avaliadas as variações máxima e média da pressão (recorrendo à UBFP) e do deslocamento do COPml (através da plataforma de forças), bem como a atividade muscular abdominal, bilateralmente, com recurso à eletromiografia de superfície. Estatisticamente recorreu-se à correlação de Spearman e ao teste Mann-Whitney U, ambos com um nível de significância de 0,05. Resultados: No GCD, ao contrário do GSD, não foi verificada uma relação entre a UBFP e a atividade do transverso abdominal/obliquo interno (TrA/OI) contra-lateral. Correlações moderadas, mas com sentidos opostos, foram evidenciadas em ambos os grupos, entre o deslocamento do COPml e a atividade do TrA/OI contra-lateral. Em ambos os grupos, a UBFP demonstrou estar fortemente correlacionada com o COPml. Não foram observadas diferenças significativas entre os grupos nas variáveis avaliadas. Conclusão: A UBFP, no GCD, não se apresentou relacionada com a atividade do TrA/OI. Contudo, demonstrou uma relação com o deslocamento do COPml, em ambos os grupos, sendo portanto um indicador de estabilidade do tronco e assim, uma ferramenta útil em ambiente clínico. No GCD observou-se que uma maior atividade muscular TrA/OI pressupõe maior deslocamento do COPml, sendo uma relação contrária à verificada no GSD, podendo ser um indicador da perda da sua ação tónica.

Categorical data clustering using a minimum message length criterion

Research on cluster analysis for categorical data continues to develop, new clustering algorithms being proposed. However, in this context, the determination of the number of clusters is rarely addressed. We propose a new approach in which clustering and the estimation of the number of clusters is done simultaneously for categorical data. We assume that the data originate from a finite mixture of multinomial distributions and use a minimum message length criterion (MML) to select the number of clusters (Wallace and Bolton, 1986). For this purpose, we implement an EM-type algorithm (Silvestre et al., 2008) based on the (Figueiredo and Jain, 2002) approach. The novelty of the approach rests on the integration of the model estimation and selection of the number of clusters in a single algorithm, rather than selecting this number based on a set of pre-estimated candidate models. The performance of our approach is compared with the use of Bayesian Information Criterion (BIC) (Schwarz, 1978) and Integrated Completed Likelihood (ICL) (Biernacki et al., 2000) using synthetic data. The obtained results illustrate the capacity of the proposed algorithm to attain the true number of cluster while outperforming BIC and ICL since it is faster, which is especially relevant when dealing with large data sets.

Contribution à l’étude du nannoplacton du leg 12, «sites» 118-119 (Ocean Atlantique)

This paper reports a Scanning Electron Microscopy study of some samples from the leg 12, Deep Sea Drilling Project, sites 118 and 119. The chronostratigraphic distribution, the frequency of the species identified and the datation of the samples studied are presented. In accordance with the calcareous nannofossil zonation proposed by E. MARTINI (1971) the samples from site 118 are ascribed to the Upper Miocene while the samples from site 119 are located between the Lower (NN1) and the Upper Miocene (NN10).