Hypoxia: a window into Mycobacterium tuberculosis latency

Tuberculosis is a massive public health problem on a global scale and the success of Mycobacterium tuberculosis is linked to its ability to persist within humans for long periods without causing any overt disease symptoms. Hypoxia is predicted to be a key host-induced stress limiting growth of the pathogen in vivo . However, multiple studies in vitro and in vivo indicate that M. tuberculosis adapts to oxygen limitation by entering into a metabolically altered state, while awaiting the opportunity to reactivate. Molecular signatures of bacteria adapted to hypoxia in vitro are accumulating, although correlations to human disease are only now being established. Similarly, defining the mechanisms that control this adaptation is an active area of research. In this review we discuss the historical precedents linking hypoxia and latency, and the gathering knowledge of M. tuberculosis hypoxic responses. We also examine the role of these responses in tuberculosis latency, and identify promising avenues for future studies.     

Radiation-induced fluctuation of the polysome content in adequately oxygenated tetrahymena pyriformis.

The polysome content of the ciliate protozoan Tetrahymena pyriformis is transiently reduced by gamma-irradiation. In order to test whether this is a result of a respiration-produced or radiation-produced hypoxia or some other mechanism, the oxygen content of the culture was determined during and after irradiation, and the polysome contents and rates of amino acid incorporation were measured with and without air bubbling. Irradiation (40 krad at approximately 3 krad/min) produced approximately a 25 per cent loss in dissolved O2 content in the medium. This decrease is not sufficient to affect the polysome level, since (a) the same radiation-induced loss of polysomes and inhibiition of amino acid incorporation was observed whether or not the culture was bubbled with air during the irradiation and (b) bubbling unirradiated cultures with gas mixtures containing as little as 17 per cent of the normal O2 content did not influence the polysome level. As long as the cells are irradiated as a shallow layer in open flasks, replacement of O2 from the gas phase appears adequate, and neither respiration-induced nor radiation-induced hypoxia masks the effects of the radiation.     

The action of phospholipase C and lipase on black film bilayer membranes

Phospholipase C(C. perfringens) added to one side of a phosphatidyl choline—phosphatidyl serine—cholesterol (0.7: 0.3: 1.0, w/w/w) bilayer membrane resulted in a rapid decrease in membrane resistance which leveled off after several minutes at about one-half its initial value. Pancreatic lipase added at this time resulted in a rapid increase in membrane resistance to a value somewhat higher than the initial. This effect was independent of the side to which the lipase was added, indicating that diglyceride is rapidly equilibrated across both sides of the membrane. When both phospholipase C and lipase were added at zero time to the same or opposite sides, the resistance decreased slightly and then increased to a value higher than the initial. Replacement of phosphatidyl choline by 2-hexadecoxy-3-octadecoxypropyl-phosphonylcholine, a phosphonate analog, resulted in an inhibition of the phospholipase C reaction which was equal to the percent analog. The analog, thus, seemed to act only as inert lipid and did not inhibit hydrolys of the active lipid.     

Ultra-low Dose Aerosol Infection of Mice with Mycobacterium tuberculosis More Closely Models Human Tuberculosis

1. Download : Download high-res image (176KB)
2. Download : Download full-size image

     

Influence of Transglutaminase (TGase) Enzyme on Mechanical and Bioactive Properties of Crayfish Protein Gels

Functional and bioactive properties of crayfish meat convert their surpluses in an excellent alternative for the development of food products. Thus, protein dispersions were subjected to a thermal treatment, obtaining a protein-based gel. Rheological and antioxidant properties were studied at three different pH values (2.0, 6.5 and 8.0) when the TGase enzyme was used. The pH value exerted a strong influence on the gelation behaviour, as well as on the functional properties and the antioxidant activity of the final gels. The activity of the TGase enzyme is highly influenced by the pH of the protein dispersions. The highest antioxidant activity was obtained against ABTS and the lowest when FC reagent was used, whereas the activity against DPPH was also remarkable. TGase enzyme can be used during the thermal treatment to increase the mechanical properties, which were lost when hydrolysate systems were used.     

On the origins of esterases

Comparisons among the primary sequences of five cloned eukaryotic esterases reveal two distinct lineages, neither bearing any significant overall sequence similarity to the functionally related serine protease multigene family. We have not eliminated the possibility that the esterases may have residual conformational similarities to the serine proteases. However, our profile analysis and analyses of the predicted conformations of the esterases reveal little similarity to the serine proteases. Four of the esterase proteins share 27%-53% overall sequence similarity and evidence of a catalytic mechanism involving the same Arg- Asp-Ser or His-Asp-Ser charge relay. We propose that these four esterases, three of them cholinesterases, form part of a multigene family essentially separate from the serine proteases.