The Boron Requirement and Cell Wall Properties of Growing and Stationary Suspension-Cultured Chenopodium album L. Cells1

Suspension-cultured Chenopodium album L. cells are capable of continuous, long-term growth on a boron-deficient medium. Compared with cultures grown with boron, these cultures contained more enlarged and detached cells, had increased turbidity due to the rupture of a small number of cells, and contained cells with an increased cell wall pore size. These characteristics were reversed by the addition of boric acid (≥7 μm) to the boron-deficient cells. C. album cells grown in the presence of 100 μm boric acid entered the stationary phase when they were not subcultured, and remained viable for at least 3 weeks. The transition from the growth phase to the stationary phase was accompanied by a decrease in the wall pore size. Cells grown without boric acid or with 7 μm boric acid were not able to reduce their wall pore size at the transition to the stationary phase. These cells could not be kept viable in the stationary phase, because they continued to expand and died as a result of wall rupture. The addition of 100 μm boric acid prevented wall rupture and the wall pore size was reduced to normal values. We conclude that boron is required to maintain the normal pore structure of the wall matrix and to mechanically stabilize the wall at growth termination.

Mutations affecting conserved cysteine residues within the extracellular domain of Neu promote receptor dimerization and activation.

Overexpression of the Neu/ErbB-2 receptor tyrosine kinase has been implicated in the genesis of human breast cancer. Indeed, expression of either activated or wild-type neu in the mammary epithelium of transgenic mice results in the induction of mammary tumors. Previously, we have shown that many of the mammary tumors arising in transgenic mice expressing wild-type neu occur through somatic activating mutations within the neu transgene itself. Here we demonstrate that these mutations promote dimerization of the Neu receptor through the formation of disulfide bonds, resulting in its constitutive activation. To explore the role of conserved cysteine residues within the region deleted in these altered Neu proteins, we examined the transforming potential of a series of Neu receptors in which the individual cysteine residues were mutated. These analyses indicated that mutation of certain cysteine residues resulted in the oncogenic activation of Neu. The increased transforming activity displayed by the altered receptors correlated with constitutive dimerization that occurred in a disulfide bond-dependent manner. We further demonstrate that addition of 2-mercaptoethanol to the culture medium interfered with the specific transforming activity of the mutant Neu receptors. These observations suggest that oncogenic activation of Neu results from constitutive disulfide bond-dependent dimerization.

Ascorbic acid is essential for the release of insulin from scorbutic guinea pig pancreatic islets.

Pancreatic islets from young normal and scorbutic male guinea pigs were examined for their ability to release insulin when stimulated with elevated D-glucose. Islets from normal guinea pigs released insulin in a D-glucose-dependent manner showing a rapid initial secretion phase and three secondary secretion waves during a 120-min period. Islets from scorbutic guinea pigs failed to release insulin during the immediate period, and only delayed and decreased responses were observed over the 40-60 min after D-glucose elevation. Insulin release from scorbutic islets was greatly elevated if 5 mM L-ascorbic acid 2-phosphate was supplemented in the perifusion medium during the last 60 min of perifusion. When 5 mM L-ascorbic acid 2-phosphate was added to the perifusion medium concurrently with elevation of medium D-glucose, islets from scorbutic guinea pigs released insulin as rapidly as control guinea pig islets and to a somewhat greater extent. L-Ascorbic acid 2-phosphate without elevated D-glucose had no effect on insulin release by islets from normal or scorbutic guinea pigs. The pancreas from scorbutic guinea pigs contained 2.4 times more insulin than that from control guinea pigs, suggesting that the decreased insulin release from the scorbutic islets was not due to decreased insulin synthesis but due to abnormal insulin secretion.

Rapid acquisition of dendritic spines by visual thalamic neurons after blockade of N-methyl-D-aspartate receptors.

N-Methyl-D-aspartate (NMDA) receptors play an important role in the development of retinal axon arbors in the mammalian lateral geniculate nucleus (LGN). We investigated whether blockade of NMDA receptors in vivo or in vitro affects the dendritic development of LGN neurons during the period that retinogeniculate axons segregate into on-center and off-center sublaminae. Osmotic minipumps containing either the NMDA receptor antagonist D-2-amino-5-phosphonovaleric acid (D-APV) or saline were implanted in ferret kits at postnatal day 14. After 1 week, LGN neurons were intracellularly injected with Lucifer yellow. Infusion of D-APV in vivo led to an increase in the number of branch points and in the density of dendritic spines compared with age-matched normal or saline-treated animals. To examine the time course of spine formation, crystals of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate were placed in the LGN in brain slices from 14- to 18-day-old ferrets. Labeled LGN cell dendrites were imaged on-line in living slices by confocal microscopy, with slices maintained either in normal perfusion medium or with the addition of D-APV or NMDA to the medium. Addition of D-APV in vitro at doses specific for blocking NMDA receptors led to a > 6-fold net increase in spine density compared with control or NMDA-treated slices. Spines appeared within a few hours of NMDA receptor blockade, indicating a rapid local response by LGN cells in the absence of NMDA receptor activation. Thus, activity-dependent structural changes in postsynaptic cells act together with changes in presynaptic arbors to shape projection patterns and specific retinogeniculate connections.

Fisiologia e formação de partículas lipídicas durante o crescimento da levedura Yarrowia lipolytica IMUFRJ 50682.

A levedura Yarrowia lipolytica tem sido muito investigada, especialmente por ser um microrganismo oleaginoso, ou seja, capaz de acumular grandes quantidades de lipídios, o que ocorre majoritariamente em organelas denominadas partículas lipídicas. Estes lipídios apresentam várias potenciais aplicações biotecnológicas, como por exemplo na produção de óleo microbiano (single cell oil) e na produção de biodiesel. Durante este projeto de mestrado, objetivou-se estudar a fisiologia de duas linhagens da levedura Y. lipolytica, sendo uma tradicionalmente estudada pela comunidade científica internacional (linhagem w29) e outra isolada da Baía da Guanabara, no Rio de Janeiro (linhagem IMUFRJ 50682). Foram realizados cultivos em frascos agitados tipo Erlenmeyer com defletores tampados com algodão (volume total 500 mL, volume de meio 100 mL, 28 oC e 200 rotações por minuto), durante os quais foi possível: 1) escolher um meio de cultivo de composição totalmente definida, com tiamina como único fator de crescimento, adequado a estudos de fisiologia quantitativa com esta levedura; 2) verificar que Y. lipolytica não é capaz de crescer com sacarose ou xilose como única fonte de carbono; 3) verificar que Y. lipolytica cresce com velocidade específica de crescimento máxima (Máx) de 0,49 h-1 num meio complexo contendo glicose, extrato de levedura e peptona (meio YPD), 0,31 h-1 em meio definido com glicose como única fonte de carbono e 0,35 h-1 no mesmo meio, mas com glicerol como única fonte de carbono, sem excreção de metabólitos para o meio de cultivo; 4) verificar que ocorreu limitação por oxigênio nos cultivos em frasco agitado, sendo este o motivo pelo qual as células deixaram de crescer exponencialmente; 5) verificar que o uso de ureia, em vez de sulfato de amônio, como fonte de nitrogênio, contribui para uma variação menor do pH durante os cultivos, sem prejuízo ao crescimento da levedura; 6) observar que, ao se restringir a oferta de nitrogênio à levedura (aumento da relação C/N inicial no meio de 12,6 para 126), as células têm sua morfologia alterada e apresentam maior quantidade de partículas lipídicas; 7) determinar uma composição elementar para a biomassa de Y. lipolytica (CH1,98O0,58N0,13), em que os átomos de carbono encontram-se em média mais reduzidos do que na biomassa de leveduras como Saccharomyces cerevisiae e Dekkera bruxellensis. Foram também realizados cultivos em biorreator em batelada (1 L de volume útil, 28 oC, aerobiose plena e pH controlado em 5,0), durante os quais foi possível: a) estabelecer um protocolo de cultivo para Y. lipolytica em biorreator (que envolvem agitação mecânica, aeração e uso de anti-espumante, entre outras diferenças em relação aos cultivos em frasco); b) confirmar os valores dos principais parâmetros fisiológicos apresentados por esta levedura, anteriormente obtidos a partir de cultivos em frasco; c) confirmar que o fator de conversão de substrato a células (Yx/s) é maior para cultivos realizados com glicerol como fonte única de carbono (0,53 g/g para a linhagem IMUFRJ 50682), do que com glicose (0,48 g/g para a mesma linhagem). Finalmente, cultivos realizados em quimiostato com glicerol como fonte de carbono e energia, limitados por amônio (fonte de nitrogênio, relação C/N 126), às vazões específicas de 0,25 h-1 e 0,15 h-1, permitiram observar que o número de partículas lipídicas por célula de Y. lipolytica permaneceu em torno de 2 em ambas as situações e houve uma diminuição no teor de nitrogênio nas células quando a velocidade específica de crescimento diminuiu de 0,25 para 0,15 h-1.

Propostes d’aula per a un enfocament motivador de la literatura

Els textos literaris, a banda de ser un terreny molt òptim per a desenvolupar la sensibilitat i ensenyar a sentir, són un recurs útil també per a millorar la competència lingüística de l’alumnat de manera motivadora i divertida. Aquests textos, junt a altres, formen part del treball diari a l’aula del Portafolis de Valencià on l’alumnat reflexiona sobre com millorar el seu nivell en aquesta llengua. La nostra proposta presenta seqüències didàctiques d’activitats on es desenvolupen tant les habilitats productives com receptives a traves del treball de textos literaris com l’endevinalla, les nadaletes, els textos narratius amb imatge (curt, fotonovel·la, còmic), el rap, els contes, la poesia...

Kosowo - przed ostatecznym rozwiazaniem. Proces uregulowania statusu miedzynarodowego = uwarunkowania polityczne i historyczne perspektywy rozwoju sytuacji. Prace OSW 2008. Kosovo before the final decision. Regulation Kosovo's international status - historical and political conditions and prospects for future developments. OSW Study 27/2008

The question of Kosovo's status is currently one of the most important issues in international politics. Since 1999, Kosovo has been an international protectorate which was created in the aftermath of the NATO intervention to stop the brutal pacification of the Albanian insurgency by Serb forces. The province has since de facto become independent of Serbia. Resolution 1244 of the UN Security Council, which established the protectorate, does not preclude any possible outcome as regards its status. Aware that after the crimes of 1999, any attempt to re-integrate Kosovo into Serbia would lead to a massive Albanian uprising, the West has decided that the best solution would be to award Kosovo internationally supervised independence, while at the same time granting very wide autonomy to the Kosovo Serbs. Serbia and Russia rejected the solution proposed by the West, and so Kosovo became an arena of international rivalry for influence in the Western Balkans as well as another element of rivalry, transcending the regional dimension, between Russia and the West. Russia has been using the Kosovo case to build a new model of its relations with the United States and the EU. Since there is a group of countries sceptical about, or even opposed to, Kosovo's independence within the EU, the Kosovo settlement will be a test of the EU's ability to speak with one voice with regard to its external policy.

The Financial Crisis and the Future of the Eurozone. Bruges European Economic Policy (BEEP) Briefing 21/2010

The government debt crisis, erupted in the Eurozone in 2009, nearly led to the collapse of European monetary union. Now that this has been averted, the question is what should be done to make the Eurozone sustainable in the long run. The survival of the Eurozone hinges on the capacity of its leaders to improve the eurozone's governance. With the exception of Greece, the root cause of the government debt crisis has little to do with the poor performance of the SGP, rather, with unsustainable debt accumulation by private actors. Also, the method of convergence implicit in the SGP has not worked well – macroeconomic divergences have stubbornly remained for nearly a decade and several countries experienced boom and bust dynamics. Although strong declines in real interest rates may explain part of the story (but e.g. Italy did not experience boom & bust), self-fulfilling waves of optimism and pessimism which might be called 'animal spirits' and are of mainly national origin, seem a good candidate for explanation. These national animal spirits endogenously trigger credit expansion and contraction. It follows that (national) movements of credit ought to be under much firmer control and this is up to the monetary authorities, including the ECB. Critical recommendations for better governance of the Eurozone should therefore combine credible measures to maintain fiscal discipline over the medium term with such instruments as minimum reserve requirements to control the growth of bank credit as well as minimum reserve requirements in different national banking systems. Finally, the idea of adding more sanctions to the SGP may be ill-conceived since, in future, it might pre-empt national governments to come to the rescue of banks (under credible threats of contagion) and/or prevent a downward spiral in economic activity.

Sound! Trumpet! = [music] : (Squillino le trombe!) /

First line of text: Fratelli avvampa la patria.

Scotch phantasy, for violin and piano, op.46.

Mode of access: Internet.

Vocalises. [music] /

v. 1. Twenty daily exercises -- v. 2. Twenty operatic vocalises.

Twenty-four vocalises for soprano. [music] : op. 3 /

Piano acc.

Sammlung von Schweizer-Kühreihen und Volksliedern Receuil [sic] de ranz de vaches et chansons nationales de la Suisse.

Songs for voice and keyboard instrument and/or guitar; instrumental music principally for keyboard instrument.

Suite aus "Studenten Music" 1654, für 2 Violinen, 2 Violen, Violoncello (Kontrabass) und Klavier. [Für den prakt. Gebrauch

Originally for 5 instruments with continuo; the bass is realized for piano. The continuo part realized for keyboard instrument.

Fantasia on Hungarian folk-melodies. [music] : for pianoforte : two pianos, four hands (in score) /

"Piano II arr. from the orchestral score by Hans von Bülow".