An axoplasmic myosin with a calmodulin-like light chain.

Organelles in the axoplasm from the squid giant axon move along exogenous actin filaments toward their barbed ends. An approximately 235-kDa protein, the only band recognized by a pan-myosin antibody in Western blots of isolated axoplasmic organelles, has been previously proposed to be a motor for these movements. Here, we purify this approximately 235-kDa protein (p235) from axoplasm and demonstrate that it is a myosin, because it is recognized by a pan-myosin antibody and has an actin-activated Mg-ATPase activity per mg of protein 40-fold higher than that of axoplasm. By low-angle rotary shadowing, p235 differs from myosin II and it does not form bipolar filaments in low salt. The amino acid sequence of a 17-kDa protein that copurifies with p235 shows that it is a squid optic lobe calcium-binding protein, which is more similar by amino acid sequence to calmodulin (69% identity) than to the light chains of myosin II (33% identity). A polyclonal antibody to this light chain was raised by using a synthetic peptide representing the calcium binding domain least similar to calmodulin. We then cloned this light chain by reverse transcriptase-PCR and showed that this antibody recognizes the bacterially expressed protein but not brain calmodulin. In Western blots of sucrose gradient fractions, the 17-kDa protein is found in the organelle fraction, suggesting that it is a light chain of the p235 myosin that is also associated with organelles.

Variação sazonal do metabolismo energético no primeiro ciclo anual de lagartos teiú Tupinambis merianae: correlatos com atividades diárias, adiposidade e proteção antioxidante

Lagartos teiú eclodem no verão e enfrentam o desafio de crescer e armazenar substratos em um curto período de tempo, antes do início do período de jejum e depressão metabólica (≈80%) a temperaturas amenas durante o inverno (≈17 °C). No despertar, o aumento do metabolismo e a reperfusão de órgãos favoreceriam a ocorrência de estresse oxidativo. Na primeira parte do presente estudo investigou−se os ajustes que compatibilizam as demandas em teiús neonatos, especialmente na pré-hibernação, por meio da gravação do comportamento em vídeo e da análise da massa dos corpos gordurosos abdominais e do nível plasmático de corticosterona (CORT) durante o primeiro ciclo anual. No início do outono a massa corpórea dos teiús foi 27 g e o comprimento rostro−cloacal 9,3 cm e aumentaram 40% e 20%, respectivamente, ao longo do outono, enquanto que as taxas diminuíram progressivamente até atingirem o valor zero no início do inverno. Na primavera, a massa corpórea dos teiús aumentou 80% em relação ao despertar e dobrou em relação ao final do verão; o comprimento acumulou um aumento de 27% em relação ao final do verão. A massa relativa dos corpos gordurosos foi 3,7% no início do outono e diminuiu nos meses subsequentes; no despertar, este estoque acumulou uma perda de 63% da sua massa. No início do outono 74% dos teiús estavam ativos por 4,7 h e permaneceram 2 h assoalhando diariamente; ao longo do outono o número de animais ativos e o tempo em atividade diminuíram até que todos se tornaram inativos. Na primavera 83% dos teiús estavam ativos por 7 h e permaneceram 4 h assoalhando. Um padrão sazonal similar foi observado na atividade locomotora e na alimentação. No outono, a alimentação cessou antes da atividade diária e os teiús tornaram−se afágicos algumas semanas antes da entrada em hibernação. Os maiores níveis de CORT foram observados no início do outono, reduzindo progressivamente até valores 75 e 86% menores na dormência e despertar, respectivamente; na primavera os níveis de CORT foram 32% menores em comparação com o início do outono. Este padrão sugere um papel da CORT nos ajustes que promovem a ingestão de alimento e a deposição de substratos energéticos no outono. A redução da atividade geral no final do outono contribuiria para a economia energética e manutenção da massa corpórea, apesar da redução da ingestão de alimento. O curso temporal das alterações fisiológicas e comportamentais em neonatos reforça a ideia de que a dormência sazonal nos teiús é o resultado da expressão de um ritmo endógeno. Na segunda parte do estudo foi investigada a hipótese de que ocorreriam ajustes das defesas antioxidantes durante a hibernação, em antecipação ao despertar. Foram analisados marcadores de estresse oxidativo e antioxidantes em vários órgãos de teiús em diferentes fases do primeiro ciclo anual. A CS, um indicador do potencial oxidante, não variou no fígado e foi menor no rim e no pulmão na hibernação. As enzimas antioxidantes revelaram (1) um efeito abrangente de redução das taxas na hibernação e despertar; por exemplo, GR e CAT foram menores em todos órgãos analisados e a GST tendeu a diminuir no fígado e no rim, embora constante no coração e no pulmão. A G6PDH no fígado e no rim não variou. (2) No fígado, a GST, a Se−GPX e o teor de TBARS foram maiores na atividade de outono em relação à primavera e a Se−GPX permaneceu elevada na hibernação. (3) No fígado, a SOD foi maior na hibernação e despertar em relação ao outono e a Mn−SOD seguiu este padrão. Em contraste, no rim, coração e pulmão a SOD foi menor na hibernação e as taxas se recuperaram no coração e pulmão no despertar. A Mn−SOD seguiu este padrão no pulmão. A concentração e o estado redox da glutationa não variaram no fígado, rim e coração; no pulmão o teor de Eq−GSH e GSH foi menor na hibernação, com tendência à recuperação no despertar. O teor de PC no rim foi maior na hibernação e diminuiu no despertar. No fígado, as alterações no jejum se assemelham às sazonais, como sugerem a inibição da CAT e GR e aumento da Se−GPX. Os efeitos do jejum na primavera no rim diferem dos efeitos sazonais, como sugerem a redução do teor de Eq−GSH e GSH e o aumento da razão GSSG:GSH, a redução da G6PDH e o aumento de PC. No conjunto, houve um efeito predominante de redução das taxas enzimáticas na hibernação e no despertar, exceto pelas taxas aumentadas da SOD e Se−GPX no fígado e pela recuperação da SOD no coração e da GR, SOD e Mn−SOD no pulmão no despertar. As elevadas taxas das enzimas antioxidantes no teiú em comparação a outros ectotermos e a ausência de evidências de estresse oxidativo no despertar sugerem que a atividade enzimática remanescente é suficiente para prevenir danos aos tecidos face às flutuações do metabolismo

(Table 6a) 137Cs concentrations in waters of the Sea of Japan in 1994 and in 2002

Sulfide, S°, and thiosulfate were determined in waters of the Baltic Sea. Microquantities of these compounds were observed in oxic waters. Concentration levels of reduced sulfur compounds in Baltic oxic waters were very close to levels of the Black Sea oxic zone. Thiosulfate and S° were predominate compounds in oxic water whereas sulfide was a predominant compound Baltic waters high in hydrogen sulfide. Conclusion was made that during sedimentation in oxic waters anaerobic microorganisms along with aerobic bacteria take part in mineralization of organic matter. They exist on surfaces and in microniches of particles of organic detritus.

Biochemical investigation of multicorer sediment profile PS74/109-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Biochemical investigation of multicorer sediment profile PS74/113-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Biochemical investigation of multicorer sediment profile PS74/118-2

To detect and track the impact of large-scale environmental changes in a the transition zone between the northern North Atlantic and the central Arctic Ocean, and to determine experimentally the factors controlling deep-sea biodiversity, the Alfred- Wegener-Institute for Polar and Marine Research (AWI) established the deep-sea long-term observatory HAUSGARTEN, which constitutes the first, and until now only open-ocean long-term station in a polar region. Virtually undisturbed sediment samples have been taken using a video-guided multiple corer (MUC) at 13 HAUSGARTEN stations along a bathymetric (1,000 - 4,000 m water depth) and a latitudinal transect in 2,500 m water depth as well as two stations at 230 and 1,200 m water depth within the framework of the KONGHAU project. Various biogenic sediment compounds were analyzed to estimate the input of organic matter from phytodetritus sedimentation, benthic activities (e.g. bacterial exoenzymatic activity), and the total biomass of the smallest sediment-inhabiting organisms (size range: bacteria to meiofauna).

Chemical constituents associated with sewage settling velocity profiles

A methodology has been developed to measure the chemical constituents associated with the settling velocity fractions that comprise a wastewater settling velocity profile (SVP). 31 wastewater samples were collected from fifteen different catchments in England and Wales. For each catchment, settling velocity and associated chemical constituent profiles were determined. The results are mainly for Suspended Solids (SS), Chemical Oxygen Demand (COD), Phosphorus (P) and Total Kjeadahl Nitrogen (TKN), however these are supplemented by the results from 5 events for a suite of heavy metals. COD, P, Hg, Mn and Pb were found to be predominantly associated with the solid phase and TKN, Al, Cu and Fe with the liquor phase of the wastewater samples. The results in the thesis are expressed as mass of pollutant (g) per mass total SS (kg). COD and P were found to be mainly associated with the sinkers and had a particular affinity for solids with settling velocities in the range 0.9-9.03mm/sec. TKN was mainly associated with the soluble phase, however of the solids that did settle, a peak was found to be associated within the settling velocity range 0.9-9.03mm/sec. The relationships identified for COD and P were generally found to be unaffected by flow conditions and catchment characteristics. However, TKN was found to be affected by catchment type. Data on the distribution of heavy metals was limited, and no specific relationships with solids were identified. 16 mean pollutant profiles are presented in the thesis. Presentation of the data in this form will enable the results to be of use in the design of sedimentation devices to predict removal efficiencies for solids and associated pollutants. The findings of the research may also be applied to modelling tools to provide further characteristics on the solids that are modelled than is currently used. This would enhance the overall performance of tools used in integrated catchment modelling.

The topography of transmembrane segment six is altered during the catalytic cycle of P-glycoprotein

Structural evidence has demonstrated that P-glycoprotein (P-gp) undergoes considerable conformational changes during catalysis, and these alterations are important in drug interaction. Knowledge of which regions in P-gp undergo conformational alterations will provide vital information to elucidate the locations of drug binding sites and the mechanism of coupling. A number of investigations have implicated transmembrane segment six (TM6) in drug-P-gp interactions, and a cysteine-scanning mutagenesis approach was directed to this segment. Introduction of cysteine residues into TM6 did not disturb basal or drug-stimulated ATPase activity per se. Under basal conditions the hydrophobic probe coumarin maleimide readily labeled all introduced cysteine residues, whereas the hydrophilic fluorescein maleimide only labeled residue Cys-343. The amphiphilic BODIPY-maleimide displayed a more complex labeling profile. The extent of labeling with coumarin maleimide did not vary during the catalytic cycle, whereas fluorescein maleimide labeling of F343C was lost after nucleotide binding or hydrolysis. BODIPY-maleimide labeling was markedly altered during the catalytic cycle and indicated that the adenosine 5'-(beta,gamma-imino)triphosphate-bound and ADP/vanadate-trapped intermediates were conformationally distinct. Our data are reconciled with a recent atomic scale model of P-gp and are consistent with a tilting of TM6 in response to nucleotide binding and ATP hydrolysis.

Assessment of policies for innovative financing in infrastructure systems

Infrastructure systems are drivers of the economy in the nation. A dollar spent on infrastructure development yields roughly double the initial spending in ultimate economic output in the short term; and over a twenty-year period, and generalized ‘public investment’ produces an aggregated $3.21 of economic activity per $1.00 spent [1]. Thus, formulation of policies pertaining to infrastructure investment and development is of significance affecting the social and economic wellbeing of the nation. The aim of this policy brief is to evaluate innovative financing in infrastructure systems from two different perspectives: (1) through consideration of the current condition of infrastructure in the U.S., the current trends in public spending, and the emerging innovative financing tools; (2) through evaluation of the roles and interactions of different agencies in the creation and the diffusion of innovative financing tools. Then using the example of transportation financing, the policy brief provides an assessment of policy landscapes which could lead to the closure of infrastructure financing gap in the U.S and proposes strategies for citizen involvement to gain public support of innovative financing.