Cryptococcus neoformans Thermotolerance to Avian Body Temperature Is Sufficient For Extracellular Growth But Not Intracellular Survival In Macrophages

Cryptococcus neoformans is a fatal fungal pathogen of humans that efficiently parasitises macrophages. Birds can be colonised by cryptococci and can transmit cryptococcosis to humans via inhalation of inoculated bird excreta. However, colonisation of birds appears to occur in the absence of symptomatic infection. Here, using a pure population of primary bird macrophages, we demonstrate a mechanism for this relationship. We find that bird macrophages are able to suppress the growth of cryptococci seen in mammalian cells despite C. neoformans being able to grow at bird body temperature, and are able to escape from bird macrophages by vomocytosis. A small subset of cryptococci are able to adapt to the inhibitory intracellular environment of bird macrophages, exhibiting a large cell phenotype that rescues growth suppression. Thus, restriction of intracellular growth combined with survival at bird body temperature explains the ability of birds to efficiently spread C. neoformans in the environment whilst avoiding systemic disease

Daytime lidar measurements of tidal winds in the mesospheric sodium layer at Urbana, Illinois

For more than 15 years lidar systems have been used to study the chemistry and dynamics of the mesospheric sodium layer. Because the layer is an excellent tracer of atmospheric wave motions, sodium lidar has proven to be particularly useful for studying the influence of gravity waves and tides on mesospheric dynamics. These waves, which originate in the troposphere and stratosphere, propagate through the mesosphere and dissipate their energy near the mesopause making important contributions to the momentum and turbulence budget in this region of the atmosphere. Recently, the sodium lidar was modified for daytime operation so that wave phenomena and chemical effects could be monitored throughout the complete diurnal cycle. The results of continuous 24 hour lidar observations of the sodium layer structure are presented alond with measurement of the semidiurnal tidal winds

Using Flow Cytometry to Analyze Cryptococcus Infection of Macrophages.

Flow cytometry is a powerful analytical technique, which is increasingly being used to study the interaction between host cells and intracellular pathogens. Flow cytometry is capable of measuring a greater number of infected cells within a sample compared to alternative techniques such as fluorescence microscopy. This means that robust quantification of rare events during infection is possible. Our lab and others have developed flow cytometry methods to study interactions between host cells and intracellular pathogens, such as Cryptococcus neoformans, to quantify phagocytosis, intracellular replication, and non-lytic expulsion or "vomocytosis" from the phagosome. Herein we describe these methods and how they can be applied to the study of C. neoformans as well as other similar intracellular pathogens

Using Flow Cytometry to Analyze Cryptococcus Infection of Macrophages.

Flow cytometry is a powerful analytical technique, which is increasingly being used to study the interaction between host cells and intracellular pathogens. Flow cytometry is capable of measuring a greater number of infected cells within a sample compared to alternative techniques such as fluorescence microscopy. This means that robust quantification of rare events during infection is possible. Our lab and others have developed flow cytometry methods to study interactions between host cells and intracellular pathogens, such as Cryptococcus neoformans, to quantify phagocytosis, intracellular replication, and non-lytic expulsion or "vomocytosis" from the phagosome. Herein we describe these methods and how they can be applied to the study of C. neoformans as well as other similar intracellular pathogens

Vertical Structure and Color of Jovian Latitudinal Cloud Bands during the Juno Era

The identity of the coloring agent(s) in Jupiter's atmosphere and the exact structure of Jupiter's uppermost cloud deck are yet to be conclusively understood. The Cr\`{e}me Br\^ul\'ee model of Jupiter's tropospheric clouds, originally proposed by Baines et al. (2014) and expanded upon by Sromovsky et al. (2017) and Baines et al. (2019), presumes that the chromophore measured by Carlson et al. (2016) is the singular coloring agent in Jupiter's troposphere. In this work, we test the validity of the Cr\`{e}me Br\^ul\'ee model of Jupiter's uppermost cloud deck using spectra measured during the Juno spacecraft's 5$^{\mathrm{th}}$ perijove pass in March 2017. These data were obtained as part of an international ground-based observing campaign in support of the Juno mission using the NMSU Acousto-optic Imaging Camera (NAIC) at the 3.5-m telescope at Apache Point Observatory in Sunspot, NM. We find that the Cr\`{e}me Br\^ul\'ee model cloud layering scheme can reproduce Jupiter's visible spectrum both with the Carlson et al. (2016) chromophore and with modifications to its imaginary index of refraction spectrum. While the Cr\`{e}me Br\^ul\'ee model provides reasonable results for regions of Jupiter's cloud bands such as the North Equatorial Belt and Equatorial Zone, we find that it is not a safe assumption for unique weather events, such as the 2016-2017 Southern Equatorial Belt outbreak that was captured by our measurements.Comment: 38 pages, 21 figures; Accepted for publication in AAS Planetary Science Journa

Automated Analysis of Cryptococcal Macrophage Parasitism Using GFP-Tagged Cryptococci

The human fungal pathogens Cryptococcus neoformans and C. gattii cause life-threatening infections of the central nervous system. One of the major characteristics of cryptococcal disease is the ability of the pathogen to parasitise upon phagocytic immune effector cells, a phenomenon that correlates strongly with virulence in rodent models of infection. Despite the importance of phagocyte/Cryptococcus interactions to disease progression, current methods for assaying virulence in the acrophage system are both time consuming and low throughput. Here, we introduce the first stable and fully characterised GFP–expressing derivatives of two widely used cryptococcal strains: C. neoformans serotype A type strain H99 and C. gattii serotype B type strain R265. Both strains show unaltered responses to environmental and host stress conditions and no deficiency in virulence in the macrophage model system. In addition, we report the development of a method to effectively and rapidly investigate macrophage parasitism by flow cytometry, a technique that preserves the accuracy of current approaches but offers a four-fold improvement in speed

Laser Time-of-Flight Mass Spectrometry for Future In Situ Planetary Missions

Laser desorption/ionization time-of-flight mass spectrometry (LD-TOF-MS) is a versatile, low-complexity instrument class that holds significant promise for future landed in situ planetary missions that emphasize compositional analysis of surface materials. Here we describe a 5kg-class instrument that is capable of detecting and analyzing a variety of analytes directly from rock or ice samples. Through laboratory studies of a suite of representative samples, we show that detection and analysis of key mineral composition, small organics, and particularly, higher molecular weight organics are well suited to this instrument design. A mass range exceeding 100,000 Da has recently been demonstrated. We describe recent efforts in instrument prototype development and future directions that will enhance our analytical capabilities targeting organic mixtures on primitive and icy bodies. We present results on a series of standards, simulated mixtures, and meteoritic samples

"A little theatrical but mostly athletic": The mutable erotics of Miranda July's The First Bad Man

By attending to the inherent flux of sexual fantasy, Miranda July’s first novel The First Bad Man reveals a mobile and mutable erotics capable of generating an enlarged range of self-identification and relational intimacy, far from any essentialist assumptions of stable or coherent sexual identity. July focuses specifically upon role-play as the means to unpack the normative categories of hetero/homosexuality, masculinity and femininity, celebrating rather than pathologising qualities like superficiality and inconsistency. The novel touches upon many polarising issues (for example, sexual violence, sadomasochism and assisted reproductive technology), deftly avoiding the conventional language which colours perception. Both erotic and humorous, The First Bad Man helps to redefine the often highly charged discourse around sex and sexuality