Investigation of candidate division TM7, a recently recognized major lineage of the domain bacteria with no known pure-culture representatives

A molecular approach was used to investigate a recently described candidate division of the domain Bacteria, TM7, currently known only from environmental 16S ribosomal DNA sequence data, A number of TM7-specific primers and probes were designed and evaluated. Fluorescence in situ hybridization (FISH) of a laboratory scale bioreactor using two independent TM7-specific probes revealed a conspicuous sheathed-filament morphotype, fortuitously enriched in the reactor. Morphologically, the filament matched the description of the Eikelboom morphotype 0041-0675 widely associated with bulking problems in activated-sludge wastewater treatment systems. Transmission electron microscopy of the bioreactor sludge demonstrated that the sheathed-filament morphotype had a typical gram-positive cell envelope ultrastructure. Therefore, TM7 is only the third bacterial lineage recognized to have gram-positive representatives. TM7-specific FISH analysis of two full-scale wastewater treatment plant sludges, including the one used to seed the laboratory scale reactor, indicated the presence of a number of morphotypes, including sheathed filaments. TM7-specific PCR clone libraries prepared from the two full-scale sludges yielded 23 novel TM7 sequences. Three subdivisions could be defined based on these data and publicly available sequences. Environmental sequence data and TM7-specific FISH analysis indicate that members of the TM7 division are present in a variety of terrestrial, aquatic, and clinical habitats. A highly atypical base substitution (Escherichia coli position 912; C to U) for bacterial 16S rRNAs was present in almost all TM7 sequences, suggesting that TM7 bacteria, like Archaea, may be streptomycin resistant at the ribosome level.

Human melanoblasts in culture: Expression of BRN2 and synergistic regulation by fibroblast growth factor-2, stem cell factor, and endothelin-3

The BRN2 transcription factor (POU3F2, N-Oct-3) has been implicated in development of the melanocytic lineage and in melanoma. Using a low calcium medium supplemented with stem cell factor, fibroblast growth factor-2, endothelin-3 and cholera toxin, we have established and partially characterised human melanocyte precursor cells, which are unpigmented, contain immature melanosomes and lack L-dihydroxyphenylalanine reactivity. Melanoblast cultures expressed high levels of BRN2 compared to melanocytes, which decreased to a level similar to that of melanocytes when cultured in medium that contained phorbol ester but lacked endothelin-3, stem cell factor and fibroblast growth factor-2. This decrease in BRN2 accompanied a positive L-dihydroxyphenylalanine reaction and induction of melanosome maturation consistent with melanoblast differentiation seen during development. Culture of primary melanocytes in low calcium medium supplemented with stem cell factor, fibroblast growth factor-2 and endothelin-3 caused an increase in BRN2 protein levels with a concomitant change to a melanoblast-like morphology. Synergism between any two of these growth factors was required for BRN2 protein induction, whereas all three factors were required to alter melanocyte morphology and for maximal BRN2 protein expression. These finding implicate BRN2 as an early marker of melanoblasts that may contribute to the hierarchy of melanocytic gene control.

Model-based analysis of anaerobic acetate uptake by a mixed culture of polyphosphate-accumulating and glycogen-accumulating organisms

An increasing number of studies shows that the glycogen-accumulating organisms (GAOs) can survive and may indeed proliferate under the alternating anaerobic/aerobic conditions found in EBPR systems, thus forming a strong competitor of the polyphosphate-accumulating organisms (PAOs). Understanding their behaviors in a mixed PAO and GAO culture under various operational conditions is essential for developing operating strategies that disadvantage the growth of this group of unwanted organisms. A model-based data analysis method is developed in this paper for the study of the anaerobic PAO and GAO activities in a mixed PAO and GAO culture. The method primarily makes use of the hydrogen ion production rate and the carbon dioxide transfer rate resulting from the acetate uptake processes by PAOs and GAOs, measured with a recently developed titration and off-gas analysis (TOGA) sensor. The method is demonstrated using the data from a laboratory-scale sequencing batch reactor (SBR) operated under alternating anaerobic and aerobic conditions. The data analysis using the proposed method strongly indicates a coexistence of PAOs and GAOs in the system, which was independently confirmed by fluorescent in situ hybridization (FISH) measurement. The model-based analysis also allowed the identification of the respective acetate uptake rates by PAOs and GAOs, along with a number of kinetic and stoichiometric parameters involved in the PAO and GAO models. The excellent fit between the model predictions and the experimental data not involved in parameter identification shows that the parameter values found are reliable and accurate. It also demonstrates that the current anaerobic PAO and GAO models are able to accurately characterize the PAO/GAO mixed culture obtained in this study. This is of major importance as no pure culture of either PAOs or GAOs has been reported to date, and hence the current PAO and GAO models were developed for the interpretation of experimental results of mixed cultures. The proposed method is readily applicable for detailed investigations of the competition between PAOs and GAOs in enriched cultures. However, the fermentation of organic substrates carried out by ordinary heterotrophs needs to be accounted for when the method is applied to the study of PAO and GAO competition in full-scale sludges. (C) 2003 Wiley Periodicals, Inc.

Growth of papaya nodal and rooted shoot cultures as affected by ACC, STS, culture vessel size and incubation conditions

Nodal shoot cultures of 'Clone 003', a selected Australian papaya cultivar, were cultured on modified De Fossard medium supplemented with chemicals that either promote ethylene evolution or inhibit action while in culture. Nodal shoot cultures grown in the presence of 1-aminocyclopropane carboxylic acid (ACC, 1.0 mM) resulted in a significant reduction in percent fresh and dry weights, shoot length, leaf area, petiole length and chlorophyll content, but leaf development was significantly increased. In contrast, nodal cultures grown in the presence of silver thiosulphate (STS, 0.5 mM) significantly produced the highest percentage of fresh and dry weights, shoot length, leaf production, leaf area expansion, petiole length and leaf chlorophyll content. Nodal cultures and rooted whole plantlets placed in medium-sized (125 mL) culture vessels had significantly better growth than those cultures placed in small (70 mL) or in large (250 mL) vessels. Cultures grown in medium-sized vessels had higher fresh and dry weights, longer shoots, more leaves and larger leaf area than those cultures placed in smaller or larger vessels. Similarly, values for said growth parameters and for chlorophyll content of the nodal and rooted whole plantlets were higher when they were incubated under high light intensity of 120 mumol m(-2)s(-1) at a prevailing temperature of either 20+/-1 C or 25+/-1 C.

Performing race, culture, and gender in an Indigenous Australian women's music and dance classroom

One perpetual concern among Indigenous Australian peoples is authenticity of voice. Who has the right to speak for, and to make representations about, the knowledges and cultures of Indigenous Australian peoples? Whose voice is more authentic, and what happens to these ways of knowing when they make the journey into mainstream Western academic classrooms? In this paper, I examine these questions within the politics of “doing” Indigenous Australian studies by focusing on my own experiences as a lecturer in the Aboriginal and Torres Strait Islander Studies Unit at the University of Queensland. My findings suggest that representation is a matter of problematizing positionality and, from a pedagogical standpoint, being aware of, and willing to address, the ways in which power, authority, and voice are performed and negotiated as teachers and learners of Indigenous Australian studies.

Effect of time of harvest of budded virus on the selection of baculovirus FP mutants in cell culture

Rapid formation and selection of FP (few polyhedra) mutants occurs during serial passaging of Helicoverpa armigera nucleopolyhedrovirus (HaSNPV) in insect cell culture. The production of HaSNPV for use as biopesticides requires the passaging of the virus over a number of passages to produce enough virus inoculum for large-scale fermentation. During serial passaging in cell culture, FP mutants were rapidly selected, resulting in declined productivity and reduced potency of virus. Budded virus (BV) is usually harvested between 72 and 96 h postinfection (hpi) in order to obtain a high titer virus stock. In this study, the effect of tine of harvest (TOH) for BV on the selection rate of HaSNPV FP mutants during serial passaging was investigated. BV were harvested at different times postinfection, and each series was serially passaged for six passages. The productivity and percentage of FP mutants at each passage were determined. It was found that the selection of FP mutants can he reduced by employing an earlier TOH for BV. Serial passaging with BV harvested at 48 hpi showed a slower accumulation of FP mutants compared to that of BV harvested after 48 hpi. Higher cell specific yields were also maintained when BV were harvested at 48 hpi. When BV that were formed between 48 and 96 hpi were harvested and serially passaged, FP mutants quickly dominated the virus population. This suggests that the V formed and released between 48 and 96 hpi are most likely from FP mutant infected cells.

Diverse effects of renal denervation on ventricular hypertrophy and blood pressure in DOCA-salt hypertensive rats

Cardiac hypertrophy that accompanies hypertension seems to be a phenomenon of multifactorial origin whose development does not seem to depend on an increased pressure load alone, but also on local growth factors and cardioadrenergic activity. The aim of the present study was to determine if sympathetic renal denervation and its effects on arterial pressure level can prevent cardiac hypertrophy and if it can also delay the onset and attenuate the severity of deoxycorticosterone acetate (DOCA)-salt hypertension. DOCA-salt treatment was initiated in rats seven days after uninephrectomy and contralateral renal denervation or sham renal denervation. DOCA (15 mg/kg, sc) or vehicle (soybean oil, 0.25 ml per animal) was administered twice a week for two weeks. Rats treated with DOCA or vehicle (control) were provided drinking water containing 1% NaCl and 0.03% KCl. At the end of the treatment period, mean arterial pressure (MAP) and heart rate measurements were made in conscious animals. Under ether anesthesia, the heart was removed and the right and left ventricles (including the septum) were separated and weighed. DOCA-salt treatment produced a significant increase in left ventricular weight/body weight (LVW/BW) ratio (2.44 ± 0.09 mg/g) and right ventricular weight/body weight (RVW/BW) ratio (0.53 ± 0.01 mg/g) compared to control (1.92 ± 0.04 and 0.48 ± 0.01 mg/g, respectively) rats. MAP was significantly higher (39%) in DOCA-salt rats. Renal denervation prevented (P>0.05) the development of hypertension in DOCA-salt rats but did not prevent the increase in LVW/BW (2.27 ± 0.03 mg/g) and RVW/BW (0.52 ± 0.01 mg/g). We have shown that the increase in arterial pressure level is not responsible for cardiac hypertrophy, which may be more related to other events associated with DOCA-salt hypertension, such as an increase in cardiac sympathetic activity.

Neural reflex regulation of arterial pressure in pathophysiological conditions : interplay among the baroreflex, the cardiopulmonary reflexes and the chemoreflex

The maintenance of arterial pressure at levels adequate to perfuse the tissues is a basic requirement for the constancy of the internal environment and survival. The objective of the present review was to provide information about the basic reflex mechanisms that are responsible for the moment-to-moment regulation of the cardiovascular system. We demonstrate that this control is largely provided by the action of arterial and non-arterial reflexes that detect and correct changes in arterial pressure (baroreflex), blood volume or chemical composition (mechano- and chemosensitive cardiopulmonary reflexes), and changes in blood-gas composition (chemoreceptor reflex). The importance of the integration of these cardiovascular reflexes is well understood and it is clear that processing mainly occurs in the nucleus tractus solitarii, although the mechanism is poorly understood. There are several indications that the interactions of baroreflex, chemoreflex and Bezold-Jarisch reflex inputs, and the central nervous system control the activity of autonomic preganglionic neurons through parallel afferent and efferent pathways to achieve cardiovascular homeostasis. It is surprising that so little appears in the literature about the integration of these neural reflexes in cardiovascular function. Thus, our purpose was to review the interplay between peripheral neural reflex mechanisms of arterial blood pressure and blood volume regulation in physiological and pathophysiological states. Special emphasis is placed on the experimental model of arterial hypertension induced by N-nitro-L-arginine methyl ester (L-NAME) in which the interplay of these three reflexes is demonstrable.

Experiências formativas no início da docência mediadas pelo PIBID educação física da Universidade Estadual de Feira de Santana, Bahia

O presente trabalho objetiva compreender como o Coordenador, os Supervisores e os Bolsistas do PIBID Educação Física da Universidade Estadual de Feira de Santana (UEFS) no município de Feira de Santana – Bahia percebem os processos de iniciação à docência na formação inicial de professores de Educação Física no PIBID Educação Física – UEFS. Por meio da Pesquisa Narrativa, numa perspectiva de pesquisa e de formação, oportunizamos aos sujeitos da pesquisa entrevistas narrativas compostas pela reflexividade crítica necessária na constituição da docência. Analisamos a produção acadêmica sobre a temática da iniciação à docência no tempo da formação inicial nos bancos de dados da Associação Nacional de Pós-Graduação e Pesquisa em Educação (ANPED) e do Congresso Brasileiro de Ciências do Esporte (CONBRACE) e discutimos com Antonio Nóvoa, Carlos Marcelo e Molina Neto sobre as concepções de formação e as experiências vividas no início da docência e buscamos em Walter Benjamin e Jorge Larrosa ancoragens para o entendimento do conceito de experiência. Os resultados apontam para o reconhecimento do PIBID como um programa que aproxima do campo de atuação interferindo na escolha pela docência. Sinalizam que o programa possibilita a articulação entre a universidade e a escola fortalecendo a perspectiva de formação que acontece coletivamente. Confirmam que o PIBID permite a constatação dos desafios presentes na escola e da necessidade de transformação desta realidade com a articulação dos saberes acadêmicos e os escolares. Neste sentido, constatamos a afirmação da entrada na carreira, como uma experiência formativa que intensifica o desenvolvimento profissional docente e, portanto, deve ser concebida como política nacional institucionalizada de iniciação à docência.

Conteúdos de ensino da educação física escolar : da produção acadêmica às narrativas docentes

Possui como objetivo compreender a relação entre os conteúdos de ensino com a especificidade da Educação Física. Se caracteriza pela abordagem plurimetodológica qualitativa/quantitativa e foi delineada em três capítulos que dialogam entre si, considerando a natureza das fontes estudadas e os pressupostos teórico-metodológicos assumidos. Do tipo “estado do conhecimento”, o primeiro capítulo investiga as produções acadêmicas que versam sobre os conteúdos de ensino da Educação Física escolar, no período de 1981 a 2010. Os 146 artigos mapeados encontram-se distribuídos em 14 periódicos, analisados por meio de indicadores bibliométricos (MUGNAINI; CARVALHO; CAMPANATTI-OSTIZ, 2006). A pesquisa indica os autores que se constituem como referência numérica na área e sua relação com os Programas de Pós-Graduação, suas linhas de estudo e os grupos de pesquisa que oferecem suporte teórico-metodológico. O segundo capítulo, de natureza quali-quantitativa, discute a opinião de 266 professores de Educação Física do Estado do Espírito Santo sobre os processos de seleção e sistematização dos conteúdos de ensino, expressos em um questionário elaborado com perguntas fechadas e semiabertas. Dá visibilidade às aproximações dos conteúdos com a cultura local e às implicações das lógicas de escolarização na escolha do que e quando ensinar. O terceiro capítulo se caracteriza por ser uma pesquisa narrativa (CERTEAU, 2002; RIBEIRO; SOUZA, 2010) e evidencia o modo como os professores de Educação Física trabalham com os conteúdos de ensino. Possui como colaboradores 14 docentes de Educação Física atuantes na Educação Básica da Microrregião Metropolitana do Estado do Espírito Santo e, para a produção das fontes, utiliza uma entrevista semiestruturada, um grupo focal e três a cinco narrativas individuais orais. Os professores explicitam os saberes privilegiados por essa disciplina como aqueles incorporados pela imbricação do Eu na situação e pela distanciação-regulação (CHARLOT, 2000). Ao dar visibilidade aos projetos de ensino desenvolvidos pelos professores, a pesquisa apresenta possibilidades de se constituir uma integração curricular, por meio de temas que perpassam diferentes disciplinas

Cultura, Consumo e Diferença : estudo com um grupo de surdos em Vitória-ES

Tanto pessoas diferentes, como culturas diferentes, quanto percepções diferentes produzem significados de consumo particulares, sejam esses referentes a determinado grupo de pessoas, sejam relacionados a uma cultura ou a seletividade de percepção de cada um de nós. Nessa perspectiva, cultura, consumo e diferença são analisados através de situações específicas de consumo de um grupo de surdos em Vitória-ES. Tal escolha deveu-se a ausência de estudos dentro da área da Administração que se ocupem do tema surdez, uma vez que a discussão sobre a surdez vem ganhando cada vez mais espaço, dentro e fora da esfera acadêmica. A busca aqui se realiza pela reflexão acerca da ausência do sentido da audição não pelas abordagens clínicas, médicas ou terapêuticas, mas aproximando-se dos estudos sociais interpretativistas (THOMA, 2008; STROBEL, 2008; SÁ, 2006). O intento é buscar compreender de que forma esses consumidores forjam suas relações sociais através do consumo de bens e serviços. Para tal análise, realizouse pesquisa bibliográfica sobre o consumo por uma perspectiva antropológica (ROCHA, 2000, 2005; MILLER, 2002; MCCRACKEN, 2003). A partir de então, aproximou-se da perspectiva cultural proposta pelos estudos surdos, na busca pela compreensão do consumo como mediador de relações e construtor de significações. Através de inspiração etnográfica com a utilização de observação direta e entrevistas informais (MALINOWSKI, 1978; GEERTZ, 1989; MAGNANI, 2002, 2003), o campo de pesquisa se desenvolveu a partir dos eventos, encontros, discussões e passeios realizados pelo grupo de surdos. Com estes, foi possível vivenciar algumas situações de consumo que envolvem do uso de transporte coletivo à encontros informais. Viu-se que das relações sociais forjadas pelos surdos e ouvintes emergem sujeitos-chaves que atuam na construção coletiva do grupo. Também identificou-se possibilidades de compreensão do consumidor através das abordagens política, médico/terapêutica e religiosa, além de artefatos que permeiam o grupo estudado. Deixou-se como sugestão para pesquisas futuras, dentre outras, a busca pela utilização da etnografia para a compreensão de grupos de consumidores, pela compreensão da relação entre a surdez e bens específicos de consumo, além da possibilidade de utilizar as perspectivas elencadas nesse trabalho em outras situações, com o intuito de alargar o universo de possibilidade de compreensão dos surdos.