Influence of Cytoplasmatic Folding on Mitochondrial Import.

The eukaryotic cell, with its organelle organization, represents a challenge for protein traffic. Contrary to what occurs in the endoplasmic reticulum, mitochondrial protein import is proposed to occur postranslationaly, as proteins are synthesized in cytoplasmic ribosomes and only then imported to the organelle. Because the diameter of the Tom and Tim pores is too narrow for the passage of a folded protein, it is assumed that polypeptides must be already in an unfolded, import competent, state for organelle entry. However, it has been suggested that mitochondria might be able to actively unfold proteins itself at the outer membrane. Here we discuss the influence of cytoplasmatic protein folding on mitochondrial import. Despite the contribution of active mitochondrial unfolding to protein import is not excluded, this mechanism is inconsistent with a number of experimental evidences. Accordingly, other alternative models for mitochondrial import are here discussed. Understanding the molecular constraints regulating this process is of crucial importance, since its failure can lead to a number of pathological situations.

Future scenarios for viticultural zoning in Europe: ensemble projections and uncertainties.

Optimum climate conditions for grapevine growth are limited geographically and may be further challenged by a changing climate. Due to the importance of the winemaking sector in Europe, the assessment of future scenarios for European viticulture is of foremost relevance. A 16-member ensemble of model transient experiments (generated by the ENSEMBLES project) under a greenhouse gas emission scenario and for two future periods (2011-2040 and 2041-2070) is used in assessing climate change projections for six viticultural zoning indices. After model data calibration/validation using an observational gridded daily dataset, changes in their ensemble means and inter-annual variability are discussed, also taking into account the model uncertainties. Over southern Europe, the projected warming combined with severe dryness in the growing season is expected to have detrimental impacts on the grapevine development and wine quality, requiring measures to cope with heat and water stress. Furthermore, the expected warming and the maintenance of moderately wet growing seasons over most of the central European winemaking regions may require a selection of new grapevine varieties, as well as an enhancement of pest/disease control. New winemaking regions may arise over northern Europe and high altitude areas, when considering climatic factors only. An enhanced inter-annual variability is also projected over most of Europe. All these future changes pose new challenges for the European winemaking sector.

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity.

Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 +/- 3.43 nmol p-nitrophenol.mg protein-1.min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84%, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11%, respectively). beta-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36%, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18%) and inhibited (13%) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity

Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 ± 3.43 nmol p-nitrophenol·mg protein-1·min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84 percent, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11 percent, respectively). â-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36 percent, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18 percent) and inhibited (13 percent) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.