A nonnatural transcriptional coactivator

In eukaryotes, sequence-specific DNA-binding proteins activate gene expression by recruiting the transcriptional apparatus and chromatin remodeling proteins to the promoter through protein-protein contacts. In many instances, the connection between DNA-binding proteins and the transcriptional apparatus is established through the intermediacy of adapter proteins known as coactivators. Here we describe synthetic molecules with low molecular weight that act as transcriptional coactivators. We demonstrate that a completely nonnatural activation domain in one such molecule is capable of stimulating transcription in vitro and in vivo. The present strategy provides a means of gaining external control over gene activation through intervention using small molecules.

Intracellular Localization of Phospholipase D1 in Mammalian Cells

Phospholipase D (PLD) hydrolyzes phosphatidylcholine to generate phosphatidic acid. In mammalian cells this reaction has been implicated in the recruitment of coatomer to Golgi membranes and release of nascent secretory vesicles from the trans-Golgi network. These observations suggest that PLD is associated with the Golgi complex; however, to date, because of its low abundance, the intracellular localization of PLD has been characterized only indirectly through overexpression of chimeric proteins. We have used highly sensitive antibodies to PLD1 together with immunofluorescence and immunogold electron microscopy as well as cell fractionation to identify the intracellular localization of endogenous PLD1 in several cell types. Although PLD1 had a diffuse staining pattern, it was enriched significantly in the Golgi apparatus and was also present in cell nuclei. On fragmentation of the Golgi apparatus by treatment with nocodazole, PLD1 closely associated with membrane fragments, whereas after inhibition of PA synthesis, PLD1 dissociated from the membranes. Overexpression of an hemagglutinin-tagged form of PLD1 resulted in displacement of the endogenous enzyme from its perinuclear localization to large vesicular structures. Surprisingly, when the Golgi apparatus collapsed in response to brefeldin A, the nuclear localization of PLD1 was enhanced significantly. Our data show that the intracellular localization of PLD1 is consistent with a role in vesicle trafficking from the Golgi apparatus and suggest that it also functions in the cell nucleus.

Transdifferentiation of Mature Cortical Cells to Functional Abscission Cells in Bean1

Abscission explants of bean (Phaseolus vulgaris L.) were treated with ethylene to induce cell separation at the primary abscission zone. After several days of further incubation of the remaining petiole in endogenously produced ethylene, the distal two-thirds of the petiole became senescent, and the remaining (proximal) portion stayed green. Cell-to-cell separation (secondary abscission) takes place precisely at the interface between the senescing yellow and the enlarging green cells. The expression of the abscission-associated isoform of β-1,4-glucanhydrolase, the activation of the Golgi apparatus, and enhanced vesicle formation occurred only in the enlarging cortical cells on the green side. These changes were indistinguishable from those that occur in normal abscission cells and confirm the conversion of the cortical cells to abscission-type cells. Secondary abscission cells were also induced by applying auxin to the exposed primary abscission surface after the pulvinus was shed, provided ethylene was added. Then, the orientation of development of green and yellow tissue was reversed; the distal tissue remained green and the proximal tissue yellowed. Nevertheless, separation still occurred at the junction between green and yellow cells and, again, it was one to two cell layers of the green side that enlarged and separated from their senescing neighbors. Evaluation of Feulgen-stained tissue establishes that, although nuclear changes occur, the conversion of the cortical cell to an abscission zone cell is a true transdifferentiation event, occurring in the absence of cell division.

Thapsigargin-induced transport of cholera toxin to the endoplasmic reticulum.

Cholera toxin is normally observed only in the Golgi apparatus and not in the endoplasmic reticulum (ER) although the enzymatically active A subunit of cholera toxin has a KDEL sequence. Here we demonstrate transport of horseradish peroxidase-labeled cholera toxin to the ER by electron microscopy in thapsigargin-treated A431 cells. Thapsigargin treatment strongly increased cholera toxin-induced cAMP production, and the formation of the catalytically active A1 fragment was somewhat increased. Binding of cholera toxin to the cell surface and transport of toxin to the Golgi apparatus were not changed in thapsigargin-treated cells, suggesting increased retrograde transport of cholera toxin from the Golgi apparatus to the ER. The data demonstrate that retrograde transport of cholera toxin can take place and that the transport is under regulation. The results are consistent with the idea that retrograde transport can be important for the action of cholera toxin.

Sphingolipid synthesis as a target for chemotherapy against malaria parasites.

The human malaria parasite Plasmodium falciparum contains sphingomyelin synthase in its Golgi apparatus and in a network of tubovesicular membranes in the cytoplasm of the infected erythrocyte. Palmitoyl and decanoyl analogues of 1-phenyl-2-acylamino-3-morpholino-1-propanol inhibit the enzyme activity in infected erythrocytes. An average of 35% of the activity is extremely sensitive to these drugs and undergoes a rapid, linear decrease at drug concentrations of 0.05-1 microM. The remaining 65% suffers a slower linear inhibition at drug concentrations ranging from 25 to 500 microM. Evidence is presented that inhibition of the sensitive fraction alone selectively disrupts the appearance of the interconnected tubular network in the host cell cytoplasm, without blocking secretory development at the parasite plasma membrane or in organelles within the parasite, such as the Golgi and the digestive food vacuole. This inhibition also blocks parasite proliferation in culture, indicating that the sensitive sphingomyelin synthase activity as well as the tubovesicular network may provide rational targets for drugs against malaria.

Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion.

Chlamydia trachomatis undergoes its entire life cycle within an uncharacterized intracellular vesicle that does not fuse with lysosomes. We used a fluorescent Golgi-specific probe, (N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]) aminocaproylsphingosine (C6-NBD-Cer), in conjunction with conventional fluorescence or confocal microscopy to identify interactions between the Golgi apparatus and the chlamydial inclusion. We observed not only a close physical association between the Golgi apparatus and the chlamydial inclusion but the eventual presence of a metabolite of this fluorescent probe associated with the chlamydiae themselves. Sphingomyelin, endogenously synthesized from C6-NBD-Cer, was specifically transported to the inclusion and incorporated into the cell wall of the intracellular chlamydiae. Incorporation of the fluorescent sphingolipid by chlamydiae was inhibited by brefeldin A. Chlamydiae therefore occupy a vesicle distal to the Golgi apparatus that receives anterograde vesicular traffic from the Golgi normally bound for the plasma membrane. Collectively, the data suggest that the chlamydial inclusion may represent a unique compartment within the trans-Golgi network.

O conceito de equilíbrio hidráulico e termodinâmico da água no solo aplicado ao estudo da curva de retenção e murchamento de plantas

Estimativas sobre alterações climáticas globais têm aumentando a demanda por estudos sobre propriedades dos solos relativamente secos e limitações impostas à absorção de água pelas plantas em condições de escassez hídrica. Neste estudo, fatores que influenciam a retenção da água no solo e o murchamento de plantas foram avaliados com base no conceito de equilíbrio da água no solo. Objetivou-se com este estudo: (i) avaliar a confiabilidade de medições do conteúdo de água no solo sob altas sucções matriciais em câmaras de pressão, usando como referência a técnica de ponto de orvalho (ii) avaliar as interações entre espécies de plantas e solos com diferentes classes texturais no ponto de murcha permanente (iii) investigar as relações entre equilíbrio hidráulico da água no solo e murchamento de plantas a partir do conceito de corte hidráulico. Para tanto, um experimento para avaliar a influência dos tipos de solos e espécies de plantas, no ponto de murcha permanente foi conduzido em casa de vegetação da Escola Superior de Agricultura \"Luiz de Queiroz\" da Universidade de São Paulo, Piracicaba, São Paulo. Avaliou-se o murchamento de plantas de girassol (Helianthus annuus L.), milho (Zea mays L.) e soja (Glycine max L.). Os solos utilizados no estudo foram coletados na camada superficial (0-10 cm) em quatro áreas, selecionadas com o objetivo de obter classes texturais contrastantes, localizadas no município de Piracicaba, São Paulo, Brasil. Sub-amostras foram utilizadas para determinação da distribuição do tamanho de partículas e atributos químicos. Amostras indeformadas foram coletadas para a determinação da curva de retenção da água no solo pela técnica de câmaras de pressão. Adicionalmente, amostras deformadas foram utilizadas para determinação das características de retenção da água no solo pela técnica do ponto de orvalho em altos valores de sucções matriciais. Os dados de retenção de água no solo foram ajustados a modelos empíricos para estimativas da sucção matricial e conteúdo de água relacionada à água em equilíbrio hidráulico (água residual). Foram observadas similaridades nas determinações das características de retenção da água no solo entre as técnicas de câmaras de pressão e ponto de orvalho, sugerindo a boa drenagem das amostras de solo em câmaras de pressão. Interações significativas foram observadas entre os tipos de solos e espécies de plantas no ponto de murcha permanente, indicando que o movimento de água no contínuo solo-planta-atmosfera foi dependente de resistências relacionadas tanto ao solo quanto às plantas. Ou seja, tanto à capacidade do solo em transportar água até raízes, quanto à habilidade das plantas em absorver a água transportada, assim como, aos processos de regulação de água que ocorrem nas plantas. A abordagem baseada no conteúdo de água residual para o intervalo de sucções matriciais de 0 a 15.000 hPa não foi adequada para ilustrar a condição de equilíbrio hidráulico da água no solo, definidos pelo corte hidráulico, e relações com as sucções matriciais em ocorre o murchamento de plantas.

Influência da configuração da célula combustível microbiana na geração de energia elétrica a partir da degradação de compostos orgânicos

Nessa pesquisa foram testadas tanto CCM com membranas e sem membranas que tinham por característica reproduzir sistemas de tratamento de esgoto sanitário. A etapa experimental desse trabalho foi dividida entre o Brasil (ensaios com CCM sem a MTP e utilizando esgoto sanitário) e Portugal (ensaios com CCM tradicionais de uma e duas câmaras, utilizando água residuária sintética e a bactéria Lactobacillus pentosus). A execução em dois locais diferentes resultou em um maior aprofundamento e desenvolvimento da pesquisa. As CCM foram avaliadas principalmente quanto ao potencial elétrico e eficiência da degradação de compostos orgânicos (esgoto sanitário e água residuária sintética). Para os dados obtidos no Brasil, as três configurações apresentaram maior diferença na potência em função do modo de operação. A operação intermitente apresentou a maior potência (11 mW/m2) para a CCM cilíndrica de fluxo ascendente, enquanto que operação continua a maior potência (4,2 mW/m2) foi obtida para a CCM retangular de fluxo horizontal, a qual também apresentava uma maior facilidade na manutenção quanto aos eletrodos (adição/remoção). A CCM cúbica de fluxo ascendente devido a sua concepção simples demandava um sistema complementar para o aumento da remoção de DQO. Apesar da baixa potência mensurada para os ensaios realizados no Brasil há de se pontuar que os mesmos foram obtidos para reatores sem membranas e utilizando o esgoto sanitário, o qual apresentou grande sazonalidade. Para a etapa realizada em Portugal, foi possível realizar quinze diferentes ensaios e mais um ensaio específico de crescimento. A maior potência (10,37 mW/m2) foi obtida para CCM de câmara dupla operada de modo contínuo para um tempo de detenção hidráulico (TDH) de 20 horas. A maior potência obtida para a CCM de câmara única foi de 5,53 mW/m2 quando houve a adição do extrato de levedura (função teórica de mediador). A potência da CCM, na maioria das vezes, esteve relacionada à proporção de sólidos voláteis e totais, SV/ST, quantidade de bactérias, pH, características de operação e por fim a configuração da CCM. O ensaio de crescimento revelou a correlação da potência em função da quantidade de bactérias inseridas da massa do biofilme (SV) e mostra-se como uma ferramenta na avaliação da potência das CCM.

Fault location in power distribution networks with distributed generation.

This research presents the development and implementation of fault location algorithms in power distribution networks with distributed generation units installed along their feeders. The proposed algorithms are capable of locating the fault based on voltage and current signals recorded by intelligent electronic devices installed at the end of the feeder sections, information to compute the loads connected to these feeders and their electric characteristics, and the operating status of the network. In addition, this work presents the study of analytical models of distributed generation and load technologies that could contribute to the performance of the proposed fault location algorithms. The validation of the algorithms was based on computer simulations using network models implemented in ATP, whereas the algorithms were implemented in MATLAB.

Modelagem de sinais neuronais utilizando filtros lineares de tempo discreto.

A aquisição experimental de sinais neuronais é um dos principais avanços da neurociência. Por meio de observações da corrente e do potencial elétricos em uma região cerebral, é possível entender os processos fisiológicos envolvidos na geração do potencial de ação, e produzir modelos matemáticos capazes de simular o comportamento de uma célula neuronal. Uma prática comum nesse tipo de experimento é obter leituras a partir de um arranjo de eletrodos posicionado em um meio compartilhado por diversos neurônios, o que resulta em uma mistura de sinais neuronais em uma mesma série temporal. Este trabalho propõe um modelo linear de tempo discreto para o sinal produzido durante o disparo do neurônio. Os coeficientes desse modelo são calculados utilizando-se amostras reais dos sinais neuronais obtidas in vivo. O processo de modelagem concebido emprega técnicas de identificação de sistemas e processamento de sinais, e é dissociado de considerações sobre o funcionamento biofísico da célula, fornecendo uma alternativa de baixa complexidade para a modelagem do disparo neuronal. Além disso, a representação por meio de sistemas lineares permite idealizar um sistema inverso, cuja função é recuperar o sinal original de cada neurônio ativo em uma mistura extracelular. Nesse contexto, são discutidas algumas soluções baseadas em filtros adaptativos para a simulação do sistema inverso, introduzindo uma nova abordagem para o problema de separação de spikes neuronais.

Alternative Energy at a Steel Mill

More than ever alternative energy solutions are being discussed including potential legislative action. With the use of fossil fuels being the most abundant and most controversial sources of energy, new solutions must be found. This paper looks at the economic and technical feasibility of different types of alternative energy and cogeneration applications within a steel mill. This paper examines alternative energy systems. The systems examined include Solar systems, thermal electric materials, and an Organic Rankine Cycle cogeneration project. None of the projects has limiting technical feasibility issues however each of the projects face economically limiting factors. Taking into account tangible and non-tangible factors the solar system and Organic Rankine Cycle cogeneration project are recommended for further study and potential installation.

Mr. Marsh's catalogue, ca. 1764

A half-page handwritten list of books with the author's surname, title, and location in the old Harvard Library, signed "Mr. Marsh." The list includes the note, "Shuckford's Connection is charged to you." The document is undated but presumably was created following the Harvard Hall Fire of 1764 as part of the College's efforts to inventory volumes that were spared because they were checked out at the time of the fire. Many of the books are listed in a charging record for Thomas Marsh recorded in a Harvard library account book (UAIII 50.15.60, Volume 1, Box 95), including "Shuckford's connection" which was charged to Marsh on September 23, 1763.

Optimum Design of Electromagnets for Magnetic Levitation of Transport Systems based on the Inverse Problem Solutions

Article is devoted to design of optimum electromagnets for magnetic levitation of transport systems. The method of electromagnets design based on the inverse problem solution of electrical equipment is offered. The method differs from known by introducing a stage of minimization the target functions providing the stated levitation force and magnetic induction in a gap, and also the mass of an electromagnet. Initial values of parameters are received, using approximate formulas of the theory of electric devices and electrical equipment. The example of realization of a method is given. The received results show its high efficiency at design. It is practical to use the offered method and the computer program realizing it as a part of system of the automated design of electric equipment for transport with a magnetic levitation.

(Table 2) Concentrations of dissolved aluminum in interstitial water from sediments of the Indian Ocean

Results are presented of application of laser stepwise photoionization of atoms in combination with thermal atomization of matter in vacuum for direct determination of aluminum dissolved in sea and interstitial waters. Dry residue from evaporation of 40 ?l sea water was atomized in a crucible at 1800°C, and aluminum atoms in the beam thus formed were energized into Rydberg state in two steps by two tunable dye laser beams; the atoms were then ionized by an electric pulse and resulting ions were recorded by secondary emission electron multiplier (ion detector). Ionic signal dependence on sample vaporization time was studied. The procedure is suggested for separating out a selective signal in a single measurement. Dissolved aluminum concentrations in interstitial waters of the Indian Ocean and in waters of the river-sea zone were determined using preliminarily plotted calibration characteristics for aluminum solutions in deionized and sea waters. The minimum detectable Al concentration in seawater was 1 ?g/l that corresponds to 40 pg of Al in a sample.

(Table 1) Air-earth current density investigated during Atlantic Expedition 1973 of METEOR

One main point of our atmospheric-electric measurements over the Atlantic Ocean 1973 was the investigation of the air-earth current density above the sea. In addition to direct measurements at the water surface with a floating net, we calculated the air-earth current density from the electric field and the air conductivity measured simultaneously on board of the ship and during particular ascents in the free atmosphere. During all five ascents the air-earth current density did not change with altitude. For pure maritime air-conditions, the mean air-earth current density was found to be 2.9 pA/m**2. The mean hourly air-earth current density over the Atlantic shows nearly the same 24-hour pattern as measured by Cobb (1977) at the South Pole at the same time. When dust-loaden air masses of African origin reached the ship as well as under continental influence the mean air-earth current density was reduced to 2.1 pA/m**2. The global 24-hour pattern was modified by this continental influences. Finally, it is shown that the values of the air conductivity measured on board R. V. "Meteor" during our earlier expeditions have been influenced by the exhaust of the ship and must therefore be corrected. With this correction, our new mean values of the air-earth current density over the Atlantic are 2.6 pA/m**2 in 1965 and 2.0 pA/m**2 in 1969. From all measurements, the global air-earth current is estimated to be about 1250 A.