The role of the Mycobacterium tuberculosis SecA2 protein export pathway in virulence

Mycobacterium tuberculosis is an intracellular bacterial pathogen that replicates in macrophages in the lung of the host. The ability to replicate in these cells of the immune system is critical to the virulence of this important pathogen. M. tuberculosis is thought to maintain a hospitable niche in the host through several mechanisms. M. tuberculosis suppresses the host innate immune response by dampening cytokine secretion and production of reactive nitrogen and oxygen species that can be toxic to bacteria. Additionally, M. tuberculosis manipulates macrophages by arresting the normal process of phagosome maturation into acidified and hydrolytic phagolysosomes. The process of each of these immunosuppressive functions by M. tuberculosis is not fully understood. Other intracellular pathogens that control the host immune response use specialized protein export systems to deliver effectors to the host cell. In M. tuberculosis, the accessory SecA2 system is a specialized protein export system that is required for intracellular growth in macrophages. However, we do not understand the role of SecA2 in promoting growth in macrophages. The SecA2 system has a role in dampening the host cytokine and reactive nitrogen response. However, we show that the role of SecA2 in dampening these inflammatory responses cannot explain the intracellular growth defect. In this study we discovered that SecA2 is also required for blocking phagosome maturation. We showed that inhibitors of phagosome acidification rescued the intracellular growth defect of the [delta]secA2 mutant, which demonstrated that the phagosome maturation arrest defect of the [delta]secA2 mutant is responsible for the intracellular growth defect. Our data suggests there are effectors of phagosome maturation that are exported into the host environment by the accessory SecA2 system. Thus, we tested a set of putative effectors of M. tuberculosis phagosome maturation for SecA2 dependent export. We found that the level of one of these proteins, the secreted acid phosphatase (SapM), was reduced in the [delta]secA2 mutant. The research presented in this thesis establishes a role for SecA2 in promoting M. tuberculosis growth in macrophages. Additionally, we demonstrate an important causal link between phagosome maturation and arrest of M. tuberculosis intracellular replication

Psychoactive substances and the political ecology of mental distress

The goal of this paper is to both understand and depathologize clinically significant mental distress related to criminalized contact with psychoactive biotic substances by employing a framework known as critical political ecology of health and disease from the subdiscipline of medical geography. The political ecology of disease framework joins disease ecology with the power-calculus of political economy and calls for situating health-related phenomena in their broad social and economic context, demonstrating how large-scale global processes are at work at the local level, and giving due attention to historical analysis in understanding the relevant human-environment relations. Critical approaches to the political ecology of health and disease have the potential to incorporate ever-broadening social, political, economic, and cultural factors to challenge traditional causes, definitions, and sociomedical understandings of disease. Inspired by the patient-centered medical diagnosis critiques in medical geography, this paper will use a critical political ecology of disease approach to challenge certain prevailing sociomedical interpretations of disease, or more specifically, mental disorder, found in the field of substance abuse diagnostics and the related American punitive public policy regimes of substance abuse prevention and control, with regards to the use of biotic substances. It will do this by first critically interrogating the concept of "substances" and grounding them in an ecological context, reviewing the history of both the development of modern substance control laws and modern substance abuse diagnostics, and understanding the biogeographic dimensions of such approaches. It closes with proposing a non-criminalizing public health approach for regulating human close contact with psychoactive substances using the example of cannabis use

Malaysia Airlines flight MH370 search data reveal geomorphology and seafloor processes in the remote southeast Indian Ocean

© The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Marine Geology 395 (2018): 301-319, doi:10.1016/j.margeo.2017.10.014.A high-resolution multibeam echosounder (MBES) dataset covering over 279,000 km2 was acquired in the southeastern Indian Ocean to assist the search for Malaysia Airlines Flight 370 (MH370) that disappeared on 8 March 2014. The data provided an essential geospatial framework for the search and is the first large-scale coverage of MBES data in this region. Here we report on geomorphic analyses of the new MBES data, including a comparison with the Global Seafloor Geomorphic Features Map (GSFM) that is based on coarser resolution satellite altimetry data, and the insights the new data provide into geological processes that have formed and are currently shaping this remote deepsea area. Our comparison between the new MBES bathymetric model and the latest global topographic/bathymetric model (SRTM15_plus) reveals that 62% of the satellite-derived data points for the study area are comparable with MBES measurements within the estimated vertical uncertainty of the SRTM15_plus model (± 100 m). However, > 38% of the SRTM15_plus depth estimates disagree with the MBES data by > 100 m, in places by up to 1900 m. The new MBES data show that abyssal plains and basins in the study area are significantly more rugged than their representation in the GSFM, with a 20% increase in the extent of hills and mountains. The new model also reveals four times more seamounts than presented in the GSFM, suggesting more of these features than previously estimated for the broader region. This is important considering the ecological significance of high-relief structures on the seabed, such as hosting high levels of biodiversity. Analyses of the new data also enabled sea knolls, fans, valleys, canyons, troughs, and holes to be identified, doubling the number of discrete features mapped. Importantly, mapping the study area using MBES data improves our understanding of the geological evolution of the region and reveals a range of modern sedimentary processes. For example, a large series of ridges extending over approximately 20% of the mapped area, in places capped by sea knolls, highlight the preserved seafloor spreading fabric and provide valuable insights into Southeast Indian Ridge seafloor spreading processes, especially volcanism. Rifting is also recorded along the Broken Ridge – Diamantina Escarpment, with rift blocks and well-bedded sedimentary bedrock outcrops discernible down to 2400 m water depth. Modern ocean floor sedimentary processes are documented by sediment mass transport features, especially along the northern margin of Broken Ridge, and in pockmarks (the finest-scale features mapped), which are numerous south of Diamantina Trench and appear to record gas and/or fluid discharge from underlying marine sediments. The new MBES data highlight the complexity of the search area and serve to demonstrate how little we know about the vast areas of the ocean that have not been mapped with MBES. The availability of high-resolution and accurate maps of the ocean floor can clearly provide new insights into the Earth's geological evolution, modern ocean floor processes, and the location of sites that are likely to have relatively high biodiversity

Life in Hot Carbon Monoxide: The Complete Genome Sequence of Carboxydothermus hydrogenoformans Z-2901

We report here the sequencing and analysis of the genome of the thermophilic bacterium Carboxydothermus hydrogenoformans Z-2901. This species is a model for studies of hydrogenogens, which are diverse bacteria and archaea that grow anaerobically utilizing carbon monoxide (CO) as their sole carbon source and water as an electron acceptor, producing carbon dioxide and hydrogen as waste products. Organisms that make use of CO do so through carbon monoxide dehydrogenase complexes. Remarkably, analysis of the genome of C. hydrogenoformans reveals the presence of at least five highly differentiated anaerobic carbon monoxide dehydrogenase complexes, which may in part explain how this species is able to grow so much more rapidly on CO than many other species. Analysis of the genome also has provided many general insights into the metabolism of this organism which should make it easier to use it as a source of biologically produced hydrogen gas. One surprising finding is the presence of many genes previously found only in sporulating species in the Firmicutes Phylum. Although this species is also a Firmicutes, it was not known to sporulate previously. Here we show that it does sporulate and because it is missing many of the genes involved in sporulation in other species, this organism may serve as a “minimal” model for sporulation studies. In addition, using phylogenetic profile analysis, we have identified many uncharacterized gene families found in all known sporulating Firmicutes, but not in any non-sporulating bacteria, including a sigma factor not known to be involved in sporulation previously

TgPRELID, a Mitochondrial Protein Linked to Multidrug Resistance in the Parasite Toxoplasma gondii

New drugs to control infection with the protozoan parasite Toxoplasma gondii are needed as current treatments exert toxic side effects on patients. Approaches to develop novel compounds for drug development include screening of compound libraries and targeted inhibition of essential cellular pathways. We identified two distinct compounds that display inhibitory activity against the parasite's replicative stage: F3215-0002, which we previously identified during a compound library screen, and I-BET151, an inhibitor of bromodomains, the "reader" module of acetylated lysines. In independent studies, we sought to determine the targets of these two compounds using forward genetics, generating resistant mutants and identifying the determinants of resistance with comparative genome sequencing. Despite the dissimilarity of the two compounds, we recovered resistant mutants with nonsynonymous mutations in the same domain of the same gene, TGGT1_254250, which we found encodes a protein that localizes to the parasite mitochondrion (designated TgPRELID after the name of said domain). We found that mutants selected with one compound were cross resistant to the other compound, suggesting a common mechanism of resistance. To further support our hypothesis that TgPRELID mutations facilitate resistance to both I-BET151 and F3215-0002, CRISPR (clustered regularly interspaced short palindromic repeat)/CAS9-mediated mutation of TgPRELID directly led to increased F3215-0002 resistance. Finally, all resistance mutations clustered in the same subdomain of TgPRELID. These findings suggest that TgPRELID may encode a multidrug resistance factor or that I-BET151 and F3215-0002 have the same target(s) despite their distinct chemical structures. IMPORTANCE We report the discovery of TgPRELID, a previously uncharacterized mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. Drug resistance remains a major problem in the battle against parasitic infection, and understanding how TgPRELID mutations augment resistance to multiple, distinct compounds will reveal needed insights into the development of new therapies for toxoplasmosis and other related parasitic diseases

Same-sign W pair production as a probe of double parton scattering at the LHC

We study the production of same-sign W boson pairs at the LHC in double parton interactions. Compared with simple factorised double parton distributions (dPDFs), we show that the recently developed dPDFs, GS09, lead to non-trivial kinematic correlations between the W bosons. A numerical study of the prospects for observing this process using same-sign dilepton signatures, including same-sign WWjj, di-boson and heavy flavour backgrounds, at 14 TeV centre-of-mass energy is then performed. It is shown that a small excess of same-sign dilepton events from double parton scattering over a background dominated by single scattering WZ(gamma*) production could be observed at the LHC.Comment: 14 pages, 8 figures. Added references, slight changes in the text

Characterization of NLRP12 during the In Vivo Host Immune Response to Klebsiella pneumoniae and Mycobacterium tuberculosis

The majority of nucleotide binding domain leucine rich repeats-containing (NLR) family members has yet to be functionally characterized. Of the described NLRs, most are considered to be proinflammatory and facilitate IL-1β production. However, a newly defined sub-group of NLRs that function as negative regulators of inflammation have been identified based on their abilities to attenuate NF-κB signaling. NLRP12 (Monarch-1) is a prototypical member of this sub-group that negatively regulates both canonical and noncanonical NF-κB signaling in biochemical assays and in colitis and colon cancer models. The role of NLRP12 in infectious diseases has not been extensively studied. Here, we characterized the innate immune response of Nlrp12−/− mice following airway exposure to LPS, Klebsiella pneumoniae and Mycobacterium tuberculosis. In response to E. coli LPS, Nlrp12−/− mice showed a slight decrease in IL-1β and increase in IL-6 production, but these levels were not statistically significant. During K. pneumoniae infection, we observed subtle differences in cytokine levels and significantly reduced numbers of monocytes and lymphocytes in Nlrp12−/− mice. However, the physiological relevance of these findings is unclear as no overt differences in the development of lung disease were observed in the Nlrp12−/− mice. Likewise, Nlrp12−/− mice demonstrated pathologies similar to those observed in the wild type mice following M. tuberculosis infection. Together, these data suggest that NLRP12 does not significantly contribute to the in vivo host innate immune response to LPS stimulation, Klebsiella pneumonia infection or Mycobacterium tuberculosis

PTF11eon/SN2011dh: Discovery of a Type IIb Supernova From a Compact Progenitor in the Nearby Galaxy M51

On May 31, 2011 UT a supernova (SN) exploded in the nearby galaxy M51 (the Whirlpool Galaxy). We discovered this event using small telescopes equipped with CCD cameras, as well as by the Palomar Transient Factory (PTF) survey, and rapidly confirmed it to be a Type II supernova. Our early light curve and spectroscopy indicates that PTF11eon resulted from the explosion of a relatively compact progenitor star as evidenced by the rapid shock-breakout cooling seen in the light curve, the relatively low temperature in early-time spectra and the prompt appearance of low-ionization spectral features. The spectra of PTF11eon are dominated by H lines out to day 10 after explosion, but initial signs of He appear to be present. Assuming that He lines continue to develop in the near future, this SN is likely a member of the cIIb (compact IIb; Chevalier and Soderberg 2010) class, with progenitor radius larger than that of SN 2008ax and smaller than the eIIb (extended IIb) SN 1993J progenitor. Our data imply that the object identified in pre-explosion Hubble Space Telescope images at the SN location is possibly a companion to the progenitor or a blended source, and not the progenitor star itself, as its radius (~10^13 cm) would be highly inconsistent with constraints from our post-explosion photometric and spectroscopic data

Compactifications and algebraic completions of Limit groups

In this paper we consider the existence of dense embeddings of Limit groups in locally compact groups generalizing earlier work of Breuillard, Gelander, Souto and Storm [GBSS] where surface groups were considered. Our main results are proved in the context of compact groups and algebraic groups over local fields. In addition we prove a generalization of the classical Baumslag lemma which is a useful tool for generating eventually faithful sequences of homomorphisms. The last section is dedicated to correct a mistake from [BGSS] and to get rid of the even genus assumption.Comment: v2: Substantial changes to sections 7 and 8.2. Typos corrected. References added. v3: Acknowledgement correcte