Pseudomonas Aeruginosa-Induced Bleb-Niche Formation in Epithelial Cells Is Independent of Actinomyosin Contraction and Enhanced by Loss of Cystic Fibrosis Transmembrane-Conductance Regulator Osmoregulatory Function

The opportunistic pathogen Pseudomonas aeruginosa can infect almost any site in the body but most often targets epithelial cell-lined tissues such as the airways, skin, and the cornea of the eye. A common predisposing factor is cystic fibrosis (CF), caused by defects in the cystic fibrosis transmembrane-conductance regulator (CFTR). Previously, we showed that when P. aeruginosa enters epithelial cells it replicates intracellularly and occupies plasma membrane blebs. This phenotype is dependent on the type 3 secretion system (T3SS) effector ExoS, shown by others to induce host cell apoptosis. Here, we examined mechanisms for P. aeruginosa-induced bleb formation, focusing on its relationship to apoptosis and the CFTR. The data showed that P. aeruginosa-induced blebbing in epithelial cells is independent of actin contraction and is inhibited by hyperosmotic media (400 to 600 mOsM), distinguishing bacterially induced blebs from apoptotic blebs. Cells with defective CFTR displayed enhanced bleb formation upon infection, as demonstrated using bronchial epithelial cells from a patient with cystic fibrosis and a CFTR inhibitor, CFTR(Inh)-172. The defect was found to be correctable either by incubation in hyperosmotic media or by complementation with CFTR (pGFP-CFTR), suggesting that the osmoregulatory function of CFTR counters P. aeruginosa-induced bleb-niche formation. Accordingly, and despite their reduced capacity for bacterial internalization, CFTR-deficient cells showed greater bacterial occupation of blebs and enhanced intracellular replication. Together, these data suggest that P. aeruginosa bleb niches are distinct from apoptotic blebs, are driven by osmotic forces countered by CFTR, and could provide a novel mechanism for bacterial persistence in the host. IMPORTANCE: Pseudomonas aeruginosa is an opportunistic pathogen problematic in hospitalized patients and those with cystic fibrosis (CF). Previously, we showed that P. aeruginosa can enter epithelial cells and replicate within them and traffics to the membrane blebs that it induces. This “bleb-niche” formation requires ExoS, previously shown to cause apoptosis. Here, we show that the driving force for bleb-niche formation is osmotic pressure, differentiating P. aeruginosa-induced blebs from apoptotic blebs. Either CFTR inhibition or CFTR mutation (as seen in people with CF) causes P. aeruginosa to make more bleb niches and provides an osmotic driving force for blebbing. CFTR inhibition also enhances bacterial occupation of blebs and intracellular replication. Since CFTR is targeted for removal from the plasma membrane when P. aeruginosa invades a healthy cell, these findings could relate to pathogenesis in both CF and healthy patient populations

Towards Efficient Detection of Small Near-Earth Asteroids Using the Zwicky Transient Facility (ZTF)

We describe ZStreak, a semi-real-time pipeline specialized in detecting small, fast-moving near-Earth asteroids (NEAs) that is currently operating on the data from the newly-commissioned Zwicky Transient Facility (ZTF) survey. Based on a prototype originally developed by Waszczak et al. (2017) for the Palomar Transient Factory (PTF), the predecessor of ZTF, ZStreak features an improved machine-learning model that can cope with the $10\times$ data rate increment between PTF and ZTF. Since its first discovery on 2018 February 5 (2018 CL), ZTF/ZStreak has discovered $45$ confirmed new NEAs over a total of 232 observable nights until 2018 December 31. Most of the discoveries are small NEAs, with diameters less than $\sim100$ m. By analyzing the discovery circumstances, we find that objects having the first to last detection time interval under 2 hr are at risk of being lost. We will further improve real-time follow-up capabilities, and work on suppressing false positives using deep learning.Comment: PASP in pres

Overview of the Design, Fabrication and Performance Requirements of Micro-Spec, an Integrated Submillimeter Spectrometer

Micro-Spec is a compact submillimeter (350-700 GHz) spectrometer which uses low loss superconducting niobium microstrip transmission lines and a single-crystal silicon dielectric to integrate all of the components of a grating-analog spectrometer onto a single chip. Here we present details of the fabrication and design of a prototype Micro-Spec spectrometer with resolution, R64, where we use a high-yield single-flip wafer bonding process to realize instrument components on a 0.45 m single-crystal silicon dielectric. We discuss some of the electromagnetic design concerns (such as loss, stray-light, cross-talk, and fabrication tolerances) for each of the spectrometer components and their integration into the instrument as a whole. These components include a slot antenna with a silicon lens for optical coupling, a phase delay transmission line network, parallel plate waveguide interference region, and aluminum microstrip transmission line kinetic inductance detectors with extremely low cross-talk and immunity to stray light. We have demonstrated this prototype spectrometer with design resolution of R64. Given the optical performance of this prototype, we will also discuss the extension of this design to higher resolutions suitable for balloon-flight

“I Think That’s the Most Beneficial Change That WIC Has Made in a Really Long Time”: Perceptions and Awareness of an Increase in the WIC Cash Value Benefit

During the COVID-19 pandemic, the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) Cash Value Benefit (CVB) for fruits and vegetables increased by roughly USD 25/month/person. We sought to understand WIC participant perceptions of this change and barriers and facilitators to using the CVB. We conducted 10 virtual focus groups (5 rural, 5 urban/suburban) with WIC participants (n = 55) in North Carolina in March 2022. Focus groups were recorded and transcribed. We open-coded the content and used thematic analysis to uncover consistencies within and between sampled groups. Participants expressed favorable perceptions of the CVB increase and stated the pre-pandemic CVB amount was insufficient. Barriers to using the increased CVB were identifying WIC-approved fruits and vegetables in stores and insufficient supply of fruits and vegetables. Barriers were more pronounced in rural groups. Facilitators of CVB use were existing household preferences for fruits and vegetables and the variety of products that can be purchased with CVB relative to other components of the WIC food package. Participants felt the CVB increase allowed their families to eat a wider variety of fruits and vegetables. The CVB increase may improve fruit and vegetable intake, particularly if made permanent, but barriers to CVB and WIC benefit use may limit the potential impact

GRB 090426: The Environment of a Rest-Frame 0.35-second Gamma-Ray Burst at Redshift z=2.609

We present the discovery of an absorption-line redshift of z = 2.609 for GRB 090426, establishing the first firm lower limit to a redshift for a gamma-ray burst with an observed duration of <2 s. With a rest-frame burst duration of T_90z = 0.35 s and a detailed examination of the peak energy of the event, we suggest that this is likely (at >90% confidence) a member of the short/hard phenomenological class of GRBs. From analysis of the optical-afterglow spectrum we find that the burst originated along a very low HI column density sightline, with N_HI < 3.2 x 10^19 cm^-2. Our GRB 090426 afterglow spectrum also appears to have weaker low-ionisation absorption (Si II, C II) than ~95% of previous afterglow spectra. Finally, we also report the discovery of a blue, very luminous, star-forming putative host galaxy (~2 L*) at a small angular offset from the location of the optical afterglow. We consider the implications of this unique GRB in the context of burst duration classification and our understanding of GRB progenitor scenarios.Comment: Submitted to MNRA

Interconnected Microphysiological Systems for Quantitative Biology and Pharmacology Studies

Microphysiological systems (MPSs) are in vitro models that capture facets of in vivo organ function through use of specialized culture microenvironments, including 3D matrices and microperfusion. Here, we report an approach to co-culture multiple different MPSs linked together physiologically on re-useable, open-system microfluidic platforms that are compatible with the quantitative study of a range of compounds, including lipophilic drugs. We describe three different platform designs - "4-way", "7-way", and "10-way" - each accommodating a mixing chamber and up to 4, 7, or 10 MPSs. Platforms accommodate multiple different MPS flow configurations, each with internal re-circulation to enhance molecular exchange, and feature on-board pneumatically-driven pumps with independently programmable flow rates to provide precise control over both intra- and inter-MPS flow partitioning and drug distribution. We first developed a 4-MPS system, showing accurate prediction of secreted liver protein distribution and 2-week maintenance of phenotypic markers. We then developed 7-MPS and 10-MPS platforms, demonstrating reliable, robust operation and maintenance of MPS phenotypic function for 3 weeks (7-way) and 4 weeks (10-way) of continuous interaction, as well as PK analysis of diclofenac metabolism. This study illustrates several generalizable design and operational principles for implementing multi-MPS "physiome-on-a-chip" approaches in drug discovery.United States. Army Research Office (Grant W911NF-12-2-0039

Zfp206 regulates ES cell gene expression and differentiation

Understanding transcriptional regulation in early developmental stages is fundamental to understanding mammalian development and embryonic stem (ES) cell properties. Expression surveys suggest that the putative SCAN-Zinc finger transcription factor Zfp206 is expressed specifically in ES cells [Zhang,W., Morris,Q.D., Chang,R., Shai,O., Bakowski,M.A., Mitsakakis,N., Mohammad,N., Robinson,M.D., Zirngibl,R., Somogyi,E. et al., (2004) J. Biol., 3, 21; Brandenberger,R., Wei,H., Zhang,S., Lei,S., Murage,J., Fisk,G.J., Li,Y., Xu,C., Fang,R., Guegler,K. et al., (2004) Nat. Biotechnol., 22, 707–716]. Here, we confirm this observation, and we show that ZFP206 expression decreases rapidly upon differentiation of cultured mouse ES cells, and during development of mouse embryos. We find that there are at least six isoforms of the ZFP206 transcript, the longest being predominant. Overexpression and depletion experiments show that Zfp206 promotes formation of undifferentiated ES cell clones, and positively regulates abundance of a very small set of transcripts whose expression is also specific to ES cells and the two- to four-cell stages of preimplantation embryos. This set includes members of the Zscan4, Thoc4, Tcstv1 and eIF-1A gene families, none of which have been functionally characterized in vivo but whose members include apparent transcription factors, RNA-binding proteins and translation factors. Together, these data indicate that Zfp206 is a regulator of ES cell differentiation that controls a set of genes expressed very early in development, most of which themselves appear to be regulators

Snowballs in Euclid and WFIRST Detectors

Snowballs are transient events observed in HgCdTe detectors with a sudden increase of charge in a few pixels. They appear between consecutive reads of the detector, after which the affected pixels return to their normal behavior. The origin of the snowballs is unknown, but it was speculated that they could be the result of alpha decay of naturally radioactive contaminants in the detectors, but a cosmic ray origin cannot be ruled out. Even though previous studies predicted a low rate of occurrence of these events, and consequently, a minimal impact on science, it is interesting to investigate the cause or causes that may generate snowballs and their impact in detectors designed for future missions. We searched for the presence of snowballs in the dark current data in Euclid and Wide Field Infrared Survey Telescope (WFIRST) detectors tested in the Detector Characterization Laboratory at Goddard Space Flight Center. Our investigation shows that for Euclid and WFIRST detectors, there are snowballs that appear only one time, and others that repeat in the same spatial localization. For Euclid detectors, there is a correlation between the snowballs that repeat and bad pixels in the operational masks (pixels that do not fulfill the requirements to pass spectroscopy noise, photometry noise, quantum efficiency, and/or linearity). The rate of occurrence for a snowball event is about 0.9 snowballs/hr. in Euclid detectors (for the ones that do not have associated bad pixels in the mask), and about 0.7 snowballs/hr. in PV3 Full Array Lot WFIRST detectors