38 research outputs found

    Biochemical properties of Paracoccus denitrificans FnrP:Reactions with molecular oxygen and nitric oxide

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    In Paracoccus denitrificans, three CRP/FNR family regulatory proteins, NarR, NnrR and FnrP, control the switch between aerobic and anaerobic (denitrification) respiration. FnrP is a [4Fe-4S] cluster containing homologue of the archetypal O2 sensor FNR from E. coli and accordingly regulates genes encoding aerobic and anaerobic respiratory enzymes in response to O2, and also NO, availability. Here we show that FnrP undergoes O2-driven [4Fe-4S] to [2Fe-2S] cluster conversion that involves up to 2 O2 per cluster, with significant oxidation of released cluster sulfide to sulfane observed at higher O2 concentrations. The rate of the cluster reaction was found to be ~6-fold lower than that of E. coli FNR, suggesting that FnrP can remain transcriptionally active under microaerobic conditions. This is consistent with a role for FnrP in activating expression of the high O2 affinity cytochrome c oxidase under microaerobic conditions. Cluster conversion resulted in dissociation of the transcriptionally active FnrP dimer into monomers. Therefore, along with E. coli FNR, FnrP belongs to the subset of FNR proteins in which cluster type is correlated with association state. Interestingly, two key charged residues, Arg140 and Asp154, that have been shown to play key roles in the monomer-dimer equilibrium in E. coli FNR are not conserved in FnrP, indicating that different protomer interactions are important for this equilibrium. Finally, the FnrP [4Fe-4S] cluster is shown to undergo reaction with multiple NO molecules, resulting in iron nitrosyl species and dissociation into monomers

    Characterization of a putative NsrR homologue in Streptomyces venezuelae reveals a new member of the Rrf2 superfamily

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    Members of the Rrf2 superfamily of transcription factors are widespread in bacteria but their functions are largely unexplored. The few that have been characterized in detail sense nitric oxide (NsrR), iron limitation (RirA), cysteine availability (CymR) and the iron sulfur (Fe-S) cluster status of the cell (IscR). In this study we combined ChIP-seq with in vitro biochemistry to characterize a putative NsrR homologue in the model organism Streptomyces venezuelae. ChIP seq analysis revealed that rather than regulating the nitrosative stress response like NsrR, Sven6563 binds to a different, much larger regulon of genes with a diverse range of functions, including a range of regulators, genes required for glutamine synthesis, NADH/NAD(P)H metabolism, as well as general DNA/RNA and amino acid/protein turn over. Our biochemical experiments further show that Sven6563 has a [2Fe-2S] cluster and that the switch between oxidized and reduced cluster controls its DNA binding activity in vitro. To our knowledge, both the sensing domain and the target gene regulon are novel for an Rrf2 protein, suggesting Sven6563 represents a new member of the Rrf2 superfamily. Given the redox sensitivity of its Fe-S cluster we have tentatively named the protein RsrR for Redox sensitive response Regulator

    The prevalence and experience of oral diseases in Adelaide nursing home residents

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    The document attached has been archived with permission from the Australian Dental Association. An external link to the publisher’s copy is included.Background: The twenty-first century will see the evolution of a population of dentate older Australians with dental needs very different from those of older adults in past years. This study provided comprehensive information concerning oral disease prevalence in older South Australian nursing home residents. Methods: This paper presents cross-sectional baseline results. Results: Most of the 224 residents, from seven randomly selected nursing homes, were functionally dependent, medically compromised, cognitively impaired and behaviourally difficult older adults who presented many complex challenges to carers and to dental professionals. Two-thirds (66 per cent) were edentulous with many dental problems and treatment needs. Dentate residents had a mean of 11.9 teeth present, higher than previously reported. The prevalence and experience of coronal and root caries and plaque accumulation was very high in dentate residents; especially males, those admitted more than three years previously, those who ate fewer food types and those who were severely cognitively impaired. These residents had more retained roots, decayed teeth and missing teeth, and fewer filled teeth when compared with data for community-dwelling older adults. Conclusions: This study highlighted the poor oral health status of these nursing home residents and the great impact of dementia on their high levels of oral diseases.JM Chalmers, C Hodge, JM Fuss, AJ Spencer, KD Carte

    Opinions of dentists and directors of nursing concerning dental care provision for Adelaide nursing homes

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    The document attached has been archived with permission from the Australian Dental Association (7th Jan 2007). An external link to the publisher’s copy is included.Background: The complex oral health problems of nursing home residents have been well documented. However, the influences on residents’ oral health status, including opinions and experiences of dental professionals and nursing home staff, have not yet been adequately investigated. Methods: The baseline questionnaire component of this longitudinal study was mailed to all registered dentists practising in Adelaide and Adelaide nursing home directors of nursing (DONs). Results: 413 dentists and 97 DONs indicated that Adelaide dentists’ interest and training in nursing home dentistry was low. Dental service provision for nursing home residents was very low and dentists preferred to provide treatment at their dental practices. Few dental hygienists were working in nursing homes and dental professionals provided little educational assistance for nursing home staff. Dentists and DONs held several common and many varying perceptions of the problems associated with dental care provision in nursing homes. Both identified a group of nursing home environmental constraints and a lack of portable dental equipment. DONs further identified a group of resident related problems, and dentists a group of dental practicerelated problems. Conclusions: These study results provide important information concerning problems with nursing home dentistry for dental service providers, educators, policy-makers, administrators and nursing home staff.JM Chalmers, C Hodge, JM Fuss, AJ Spencer, KD Carter and R Mathe

    Drosophila Ribosomal Protein Mutants Control Tissue Growth Non-Autonomously via Effects on the Prothoracic Gland and Ecdysone

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    The ribosome is critical for all aspects of cell growth due to its essential role in protein synthesis. Paradoxically, many Ribosomal proteins (Rps) act as tumour suppressors in Drosophila and vertebrates. To examine how reductions in Rps could lead to tissue overgrowth, we took advantage of the observation that an RpS6 mutant dominantly suppresses the small rough eye phenotype in a cyclin E hypomorphic mutant (cycEJP). We demonstrated that the suppression of cycEJP by the RpS6 mutant is not a consequence of restoring CycE protein levels or activity in the eye imaginal tissue. Rather, the use of UAS-RpS6 RNAi transgenics revealed that the suppression of cycEJP is exerted via a mechanism extrinsic to the eye, whereby reduced Rp levels in the prothoracic gland decreases the activity of ecdysone, the steroid hormone, delaying developmental timing and hence allowing time for tissue and organ overgrowth. These data provide for the first time a rationale to explain the counter-intuitive organ overgrowth phenotypes observed for certain members of the Minute class of Drosophila Rp mutants. They also demonstrate how Rp mutants can affect growth and development cell non-autonomously

    The oral health of Adelaide nursing home residents: longitudinal study

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    The definitive version is available at www.blackwell-synergy.comObjective: The Adelaide Dental Study of Nursing Homes aimed to quantify oral disease experience, incidence and increments in Adelaide nursing home residents. Methods: Questionnaires and dental inspections were completed at baseline and at 1-year for residents from randomly selected Adelaide nursing homes. Results: The residents were very functionally dependent, cognitively impaired and behaviourally difficult older adults with complex oral problems and dental treatment needs. The prevalence of edentulism (total tooth loss) (63%) decreased and more residents were retaining natural teeth. Existing residents had a mean of 10.8 teeth present and new residents had a mean of 12.7 teeth present. Residents’ previous experiences of caries (decay) were high – existing residents had a mean of 1.2 decayed teeth and new residents had a mean of 0.8 decayed teeth. Residents’ caries increments (new decay) over the 1-year period were high (coronal = 2.5 surfaces; root = 1.0 surfaces), especially in those who had lost weight and who could eat fewer food types. These levels of caries were many times greater than had been reported for communitydwelling older adults. Large accumulations of plaque, calculus and debris (food) were evident on residents’ natural teeth and dentures, especially those with dementia. Up to 25% of residents owned dentures that were not worn. Residents with dementia gave their carers complex and challenging oral hygiene care problems. Existing and new residents had similar general health and oral health characteristics, with the exceptions that new residents had significantly more filled tooth surfaces, and fewer decayed retained roots. Conclusion: New residents were being admitted to the nursing homes with a compromised oral health status or developed severe oral diseases and conditions within several months of their admittance. Residents’ oral diseases, especially coronal and root caries, rapidly progressed during their stay in residential care.JM Chalmers, KD Carter and AJ Spence
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