Influence of salinity, nutrients and light on the germination and growth of Enteromorpha sp. spores

In many shallow coastal areas worldwide, several species of opportunistic macroalgae (mainly Chlorophyta) have an excessive growth, as a consequence of eutrophication processes. Therefore, bloom-forming macroalgae become the dominant primary producers within these coastal systems. However, frequently the ecology and the ecophysiology of adult macroalgae have been insufficient to explain their seasonal abundances. Thus, it is essential to understand the factors that regulate the germination and growth of spores of opportunistic green macroalgae. In the present work, we assessed the effects of nutrients (N and P), salinity and light on the germination and growth of Enteromorpha spores. Overall, the results highlight the fact that, such as for adult macroalgae, spore germination and growth are adversely affected by low salinities. Growth of the spores is significantly decreased at 5 psu, while salinities of 20 and, especially of 35 psu, clearly promote the spore growth. Additionally, Enteromorpha spores seem to be particularly sensitive to PO₄-P limitation and to NH₄-N toxicity, which suggests a higher sensitivity to the variation of external nutrient concentrations than adult macroalgae. The present results contribute to increase the understanding about the factors that control macroalgal growth at its early phases of development. In particular, the results suggest that the growth of spores from opportunistic green macroalgae is strongly salinity-dependent. Consequently, in highly hydrodynamic systems such as most shallow estuaries, salinity variations may play a determinant role in the yearly abundances of green macroalgae, since it controls macroalgal growth from the spores to the adults.     

Heterologous expression and characterization of a new heme-catalase in <Emphasis Type="Italic">Bacillus subtilis</Emphasis> 168

Reactive oxygen species (ROS) is an inherent consequence to all aerobically living organisms that might lead to the cells being lethal and susceptible to oxidative stress. Bacillus pumilus is characterized by high-resistance oxidative stress that stimulated our interest to investigate the heterologous expression and characterization of heme-catalase as potential biocatalyst. Results indicated that recombinant enzyme significantly exhibited the high catalytic activity of 55,784 U/mg expressed in Bacillus subtilis 168 and 98.097 µmol/min/mg peroxidatic activity, the apparent K _m of catalytic activity was 59.6 ± 13 mM with higher turnover rate (K _cat = 322.651 × 10³ s^?1). The pH dependence of catalatic and peroxidatic activity was pH 7.0 and pH 4.5 respectively with temperature dependence of 40 °C and the recombinant heme-catalase exhibited a strong Fe²⁺ preference. It was further revealed that catalase KatX2 improved the resistance oxidative stress of B. subtilis. These findings suggest that this B. pumilus heme-catalase can be considered among the industrially relevant biocatalysts due to its exceptional catalytic rate and high stability and it can be a potential candidate for the improvement of oxidative resistance of industrially produced strains.     

Seeing Touches Early in Life

The sense of touch provides fundamental information about the surrounding world, and feedback about our own actions. Although touch is very important during the earliest stages of life, to date no study has investigated infants’ abilities to process visual stimuli implying touch. This study explores the developmental origins of the ability to visually recognize touching gestures involving others. Looking times and orienting responses were measured in a visual preference task, in which participants were simultaneously presented with two videos depicting a touching and a no-touching gesture involving human body parts (face, hand) and/or an object (spoon). In Experiment 1, 2-day-old newborns and 3-month-old infants viewed two videos: in one video a moving hand touched a static face, in the other the moving hand stopped before touching it. Results showed that only 3-month-olds, but not newborns, differentiated the touching from the no-touching gesture, displaying a preference for the former over the latter. To test whether newborns could manifest a preferential visual response when the touched body part is different from the face, in Experiment 2 newborns were presented with touching/no-touching gestures in which a hand or an inanimate object—i.e., a spoon- moved towards a static hand. Newborns were able to discriminate a hand-to-hand touching gesture, but they did not manifest any preference for the object-to-hand touch. The present findings speak in favour of an early ability to visually recognize touching gestures involving the interaction between human body parts.     

Haying and grazing effects on the butterfly communities of two Mediterranean-area grasslands

Grasslands are usually the most suitable environment for butterflies, but have been also traditionally used for productive activities. This paper compares the impact of mowing and grazing on butterfly biodiversity in two S Italian (Campania) grasslands, at Campo Somma (CS) and Pianoro di Prada (PP) located at an identical altitude of 850 m. These grasslands have an area of approximately 6 ha each and are at a 3 km distance from each other. They share similar climate and are both surrounded by woods, mainly of sweet chestnut. CS is managed for haying and is mown once a year, in June. PP is used for sheep and cattle grazing. Weekly transects were made from April to September in 2008 and 2009, during the butterfly flight activity. We analysed data from eight monthly transects by Kruskal–Wallis and Mann–Whitney tests. We observed 45 butterfly species at PP, and 28 at CS. Diversity indexes were significantly higher at PP, whereas evenness was similar. The monthly comparisons of species richness showed that, except for April, PP values were always significantly higher. Species abundance was significantly higher at PP in June, July and August. In all months, except in April, the Shannon–Wiener index was also significantly higher at PP, as was Simpson’s index in May, June and September. Dominance index differed significantly only in May, June and September, whereas evenness was never significantly different between the two grasslands. These data show that, as concerns butterflies, the impact of mowing was much stronger than that of grazing.     

Tyrosine phosphorylation turns alkaline transition into a biologically relevant process and makes human cytochrome <Emphasis Type="Italic">c</Emphasis> behave as an anti-apoptotic switch

Cytochrome c (Cc) is a key protein in cell life (respiration) and cell death (apoptosis). On the one hand, it serves as a mitochondrial redox carrier, transferring electrons between the membrane-embedded complexes III and IV. On the other hand, it acts as a cytoplasmic apoptosis-triggering agent, forming the apoptosome with apoptosis protease-activating factor-1 (Apaf-1) and activating the caspase cascade. The two functions of cytochrome c are finely tuned by the phosphorylation of tyrosines and, in particular, those located at positions 48 and 97. However, the specific cytochrome c-phosphorylating kinase is still unknown. To study the structural and functional changes induced by tyrosine phosphorylation in cytochrome c, we studied the two phosphomimetic mutants Y48E and Y97E, in which each tyrosine residue is replaced by glutamate. Such substitutions alter both the physicochemical features and the function of each mutant compared with the native protein. Y97E is significantly less stable than the WT species, whereas Y48E not only exhibits lower values for the alkaline transition pK _a and the midpoint redox potential, but it also impairs Apaf-1-mediated caspase activation. Altogether, these findings suggest that the specific phosphorylation of Tyr48 makes cytochrome c act as an anti-apoptotic switch.     

Development of the downstream process in the production of the recombinant histone H1.3 variant

It was shown that H1 type histones possess anticancer activity and could be utilized in therapy of acute myeloid leukemia. We developed an experimental pilot-scale technology of the recombinant histone H1.3 variant production. The downstream process includes acidic extraction of the target protein from the culture broth, ion-exchange, reversed-phase and size-exclusion chromatography, and freeze-drying. The accent was made on the reduction of bacterial endotoxin contamination of the active pharmaceutical ingredient. Highly efficient downstream strategy of the target protein depyrogenation is discussed in the paper. The developed technology allows the production of the protein with high yield (approx. 110 g per 1000 L of the culture broth) and of high purity (estimated productivity is 75–100 g/month). The activity and purity of the active pharmaceutical ingredients produced for clinical trials were confirmed by different tests including ultra performance liquid chromatography, size exclusion chromatography, ELISA, SDS–PAGE, gel-clot LAL-test. Clinical trials were started in the Russian Federation.     

Modulation of Plasma Membrane Ca2+-ATPase by Neutral Phospholipids: EFFECT OF THE MICELLE-VESICLE TRANSITION AND THE BILAYER THICKNESS*

The effects of lipids on membrane proteins are likely to be complex and unique for each membrane protein. Here we studied different detergent/phosphatidylcholine reconstitution media and tested their effects on plasma membrane Ca²⁺ pump (PMCA). We found that Ca²⁺-ATPase activity shows a biphasic behavior with respect to the detergent/phosphatidylcholine ratio. Moreover, the maximal Ca²⁺-ATPase activity largely depends on the length and the unsaturation degree of the hydrocarbon chain. Using static light scattering and fluorescence correlation spectroscopy, we monitored the changes in hydrodynamic radius of detergent/phosphatidylcholine particles during the micelle-vesicle transition. We found that, when PMCA is reconstituted in mixed micelles, neutral phospholipids increase the enzyme turnover. The biophysical changes associated with the transition from mixed micelles to bicelles increase the time of residence of the phosphorylated intermediate (EP), decreasing the enzyme turnover. Molecular dynamics simulations analysis of the interactions between PMCA and the phospholipid bilayer in which it is embedded show that in the 1,2-dioleoyl-sn-glycero-3-phosphocholine bilayer, charged residues of the protein are trapped in the hydrophobic core. Conversely, in the 1,2-dimyristoyl-sn-glycero-3-phosphocholine bilayer, the overall hydrophobic-hydrophilic requirements of the protein surface are fulfilled the best, reducing the thermodynamic cost of exposing charged residues to the hydrophobic core. The apparent mismatch produced by a 1,2-dioleoyl-sn-glycero-3-phosphocholine thicker bilayer could be a structural foundation to explain its functional effect on PMCA.