881 research outputs found

    Effect of Tryptophan Analogs on Derepression of the \u3cem\u3eEscherichia coli\u3c/em\u3e Tryptophan Operon by Indole-3-Propionic Acid

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    The abilities of 14 tryptophan analogs to repress the tryptophan (trp) operon have been studied in Escherichia coli cells derepressed by incubation with 0.25 mM indole-3-propionic acid (IPA). trp operon expression was monitored by measuring the specific activities of anthranilate synthase (EC 4.1.3.27) and the tryptophan synthase (EC 4.2.1.20) β subunit. Analogs characterized by modification or removal of the α-amino group or the α-carboxyl group did not repress the trp operon. The only analogs among this group that appeared to interact with the trp aporepressor were IPA, which derepressed the trp operon, and d-tryptophan. Analogs with modifications of the indole ring repressed the trp operon to various degrees. 7-Methyl-tryptophan inhibited anthranilate synthase activity and consequently derepressed the trp operon. Additionally, 7-methyltryptophan prevented IPA-mediated derepression but, unlike tryptophan, did so in a non-coordinate manner, with the later enzymes of the operon being relatively more repressed than the early enzymes. The effect of 7-methyltryptophan on IPA-mediated derepression was likely not due to the interaction of IPA with the allosteric site of anthranilate synthase, even though feedback-resistant mutants of anthranilate synthase were partially resistant to derepression by IPA. The effect of 7-methyltryptophan on derepression by IPA was probably due to the effect of the analog-aporepressor complex on trp operon expression

    Douglas-Fir Beetle (Dendroctonus pseudotsugae Hopkins, Coleoptera : Scolytidae) Brood Production on Douglas-Fir Defoliated by Western Spruce Budworm (Choristoneura occidentalis Freeman, Lepidoptera : Tortricidae) in Logan Canyon, Utah

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    Douglas-fir beetle brood production was studied on Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) trees defoliated by the western spruce budworm between 1983 and 1985. Tallies were made of the number of attacks, total length and number of egg galleries, number of eggs deposited, number of larval tunnels, number of pupal chambers, and number of emerging beetles (per female and per unit area). Data analysis showed no significant differences among the three years studied. The number of emerging beetles per female parent was 0.59, and emergence per 90 cm2 was 2.32 beetles. Egg, larval, and pupal survivals were 47.5%, 30.0%, and 15.5%, respectively

    Rhinosporidium seeberi: a human pathogen from a novel group of aquatic protistan parasites.

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    Rhinosporidium seeberi, a microorganism that can infect the mucosal surfaces of humans and animals, has been classified as a fungus on the basis of morphologic and histochemical characteristics. Using consensus polymerase chain reaction (PCR), we amplified a portion of the R. seeberi 18S rRNA gene directly from infected tissue. Analysis of the aligned sequence and inference of phylogenetic relationships showed that R. seeberi is a protist from a novel clade of parasites that infect fish and amphibians. Fluorescence in situ hybridization and R. seeberi- specific PCR showed that this unique 18S rRNA sequence is also present in other tissues infected with R. seeberi. Our data support the R. seeberi phylogeny recently suggested by another group. R. seeberi is not a classic fungus, but rather the first known human pathogen from the DRIPs clade, a novel clade of aquatic protistan parasites (Ichthyosporea)

    Comparison of oral and vaginal metronidazole for treatment of bacterial vaginosis in pregnancy: impact on fastidious bacteria

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    <p>Abstract</p> <p>Background</p> <p>Bacterial vaginosis (BV) is a common condition that is associated with preterm birth and acquisition of complex communities of vaginal bacteria that include several fastidious species. Treatment of BV in pregnancy has mixed effects on the risk of preterm delivery, which some hypothesize is due to variable antibiotic efficacy for the fastidious bacteria. Both oral and intravaginal metronidazole can be used to treat bacterial vaginosis in pregnancy, but little is known about the impact of different routes of antibiotic administration on concentrations of fastidious vaginal bacteria.</p> <p>Methods</p> <p>This was a sub-study of a larger randomized trial of oral versus vaginal metronidazole for treatment of BV in pregnancy. Fifty-three women were evaluated, including 30 women who received oral metronidazole and 23 who received intravaginal metronidazole. Bacterial taxon-specific quantitative PCR assays were used to measure concentrations of bacterial vaginosis associated bacterium (BVAB) 1, 2, and 3, <it>Gardnerella vaginalis, Atopobium </it>species, <it>Leptotrichia/Sneathia </it>species, <it>Megasphaera </it>species, and <it>Lactobacillus crispatus </it>before and after antibiotic treatment.</p> <p>Results</p> <p>Concentrations of <it>Leptotrichia </it>and <it>Sneathia </it>spp. and the fastidious Clostridia-like bacterium designated BVAB1 decreased significantly with oral (p = .002, p = .02) but not vaginal therapy (p = .141, p = .126). The fastidious bacterium BVAB3 did not significantly decrease with either treatment. Concentrations of <it>Atopobium </it>spp., reportedly resistant to metronidazole <it>in vitro</it>, dropped significantly with oral (p = .002) and vaginal (p = .001) treatment. There was no significant difference in the magnitude of change in bacterial concentrations between oral and vaginal treatment arms for any of the bacterial species. <it>Lactobacillus crispatus </it>concentrations did not change.</p> <p>Conclusion</p> <p>Both oral and vaginal metronidazole therapy in pregnant women result in a significant decrease in concentrations of most BV-associated anaerobic bacteria, with the exception that <it>Leptotrichia, Sneathia </it>and BVAB1 do not significantly decrease with vaginal metronidazole therapy. These data suggest that the route of antibiotic administration has a minor impact on bacterial eradication in pregnant women with BV.</p> <p>Trail Registration</p> <p>This trial is registered with ClinicalTrials.gov, number NCT00153517</p

    The impact of parent-created motivational climate on adolescent athletes' perceptions of physical self-concept

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    This is a preliminary version of this article. The official published version can be obtained from the link below.Grounded in expectancy-value model (Eccles, 1993) and achievement goal theory (Nicholls, 1989), this study examined the perceived parental climate and its impact on athletes' perceptions of competence and ability. Hierarchical regression analyses with a sample of 237 British adolescent athletes revealed that mothers and fathers' task- and ego-involving climate predicted their son's physical self-concept; the father in particular is the strongest influence in shaping a son's physical self-concept positively and negatively. It was also found that the self-concept of the young adolescent athlete is more strongly affected by the perceived parental-created motivational climate (both task and ego) than the older adolescent athlete's self-concept. These findings support the expectancy-value model assumptions related to the role of parents as important socializing agents, the existence of gender-stereotyping, and the heavy reliance younger children place on parents' feedback

    Comparison of quantitative real time PCR with Sequencing and ribosomal RNA-FISH for the identification of fungi in Formalin fixed, paraffin-embedded tissue specimens

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    Background: Identification of the causative agents of invasive fungal infections (IFI) is critical for guiding antifungal therapy. Cultures remain negative in a substantial number of IFI cases. Accordingly, species identification from formalin fixed, paraffin embedded (FFPE) tissue specimens by molecular methods such as fluorescence in situ hybridisation (FISH) and PCR provides an appealing approach to improve management of patients. Methods: We designed FISH probes targeting the 28S rRNA of Aspergillus and Candida and evaluated them with type strains. Fluorescence microscopy (FM), using FISH probes and quantitative broadrange fungal PCR targeting the rRNA gene were applied to FFPE tissue specimens from patients with proven IFI in order to explore benefits and limitations of each approach. Results: PCR followed by sequencing identified a broad spectrum of pathogenic fungi in 28 of 40 evaluable samples (70%). Hybridisation of FISH probes to fungal rRNA was documented in 19 of 40 tissue samples (47.5%), including 3 PCR negative samples with low fungal burden. The use of FISH was highly sensitive in invasive yeast infections, but less sensitive for moulds. In samples with hyphal elements, the evaluation of hybridisation was impaired due to autofluorescence of hyphae and necrotic tissue background. Conclusions: While PCR appears to be more sensitive in identifying the causative agents of IFI, some PCR negative and FISH positive samples suggest that FISH has some potential in the rapid identification of fungi from FFPE tissue samples

    Temporal Variability of Human Vaginal Bacteria and Relationship with Bacterial Vaginosis

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    Little is known about short-term bacterial fluctuations in the human vagina. This study used PCR to assess the variability in concentrations of key vaginal bacteria in healthy women and the immediate response to antibiotic treatment in women with bacterial vaginosis (BV).Twenty-two women assessed for BV using Amsel's criteria were evaluated daily for 7 or 14 days, then at 2, 3 and 4 weeks, using a panel of 11 bacterium-specific quantitative PCR assays. Participants with BV were treated with 5 days of intravaginal metronidazole. Participants without BV had vaginal biotas dominated by lactobacilli, whose levels fluctuated with menses. With onset of menstruation, quantities of Lactobacillus jensenii and Lactobacillus crispatus decreased and were found to be inversely related to Gardnerella vaginalis concentrations (p<0.001). Women with BV had a variety of fastidious bacteria whose concentrations dropped below detection thresholds 1-5 days after starting metronidazole. Recurrent BV was characterized by initial profound decreases of BV-associated bacteria after treatment followed by subsequent increases at relapse.The microbiota of the human vagina can be highly dynamic. Healthy women are colonized with Lactobacillus species, but levels can change dramatically over a month. Marked increases in G. vaginalis were observed during menses. Participants with BV have diverse communities of fastidious bacteria that are depleted by vaginal metronidazole therapy. Women with recurrent BV initially respond to antibiotic treatment with steep declines in bacterial concentrations, but these bacteria later reemerge, suggesting that antibiotic resistance in these bacteria is not an important factor mediating BV recurrence

    Data-Driven Analysis of Engagement in Gamified Learning Environments: A Methodology for Real-Time Measurement of MOOCs

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    Welfare and economic development is directly dependent on the availability of highly skilled and educated individuals in society. In the UK, higher education is accessed by a large percentage of high school graduates (50% in 2017). Still, in Brazil, a limited number of pupils leaving high schools continue their education (up to 20%). Initial pioneering efforts of universities and companies to support pupils from underprivileged backgrounds, to be able to succeed in being accepted by universities include personalised learning solutions. However, initial findings show that typical distance learning problems occur with the pupil population: isolation, demotivation, and lack of engagement. Thus, researchers and companies proposed gamification. However, gamification design is traditionally exclusively based on theory-driven approaches and usually ignore the data itself. This paper takes a different approach, presenting a large-scale study that analysed, statistically and via machine learning (deep and shallow), the first batch of students trained with a Brazilian gamified intelligent learning software (called CamaleOn), to establish, via a grassroots method based on learning analytics, how gamification elements impact on student engagement. The exercise results in a novel proposal for real-time measurement on Massive Open Online Courses (MOOCs), potentially leading to iterative improvements of student support. It also specifically analyses the engagement patterns of an underserved community
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