Use of Image Cytometry for Quantification of Pathogenic Fungi in Association with Host Cells

Studies of the cellular pathogenesis mechanisms of pathogenic yeasts such as Candida albicans, Histoplasma capsulatum, and Cryptococcus neoformans commonly employ infection of mammalian hosts or host cells (i.e. macrophages) followed by yeast quantification using colony forming unit analysis or flow cytometry. While colony forming unit enumeration has been the most commonly used method in the field, this technique has disadvantages and limitations, including slow growth of some fungal species on solid media and low and/or variable plating efficiencies, which is of particular concern when comparing growth of wild-type and mutant strains. Flow cytometry can provide rapid quantitative information regarding yeast viability, however, adoption of flow cytometric detection for pathogenic yeasts has been limited for a number of practical reasons including its high cost and biosafety considerations. Here, we demonstrate an image-based cytometric methodology using the Cellometer Vision (Nexcelom Bioscience, LLC) for the quantification of viable pathogenic yeasts in co-culture with macrophages. Our studies focus on detection of two human fungal pathogens: Histoplasma capsulatum and Candida albicans. H. capsulatum colonizes alveolar macrophages by replicating within the macrophage phagosome, and here, we quantitatively assess the growth of H. capsulatum yeasts in RAW 264.7 macrophages using acridine orange/propidium iodide staining in combination with image cytometry. Our method faithfully recapitulates growth trends as measured by traditional colony forming unit enumeration, but with significantly increased sensitivity. Additionally, we directly assess infection of live macrophages with a GFP-expressing strain of C. albicans. Our methodology offers a rapid, accurate, and economical means for detection and quantification of important human fungal pathogens in association with host cells

Image cytometry as an alternative to flow cytometry for the transplant histocompatibility crossmatch assay

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/149300/1/tan13515_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/149300/2/tan13515.pd

Dust Emission from Evolved and Unevolved HII Regions in the Large Magellanic Cloud

We present a study of the dust properties of 12 classical and superbubble HII regions in the Large Magellanic Cloud. We use infrared photometry from Spitzer (8, 24, 70, and 160 \mum bands), obtained as part of the Surveying the Agents of a Galaxy's Evolution (SAGE) program, along with archival spectroscopic classifications of the ionizing stars to examine the role of stellar sources on dust heating and processing. Our infrared observations show surprisingly little correlation between the emission properties of the dust and the effective temperatures or bolometric magnitudes of stars in the HII regions, suggesting that the HII region evolutionary timescale is not on the order of the dust processing timescale. We find that the infrared emission of superbubbles and classical HII regions shows little differentiation between the two classes, despite the significant differences in age and morphology. We do detect a correlation of the 24 \mum emission from hot dust with the ratio of 70 to 160 \mum flux. This correlation can be modeled as a trend in the temperature of a minority hot dust component, while a majority of the dust remains significantly cooler.Comment: 15 pages, 5 figures. Accepted to Ap

The global dust SED: Tracing the nature and evolution of dust with DustEM

The Planck and Herschel missions are currently measuring the farIR-mm emission of dust, which combined with existing IR data, will for the first time provide the full SED of the galactic ISM dust emission with an unprecedented sensitivity and angular resolution. It will allow a systematic study of the dust evolution processes that affect the SED. Here we present a versatile numerical tool, DustEM, that predicts the emission and extinction of dust given their size distribution and their optical and thermal properties. In order to model dust evolution, DustEM has been designed to deal with a variety of grain types, structures and size distributions and to be able to easily include new dust physics. We use DustEM to model the dust SED and extinction in the diffuse interstellar medium at high-galactic latitude (DHGL), a natural reference SED. We present a coherent set of observations for the DHGL SED. The dust components in our DHGL model are (i) PAHs, (ii) amorphous carbon and (iii) amorphous silicates. We use amorphous carbon dust, rather than graphite, because it better explains the observed high abundances of gas-phase carbon in shocked regions of the interstellar medium. Using the DustEM model, we illustrate how, in the optically thin limit, the IRAS/Planck HFI (and likewise Spitzer/Herschel for smaller spatial scales) photometric band ratios of the dust SED can disentangle the influence of the exciting radiation field intensity and constrain the abundance of small grains relative to the larger grains. We also discuss the contributions of the different grain populations to the IRAS, Planck and Herschel channels. Such information is required to enable a study of the evolution of dust as well as to systematically extract the dust thermal emission from CMB data and to analyze the emission in the Planck polarized channels. The DustEM code described in this paper is publically available.Comment: accepted for publication in A&

Shattering by turbulence as a production source of very small grains

The origin of grain size distribution in the interstellar medium is one of the most fundamental problems in the interstellar physics. In the Milky Way, smaller grains are more abundant in number, but their origins are not necessarily specified and quantified. One of the most efficient drivers of small grain production is interstellar turbulence, in which dust grains can acquire relative velocities large enough to be shattered. Applying the framework of shattering developed in previous papers, we show that small (a\la 0.01~\micron) grains reach the abundance level observed in the Milky Way in $\sim 10^8$ yr (i.e. within the grain lifetime) by shattering in warm neutral medium. We also show that if part of grains experience additional shattering in warm ionized medium, carbonaceous grains with a\sim 0.01~\micron are redistributed into smaller sizes. This could explain the relative enhancement of very small carbonaceous grains with $a\sim 3$--100 \AA. Our theory also explains the ubiquitous association between large grains and very small grains naturally. Some tests for our theory are proposed in terms of the metallicity dependence.Comment: 5 pages, 2 figures, accepted for publication in MNRAS Letter

Submillimeter to centimeter excess emission from the Magellanic Clouds. II. On the nature of the excess

Dust emission at submm to cm wavelengths is often simply the Rayleigh-Jeans tail of dust particles at thermal equilibrium and is used as a cold mass tracer in various environments including nearby galaxies. However, well-sampled spectral energy distributions of the nearby, star-forming Magellanic Clouds have a pronounced (sub-)millimeter excess (Israel et al., 2010). This study attempts to confirm the existence of such a millimeter excess above expected dust, free-free and synchrotron emission and to explore different possibilities for its origin. We model NIR to radio spectral energy distributions of the Magellanic Clouds with dust, free-free and synchrotron emission. A millimeter excess emission is confirmed above these components and its spectral shape and intensity are analysed in light of different scenarios: very cold dust, Cosmic Microwave Background (CMB) fluctuations, a change of the dust spectral index and spinning dust emission. We show that very cold dust or CMB fluctuations are very unlikely explanations for the observed excess in these two galaxies. The excess in the LMC can be satisfactorily explained either by a change of the spectral index due to intrinsic properties of amorphous grains, or by spinning dust emission. In the SMC however, due to the importance of the excess, the dust grain model including TLS/DCD effects cannot reproduce the observed emission in a simple way. A possible solution was achieved with spinning dust emission, but many assumptions on the physical state of the interstellar medium had to be made. Further studies, using higher resolution data from Planck and Herschel, are needed to probe the origin of this observed submm-cm excess more definitely. Our study shows that the different possible origins will be best distinguished where the excess is the highest, as is the case in the SMC.Comment: 7 pages, 6 figures; accepted in A&

Prevalence of Obesity and Its Association with Cardiovascular Disease Risk Factors in Adolescent Girls from a College in Central Taiwan

Although obesity is associated with important hemodynamic disturbances, little data exists on population-wide cardiovascular risk factors in obese adolescent girls in Taiwan. This study measured the prevalence of overweight/obesity and related cardiovascular disease risk factors in adolescent females. This was a school-based survey of a representative sample of 291 females aged 15 and 18 years in a public college in Central Taiwan. The main measures were height, body weight, systolic (SBP) and diastolic blood pressure (DBP), uric acid, cholesterol, triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C). Obese (body mass index [BMI]≥25.3) and overweight (22.7≤BMI≤25.2) individuals were combined and labeled as overweight (BMI ≥22.7) to make communication of results clearer. Data gleaned from freshmen's health examinations were analyzed. The prevalence of obesity (BMI≥25.3) was 9.28% and of overweight (BMI≥22.7) was 21.31%. Being overweight was associated with higher SBP, DBP, uric acid and TG, and lower levels of HDL-C, but was not associated with cholesterol. The 15-year-old group showed higher mean levels of uric acid, total cholesterol, TG and HDL-C than the 18-year-old group (p < 0.05). All told, 3.1%, 15.12% and 2.1% of the girls showed abnormally elevated levels of uric acid, cholesterol and TG, respectively. In addition, 5.84% had abnormally lower HDL-C levels, indicating that interventions should focus on reducing obesity and encouraging proper dietary habits and sufficient exercise, especially in subjects with lower HDL-C levels and higher levels of cholesterol, TG and uric acid

The Spatial Distribution of Dust and Stellar Emission of the Magellanic Clouds

We study the emission by dust and stars in the Large and Small Magellanic Clouds, a pair of low-metallicity nearby galaxies, as traced by their spatially resolved spectral energy distributions (SEDs). This project combines Herschel Space Observatory PACS and SPIRE far-infrared photometry with other data at infrared and optical wavelengths. We build maps of dust and stellar luminosity and mass of both Magellanic Clouds, and analyze the spatial distribution of dust/stellar luminosity and mass ratios. These ratios vary considerably throughout the galaxies, generally between the range $0.01\leq L_{\rm dust}/L_\ast\leq 0.6$ and $10^{-4}\leq M_{\rm dust}/M_\ast\leq 4\times10^{-3}$. We observe that the dust/stellar ratios depend on the interstellar medium (ISM) environment, such as the distance from currently or previously star-forming regions, and on the intensity of the interstellar radiation field (ISRF). In addition, we construct star formation rate (SFR) maps, and find that the SFR is correlated with the dust/stellar luminosity and dust temperature in both galaxies, demonstrating the relation between star formation, dust emission and heating, though these correlations exhibit substantial scatter.Comment: 15 pages, 18 figures; ApJ, in press; version published in the journal will have higher-resolution figure

TSPO: kaleidoscopic 18-kDa amid biochemical pharmacology, control and targeting of mitochondria

The 18-kDa translocator protein (TSPO) localizes in the outer mitochondrial membrane (OMM) of cells and is readily up-regulated under various pathological conditions such as cancer, inflammation, mechanical lesions and neurological diseases. Able to bind with high affinity synthetic and endogenous ligands, its core biochemical function resides in the translocation of cholesterol into the mitochondria influencing the subsequent steps of (neuro-)steroid synthesis and systemic endocrine regulation. Over the years, however, TSPO has also been linked to core cellular processes such as apoptosis and autophagy. It interacts and forms complexes with other mitochondrial proteins such as the voltage-dependent anion channel (VDAC) via which signalling and regulatory transduction of these core cellular events may be influenced. Despite nearly 40 years of study, the precise functional role of TSPO beyond cholesterol trafficking remains elusive even though the recent breakthroughs on its high-resolution crystal structure and contribution to quality-control signalling of mitochondria. All this along with a captivating pharmacological profile provides novel opportunities to investigate and understand the significance of this highly conserved protein as well as contribute the development of specific therapeutics as presented and discussed in the present review