Identification and comparison of aerobic and denitrifying polyphosphate-accumulating organisms

Two laboratory-scale sequencing batch reactors (SBRs) were operated for enhanced biological phosphorus removal (EBPR) in alternating anaerobic-aerobic or alternating anaerobic-anoxic modes, respectively. Polyphosphate-accumulating organisms (PAOs) were enriched in the anaerobic-aerobic SBR and denitrifying PAOs (DPAOs) were enriched in the anaerobic-aerobic SBR. Fluorescence in situ hybridization (FISH) demonstrated that the well-known PAO, Candidatus Accumulibacter phosphatis was abundant in both SBRs, and post-FISH chemical staining with 4,6-diamidino-2-phenylindol (DAPI) confirmed that they accumulated polyphosphate. When the anaerobic-anoxic SBR enriched for DPAOs was converted to anaerobic-aerobic operation, aerobic uptake of phosphorus by the resident microbial community occurred immediately. However, when the anaerobic-aerobic SBR enriched for PAOs was exposed to one cycle with anoxic rather than aerobic conditions, a 5-h lag period elapsed before phosphorus uptake proceeded. This anoxic phosphorus-uptake lag phase was not observed in the subsequent anaerobic-aerobic cycle. These results demonstrate that the PAOs that dominated the anaerobic-aerobic SBR biomass were the same organisms as the DPAOs enriched under anaerobic-anoxic conditions. (C) 2003 Wiley Periodicals, Inc.

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.

Identification of slow and fast-acting toxins in a highly ciguatoxic barracuda (Sphyraena barracuda) by HPLC/MS and radiolabelled ligand binding

A barracuda implicated in ciguatera fish poisoning in Guadeloupe was estimated to have an overall flesh toxicity of 15 MUg/g using mouse bioassay. A lipid soluble extract was separated into two toxic fractions, FrA and FrB, on a LH20 Sephadex column eluted with dichloromethane/methanol (1:1). When intraperitoneal injected into mice, FrA provoked symptoms characteristic of slow-acting ciguatoxins, whereas FrB produced symptoms indicative of fast-acting toxins (FAT). High performance liquid chromatography/mass spectrometry/radio-ligand binding (HPLC/MS/RLB) analysis confirmed the two fractions were distinct, because only a weak overlap of some compounds was observed. HPLC/MS/RLB analysis revealed C-CTX-1 as the potent toxin present in FrA, and two coeluting active compounds at m/z 809.43 and 857.42 in FrB, all displaying the characteristic pattern of ion formation for hydroxy-polyethers. Other C-CTX congeners and putative hydroxy-polyether-like compounds were detected in both fractions, however, the RLB found them inactive. C-CTX-1 accounted for >90% of total toxicity in this barracuda and was confirmed to be a competitive inhibitor of brevetoxin binding to voltage-sensitive sodium channels (VSSCs) with a potency two-times lower than P-CTX-1. However, FAT active on VSSCs and

Identification of cholinoceptive glycinergic neurons in the mammalian retina

The light-evoked release of acetylcholine (ACh) affects the responses of many retinal ganglion cells, in part via nicotinic acetylcholine receptors (nAChRs). nAChRs that contain beta2alpha3 neuronal nicotinic acetylcholine receptors have been identified and localized in the rabbit retina; these nAChRs are recognized by the monoclonal antibody mAb210. We have examined the expression of beta2alpha3 nAChRs by glycinergic amacrine cells in the rabbit retina and have identified different subpopulations of nicotinic cholinoceptive glycinergic cells using double and triple immunohistochemistry with quantitative analysis. Here we demonstrate that about 70% of the cholinoceptive amacrine cells in rabbit retina are glycinergic cells. At least three nonoverlapping subpopulations of mAb210 glycine-immunoreactive cells can be distinguished with antibodies against calretinin, calbindin, and gamma-aminobutyric acid (GABA)(A) receptors. The cholinergic cells in rabbit retina are thought to synapse only on other cholinergic cells and ganglion cells. Thus, the expression of beta2alpha3 nAChRs on diverse populations of glycinergic cells is puzzling. To explore this finding, the subcellular localization of beta2alpha3 was studied at the electron microscopic level. mAb210 immunoreactivity was localized on the dendrites of amacrines and ganglion cells throughout the inner plexiform layer, and much of the labeling was not associated with recognizable synapses. Thus, our findings indicate that ACh in the mammalian retina may modulate glycinergic circuits via extrasynaptic beta2alpha3 nAChRs. (C) 2002 Wiley-Liss, Inc.

Identification and characterization of pptA: a gene involved in the phase-variable expression of phosphorylcholine on pili of Neisseria meningitidis

Pili of pathogenic Neisseria are major virulence factors associated with adhesion, cytotoxicity, twitching motility, autoaggregation, and DNA transformation. Pili are modified posttranslationally by the addition of phosphorylcholine. However, no genes involved in either the biosynthesis or the transfer of phosphorylcholine in Neisseria meningitidis have been identified. In this study, we identified five candidate open reading frames (ORFs) potentially involved in the biosynthesis or transfer of phosphorylcholine to pilin in N. meningitidis. Insertional mutants were constructed for each ORF in N. meningitidis strain C311#3 to determine their effect on phosphorylcholine expression. The effect of the mutant ORFs on the modification by phosphorylcholine was analyzed by Western analysis with phosphorylcholine-specific monoclonal antibody TEPC-15. Analysis of the mutants showed that ORF NMB0415, now defined as pptA (pilin phosphorylcholine transferase A), is involved in the addition of phosphorylcholine to pilin in N. meningitidis. Additionally, the phase variation (high frequency on-off switching of expression) of phosphorylcholine on pilin is due to changes in a homopolymeric guanosine tract in pptA.

Um (Des)caso do planejamento estrat??gico em ??rg??o da Administra????o P??blica

O estudo de caso descreve o processo de planejamento estrat??gico ocorrido em uma organiza????o p??blica e os resultados alcan??ados ap??s a sua implanta????o. A organiza????o, com o nome fict??cio de Companhia de Defesa, desenvolveu seu planejamento estrat??gico com inten????o de aderir ?? administra????o estrat??gica. Os principais problemas reconhecidos pela organiza????o s??o a morosidade, a falta de transpar??ncia e a inefici??ncia de suas a????es. Atuaram no planejamento estrat??gico a alta administra????o, coordenadores nomeados e representantes regionais e uma consultoria especializada. O ambiente organizacional existente ?? ??poca n??o foi prop??cio ao planejamento estrat??gico: mudan??as de dire????o, pouca capacita????o funcional, capilaridade da institui????o e dificuldade de consenso entre planejadores e executores. ?? poss??vel, ??queles que estudarem o caso, contrapor a teoria e a pr??tica do planejamento estrat??gico, identificando boas a????es realizadas e erros cometidos

??cole Nationale d'Administration Publique: d??velopper les comp??tences des fonctionnaires pour augmenter la capacit?? de L'Etat dans la gestion des politiques publiques

Plus d???une vingtaine d???ann??es d???existence d??di??es ?? la mission de d??velopper les comp??tences des fonctionnaires pour augmenter la capacit?? de l?????tat dans la gestion des politiques publiques fait de l???ENAP un mod??le dans le domaine de la formation. Rattach??e au Minist??re du Plan, du Budget et de la Gestion, l?????cole, d??s sa cr??ation en 1986, a re??ue 245 mille fonctionnaires de tout le pays. En 2007, 26 mille fonctionnaires de l???administration f??d??rale ont attendu un de nos 60 stages offerts.

Conselhos locais de saúde : caminhos e (des)caminhos da participação social no Sistema Único de Saúde

A partir da descentralização, novas instâncias de negociação e novas alternativas de ordenamento da estrutura organizacional do Sistema Único de Saúde (SUS) foram criadas. Dentre estas alternativas, podemos citar os conselhos de saúde, importantes canais de participação social. Todavia, frente às limitações destes canais tradicionais de articulação entre Estado e sociedade, destacamos os ideais da gestão participativa e os Conselhos Locais de Saúde (CLS) como alternativa de renovação e criação de instâncias mais flexíveis, porosas e efetivas às complexas demandas sociais. Neste sentido, buscamos analisar o processo de criação e implementação dos CLS do município de Anchieta/ES, a partir de uma abordagem quali-quantativa. Inicialmente, traçamos o perfil socioeconômico e político dos conselheiros eleitos, a partir de um questionário aplicado a uma amostra de 54 conselheiros; dados que foram categorizados e analisados por meio do emprego de estatísticas descritivas. Em seguida, entrevistamos treze conselheiros, de dois conselhos distintos do município, procedendo à análise de conteúdo do material, a partir dos ideais de Bardin (2000). Os resultados demonstraram que os conselhos foram criados a partir da iniciativa da gestão municipal em 2011, e que simplesmente institucionalizá-los como espaço de participação social não foi suficiente para promover a mobilização social e o envolvimento comunitário. Quanto ao perfil dos conselheiros locais, 78% são mulheres, com predominância de raça/cor branca, idade entre os 20 e 39 anos e funcionárias públicas; 57% possuem Ensino Médio e participaram como conselheiro por dois anos, e 60% destes já tiveram outras experiências de participação similares aos CLS. Do material oriundo das entrevistas, emergiram quatro categorias de análise, a saber: 1) Ser ou não ser conselheiro de saúde? Eis a questão!; 2) O não pertencimento e a não-participação; 3) Conselhos Locais de Saúde: elos, meios e mediações; e 4) A exogenia da administração e os obstáculos à participação social. Os entraves ao funcionamento dos conselhos de saúde, mesmo em nível local, ainda são desafios a serem superados, para que estas instâncias sejam mais influentes na gestão pública, conforme os princípios de sua criação. A participação social e a democracia são fundamentais para a construção de políticas de saúde que correspondam às reais demandas da comunidade. Contudo, para garantir a democracia na sociedade não basta promover a descentralização. É necessário que os sujeitos políticos resistam às relações de dominação, opressão e subordinação. Para isso, torna-se imprescindível os programas de educação para cidadania dos sujeitos envolvidos nestes fóruns de participação. O que nos motiva, enfim, é notarmos a existência, entre os conselheiros eleitos, de sujeitos protagonistas de seu próprio devir; sujeitos que atuam como agentes transformadores, motivadores de sonhos e projetos em prol da saúde pública e de sua comunidade.

Eu bebo sim e estou vivendo : qualidade de vida, masculinidade, padrões de consumo de álcool e representações sociais de bebida alcoólica para homens usuários de unidade de saúde da família

A Política Nacional de Atenção Integral à Saúde do Homem propõe formas diferenciadas de atuação da equipe de saúde no atendimento da população masculina, uma vez que este público demanda estratégias diferenciadas de serviço. Ao se deparar com a realidade vivenciada pelos homens, surgem várias questões acerca do estereótipo social construído acerca das características masculinas e suas vivências. O objetivo da pesquisa foi compreender alguns aspectos relevantes para as práticas de saúde de homens usuários de Unidade de Saúde da Família, como qualidade de vida, consumo de álcool, representações sociais da bebida alcoólica e características de masculinidade. Foi utilizada uma amostra de 300 homens, frequentadores de Unidade de Saúde da Família, e aplicado um questionário contendo os dados sociodemográficos, o World Health Organization Quality of Life (Whoqol-bref), o Bem Sex-Role Inventory (BSRI), um exercício de evocação sobre bebida alcoólica, o Alcohol Use Disorders Identification Test (Audit) e um bloco para verificar os problemas ocasionados pelo consumo de álcool e a procura por tratamento. Os dados dos instrumentos quantitativos foram analisados com testes estatísticos de comparação de médias e de correlação. Os dados das evocações foram analisados com o software EVOC (Ensemble de Programmes Permettant l’Analyse des Évocations). Na primeira análise, constatou-se adesão mais alta a características femininas, alta percepção de qualidade de vida e padrões de consumo de álcool semelhantes às médias nacionais. Homens que declararam praticar sua religião apresentaram média significativamente menor de consumo de álcool. Apresentaram correlação inversamente proporcional ao consumo de álcool as características femininas de gênero, os domínios físico, social, psicológico e percepção global de qualidade de vida. Na análise das evocações, constatou-se que os elementos com tendência à centralidade são, em sua maioria, de cunho negativo. Os dados da população geral apresentaram o termo gosto como um aspecto positivo e central da bebida alcoólica. O grupo de abstinentes não apresentou avaliação positiva do termo e o grupo de bebedores apresentou o termo diversão na primeira periferia, referindo-se aos aspectos positivos e de socialização da bebida alcoólica. Os resultados indicaram uma qualidade de vida satisfatória, a religião e as características femininas destacaram-se como um fator de proteção ao uso de bebida alcoólica. Apresentaram, ainda, percepção dos problemas associados ao próprio consumo. Apesar de a maioria dos termos relacionados à bebida alcoólica ser negativo, este consumo ainda se dá em um nível considerável. Por isso, se faz necessária a construção de vínculo entre o profissional e o usuário do serviço de saúde a fim de dar oportunidade para que as reais práticas sobre a bebida alcoólica sejam evidenciadas. Esses dados podem ajudar profissionais de Saúde da Família a refletirem sobre as representações sociais que constroem acerca dos homens de classe popular usuários do serviço

Automated traffic route identification through the shared nearest neighbour algorithm

Many organisations need to extract useful information from huge amounts of movement data. One example is found in maritime transportation, where the automated identification of a diverse range of traffic routes is a key management issue for improving the maintenance of ports and ocean routes, and accelerating ship traffic. This paper addresses, in a first stage, the research challenge of developing an approach for the automated identification of traffic routes based on clustering motion vectors rather than reconstructed trajectories. The immediate benefit of the proposed approach is to avoid the reconstruction of trajectories in terms of their geometric shape of the path, their position in space, their life span, and changes of speed, direction and other attributes over time. For clustering the moving objects, an adapted version of the Shared Nearest Neighbour algorithm is used. The motion vectors, with a position and a direction, are analysed in order to identify clusters of vectors that are moving towards the same direction. These clusters represent traffic routes and the preliminary results have shown to be promising for the automated identification of traffic routes with different shapes and densities, as well as for handling noise data.

Component identification through program slicing

This paper reports on the development of specific slicing techniques for functional programs and their use for the identification of possible coherent components from monolithic code. An associated tool is also introduced. This piece of research is part of a broader project on program understanding and re-engineering of legacy code supported by formal methods

Cytogenetic and identification of the nucleolus organizer region in Heliconia bihai (L.) L.

The genus Heliconia is not much studied and the number of existing species in this genus is still uncertain. It is known that this number relies between 150 to 250 species. In Brazil, about 40 species are native and known by many different names. The objective of this paper was to characterize morphometrically and to identify the NOR (active nucleolus organizer regions) by Ag-NOR banding of chromosomes of Heliconia bihai (L) L. Root meristems were submitted to blocking treatment in an amiprofos-methyl (APM) solution, fixed in methanol-acetic acid solution for 24 hours, at least. The meristems were washed in distilled water and submitted to enzymatic digestion with pectinase enzyme. The slides were prepared by dissociation of the root meristem, dried in the air and also on hot plate at 50°C. Subsequently, some slides were submitted to 5% Giemsa stain for karyotype construction and to a solution of silver nitrate (AgNO3) 50% for Ag-NOR banding. The species H. bihai has 2n = 22 chromosomes, 4 pairs of submetacentric chromosomes and 7 pairs of metacentric chromosomes, and graded medium to short (3.96 to 0.67 μM), with the presence of active NOR in pairs 1 and 2 and interphase cells with 2 nucleoli. These are the features of a diploid species.

Identification of 'Ubá' mango tree zygotic and nucellar seedlings using ISSR markers

Polyembryonic seeds are characterized by the development of over one embryo in the same seed, which can be zygotic and nucellar. The objective of this work was to identify the genetic origin, whether zygotic or nucellar, of seedlings of polyembryonic seeds of 'Ubá' mango tree using ISSR markers, and relating them with the vigor of the seedlings. Thus, mangos were harvested in Visconde do Rio Branco (accession 102) and Ubá (accessions 112, 138, 152 and 159), whose seeds were germinated in plastic trays filled with washed sand. Fifty days after sowing, seedlings from five seeds of each one of the accessions 102, 112, 138, 159 and from 10 seeds of the accession 152, were analyzed. These sseedlings were characterized and evaluated for plant height, stem circumference and mass of fresh aerial part and the most vigorous seedling was the one displaying at least two of these traits higher than the other seedlings from seed. Leaves were collected for genomic DNA extraction, which was amplified using seven ISSR primers previously selected based on the amplification profile and considering the number and resolution of fragments. Zygotic seedlings were found in 18 seeds, which were the most vigorous in six seeds. The results evidenced the existence of genetic variability in orchards using seedlings grown from seeds, because the farmer usually uses the most vigorous ones, assuming that this is of nucellar origin. These results also indicate that the most vigorous seedling are not always nucellar, inasmuch as of 20% of the total seeds evaluated, the zygotic seedling was the most vigorous.