Distribution and ecological relations among the alien crab, <Emphasis Type="Italic">Percnon gibbesi</Emphasis> (H. Milne-Edwards 1853) and autochthonous species,in and out of an SW Mediterranean MPA

The distribution of the allochthonous crab Percnon gibbesi and its relationships with other benthic invertebrate species was assessed inside the marine reserve of Cabo de Palos—Islas Hormigas (Mediterranean sea, Spain) and neighbouring non-protected sites. Although a significant spatial variability was detected at finer spatial scale, there was no influence of protection measures or insularity on the abundance of P. gibbesi. The presence of small holes, encrusting algae and low slope favour the colonization success of this crustacean, and the spatial distribution of these habitat features could explain the observed pattern. The abundance of P. gibbesi was similar to that of native crab species; however, a non-significant negative relationship between the abundance of P. gibbesi and native crabs (Pachygrapsus marmoratus and Eriphia verrucosa), urchins (Arbacia lixula and Paracentrotus lividus) and a snail (Phorcus turbinatus) was observed. This work highlights the importance of monitoring alien crab population densities taking structural habitat and other potentially influential factors into account and the likely effect of this alien species on the native ones.     

Crystal Structure and Site-Directed Mutagenesis Analyses of Haloalkane Dehalogenase LinB from Sphingobium sp. Strain MI1205

The enzymes LinB_UT and LinB_MI (LinB from Sphingobium japonicum UT26 and Sphingobium sp. MI1205, respectively) catalyze the hydrolytic dechlorination of β-hexachlorocyclohexane (β-HCH) and yield different products, 2,3,4,5,6-pentachlorocyclohexanol (PCHL) and 2,3,5,6-tetrachlorocyclohexane-1,4-diol (TCDL), respectively, despite their 98% identity in amino acid sequence. To reveal the structural basis of their different enzymatic properties, we performed site-directed mutagenesis and X-ray crystallographic studies of LinB_MI and its seven point mutants. The mutation analysis revealed that the seven amino acid residues uniquely found in LinB_MI were categorized into three groups based on the efficiency of the first-step (from β-HCH to PCHL) and second-step (from PCHL to TCDL) conversions. Crystal structure analyses of wild-type LinB_MI and its seven point mutants indicated how each mutated residue contributed to the first- and second-step conversions by LinB_MI. The dynamics simulation analyses of wild-type LinB_MI and LinB_UT revealed that the entrance of the substrate access tunnel of LinB_UT was more flexible than that of LinB_MI, which could lead to the different efficiencies of dehalogenation activity between these dehalogenases.     

Polymer--drug conjugates as nano-sized medicines

Polymer Therapeutics have enormously evolved in the past decades. Several polymeric drugs as well as polymer-protein conjugates have been in the market since the 90s, but although polymer-drug conjugates are already in clinical trials they still need to reach this final goal. There are four main convergent strategies to move this platform technology further. First, exploitation of new molecular targets in cancer therapy and design of polymer-drug conjugates as treatments for other diseases. Second, the development of combination therapy. Third, attempts to improve polymer chemistry, including the use of new well-defined architectures and the optimization of the advanced characterization techniques essential to transform a promising conjugate into a candidate for clinical evaluation. Finally, increased understanding of polymer conjugate features that govern clinical risk-benefit is leading to an appreciation of clinical biomarkers that will open new possibilities for personalized therapy.     

Bcl-2 down modulation in WEHI-3B/CTRES cells resistant to Cholera Toxin （CT）-induced apoptosis

The very different effects of Cholera Toxin （CT） on cell growth and proliferation may depend on the type of ganglioside receptors in cell membranes and different signal transduction mechanisms triggered, but other functions related to the drug resistance mechanisms can not be excluded. The effect of CT treatment on the ＂in vitro＂ clonogenicity, the Population Doubling Time （PDT）, apoptosis, PKA activation and Bax and Bcl-2 expression was evaluated in WEHI-3B cell line and its CT-resistant subclone （WEHI-3B/CTRES）. In WEHI-3B parental cells the dramatic accumulation of cAMP induced by CT correlated well with PKA activation, increased PDT value, inhibition of clonogenicity and apoptosis. H-89 treatment inhibited PKA activation by CT but did not protect the cells from apoptosis and growth inhibition. In WEHI-3B/CTRES no significant CT-dependent accumulation of cAMP occurred with any increase of PKA activity and PDT. In CT resistant cells （WEHI-3B/CTRES）, Bcl-2 expression was down regulated by both CT or drug treatment （eg, ciprofloxacin, CPX） although these cells were protected from CT-dependent apoptosis but not from drug-induced apoptosis. Differently from other cell models described, down regulation of Bcl-2 is proved to be independent on cAMP accumulation and PKA activation. Our observations support the implication of cAMP dependent kinase （PKA） in the inhibition of WEHI-3B cells growth and suggest that, in WEHI-3B/CTRES, Bcl-2 expression could be modulated by CT in the absence of cAMP accumulation. Also in consideration of many contradictory data reported in literature, our cell models （of one sensitive parental cell strain and two clones with different uncrossed specific resistance to CT and CPX） provides a new and interesting tool for better investigating the relationship between the CT signal transduction mechanisms and Bcl-2 expression and function.     

Correlations in thermal comfort and natural wind

Many field surveys have shown that naturally ventilated buildings are favorable to human thermal comfort and may allow higher cooling temperatures than air-conditioned buildings. Recreating natural wind characteristics with a mechanical cooling system may diminish the drawbacks of conventional cooling systems such as drafts and high energy demands.Natural wind characteristics (wind velocity, direction, turbulent intensity, temperature and relative humidity) were recorded in a mountain environment and correlated with the human thermal sensation of 48 subjects. Natural wind fluctuation characteristics were analyzed using the Fast Fourier Transform (FFT) analysis. The dynamic characteristics of natural wind were averaged through the power spectrum exponent (β−value), which represents the energy distribution of the turbulent flow of natural wind. The power spectrum exponent (β−value) of the natural wind will decrease when the mean velocity increases, while it will increase when the turbulent intensity increases. The power spectrum exponent (β−value) was correlated (Spearman's rank coefficient=0.56, p<0.001) with thermal comfort. The power spectrum exponent (β-value) for people feeling comfortable has a median value of 1.62 [1.41–1.80 for the first and third quartiles, respectively] and the β−value for people feeling uncomfortable has a median value of 1.10 [0.97–1.25].     

Expression of functional recombinant human factor IX in milk of mice

Human factor IX is synthesized in the liver and secreted in the blood, where it participates in a group of reactions involving coagulation factors and proteins that permit sanguinary coagulation. In this work two lines of transgenic mice were developed to express the FIX gene in the mammalian glands under control of milk β-casein promoter. The founding females secreted the FIX in their milk (3% total soluble protein). The stable integration of transgene was confirmed by southern blot analysis. The presence of the FIX recombinant protein in the milk of transgenic females was confirmed by western blot and the clotting activity was revealed in blood-clotting assays. The coagulation activity in human blood treated with recombinant FIX increased while the time of coagulation decreased. Our results confirm the production of a large amount of recombinant biologically active FIX in the mammary gland of transgenic mice.