Inhibition of blood platelet aggregation by dioxo-ene compounds

Compounds with a conjugated oxo-ene-oxo system were tested for inhibition of blood platelet aggregation. All compounds with this structure in trans configuration were effective inhibitors of aggregation induced by thrombin and by arachidonic acid. While the oxo-trans-ene-oxo system is prerequisite for such activity, other structural features of the compounds may be varied without loss of activity. Inhibition is exemplified by 9,12-dioxo-trans-10- and 10,13-dioxo-trans-11-octadecenoic acids and their methyl esters, by 11,14-dioxo-trans-12- eicosenoic acid, by 4,7-dioxo-trans-5- decene and by trans- dibenzoylethylene . The half-inhibition concentrations are in the order of 2-6 microM, with complete inhibition at 8-20 microM. According to experiments with the inhibiting 9,12-dioxo-trans-10-octadecenoic acid, the normal oxygenation of exogenous arachidonic acid by platelets is not affected but the thrombin-induced internal release of this acid seems to be abolished by the inhibitor. The inhibition of aggregation in the presence of exogenous arachidonic acid and its products suggests that the inhibitor also interferes with other events leading to aggregation. By implication from other properties of the oxo-trans-ene-oxo system, reaction with SH groups may be a mechanism for inhibition.     

Preparation of Phgsphatioyll2-3Hjinositgl from Yeast Grown in Medium Containing N;Yol2-3Hjinositol

Phosphatidy[2-³]jinositol was prepared from Saccharoniycts cerevisiae (YSC-2), grown in synthetic meaiurn containing myo[2-³H]inositol. Over 44 μCi (or 81 %) of the racio-labeleo inositol was taken up by the organism, with 34 yCi incorporated into phospnatiaylinositol. Upon purification d) silicic acia-meaium pressure liquia chrcnatography (MPLC), a final yield of 24 to 2b μCi of phosphatiayl[2-³h]inositot with a specific radioactivity of 40 ± 10³ apm/nmoie wäs obtained. The purified phosphatiuyl[2-³H] inositol was founo to be a suitable substrate for phospholipase C from human platelets     

Impact of rainfall manipulations and biotic controls on soil respiration in Mediterranean and desert ecosystems along an aridity gradient

Spatially heterogeneous ecosystems form a majority of land types in the vast drylands of the globe. To evaluate climate‐change effects on CO₂ fluxes in such ecosystems, it is critical to understand the relative responses of each ecosystem component (microsite). We investigated soil respiration (R_s) at four sites along an aridity gradient (90–780 mm mean annual precipitation, MAP) during almost 2 years. In addition, R_s was measured in rainfall manipulations plots at the two central sites where ～30% droughting and ～30% water supplementation treatments were used over 5 years. Annual R_s was higher by 23% under shrub canopies compared with herbaceous gaps between shrubs, but R_s at both microsites responded similarly to rainfall reduction. Decreasing precipitation and soil water content along the aridity gradient and across rainfall manipulations resulted in a progressive decline in R_s at both microsites, i.e. the drier the conditions, the larger was the effect of reduction in water availability on R_s. Annual R_s on the ecosystem scale decreased at a slope of 256/MAP g C m^?2 yr^?1 mm^?1 (r²=0.97). The reduction in R_s amounted to 77% along the aridity gradient and to 16% across rainfall manipulations. Soil organic carbon (SOC) decreased with declining precipitation, and variation in SOC stocks explained 77% of the variation in annual R_s across sites, rainfall manipulations and microsites. This study shows that rainfall manipulations over several years are a useful tool for experimentally predicting climate‐change effects on CO₂ fluxes for time scales (such as approximated by aridity gradients) that are beyond common research periods. Rainfall reduction decreases rates of R_s not only by lowering biological activity, but also by drastically reducing shrub cover. We postulate that future climate change in heterogeneous ecosystems, such as Mediterranean and deserts shrublands will have a major impact on R_s by feedbacks through changes in vegetation structure.     

Dissolved carbon leaching from soil is a crucial component of the net ecosystem carbon balance

Estimates of carbon leaching losses from different land use systems are few and their contribution to the net ecosystem carbon balance is uncertain. We investigated leaching of dissolved organic carbon (DOC), dissolved inorganic carbon (DIC), and dissolved methane (CH₄), at forests, grasslands, and croplands across Europe. Biogenic contributions to DIC were estimated by means of its δ¹³C signature. Leaching of biogenic DIC was 8.3±4.9 g m^?2 yr^?1 for forests, 24.1±7.2 g m^?2 yr^?1 for grasslands, and 14.6±4.8 g m^?2 yr^?1 for croplands. DOC leaching equalled 3.5±1.3 g m^?2 yr^?1 for forests, 5.3±2.0 g m^?2 yr^?1 for grasslands, and 4.1±1.3 g m^?2 yr^?1 for croplands. The average flux of total biogenic carbon across land use systems was 19.4±4.0 g C m^?2 yr^?1. Production of DOC in topsoils was positively related to their C/N ratio and DOC retention in subsoils was inversely related to the ratio of organic carbon to iron plus aluminium (hydr)oxides. Partial pressures of CO₂ in soil air and soil pH determined DIC concentrations and fluxes, but soil solutions were often supersaturated with DIC relative to soil air CO₂. Leaching losses of biogenic carbon (DOC plus biogenic DIC) from grasslands equalled 5–98% (median: 22%) of net ecosystem exchange (NEE) plus carbon inputs with fertilization minus carbon removal with harvest. Carbon leaching increased the net losses from cropland soils by 24–105% (median: 25%). For the majority of forest sites, leaching hardly affected actual net ecosystem carbon balances because of the small solubility of CO₂ in acidic forest soil solutions and large NEE. Leaching of CH₄ proved to be insignificant compared with other fluxes of carbon. Overall, our results show that leaching losses are particularly important for the carbon balance of agricultural systems.     

Targeting Eukaryotic Translation in Mesothelioma Cells with an eIF4E-Specific Antisense Oligonucleotide

Background

Aberrant cap-dependent translation is implicated in tumorigenesis in multiple tumor types including mesothelioma. In this study, disabling the eIF4F complex by targeting eIF4E with eIF4E-specific antisense oligonucleotide (4EASO) is assessed as a therapy for mesothelioma.

Methods

Mesothelioma cells were transfected with 4EASO, designed to target eIF4E mRNA, or mismatch-ASO control. Cell survival was measured in mesothelioma treated with 4EASO alone or combined with either gemcitabine or pemetrexed. Levels of eIF4E, ODC, Bcl-2 and β-actin were assessed following treatment. Binding to a synthetic cap-analogue was used to study the strength of eIF4F complex activation following treatment.

Results

eIF4E level and the formation of eIF4F cap-complex decreased in response to 4EASO, but not mismatch control ASO, resulting in cleavage of PARP indicating apoptosis. 4EASO treatment resulted in dose dependent decrease in eIF4E levels, which corresponded to cytotoxicity of mesothelioma cells. 4EASO resulted in decreased levels of eIF4E in non-malignant LP9 cells, but this did not correspond to increased cytotoxicity. Proteins thought to be regulated by cap-dependent translation, Bcl-2 and ODC, were decreased upon treatment with 4EASO. Combination therapy of 4EASO with pemetrexed or gemcitabine further reduced cell number.

Conclusion

4EASO is a novel drug that causes apoptosis and selectively reduces eIF4E levels, eIF4F complex formation, and proliferation of mesothelioma cells. eIF4E knockdown results in decreased expression of anti-apoptotic and pro-growth proteins and enhances chemosensitivity.     

Exclusion of ground predators improves Northern Lapwing Vanellus vanellus chick survival

Many farmland‐breeding wader species have declined across Europe, probably due to reductions in reproductive output caused by high nest losses as a result of agriculture or predation, or low chick survival between hatching and fledging. Most studies have focused on nest failures, and the factors affecting post‐hatching survival of chicks are poorly known. In an experimental approach, we fenced parts of the arable foraging areas of Northern Lapwing Vanellus vanellus families to quantify chick survival simultaneously in the presence and absence of ground predators. Lapwing chicks were radiotagged to estimate survival probabilities by daily locations, applying multistate capture–recapture models. During the night, chick survival was considerably lower outside fenced plots than within. During the day, chick survival was higher than at night and did not differ between protected and unprotected plots. This suggests that nocturnal ground predators such as Red Foxes Vulpes vulpes were responsible for a significant proportion of chick mortality. Cumulative survival probability from hatching to fledging was 0.24 in chicks within fenced plots, but virtually zero in chicks outside fenced plots. In farmland, temporary electric fences can be effective in minimizing the impact of ground predators and offer a promising short‐term method to increase fledging success of precocial birds.