Identification of residues and domains of Raf important for function in vivo and in vitro

Random mutagenesis and genetic screens for impaired Raf function in Caenorhabditis elegans were used to identify six loss-of-function alleles of lin-45 raf that result in a substitution of a single amino acid. The mutations were classified as weak, intermediate, and strong based on phenotypic severity. We engineered these mutations into the homologous residues of vertebrate Raf-1 and analyzed the mutant proteins for their underlying biochemical defects. Surprisingly, phenotype strength did not correlate with the catalytic activity of the mutant proteins. Amino acid substitutions Val-589 and Ser-619 severely compromised Raf kinase activity, yet these mutants displayed weak phenotypes in the genetic screen. Interestingly, this is because these mutant Raf proteins efficiently activate the MAPK (mitogen-activated protein kinase) cascade in living cells, a result that may inform the analysis of knockout mice. Equally intriguing was the observation that mutant proteins with non-functional Ras-binding domains, and thereby deficient in Ras-mediated membrane recruitment, displayed only intermediate strength phenotypes. This confirms that secondary mechanisms exist to couple Ras to Raf in vivo. The strongest phenotype in the genetic screens was displayed by a S508N mutation that again did not correlate with a significant loss of kinase activity or membrane recruitment by oncogenic Ras in biochemical assays. Ser-508 lies within the Raf-1 activation loop, and mutation of this residue in Raf-1 and the equivalent Ser-615 in B-Raf revealed that this residue regulates Raf binding to MEK. Further characterization revealed that in response to activation by epidermal growth factor, the Raf-S508N mutant protein displayed both reduced catalytic activity and aberrant activation kinetics: characteristics that may explain the C. elegans phenotype.

gl(2 vertical bar 2) current superalgebra and non-unitary conformal field theory

Motivated by application of current superalgebras in the study of disordered systems such as the random XY and Dirac models, we investigate gl(2\2) current superalgebra at general level k. We construct its free field representation and corresponding Sugawara energy-momentum tensor in the non-standard basis. Three screen currents of the first kind are also presented. (C) 2003 Elsevier B.V. All rights reserved.

An energy-based model for swing hammer mills

An energy-based swing hammer mill model has been developed for coke oven feed preparation. it comprises a mechanistic power model to determine the dynamic internal recirculation and a perfect mixing mill model with a dual-classification function to mimic the operations of crusher and screen. The model parameters were calibrated using a pilot-scale swing hammer mill at various operating conditions. The effects of the underscreen configurations and the feed sizes on hammer mill operations were demonstrated through the fitted model parameters. Relationships between the model parameters and the machine configurations were established. The model was validated using the independent experimental data of single lithotype coal tests with the same BJD pilot-scale hammer mill and full operation audit data of an industrial hammer mill. The outcome of the energy-based swing hammer mill model is the capability to simulate the impact of changing blends of coal or mill configurations and operating conditions on product size distribution. Alternatively, the model can be used to select the machine settings required to achieve a desired product. (C) 2003 Elsevier Science B.V. All rights reserved.

Media Convergence's Third Wave Content Streaming

Movies, pieces of music, books, or newspapers can all be expressed in the same binary code. Discrete forms of analogue media are just different dialects of the language of computerese. Content is becoming a very liquid asset. To take Marshall McLuhan's famed dictum a step further: The message is now independent of the medium

Vps20p and Vta1p interact with Vps4p and function in multivesicular body sorting and endosomal transport in Saccharomyces cerevisiae

Vps4p (End13p) is an AAA-family ATPase that functions in membrane transport through endosomes, sorting of soluble vacuolar proteins to the vacuole, and multivesicular body (MVB) sorting of membrane proteins to the vacuole lumen. In a yeast two-hybrid screen with Vps4p as bait we isolated VPS20 (YMR077c) and the novel open reading frame YLA181c, for which the name VTA1 has recently been assigned (Saccharomyces Genome Database). Vps4p directly binds Vps20p and Vta1p in vitro and binding is not dependent on ATP-conversely, Vps4p binding to Vps20p is partially sensitive to ATP hydrolysis. Both ATP binding [Vps4p-(K179A)] and ATP hydrolysis [Vps4p-(E233Q)] mutant proteins exhibit enhanced binding to Vps20p and Vta1p in vitro. The Vps4p-Vps20p interaction involves the coiled-coil domain of each protein, whereas the Vps4p-Vta1p interaction involves the (non-coiled-coil) C-terminus of each protein. Deletion of either VPS20 (vps20Delta) or VTA1 (vta1Delta) leads to similar class E Vps(-) phenotypes resembling those of vps4Delta, including carboxypeptidase Y (CPY) secretion, a block in ubiquitin-dependent MVB sorting, and a delay in both post-internalisation endocytic transport and biosynthetic transport to the vacuole. The vacuole resident membrane protein Sna3p (whose MVB sorting is ubiquitin-independent) does not appear to exit the class E compartment or reach the vacuole in cells lacking Vps20p, Vta1p or Vps4p, in contrast to other proteins whose delivery to the vacuole is only delayed. We propose that Vps20p and Vta1p regulate Vps4p function in vivo.

Actively shielded multi-layer gradient coil designs with improved cooling properties

In standard cylindrical gradient coils consisting of a single layer of wires, a limiting factor in achieving very large magnetic field gradients is the rapid increase in coil resistance with efficiency. This is a particular problem in small-bore scanners, such as those used for MR microscopy. By adopting a multi-layer design in which the coil wires are allowed to spread out into multiple layers wound at increasing radii, a more favourable scaling of resistance with efficiency is achieved, thus allowing the design of more powerful gradient coils with acceptable resistance values. Previously this approach has been applied to the design of unshielded, longitudinal, and transverse gradient coils. Here, the multi-layer approach has been extended to allow the design of actively shielded multi-layer gradient coils, and also to produce coils exhibiting enhanced cooling characteristics. An iterative approach to modelling the steady-state temperature distribution within the coil has also been developed. Results indicate that a good level of screening can be achieved in multi-layer coils, that small versions of such coils can yield higher efficiencies at fixed resistance than conventional two-layer (primary and screen) coils, and that performance improves as the number of layers of increases. Simulations show that by optimising multi-layer coils for cooling it is possible to achieve significantly higher gradient strengths at a fixed maximum operating temperature. A four-layer coil of 8 mm inner diameter has been constructed and used to test the steady-state temperature model. (C) 2003 Elsevier Inc. All rights reserved.

Normal values for pelvic organ descent in health nulligravid young caucasian women

Translabial ultrasound is increasingly being used for the assessment of women presenting with pelvic floor dysfunction and incontinence (1,2). However, there is little information on normal values for bladder neck descent, with the two available studies disagreeing widely (3,4). No data has so far been published on mobility of the central and posterior compartment which can now also be assessed by ultrasound (5). This study presents normal values for urethral, bladder, cervical and rectal mobility in a cohort of young, stress continent, nulliparous nonpregnant women. Methods 118 nonpregnant nulliparous Caucasian women between 18 and 23 years of age were recruited for an ongoing twin study of pelvic floor function. Translabial ultrasound assessment of pelvic organ mobility was undertaken supine and after bladder emptying (6,7). The best of at least three effective Valsalva manoeuvres was used for evaluation, with no attempts at standardization of Valsalva pressure. Parameters of anterior compartment mobility were obtained by the use of on-screen calipers; cervical and rectal descent were evaluated on printouts. All examinations were carried out under direct supervision of the first author or by personnel trained by him for at least 100 consecutive assessments. Results The median age of participants in this study was 20 (range 18- 23). Mean body mass index was 23 (range 16.9- 36.7). Of 118 women, 2 were completely unable to perform a Valsalva manoeuvre despite repeated efforts at teaching and were excluded from analysis, as were ten women who complained of urinary stress incontinence, leaving 106 datasets. Average measurements for the parameters ‘retrovesical angle at rest’ (RVA-R) and on Valsalva (RVA-S), urethral rotation, bladder neck mobility, cysto-cele descent, cervical descent and descent of the rectal ampulla are given in Table 1.

Alterações microclimáticas causadas pelo uso de tela plástica

Com o objetivo de caracterizar as alterações microclimáticas provocadas pelo uso de tela plástica na agricultura, foi desenvolvido um experimento com alface, em área de cultivo agrícola, em Alegre - ES (latitude 20º45 S, longitude 41º28 W e altitude 150 m), no período de setembro a dezembro de 1996. Foram utilizados sombreamentos de 0; 30; 50, e 70%, segundo especificação comercial. Os resultados mostraram que a tela com especificação comercial de 50% de sombreamento provocou uma atenuação média de 41% na radiação solar global, ocorrendo, no entanto, uma flutuação significativa ao longo do dia em função do movimento aparente do sol. A cobertura com a tela plástica não provocou alterações significativas da temperatura e umidade relativa do ar. Em relação à temperatura do solo, foi possível verificar uma atenuação com o aumento do sombreamento.

Síntese, aplicação e avaliação da toxicidade aguda de complexo orgânico à base de európio proposto como marcador fotoluminescente para a Identificação de resíduos de tiro

Um complexo de alta fotoluminescência é proposto como marcador óptico para a identificação de resíduos de tiro (GSR). O marcador é o complexo [Eu(PIC)3(NMK)3], de fórmula molecular Eu(C6H2N3O7)3.(C7H13NO)3, que apresenta o íon Eu3+ e os ligantes ácido pícrico (PIC) e n-metil-Ɛ-caprolactama (NMK). Foi realizada a caracterização quimicamente através de espectroscopia de emissão, espectroscopia de infravermelho com transformada de Fourier (FTIR), termogravimetria e análise térmica diferencial (TG/DTA), e espectrometria de massas com ionização por eletrospray e ressonância ciclotrônica de íons por transformada de Fourier (ESI-FT-ICR MS), e, em seguida, foram adicionadas diferentes massas do complexo a munições convencionais (de 2 a 50 mg por cartucho). Após os tiros, o GSR marcado foi visualmente e quimicamente detectado por irradiação UV (ʎ = 395 nm) e ESI-FT-ICR MS, respectivamente. Os resultados mostraram uma fotoluminescência eficiente e duradoura, sendo facilmente visível sobre a superfície do alvo, no ambiente, no cartucho deflagrado, na arma de fogo, e sobre as mãos e braços do atirador quando utilizada massa a partir de 25 mg do marcador em cartuchos .38 e 50 mg em cartuchos .40. Sua toxicidade aguda também foi avaliada empregando-se o Protocolo 423 da Organização para a Cooperação e Desenvolvimento Econômico (OECD) e apresentou DL50 de 1000 mg.kg-1, sendo classificado como de categoria 4 na escala do Sistema Globalmente Harmonizado de Classificação e Rotulagem de Produtos Químicos (GHS), considerado, portanto, de média toxicidade. O composto mostrou ser menos tóxico do que os componentes inorgânicos de munições convencionais (em especial o Pb), justificando o seu emprego como marcador de GSR.

Entre imagens cinema e imagens escola, movimentando o pensamento com a formação de professores

A presente pesquisa aborda o estudo dos movimentos do pensamento nas redes formação com professores no Ensino Fundamental. Procura problematizar a constituição de processos formativos não dogmáticos nos cotidianos escolares, pela articulação de imagens cinema e imagens escola, exibidas, compartilhadas e criadas nas redes de conversações em uma escola municipal de Vitória/ES, entre os anos de 2013 a 2014. Apresenta um campo problemático que se dedica a compreender que movimentos do pensamento são produzidos a partir do uso de imagens cinema nas redes de conversações com professores em formação continuada, a partir de uma cartografia de pesquisa que articula o mapeamento de virtualidades presentes no banco de teses da CAPES e as cinecartografias do cotidiano escolar em duas dobras do tempo. Tece um debate teórico que se compõe pelas linhas de pensamento, principalmente, de Bergson (2006a; 2006b), Carvalho (2008; 2009; 2010; 2012) Deleuze (1988; 1991; 1992; 1995; 1996; 2003; 2006; 2007; 2010; 2011), Foucault (1979; 1999. 2006a; 2006b; 2008), Guéron (2010; 2011), Pelbart (2011), Sauvagnargues (2009), dentre outros. As opções teórico-metodológicas estabelecem uma pesquisa de campo na intercessão entre os estudos com o cotidiano escolar e os da pesquisa cartográfica, tomando o plano de imanência como mundo material para a ocorrência de acontecimentos. Utiliza, para a produção de dados a observação participante, os registros em diário de campo, considerados como uma descrição cristalina, assim como, as narrativas cristalinas que emergiam nas redes de conversações com as personagens-escolas (docentes), que se configuram como imagensnarrativas de pesquisa. Organiza o debate dos dados produzidos em sete capítulos. Apresenta como considerações três principais agrupamentos do movimento do pensamento nas redes de conversações com professores provocados pelas imagens cinema: cinescola-clichês, imagensformação docente e cristaisescolas. Destaca que não há uma realidade que comporte uma só verdade sobre a formação com professores, mas o que existe são cortes, percepções, afecções e produções de verdades que entrelaçam os diferentes modos de existir e reexistir na educação brasileira. As telasescolas, assim como as telas do cinema, apresentam e nos cercam com um mundo que articula um cinescola-clichê, imagensformação docente e cristaisescolas, que para além do pensamento reto e dogmático, exibem entre imagens cinema e imagens escolas, outros movimentos do pensamento com a formação de professores, sendo capazes de potencializar as pesquisas educacionais a não ir em ‘busca de um tempo perdido, mas de um tempo a ser redescoberto’, fazendo crer ainda mais nesse mundo e na educação.

Digital radiography detectors - A technical overview: Part 1

During the last two decades screen-film (SF) systems have been replaced by digital X-ray systems. The advent of digital technologies brought a number of digital solutions based on different detector and readout technologies. Improvements in technology allowed the development of new digital technologies for projection radiography such as computed radiography (CR) and digital radiography (DR). The large number of scientific papers concerning digital X-ray systems that have been published over the last 25 years indicates the relevance of these technologies in healthcare. There are important differences among different detector technologies that may affect system performance and image quality for diagnostic purposes. Radiographers are expected to have an effective understanding of digital X-ray technologies and a high level of knowledge and awareness concerning the capabilities of these systems. Patient safety and reliable diagnostic information are intrinsically linked to these factors. In this review article - which is the first of two parts - a global overview of the digital radiography systems (both CR and DR) currently available for clinical practice is provided.

Digital radiography detectors - A technical overview: Part 2

Digital X-ray detector technologies provide several advantages when compared with screen-film (SF) systems: better diagnostic quality of the radiographic image, increased dose efficiency, better dynamic range and possible reduction of radiation exposure to the patient. The transition from traditional SF systems to digital technology-based systems highlights the importance of the discussion around technical factors such as image acquisition, themanagement of patient dose and diagnostic image quality. Radiographers should be aware of these aspects concerning their clinical practice regarding the advantages and limitations of digital detectors. Newdigital technologies require an up-to-date of scientific knowledge concerning their use in projection radiography. This is the second of a two-part review article focused on a technical overview of digital radiography detectors. This article provides a discussion about the issues related to the image acquisition requirements and advantages of digital technologies, the management of patient dose and the diagnostic image quality.