Dynamic Localization of the Swe1 Regulator Hsl7 During the Saccharomyces cerevisiae Cell Cycle

In Saccharomyces cerevisiae, entry into mitosis requires activation of the cyclin-dependent kinase Cdc28 in its cyclin B (Clb)-associated form. Clb-bound Cdc28 is susceptible to inhibitory tyrosine phosphorylation by Swe1 protein kinase. Swe1 is itself negatively regulated by Hsl1, a Nim1-related protein kinase, and by Hsl7, a presumptive protein-arginine methyltransferase. In vivo all three proteins localize to the bud neck in a septin-dependent manner, consistent with our previous proposal that formation of Hsl1-Hsl7-Swe1 complexes constitutes a checkpoint that monitors septin assembly. We show here that Hsl7 is phosphorylated by Hsl1 in immune-complex kinase assays and can physically associate in vitro with either Hsl1 or Swe1 in the absence of any other yeast proteins. With the use of both the two-hybrid method and in vitro binding assays, we found that Hsl7 contains distinct binding sites for Hsl1 and Swe1. A differential interaction trap approach was used to isolate four single-site substitution mutations in Hsl7, which cluster within a discrete region of its N-terminal domain, that are specifically defective in binding Hsl1. When expressed in hsl7Δ cells, each of these Hsl7 point mutants is unable to localize at the bud neck and cannot mediate down-regulation of Swe1, but retains other functions of Hsl7, including oligomerization and association with Swe1. GFP-fusions of these Hsl1-binding defective Hsl7 proteins localize as a bright perinuclear dot, but never localize to the bud neck; likewise, in hsl1Δ cells, a GFP-fusion to wild-type Hsl7 or native Hsl7 localizes to this dot. Cell synchronization studies showed that, normally, Hsl7 localizes to the dot, but only in cells in the G1 phase of the cell cycle. Immunofluorescence analysis and immunoelectron microscopy established that the dot corresponds to the outer plaque of the spindle pole body (SPB). These data demonstrate that association between Hsl1 and Hsl7 at the bud neck is required to alleviate Swe1-imposed G2-M delay. Hsl7 localization at the SPB during G1 may play some additional role in fine-tuning the coordination between nuclear and cortical events before mitosis.

Direct observation of fast protein folding: the initial collapse of apomyoglobin.

The rapid refolding dynamics of apomyoglobin are followed by a new temperature-jump fluorescence technique on a 15-ns to 0.5-ms time scale in vitro. The apparatus measures the protein-folding history in a single sweep in standard aqueous buffers. The earliest steps during folding to a compact state are observed and are complete in under 20 micros. Experiments on mutants and consideration of steady-state CD and fluorescence spectra indicate that the observed microsecond phase monitors assembly of an A x (H x G) helix subunit. Measurements at different viscosities indicate diffusive behavior even at low viscosities, in agreement with motions of a solvent-exposed protein during the initial collapse.

Crossover and noncrossover recombination during meiosis: timing and pathway relationships.

During meiosis, crossovers occur at a high level, but the level of noncrossover recombinants is even higher. The biological rationale for the existence of the latter events is not known. It has been suggested that a noncrossover-specific pathway exists specifically to mediate chromosome pairing. Using a physical assay that monitors both crossovers and noncrossovers in cultures of yeast undergoing synchronous meiosis, we find that both types of products appear at essentially the same time, after chromosomes are fully synapsed at pachytene. We have also analyzed a situation in which commitment to meiotic recombination and formation of the synaptonemal complex are coordinately suppressed (mer1 versus mer1 MER2++). We find that suppression is due primarily to restoration of meiosis-specific double-strand breaks, a characteristic of the major meiotic recombination pathway. Taken together, the observations presented suggest that there probably is no noncrossover-specific pathway and that restoration of intermediate events in a single pairing/recombination pathway promotes synaptonemal complex formation. The biological significant of noncrossover recombination remains to be determined, however.

A molecular anchor for stabilizing triple-helical DNA.

Molecular modeling has been used to predict that 2,6-disubstituted amidoanthraquinones, and not the 1,4 series, should preferentially interact with and stabilize triple-stranded DNA structures over duplex DNA. This is due to marked differences in the nature of chromophore-base stacking and groove accessibility for the two series. A DNA foot-printing method that monitors the extent of protection from DNase I cleavage on triplex formation has been used to examine the effects of a number of synthetic isomer compounds in the 1,4 and 2,6 series. The experimental results are in accord with the predicted behavior and confirm that the 1,4 series bind preferentially to double- rather than triple-stranded DNA, whereas the isomeric 2,6 derivatives markedly favor binding to triplex DNA.

Desenvolvimento de sistema automatizado de monitoramento de ambientes de produção animal, utilizando uma rede de sensores sem fio

As redes de sensores sem fio, aplicadas à automação do controle de ambientes representam um paradigma emergente da computação, onde múltiplos nós providos de sensores, sistemas computacionais autônomos e capacidade de comunicação sem fio, conformam uma rede cuja topologia altamente dinâmica permite adquirir informações sobre sistemas complexos sendo monitorados. Um dos fatores essenciais para obter um ganho na produtividade avícola é o controle da ambiência animal. Atualmente os métodos utilizados para o monitoramento e controle ambiental não podem considerar a grande quantidade de microambientes internos nos ambientes de produção animal e também requerem infraestruturas cabeadas complexas. Dentro desse contexto o objetivo deste trabalho foi desenvolver e testar um sistema automatizado de controle ambiental, através da utilização de sensores sem fio, que auxilie e proporcione maior segurança no controle de ambientes automatizados. O sistema monitora variáveis que influenciam na produtividade de aves, tais como temperatura e umidade e outras variáveis físico-químicas do aviário. A infraestrutura desenvolvida foi testada em um aviário experimental e resultou em um sistema seguro e com grande escalabilidade, que é capaz de controlar e monitorar o ambiente e ainda coletar e gravar dados. Foi utilizado o protocolo ZigBee® para gerenciar o fluxo de dados do sistema. Foram feitas análises da eficiência de comunicação do sistema no aviário, monitorando os pacotes de dados perdidos. Os testes demonstraram uma perda de dados de aproximadamente 2% dos pacotes enviados, demonstrando a eficiência das redes ZigBee® para gerenciar o fluxo de dados no interior do aviário. Desta forma, pode-se concluir que é possível e viável a implantação de uma rede ZigBee®, para automatizar ambientes de produção animal com coleta de dados em tempo real, utilizando um sistema integrado via internet, que compreende: instrumentação eletrônica, comunicação sem fio e engenharia de software\".

Developing a Citizen Volunteer Lake Monitoring Program for the Flathead Basin in Northwestern Montana for the early Detection and Prevention of the Introduction of Zebra Mussels and other Aquatic Invasive Species (AIS)

Preventing the introduction of aquatic invasive species (AIS) like zebra and quagga mussels in the U.S. is a high priority. This Capstone demonstrates zebra and quagga mussels are of concern as aquatic invasive species and a volunteer monitoring and intervention program is an effective means for early detection of AIS. This Capstone developed an AIS citizen volunteer lake monitoring program consistent with other programs concerned about AIS prevention and early detection. This Capstone concludes implementing such a voluntary program will help reduce the spread of zebra and quagga mussels and will provide early detection information to appropriate agencies empowered with response actions if species are found.

Triassic redbeds in the Malaguide Complex (Betic Cordillera — Spain): petrography, geochemistry and geodynamic implications

Sandstone petrography and mudstone mineralogy and geochemistry of Triassic mudstones and sandstones from continental redbeds of the Malaguide Complex (Betic Cordillera, southern Spain) provide useful information on provenance, palaeoclimate and geodynamics during the early stages of the Pangea break-up, and on their diagenetic evolution. The sandstones are quartzarenites to sub-litharenites, with minor lithic fragments and rare feldspars. The mudstone samples show a PAAS like elemental distribution. The samples likely record recycling processes from their metasedimentary basement rocks that significantly affected the weathering indices, and monitors cumulative effects, including a first cycle of weathering at the source rocks. Sandstone composition and chemical–mineralogical features of mudstones record a provenance derived from continental block and recycled orogen that were weathered under warm and episodically wet climate. Source areas were located towards the east of the present-day Malaguide outcrops, and were formed by fairly silicic rock types, made up mainly of Palaezoic metasedimentary rocks, similar to those of the Paleozoic underlying series, with subordinate contributions from magmatic–metamorphic sources, and a rare supply from mafic metavolcanic rocks. Clay-mineral distribution of mudstones is dominated by illite and illite/smectite mixed-layer that result from differences in provenance, weathering, and burial/temperature history. Illite crystallinity values, illitization of kaolinite, occurrence of typical authigenic minerals and apatite fission-track studies, coupled with a subsidence analysis of the whole Malaguide succession suggest burial depths of at least 4–6 km with temperatures of 140–160 °C, typical of the burial diagenetic stage, and confirm the Middle Miocene exhumation of the Betic Internal Domain tectonic stack topped by the Malaguide Complex.

Análisis de la carga interna en los entrenamientos de fútbol sala femenino de 1ª división nacional

El objetivo de este estudio es analizar la frecuencia cardiaca (FC) y la percepción subjetiva del esfuerzo (RPE) en los entrenamientos de fútbol sala en jugadoras (25.42±4.42 años, 166.57±7.14 cm, 56.50±4.95 kg y 22.23±0.97 IMC) de un equipo de primera división de la liga nacional española. Se registraron, mediante monitores de ritmo cardiaco y la escala de Foster, ocho sesiones de entrenamiento de tipo técnico-táctico, físico y partidos modificados, distribuidas durante la mitad de la temporada. Los resultados de la FC máxima y media obtenida son 174.03±4.15 y 128.29±0.81 ppm en sesiones técnico-tácticas, 179.75±2.05 y 132±2.12 ppm en sesiones físicas, 180.99±1.06 y 133.63±5.52 ppm en partidos modificados, respectivamente. Habiendo diferencias significativas en los resultados de la FC máxima entre las sesiones técnico-tácticas y los partidos modificados (p=0.03) y entre la sesiones físicas y las técnico-tácticas (p=0.04). En cuanto a los resultados de la carga de entrenamiento y la RPE en las sesiones técnico-tácticas son 381.22±16.42 y 6.21±0.12 rpe, en las sesiones físicas 533.25±5.59 y 7.90±0.0 rpe, y en los partidos modificados 531.18±4.21 y 7.97±0.22 rpe, respectivamente. Encontrando diferencias significativas entre los resultados de la RPE y la magnitud de la carga de entrenamiento de las sesiones técnico-tácticas y los partidos modificados (p=0.00) y entre la sesiones físicas y las técnico-tácticas (p=0.00). Estos resultados muestran que la sesión de partido modificado obtiene unos valores de frecuencia cardiaca y de percepción subjetiva del esfuerzo mayor que las sesiones técnico-tácticas y las de trabajo físico.

Disseny d'unes jornades de salut per a primària, "Aprenem amb la salut!"

Aquest treball presenta, unes jornades de salut anomenades "Aprenem amb la salut", destinades a l'alumnat d'Educació Primària. A través aquestes jornades s'incorpora en l'alumnat la promoció d'hàbits saludables, centrant-se en l'alimentació, els hàbits d'higiene, els hàbits saludables i la pràctica d'activitat física o esport. Tots aquests aspectes es treballen mitjançant jocs i tallers, entre altres. A més, durant aquestes jornades es vincula l'aprenentatge de l'alumnat amb les seus famílies per tal de formar i conscienciar a les famílies en aspectes tan importants com és l'alimentació. Durant aquesta jornada, s'utilitzen diferents estils d'ensenyament combinant tradicionals amb un participatiu. Respecte a l'estil d'ensenyament participatiu, a través del microensenyament l'alumnat de sisè d'Educació Primària col·labora en les jornades, realitzant la funció de monitors i monitores en les gymkanes. Per acabar, en aquestes jornades es pretén fomentar en l'alumnat d'Educació Primària hàbits saludables, per tal de millorar la seua salut.

Settlement of chambers, 1741-1764

The book is comprised of folio-sized pages conserved in a modern soft-cover binding. The volume consists of yearly handwritten lists of dormitory room assignments for the years 1741-1753 and 1761-1764. Students are listed by last name and building names are often abbreviated as "M" for Massachusetts Hall, "S" for Stoughton Hall, and "O" for Old College or Harvard Hall. The organizational pattern varies by list, some are alphabetical, others arranged by building and room number. The lists for 1743, 1748, 1749, 1761-1764 also note students living outside of the College and their locations. The lists for 1761-1764 also include the waiters and monitors for the academic year.

Homeostatic plasticity induced by brief activity deprivation enhances long-term potentiation in the mature rat hippocampus

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

The Special Monitoring Mission in Ukraine: a useful but flawed OSCE tool. EPC Policy Brief, 23 April 2015

The OSCE’s Special Monitoring Mission in Ukraine functions as the eyes and ears of the international community in Eastern Ukraine; it monitors, impartially, the ongoing conflict between Ukraine and Russia and observes whether or not both parties comply with the Minsk II Agreement. The success, or failure, of this Mission will not only be of crucial importance for the situation in Eastern Ukraine, but will also determine the OSCE’s entire future and ability to operate in conflict regions. Although the SMM is a flawed tool, Dennis Sammut and Joseph D’Urso argue that its deployment and the subsequent reinvigoration of OSCE is the perfect opportunity for the EU to rethink its relation to and position in the organisation, and to assume a bigger and more decisive role.

Keeping health high on the EU agenda: role for economic governance? EPC Policy Brief, 7 May 2015

In an attempt to get Europe out of the economic crisis and establish right conditions for growth, the EU coordinates and monitors member states’ economic and budgetary policies via a system called the European Semester. As member states’ spending on the health sector accounts for 10% of GDP and is expected to grow, it is no wonder that an increasing emphasis has been paid to sustainability of health systems – an area that is traditionally considered as a national competence. In this Policy Brief, Annika Hedberg and Martina Morosi reflect on the strengths and weaknesses of the European Semester and country-specific recommendations in promoting more sustainable and efficient health systems in Europe, and why the EU must continue to play a role in encouraging member states to value health and improve their spending on health.

An Arginine Deprivation Response Pathway Is Induced in Leishmania during Macrophage Invasion

Amino acid sensing is an intracellular function that supports nutrient homeostasis, largely through controlled release of amino acids from lysosomal pools. The intracellular pathogen Leishmania resides and proliferates within human macrophage phagolysosomes. Here we describe a new pathway in Leishmania that specifically senses the extracellular levels of arginine, an amino acid that is essential for the parasite. During infection, the macrophage arginine pool is depleted due to its use to produce metabolites (NO and polyamines) that constitute part of the host defense response and its suppression, respectively. We found that parasites respond to this shortage of arginine by up-regulating expression and activity of the Leishmania arginine transporter (LdAAP3), as well as several other transporters. Our analysis indicates the parasite monitors arginine levels in the environment rather than the intracellular pools. Phosphoproteomics and genetic analysis indicates that the arginine-deprivation response is mediated through a mitogen-activated protein kinase-2-dependent signaling cascade.

Changes in the vegetation and trade winds in equatorial Northwest Africa 140,000-70,000 yr B.P., as deduced from two marine pollen records

The deep-sea cores M 16415-2 and M 16416-2 at about 9°N off Sierra Leone were analysed palynologically for the time interval 140,000-70,000 yr B.P. Results were presented in absolute (pollen concentration and pollen influx) and relative diagrams (pollen percentage). In a previous study it was evidenced that in northwest Africa pollen is mainly transported to the Atlantic by wind, so that the efficiency of aeolian pollen transport (pollen flux) could be used to evaluate changes in the intensity of the northeast trade winds. The glacial episodes (represented by the oxygen isotope stages 6 and 4) are characterized by strong northeast trade winds, whereas the last interglacial (stage 5) is characterized by weak trade winds. The pollen influx diagram shows that the intensity of the trade winds increased slightly during the relatively cool intervals of stage 5 (viz. 5.4 and 5.2). Tropical forest had maximally expanded around 124,000 yr B.P. (stage 5.5), around 98,000 yr B.P. (transition of stage 5.3 to 5.2), and around 70,000 yr B.P. (first part of stage 4): an increasing delay of the response of tropical forest to global intervals with maximum temperature is apparent during the last interglacial. As tropical forests need continuous humidity, the record of tropical forest monitors changes in climatic humidity south of the Sahara. During the last interglacial, the southern boundary of the Sahara shifted only little: expansions and contractions of the tropical forest area are correlated with contra-oscillations of the grass-dominated savanna zone. Great latitudinal shifts of the desert savanna boundary, on the contrary, occurred during the penultimate glacial interglacial transition (around 128,000 yr B.P.) to the north, and during the last interglacial-glacial transition (around 65,000 yr B.P.) to the south.