Phenotypic mixing between N- and B-tropic murine leukemia viruses: infectious particles with dual sensitivity to Fv-1 restriction.

In effort to understand how N or B tropism is determined in murine leukemia virus (MuLV) particles, we analyzed the MuLV produced after dual infection of mouse cells by N- and B-tropic MuLV. The progeny MuLV from such a mixed infection are sensitive to Fv-1 restriction in both N- and B-type cells, but are still highly infectious for mouse cells which do not exhibit Fv-1 restriction. This dual sensitivity to Fv-1 restriction is a phenotypic property of MuLV produced by mixedly infected cells, since individual virus clones derived from this MuLV are either N- or B-tropic. In further experiments, we superinfected murine sarcoma virus (MSV)-transformed cells with mixtures of N- and B-tropic MuLVs. The rescued MSV is restricted in its ability to transforms both N- and B-type cells. The results suggest that N- and B-tropic MuLVs specify different determinants, which are incorporated into virions along with the viral genome and which are the recognition sites for Fv-1 restriction. The presence of a given determinant in a virion renders the virus sensitive to restriction in cells of the opposite Fv-1 type.     

Study of permeation and blocker binding in TMEM16A calcium-activated chloride channels

We studied the effects of mutations of positively charged amino acid residues in the pore of X. tropicalis TMEM16A calcium-activated chloride channels: K613E, K628E, K630E; R646E and R761E. The activation and deactivation kinetics were not affected, and only K613E showed a lower current density. K628E and R761E affect anion selectivity without affecting Na⁺ permeation, whereas K613E, R646E and the double mutant K613E + R646E affect anion selectivity and permeability to Na⁺. Furthermore, altered blockade by the chloride channel blockers anthracene-9-carboxylic acid (A-9-C), 4, 4''-Diisothiocyano-2,2''-stilbenedisulfonic acid (DIDS) and T16inh-A01 was observed. These results suggest the existence of 2 binding sites for anions within the pore at electrical distances of 0.3 and 0.5. These sites are also relevant for anion permeation and blockade.     

Investigating the most appropriate methods for attached solids determination in moving-bed biofilm reactors

Moving-bed biofilm reactors (MBBR) have been employed worldwide as an efficient technology for the treatment of a diverse set of wastewaters. Although the attached biomass represents the major fraction of solids in MBBR systems, there is still no standard for its reliable quantification. An extensive literature review indicated that several methods for attached biomass assessment are applied, hindering the comparability of results issued from different studies. Therefore, the most reported methods for biofilm quantification in the MBBR literature were compared using three different carriers. The results revealed that the performance of each method was biased depending on the carrier type and shape. Moreover, differences in total attached solids (TAS) concentrations varied from 13% up to more than 90%, depending on the employed method for a given carrier. Overall, direct weighing of the carrier containing the biofilm, accounting for the clean carrier weight, and manual extraction of the biofilm, preceded or not by sonication for at least 15 min, were the most suitable techniques for assessing TAS and the volatile/total solids ratio in non-porous medias, respectively. The results here presented may be used as a frame of reference for standardization of the methods for assessing the biofilm mass in MBBR carriers.