The transcriptome of the arbuscular mycorrhizal fungus Glomus intraradices (DAOM 197198) reveals functional tradeoffs in an obligate symbiont.

• The arbuscular mycorrhizal symbiosis is arguably the most ecologically important eukaryotic symbiosis, yet it is poorly understood at the molecular level. To provide novel insights into the molecular basis of symbiosis-associated traits, we report the first genome-wide analysis of the transcriptome from Glomus intraradices DAOM 197198. • We generated a set of 25,906 nonredundant virtual transcripts (NRVTs) transcribed in germinated spores, extraradical mycelium and symbiotic roots using Sanger and 454 sequencing. NRVTs were used to construct an oligoarray for investigating gene expression. • We identified transcripts coding for the meiotic recombination machinery, as well as meiosis-specific proteins, suggesting that the lack of a known sexual cycle in G. intraradices is not a result of major deletions of genes essential for sexual reproduction and meiosis. Induced expression of genes encoding membrane transporters and small secreted proteins in intraradical mycelium, together with the lack of expression of hydrolytic enzymes acting on plant cell wall polysaccharides, are all features of G. intraradices that are shared with ectomycorrhizal symbionts and obligate biotrophic pathogens. • Our results illuminate the genetic basis of symbiosis-related traits of the most ancient lineage of plant biotrophs, advancing future research on these agriculturally and ecologically important symbionts.

Simulating microbial processes in extraterrestrial, aqueous environments.

Finding evidence of life elsewhere in the Solar System is dependent on understanding biotic processes that could occur within potentially habitable environments. Here, we describe a suite of high-pressure flow-through reactors that have been developed to investigate biotic and abiotic processes within simulated sub-surface martian and icy moon environments.

The identification of sulfide oxidation as a potential metabolism driving primary production on late Noachian Mars.

The transition of the martian climate from the wet Noachian era to the dry Hesperian (4.1-3.0 Gya) likely resulted in saline surface waters that were rich in sulfur species. Terrestrial analogue environments that possess a similar chemistry to these proposed waters can be used to develop an understanding of the diversity of microorganisms that could have persisted on Mars under such conditions. Here, we report on the chemistry and microbial community of the highly reducing sediment of Colour Peak springs, a sulfidic and saline spring system located within the Canadian High Arctic. DNA and cDNA 16S rRNA gene profiling demonstrated that the microbial community was dominated by sulfur oxidising bacteria, suggesting that primary production in the sediment was driven by chemolithoautotrophic sulfur oxidation. It is possible that the sulfur oxidising bacteria also supported the persistence of the additional taxa. Gibbs energy values calculated for the brines, based on the chemistry of Gale crater, suggested that the oxidation of reduced sulfur species was an energetically viable metabolism for life on early Mars.

The transfer of unsterilized material from Mars to Phobos: Laboratory tests, modelling and statistical evaluation.

Sample return missions to Phobos are the subject of future exploration plans. Given the proximity of Phobos to Mars, Mars' potential to have supported life, and the possibility of material transfer from Mars to Phobos, careful consideration of planetary protection is required. If life exists, or ever existed, on Mars, there is a possibility that material carrying organisms could be present on Phobos and be collected by a sample return mission such as the Japanese Martian Moons eXplorer (MMX). Here we describe laboratory experiments, theoretical modelling and statistical analysis undertaken to quantify whether the likelihood of a sample from Phobos material containing unsterilized material transferred from Mars is less than 10-6, the threshold to transition between restricted and unrestricted sample return classification for planetary protection. We have created heat, impact and radiation sterilization models based on the Phobos environment, and through statistical analyses investigated the level of sterilization expected for martian material transferred to Phobos. These analyses indicate that radiation is the major sterilization factor, sterilizing the Phobos surface over timescales of millions of years. The specific events of most relevance in the Phobos sample return context are the 'young' cratering events on Mars that result in Zunil-sized craters, which can emplace a large mass of martian material on Phobos, in a short period of time, thus inhibiting the effects of radiation sterilization. Major unknowns that cannot yet be constrained accurately enough are found to drive the results - the most critical being the determination of exact crater ages to statistical certainty, and the initial biological loading on Mars prior to transfer. We find that, when taking a conservative perspective and assuming the best-case scenario for organism survival, for a 100 g sample of the Phobos regolith to be below the planetary protection requirement for unrestricted sample return, the initial biological loading on Mars must be <8.2 × 103cfu kg-1. For the planned MMX mission, a ∼10 g sample to be obtained from a 25-30 mm diameter core as planned would require an initial martian biological loading to be <1.6 × 104cfu kg-1, in order to remain compliant with the planetary protection threshold.

A beta-1,3 glucan sulfate induces resistance in grapevine against Plasmopara viticola through priming of defense responses, including HR-like cell death.

Sulfated laminarin (PS3) has been shown previously to be an elicitor of plant defense reactions in tobacco and Arabidopsis and to induce protection against tobacco mosaic virus. Here, we have demonstrated the efficiency of PS3 in protecting a susceptible grapevine cultivar (Vitis vinifera cv. Marselan) against downy mildew (Plasmopara viticola) under glasshouse conditions. This induced resistance was associated with potentiated H2O2 production at the infection sites, upregulation of defense-related genes, callose and phenol depositions, and hypersensitive response-like cell death. Interestingly, similar responses were observed following P. viticola inoculation in a tolerant grapevine hybrid cultivar (Solaris). A pharmacological approach led us to conclude that both callose synthesis and jasmonic acid pathway contribute to PS3-induced resistance.

Medication use in patients with restless legs syndrome compared with a control population.

Primary restless legs syndrome (RLS) is a sensorimotor disorder causing chronic sleep deprivation in those with moderate to severe symptoms. It has been associated with other medical conditions, such as high blood pressure, depression and attention deficit hyperactive disorder (ADHD). If these conditions are more prevalent for RLS patients, then it would be expected RLS patients would use relatively more of the medications treating these conditions. Current medication use was obtained from 110 RLS patients and 54 age, race and gender-matched local-community controls. Each subject was diagnosed as primary RLS or having no indications for RLS by a clinician board-certified in sleep medicine. The RLS group used more medications than the control group even when medications used for treating RLS were excluded. Significantly more of the RLS patients than controls used anti-depressants, gastro-intestinal (GI) medications and asthma/allergy medications. RLS patients compared with those without RLS are more likely to use medications not related to treating RLS. Moreover they use medications for conditions that have not previously been considered related to RLS, i.e. GI and asthma/allergy conditions.

Glomeromycota communities survive extreme levels of metal toxicity in an orphan mining site.

Abandoned tailing basins and waste heaps of orphan mining sites are of great concern since extreme metal contamination makes soil improper for any human activity and is a permanent threat for nearby surroundings. Although spontaneous revegetation can occur, the process is slow or unsuccessful and rhizostabilisation strategies to reduce dispersal of contaminated dust represent an option to rehabilitate such sites. This requires selection of plants tolerant to such conditions, and optimization of their fitness and growth. Arbuscular mycorrhizal fungi (AMF) can enhance metal tolerance in moderately polluted soils, but their ability to survive extreme levels of metal contamination has not been reported. This question was addressed in the tailing basin and nearby waste heaps of an orphan mining site in southern France, reaching in the tailing basin exceptionally high contents of zinc (ppm: 97,333 total) and lead (ppm: 31,333 total). In order to contribute to a better understanding of AMF ecology under severe abiotic stress and to identify AMF associated with plants growing under such conditions, that may be considered in future revegetation and rhizostabilisation of highly polluted areas, nine plant species were sampled at different growing seasons and AMF root colonization was determined. Glomeromycota diversity was monitored in mycorrhizal roots by sequencing of the ribosomal LSU. This first survey of AMF in such highly contaminated soils revealed the presence of several AMF ribotypes, belonging mainly to the Glomerales, with some examples from the Paraglomerales and Diversisporales. AMF diversity and root colonization in the tailing basin were lower than in the less-contaminated waste heaps. A Paraglomus species previously identified in a polish mining site was common in roots of different plants. Presence of active AMF in such an environment is an outstanding finding, which should be clearly considered for the design of efficient rhizostabilisation processes.

The Vicia faba leghemoglobin gene VfLb29 is induced in root nodules and in roots colonized by the arbuscular mycorrhizal fungus Glomus fasciculatum.

To investigate similarities between symbiotic interactions of broad bean (Vicia faba) with rhizobia and mycorrhizal fungi, plant gene expression induced by both microsymbionts was compared. We demonstrated the exclusive expression of 19 broad bean genes, including VfENOD2, VfENOD5, VfENOD12 and three different leghemoglobin genes, in root nodules. In contrast, the leghemoglobin gene VfLb29 was found to be induced not only in root nodules, but also in broad bean roots colonized by the mycorrhizal fungus Glomus fasciculatum. In uninfected roots, none of the 20 nodulin transcripts investigated was detectable. VfLb29 has an unusually low sequence homology with all other broad bean leghemoglobins as well as with leghemoglobins from other legumes. It can be regarded as a novel kind of leghemoglobin gene not described until now and the induction of which is common to symbiotic interactions of broad bean with both Rhizobium and a mycorrhizal fungus.

Infections with various types of organisms stimulate transcription from a short promoter fragment of the potato gst1 gene.

By histochemical GUS staining, we demonstrate that transcription from a short promoter fragment of the potato gst1 gene is locally induced after infection of a host plant with various types of pathogenic or symbiotic organisms. This regulatory unit is not active in noninfected tissues, except root apices and senescing leaves. Measuring the expression of a fusion between the promoter fragment and the gus gene in transgenic plants, therefore, allows comparison of the induction of defense reactions in different types of plant-microbe interactions, in one and the same plant.

Medicago truncatula ENOD11: a novel RPRP-encoding early nodulin gene expressed during mycorrhization in arbuscule-containing cells.

Leguminous plants establish endosymbiotic associations with both rhizobia (nitrogen fixation) and arbuscular mycorrhizal fungi (phosphate uptake). These associations involve controlled entry of the soil microsymbiont into the root and the coordinated differentiation of the respective partners to generate the appropriate exchange interfaces. As part of a study to evaluate analogies at the molecular level between these two plant-microbe interactions, we focused on genes from Medicago truncatula encoding putative cell wall repetitive proline-rich proteins (RPRPs) expressed during the early stages of root nodulation. Here we report that a novel RPRP-encoding gene, MtENOD11, is transcribed during preinfection and infection stages of nodulation in root and nodule tissues. By means of reverse transcription-polymerase chain reaction and a promoter-reporter gene strategy, we demonstrate that this gene is also expressed during root colonization by endomycorrhizal fungi in inner cortical cells containing recently formed arbuscules. In contrast, no activation of MtENOD11 is observed during root colonization by a nonsymbiotic, biotrophic Rhizoctonia fungal species. Analysis of transgenic Medicago spp. plants expressing pMtENOD11-gusA also revealed that this gene is transcribed in a variety of nonsymbiotic specialized cell types in the root, shoot, and developing seed, either sharing high secretion/metabolite exchange activity or subject to regulated modifications in cell shape. The potential role of early nodulins with atypical RPRP structures such as ENOD11 and ENOD12 in symbiotic and nonsymbiotic cellular contexts is discussed.

A randomized double-blind study of carbamazepine in the treatment of cocaine abuse.

A 12-week, randomized, double-blind, placebo-controlled, fixed-dose outpatient study of carbamazepine (400 mg and 800 mg) in the treatment of cocaine dependence was performed. Data were analyzed with respect to both treatment condition and carbamazepine serum levels. Outcome variables included subject retention, cocaine urinalysis, self-reported cocaine use, cocaine craving, patient and clinical global impressions, the Drug Impairment Rating Scale for Cocaine, and side effects. Compared with placebo, the 400 mg treatment condition exhibited a greater decrease in the rate of positive cocaine urinalyses and a reduction in intensity and duration of craving over the course of the study. Higher serum carbamazepine levels were associated with a lower rate of positive cocaine urinalysis, fewer days of self-reported cocaine use, briefer craving episodes, and greater subject interval retention. The clinical and methodologic implications of these findings and of the study design are discussed.

Phenytoin in the treatment of cocaine abuse: a double-blind study.

OBJECTIVES: To evaluate the effectiveness of phenytoin in the treatment of cocaine abuse. SUBJECTS AND METHODS: A 12-week, double-blind, placebo-controlled outpatient study of phenytoin in the treatment of cocaine abuse was conducted. Sixty cocaine-using subjects were randomly assigned to a daily fixed dose of 300 mg phenytoin or placebo. Forty-four subjects initiated treatment and returned for weekly visits. Primary measures of outcome included weekly quantitative and qualitative cocaine urinalysis, self-reported cocaine use, global functioning and improvement, craving intensity, and subject retention. RESULTS: Cocaine use, as measured both by weekly urinalysis and self-report, was significantly lower in the phenytoin group. The phenytoin group was also rated as significantly less impaired and more improved than the placebo group. Craving intensity was lower in the phenytoin group, but the difference was not statistically significant. Among phenytoin subjects, serum phenytoin levels above 6.0 micrograms/ml were associated with lower rates of positive cocaine urine specimens and longer cocaine-free periods. No differences were observed between groups in study retention. CONCLUSIONS: These findings suggest that phenytoin may be useful in the treatment of cocaine abuse. Further studies are needed to replicate these findings.

DEPRESSED METABOLIC ACTIVITY OF VESICULAR-ARBUSCULAR MYCORRHIZAL FUNGI AFTER FUNGICIDE APPLICATIONS.

Application to soil of either benomyl or captan significantly decreased the growth of vesicular-arbuscular mycorrhizal (VAM) onion (Allium cepa L. cv. Hyper) plants 4 weeks after treatment but non-VAM plants were not affected. Fungal colonization of the onion roots, as indicated by non-vital staining with chlorozole black E, was depressed 2 weeks after fungicide application. However, decreases in metabolically active VAM fungal tissue, revealed by a succinate dehydrogenase assay, could be detected as soon as 3 d after fungicide treatment. There was little difference between the fungicides in their effect on the VAM fungi used. The usefulness of the succinate dehydrogenase assay in predicting effects of fungicide is discussed.

Effects on growth and comparison of root tissue colonization patterns of Eucalyptus viminalis by pathogenic and nonpathogenic strains of Fusarium oxysporum.

Soilborne pathogens, especially Fusarium oxysporum, are responsible for damping-off and root necrosis in Eucalyptus nurseries. New technologies are increasingly considering strategies for plant disease control other than chemical fungicides. Among these, natural fungal antagonists, which are colonizers of the root cortex, are potential biocontrol agents. An in vitro system was used: (1) to test the pathogenic effects of F. oxysporum strain Foeu1 which was recovered from a forest nursery soil; (2) to explore the potential of the nonpathogenic F. oxysporum strain Fo47, which is known for its efficiency in biological control, to suppress damping-off of Eucalyptus seedlings; (3) to compare the patterns of root colonization and host response to invasion by the two Fusarium strains inoculated separately in a time-course study. Root inoculation of E. viminalis with F. oxysporum strain Foeu1 caused damping-off in young seedlings in vitro, whilst disease symptoms were not visible in plants inoculated with F. oxysporum strain Fo47 or when both strains (Foeu1 + Fo47) were inoculated simultaneously. Each strain showed similarities in patterns of root tissue colonization, and in the processes of root penetration and initial colonization. Differential effects on root tissue were observed with fungal development within the cortex: ingress of strain Foeu1 was accompanied by severe host-cell alterations whilst no tissue damage occurred with development of strain Fo47.

Localization of ß (1-3) glucan in walls of the endomycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerd. & Trappe and Acaulospora laevis Gerd. & Trappe during colonization of host roots.

Previous studies showed that cell walls of endomycorrhizal fungi belonging to the Acaulosporaceae and Glomaceae contain ß (1-3) glucan polymers as well as chitin. Indirect immunolabelling with monoclonal and polyclonal antibodies has been used to investigate the distribution of these structural polysaccharides in cell walls of Glomus mosseae (Nicol. & Gerd.) Gerd. & Trappe and Acaulospora laevis Gerd. & Trappe as they interact with pea and tobacco roots, respectively. The (l-3) glucans were detected in the walls of external hyphae, and of hyphal coils and intercellular hyphae developing in outer root tissues. The glucan component was alkali-insoluble but treatment with chitinase resulted in solubilization of most of the ß(1-3) glucans from the fungal wall. A decrease in immunolabelling was associated with thinning out of the hyphal wall as the fungi colonized deeper in the host root, and ß(1-3) glucans could not be detected in walls of intercellular hyphae or arbuscules in the parenchyma cortical tissue. The molecular configuration of cell walls of G. mosseae and A. laevis is discussed in relation to fungal morphogenesis and the symbiotic state in endomycorrhiza.

Common gene expression in Medicago truncatula roots in response to Pseudomonas fluorescens colonization, mycorrhiza development and nodulation.

• Beneficial rhizosphere microorganisms may share similar molecular steps during root colonization. To test this hypothesis, we compared Medicago truncatula Gaertn. gene expression in roots colonized, or not colonized, by Glomus mosseae BEG12, Pseudomonas fluorescens C7R12 or Sinorhizobium meliloti 2011. • Pseudomonas fluorescens C7R12 formed colonies on the surface of M. truncatula roots and colonized root tissues intercellularly and intracellularly in a way similar to that previously described for other plants. • Semiquantitative reverse transcriptase polymerase chain reaction of a set of 12 mycorrhiza upregulated M. truncatula genes revealed different expression profiles in roots 3 weeks after inoculation with P. fluorescens or S. meliloti. Pseudomonas fluorescens colonization activated seven of the plant genes while nodulated root systems showed increased expression in only three genes and five appeared to be downregulated. • This first report of similar gene induction by a fluorescent pseudomonad and a mycorrhizal fungus in roots supports the hypothesis that some plant cell programmes may be shared during root colonization by these beneficial microorganisms. Less similarity existed in expression of the gene set with nodulation by S. meliloti.

Work-related social skills training for people with schizophrenia in Hong Kong.

This article describes a prospective blinded outcome study of a vocational social skills training program developed in Hong Kong for people affected by chronic schizophrenia. The aim was to improve their ability to find and keep a job. Participants were randomly assigned to three groups: a social skills training group with followup support, a social skills training group without followup support, and a comparison group who received standard after-care treatment. Participants who had participated in either of the training groups statistically outperformed those in the comparison group. Those receiving the training plus followup were statistically much more successful at finding and keeping a job than participants in either of the other two groups. A comparatively small amount of followup contact (a monthly group meeting or phone call) for 3 months after the training finished had a very significant effect on participants' success rate.

Goods on which one loses: women and mental health in China.

This article is broadly divided into three sections. The first part deals with the traditional aspects of gender discrimination in China. Before the Communist government came to power in 1949, discrimination against women was institutionalized within all the usual structures of society: family, the economy, education, culture and the political system. It was one of the major policy initiatives of the Communist government to do away with unequal treatment of women. However, it is very easy to demonstrate that significant discrimination against women still exists. The Chinese government argues that this is because of 'remnants of feudal thinking'. Although this may be partly true, there are aspects of current Chinese society that encourage the continuation of this cultural tradition. The second part of the article examines what is known of the epidemiology of mental illness in China with particular reference to gender. As is the case in Western countries, depression and neurotic disorders are diagnosed more frequently in women than in men, although, overall, the prevalence rate is much lower than in Western countries. What is unusual is that schizophrenia, which is diagnosed at roughly equal rates for men and women in Western countries, is diagnosed more frequently in women in China. Despite this, women occupy fewer psychiatric hospital beds and generally receive fewer resources (e.g. health insurance) than men. Suicide rates are very much higher in China than, for instance, in America, and the suicide figures for young, rural women are particularly disturbing. The third part of the article is based on three interviews with women in a psychiatric clinic in Hubei province.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between parenting stress and social support in mothers of children with learning disabilities: a Chinese experience.

This study examined the relationship between parenting stress and social support in a random sample of 100 mothers with children at a school for children with learning disabilities, and a control group of 75 mothers with similarly aged non-handicapped children. Mothers of children at Zhi Ling school were found to have significantly more stress and significantly less support than those in the control group. Parenting stress in the control group was also found to be relatively high when compared with what are considered to be normal levels in the West. No significant correlation was found between total parenting stress and social support. It is suggested that further studies need to differentiate between satisfaction with support that is actually received and the perceived quality of available support.

Value-added outcomes: the use of advanced practice nurses in long-term care facilities.

The purpose of this study was to determine the effect on clinical outcomes for newly admitted nursing home residents when advanced practice gerontological nurses (APNs) worked with staff to implement scientifically based protocols for incontinence, pressure ulcers, depression, and aggressive behavior. Use of APNs in this manner differs from the usual way APNs have been used in nursing homes, in which their primary focus has been to augment the physician's role. The APN treatment was randomly assigned to two nursing homes and usual care was assigned to a third. Trajectories from admission to 6 months revealed that residents with APN input into their care (n = 86) experienced significantly greater improvement or less decline in incontinence, pressure ulcers, and aggressive behavior, and they had higher mean composite trajectory scores compared with residents receiving usual care (n = 111). Significantly less deterioration in affect was noted in cognitively impaired residents in the treatment group. Findings suggest that APNs can be effective links between current scientific knowledge about clinical problems and nursing home staff.