The release of membrane-bound calcium by radiation and sulfhydryl reagents

Effects of ionizing radiation and of sulfhydryl reagents on the ⁴⁵Ca binding of red cell membranes were studied. Corresponding effects of these agents on potassium leak from intact red cells were also determined. Essentially all the ⁴⁵Ca associated with the ghosts appeared to be bound. Calcium binding could be described by assuming two independent groups of binding sites with dissociation constants of about 6 × 10^?4 m and 2 × 10^?4 m. The total binding capacity was about 2.5 × 10^?4 moles/g ghost protein. Membrane calcium was decreased by radiation and by the two sulfhydryl reagents, p-chloromercuribenzoate (PCMB) and N-ethyl maleimide (NEM). The tightly bound calcium fraction appeared to be most affected by these agents. Changes in potassium leak evoked by varying doses of agents appeared to parallel effects on membrane calcium. These investigations suggest that the increased cation permeability observed after exposure or red cells to radiation or sulfhydryl reagents may be related to alterations in the calcium-binding properties of the cell membrane.     

A micrite-dominated norian carbonate platform from Northern Calabria (Southern Italy)

Summary  This paper is focused on the facies associations of an Early-Middle Norian stratigraphic succession cropping outin the Northern Calabria (Buonvicino, Cosenza). These carbonate deposits, pertaining to the Verbicaro Unit, represent a dolomitized platform characterized by a clear dominance of automicrites over skeletal metazoans. On the basis of the chronostatigraphic data and sedimentary evolution, two stratigraphic units have been distinguished: the Lower Unit (object of the present paper) and the Upper Unit. The Lower Unit consists of an Early-Middle Norian high-relief prograding carbonate platform. The Upper Unit is represented by Middle p.p.-Upper Norian basinal deposits. Three main facies associations, indicative of different depositional settings, have been identified. The inner platform facies association is characterized by automicrite (cauliflower-columnar subordinate planar stromatolites) associated with detrital carbonates (intraclastic breccia, bioclastic grainstone and packstone). Apeculiar facies is represented by megalodontid-bearing beds. Mud-cracked horizons and low-developed teepees occur quite frequently. The margin facies association is dominated by automicrite (planar to low relief stromatolites associated with thrombolitic fenestral boundstone) and detrital carbonates with subaerial exposure features. The slope facies association includes detrital carbonates (breccia/megabreccia) associated with serpulid/sphinctozoan bioconstructions and automicrite (planar stromatolites/thrombolitic boundstone). The whole carbonate body is completely dolomitized; nevertheless the morphology and microarchitecture of carbonate components (cements, grains and automicrite) is still clearly recognizable. The dolomite Mg content ranges from 40 to 48 mole%, sometimes reaching the stoichiometric value. Cements, primary and late, represent a minor component of the rock volume; they occur more frequently on the margin and upper slope setting. Quantitative tacies analyses lead to the following conclusions:

Photodynamic inactivation of recombinant bioluminescent Escherichia coli by cationic porphyrins under artificial and solar irradiation

A faster and simpler method to monitor the photoinactivation process of Escherichia coli involving the use of recombinant bioluminescent bacteria is described here. Escherichia coli cells were transformed with luxCDABE genes from the marine bioluminescent bacterium Vibrio fischeri and the recombinant bioluminescent indicator strain was used to assess, in real time, the effect of three cationic meso-substituted porphyrin derivatives on their metabolic activity, under artificial (40 W m⁻²) and solar irradiation (≈620 W m⁻²). The photoinactivation of bioluminescent E. coli is effective (>4 log bioluminescence decrease) with the three porphyrins used, the tricationic porphyrin Tri-Py⁺-Me-PF being the most efficient compound. The photoinactivation process is efficient both with solar and artificial light, for the three porphyrins tested. The results show that bioluminescence analysis is an efficient and sensitive approach being, in addition, more affordable, faster, cheaper and much less laborious than conventional methods. This approach can be used as a screening method for bacterial photoinactivation studies in vitro and also for the monitoring of the efficiency of novel photosensitizer molecules. As far as we know, this is the first study involving the use of bioluminescent bacteria to monitor the antibacterial activity of porphyrins under environmental conditions.     

Human antibody responses to the Anopheles salivary gSG6-P1 peptide: a novel tool for evaluating the efficacy of ITNs in malaria vector control

To optimize malaria control, WHO has prioritised the need for new indicators to evaluate the efficacy of malaria vector control strategies. The gSG6-P1 peptide from gSG6 protein of Anopheles gambiae salivary glands was previously designed as a specific salivary sequence of malaria vector species. It was shown that the quantification of human antibody (Ab) responses to Anopheles salivary proteins in general and especially to the gSG6-P1 peptide was a pertinent biomarker of human exposure to Anopheles. The present objective was to validate this indicator in the evaluation of the efficacy of Insecticide Treated Nets (ITNs). A longitudinal evaluation, including parasitological, entomological and immunological assessments, was conducted on children and adults from a malaria-endemic area before and after the introduction of ITNs. Significant decrease of anti-gSG6-P1 IgG response was observed just after the efficient ITNs use. Interestingly, specific IgG Ab level was especially pertinent to evaluate a short-time period of ITNs efficacy and at individual level. However, specific IgG rose back up within four months as correct ITN use waned. IgG responses to one salivary peptide could constitute a reliable biomarker for the evaluation of ITN efficacy, at short- and long-term use, and provide a valuable tool in malaria vector control based on a real measurement of human-vector contact.     

Relation between bacterial activity in the surface microlayer and estuarine hydrodynamics

Bacterial communities of the surface microlayer (SML) of the estuary Ria de Aveiro (Portugal) were characterized in terms of abundance and activity during a 2-year survey at two sites with distinct hydrodynamic properties (marine and brackish water zones). The hydrodynamic conditions were simulated using a bidimensional numerical model and related to the microbiological observations. The pattern of variation of bacterial biomass productivity (BBP) was distinct between the two sampling sites. At the outer site, BBP was significantly lower at the SML, whereas at the inner site, it was significantly enhanced at the SML. Although the total bacterial abundance was similar in the SML and underlying water (UW), the fraction of cells attached to particles was significantly higher at the SML (two to three times). The integration of microbiological results with environmental and hydrological variables shows that strong currents in the marine zone promote the vertical mixing, inhibiting the establishment of an SML bacterial community distinct from that of UW. In contrast, in the brackish water zone, lower current velocities provide conditions for enhancing the bacterial activity in the enriched SML. Estuarine dynamics influence the distribution and activity of microorganisms at the SML and in the water column, with anticipated impacts for the carbon cycle in the estuarine environment.