An ABC transporter containing a forkhead-associated domain interacts with a serine-threonine protein kinase and is required for growth of Mycobacterium tuberculosis in mice

Forkhead-associated (FHA) domains are modular phosphopeptide recognition motifs with a striking preference for phosphothreonine-containing epitopes. FHA domains have been best characterized in eukaryotic signaling pathways but have been identified in six proteins in Mycobacterium tuberculosis, the causative organism of tuberculosis. One of these, coded by gene Rv1747, is an ABC transporter and the only one to contain two such modules. A deletion mutant of Rv1747 is attenuated in a mouse intravenous injection model of tuberculosis where the bacterial load of the mutant is 10-fold lower than that of the wild type in both lungs and spleen. In addition, growth of the mutant in mouse bone marrow-derived macrophages and dendritic cells is significantly impaired. In contrast, growth of this mutant in vitro was indistinguishable from that of the wild type. The mutant phenotype was lost when the mutation was complemented by the wild-type allele, confirming that it was due to mutation of Rv1747. Using yeast two-hybrid analysis, we have shown that the Rv1747 protein interacts with the serine-threonine protein kinase PknF. This interaction appears to be phospho-dependent since it is abrogated in a kinase-dead mutant and by mutations in the presumed activation loop of PknF and in the first FHA domain of Rv1747. These results demonstrate that the protein coded by Rv1747 is required for normal virulent infection by M. tuberculosis in mice and, since it interacts with a serine-threonine protein kinase in a kinase-dependent manner, indicate that it forms part of an important phospho-dependent signaling pathway

Method for the calculation of spacecraft umbra and penumbra shadow terminator points

A method for calculating orbital shadow terminator points is presented. The current method employs the use of an iterative process which is used for an accurate determination of shadow points. This calculation methodology is required since orbital perturbation effects can introduce large errors when a spacecraft orbits a planet in a high altitude and/or highly elliptical orbit. To compensate for the required iteration methodology, all reference frame change definitions and calculations are performed with quaternions. Quaternion algebra significantly reduces the computational time required for the accurate determination of shadow terminator points

Manufacturing High-Fidelity Lunar Agglutinate Simulants

The lunar regolith is very different from many naturally occurring material on Earth because it forms in the unique, impact-dominated environment of the lunar surface. Lunar regolith is composed of five basic particle types: mineral fragments, pristine crystalline rock fragments, breccia fragments, glasses of various kinds, and agglutinates (glass-bonded aggregates). Agglutinates are abundant in the lunar regolith, especially in mature regoliths where they can be the dominant component.This presentation will discuss the technical feasibility of manufacturing-simulated agglutinate particles that match many of the unique properties of lunar agglutinates

NASA Perspective and Modeling of Thermal Runaway Propagation Mitigation in Aerospace Batteries

NASA has traditionally sought to reduce the likelihood of a single cell thermal runaway (TR) in their aerospace batteries to an absolute minimum by employing rigorous screening program of the cells. There was generally a belief that TR propagation resulting in catastrophic failure of the battery was a forgone conclusion for densely packed aerospace lithium-ion batteries. As it turns out, this may not be the case. An increasing number of purportedly TR propagation-resistant batteries are appearing among NASA partners in the commercial sector and the Department of Defense. In the recent update of the battery safety standard (JSC 20793) to address this paradigm shift, the NASA community included requirements for assessing TR severity and identifying simple, low-cost severity reduction measures. Unfortunately, there are no best-practice guidelines for this work in the Agency, so the first project team attempting to meet these requirements would have an undue burden placed upon them. A NASA engineering Safety Center (NESC) team set out to perform pathfinding activities for meeting those requirements. This presentation will provide contextual background to this effort, as well as initial results in attempting to model and simulate TR heat transfer and propagation within battery designs

Secretory vesicles are preferentially targeted to areas of low molecular SNARE density

Intercellular communication is commonly mediated by the regulated fusion, or exocytosis, of vesicles with the cell surface. SNARE (soluble N-ethymaleimide sensitive factor attachment protein receptor) proteins are the catalytic core of the secretory machinery, driving vesicle and plasma membrane merger. Plasma membrane SNAREs (tSNAREs) are proposed to reside in dense clusters containing many molecules, thus providing a concentrated reservoir to promote membrane fusion. However, biophysical experiments suggest that a small number of SNAREs are sufficient to drive a single fusion event. Here we show, using molecular imaging, that the majority of tSNARE molecules are spatially separated from secretory vesicles. Furthermore, the motilities of the individual tSNAREs are constrained in membrane micro-domains, maintaining a non-random molecular distribution and limiting the maximum number of molecules encountered by secretory vesicles. Together our results provide a new model for the molecular mechanism of regulated exocytosis and demonstrate the exquisite organization of the plasma membrane at the level of individual molecular machines

The effect of human serum in vitro on Trypanosoma (Trypanozoon) brucei species trypanosomes and its relationship to infectivity in the BIood Incubation lnfectivity test

This article presents the trypanolytic action of human serum in vitro at 37°C on different strains of salivarian trypanosomes.All attempts to identify 1 HH Species trypanosome strains by their sensitivity or resistance to human serum in vitro have so far been unsuccessful. Wet-film exaffiifiltTon of b.rn..and_T..(N±_nnei monas) congolense trypanosome strains, both before and after in vitro incubation in fresh human blood for 5 hour;Tffi°C showed that serum-sensitivity or serum-resistance is not a constant and reproducible property of a trypanosome strain and is totally unrelated to infectivity.Office of Global AIDS/US Department of State