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Age-related macular degeneration (AMD) causes blindness due to loss of retinal pigment epithelium (RPE) and photoreceptors (PRs), which comprise the two outermost layers of the retina. Given the small size of the macula and the importance of direct contact between RPE and PRs, the use of scaffolds for targeted reconstruction of the outer retina in later stage AMD and other macular dystrophies is particularly attractive. We developed microfabricated, honeycomb-patterned, biodegradable poly(glycerol sebacate) (PGS) scaffolds to deliver organized, adjacent layers of RPE and PRs to the subretinal space. Furthermore, an optimized process was developed to photocure PGS, shortening scaffold production time from days to minutes. The resulting scaffolds robustly supported the seeding of human pluripotent stem cell-derived RPE and PRs, either separately or as a dual cell-layered construct. These advanced, economical, and versatile scaffolds can accelerate retinal cell transplantation efforts and benefit patients with AMD and other retinal degenerative diseases.

Measurement of the temporal evolution of laser-produced plasma temperature is very important for many of its applications, and several plasma diagnostic tools are routinely used by researchers. However, it is very challenging to measure the properties of the plasma at the early and late times of its evolution using a single diagnostic tool. In this study, we combined emission and laser absorption spectroscopy to compare the excitation temperatures of a laser-produced uranium plasma system. Several U I transitions in the near-infrared spectral range (775–800 nm) were considered, and the Boltzmann plot method was used to measure the excitation temperatures using both emission and absorption spectroscopy. Emission spectroscopy provided early-time temperature measurements of the plasma up to times 2–20 µs, while absorption spectroscopy provided temperature measurements at late times of plasma evolution (for times 5–80 µs). The emission and absorbance of U I transitions were found to follow the Boltzmann distribution, indicating the plasma is likely in the state of local thermodynamic equilibrium even at late times of its lifetime. The emission and absorption-based time-resolved excitation temperatures demonstrated good agreement at earlier times (≤15 µs) in the overlapped temporal region, while a deviation in the measured values was seen at times (≥15 µs), and potential reasons for such a disagreement are discussed.

ABSTRACT Functional traits are characteristics that affect the fitness and metabolic function of a microorganism. There is growing interest in using high-throughput methods to characterize bacterial pathogens based on functional virulence traits. Traditional methods that phenotype a single organism for a single virulence trait can be time consuming and labor intensive. Alternatively, machine learning of whole-genome sequences (WGS) has shown some success in predicting virulence. However, relying solely on WGS can miss functional traits, particularly for organisms lacking classical virulence factors. We propose that high-throughput assays for functional virulence trait identification should become a prominent method of characterizing bacterial pathogens on a population scale. This work is critical as we move from compiling lists of bacterial species associated with disease to pathogen-agnostic approaches capable of detecting novel microbes. We discuss six key areas of functional trait testing and how advancing high-throughput methods could provide a greater understanding of pathogens.

Over 30 million people worldwide suffer from untreatable vision loss and blindness associated with childhood-onset and age-related eye diseases caused by photoreceptor (PR), retinal pigment epithelium (RPE), and choriocapillaris (CC) degeneration. Recent work suggests that RPE-based cell therapy may slow down vision loss in late stages of age-related macular degeneration (AMD), a polygenic disease induced by RPE atrophy. However, accelerated development of effective cell therapies is hampered by the lack of large-animal models that allow testing safety and efficacy of clinical doses covering the human macula (20 mm2). We developed a versatile pig model to mimic different types and stages of retinal degeneration. Using an adjustable power micropulse laser, we generated varying degrees of RPE, PR, and CC damage and confirmed the damage by longitudinal analysis of clinically relevant outcomes, including analyses by adaptive optics and optical coherence tomography/angiography, along with automated image analysis. By imparting a tunable yet targeted damage to the porcine CC and visual streak — with a structure similar to the human macula — this model is optimal for testing cell and gene therapies for outer retinal diseases including AMD, retinitis pigmentosa, Stargardt, and choroideremia. The amenability of this model to clinically relevant imaging outcomes will facilitate faster translation to patients.

We present a JWST mid-infrared (MIR) spectrum of the underluminous Type Ia Supernova (SN Ia) 2022xkq, obtained with the medium-resolution spectrometer on the Mid-Infrared Instrument (MIRI) ∼130 days post-explosion. We identify the first MIR lines beyond 14 μ m in SN Ia observations. We find features unique to underluminous SNe Ia, including the following: isolated emission of stable Ni, strong blends of [Ti ii ], and large ratios of singly ionized to doubly ionized species in both [Ar] and [Co]. Comparisons to normal-luminosity SNe Ia spectra at similar phases show a tentative trend between the width of the [Co iii ] 11.888 μ m feature and the SN light-curve shape. Using non-LTE-multi-dimensional radiation hydro simulations and the observed electron capture elements, we constrain the mass of the exploding WD. The best-fitting model shows that SN 2022xkq is consistent with an off-center delayed-detonation explosion of a near-Chandrasekhar mass WD ( ${M}_{{\rm{WD}}}$ ≈1.37 M _⊙ ) of high central density ( ρ _c ≥ 2.0 × 10 ^9 g cm ^−3 ) seen equator-on, which produced M ( ^56 Ni) =0.324 M _⊙ and M ( ^58 Ni) ≥0.06 M _⊙ . The observed line widths are consistent with the overall abundance distribution; and the narrow stable Ni lines indicate little to no mixing in the central regions, favoring central ignition of subsonic carbon burning followed by an off-center deflagration-to-detonation transition beginning at a single point. Additional observations may further constrain the physics revealing the presence of additional species including Cr and Mn. Our work demonstrates the power of using the full coverage of MIRI in combination with detailed modeling to elucidate the physics of SNe Ia at a level not previously possible.

We present the second and final release of optical spectroscopy of Type Ia supernovae (SNe Ia) obtained during the first and second phases of the Carnegie Supernova Project (CSP-I and CSP-II). The newly released data consist of 148 spectra of 30 SNe Ia observed in the course of CSP-I and 234 spectra of 127 SNe Ia obtained during CSP-II. We also present 216 optical spectra of 46 historical SNe Ia, including 53 spectra of 30 SNe Ia observed by the Calán/Tololo Supernova Survey. We combine these observations with previously published CSP data and publicly available spectra to compile a large sample of measurements of spectroscopic parameters at maximum light, consisting of pseudo-equivalent widths and expansion velocities of selected features for 232 CSP and historical SNe Ia (including more than 1000 spectra). Finally, we review some of the strongest correlations between spectroscopic and photometric properties of SNe Ia. Specifically, we define two samples: one consisting of SNe Ia discovered by targeted searches (most of them CSP-I objects) and the other composed of SNe Ia discovered by untargeted searches, which includes most of the CSP-II objects. The analyzed correlations are similar for both samples. We find a larger incidence of SNe Ia belonging to the cool and broad-line Branch subtypes among the events discovered by targeted searches, shallow-silicon SNe Ia are present with similar frequencies in both samples, while core normal SNe Ia are more frequent in untargeted searches.

We present a method of extrapolating the spectroscopic behavior of Type Ia supernovae (SNe Ia) in the near-infrared (NIR) wavelength regime up to 2.30 μ m using optical spectroscopy. Such a process is useful for accurately estimating K-corrections and other photometric quantities of SNe Ia in the NIR. A principal component analysis is performed on data consisting of Carnegie Supernova Project I & II optical and NIR FIRE spectra to produce models capable of making these extrapolations. This method differs from previous spectral template methods by not parameterizing models strictly by photometric light-curve properties of SNe Ia, allowing for more flexibility of the resulting extrapolated NIR flux. A difference of around −3.1% to −2.7% in the total integrated NIR flux between these extrapolations and the observations is seen here for most test cases including Branch core-normal and shallow-silicon subtypes. However, larger deviations from the observation are found for other tests, likely due to the limited high-velocity and broad-line SNe Ia in the training sample. Maximum-light principal components are shown to allow for spectroscopic predictions of the color-stretch light-curve parameter, s _BV , within approximately ±0.1 units of the value measured with photometry. We also show these results compare well with NIR templates, although in most cases the templates are marginally more fitting to observations, illustrating a need for more concurrent optical+NIR spectroscopic observations to truly understand the diversity of SNe Ia in the NIR.

We present an analysis of Type Ia supernovae (SNe Ia) from the Carnegie Supernova Project I and II and extend the Hubble diagram from optical to near-infrared wavelengths ( uBgVriYJH ). We calculate the Hubble constant, H _0 , using various distance calibrators: Cepheids, the tip of the red giant branch (TRGB), and surface brightness fluctuations (SBFs). Combining all methods of calibration, we derive H _0 = 71.76 ± 0.58 (stat) ± 1.19 (sys) km s ^−1 Mpc ^−1 from the B band and H _0 = 73.22 ± 0.68 (stat) ± 1.28 (sys) km s ^−1 Mpc ^−1 from the H band. By assigning equal weight to the Cepheid, TRGB, and SBF calibrators, we derive the systematic errors required for consistency in the first rung of the distance ladder, resulting in a systematic error of 1.2 ∼ 1.3 km s ^−1 Mpc ^−1 in H _0 . As a result, relative to the statistics-only uncertainty, the tension between the late-time H _0 we derive by combining the various distance calibrators and the early-time H _0 from the cosmic microwave background is reduced. The highest precision in SN Ia luminosity is found in the Y band (0.12 ± 0.01 mag), as defined by the intrinsic scatter ( σ _int ). We revisit SN Ia Hubble residual-host mass correlations and recover previous results that these correlations do not change significantly between the optical and near-infrared wavelengths. Finally, SNe Ia that explode beyond 10 kpc from their host centers exhibit smaller dispersion in their luminosity, confirming our earlier findings. A reduced effect of dust in the outskirts of hosts may be responsible for this effect.

We present an eigenfunction method to analyze 161 visual light curves (LCs) of Type Ia supernovae (SNe Ia) obtained by the Carnegie Supernova Project to characterize their diversity and host-galaxy correlations. The eigenfunctions are based on the delayed-detonation (DD) scenario using three parameters: the LC stretch s determined by the amount of deflagration burning governing the ^56 Ni production, the main-sequence mass M _MS of the progenitor white dwarf controlling the explosion energy, and its central density ρ _c shifting the ^56 Ni distribution. Our analysis tool (Supernova Parameter Analysis Tool) extracts the parameters from observations and projects them into physical space using their allowed ranges ( M _MS ≤ 8 M _⊙ , ρ _c ≤ 7–8 × 10 ^9 g cm ^−3 ). The residuals between fits and individual LC points are ≈1%–3% for ≈92% of objects. We find two distinct M _MS groups corresponding to a fast (≈4–65 Myr) and a slow(≈200–500 Myr) stellar evolution. Most underluminous SNe Ia have hosts with low star formation but high M _MS , suggesting slow evolution times of the progenitor system. 91T-like SNe show very similar LCs and high M _MS and are correlated to star formation regions, making them potentially important tracers of star formation in the early Universe out to z ≈ 4–11. Some ∼6% outliers with nonphysical parameters using DD scenarios can be attributed to superluminous SNe Ia and subluminous SNe Ia with hosts of active star formation. For deciphering the SNe Ia diversity and high-precision SNe Ia cosmology, the importance is shown for LCs covering out to ≈60 days past maximum. Finally, our method and results are discussed within the framework of multiple explosion scenarios, and in light of upcoming surveys.

We present a JWST MIRI medium-resolution spectrometer spectrum (5–27 μ m) of the Type Ia supernova (SN Ia) SN 2021aefx at +415 days past B -band maximum. The spectrum, which was obtained during the iron-dominated nebular phase, has been analyzed in combination with previous JWST observations of SN 2021aefx to provide the first JWST time series analysis of an SN Ia. We find that the temporal evolution of the [Co iii ] 11.888 μ m feature directly traces the decay of ^56 Co. The spectra, line profiles, and their evolution are analyzed with off-center delayed-detonation models. Best fits were obtained with white dwarf (WD) central densities of ρ _c = 0.9−1.1 × 10 ^9 g cm ^−3 , a WD mass of M _WD = 1.33–1.35 M _⊙ , a WD magnetic field of ≈10 ^6 G, and an off-center deflagration-to-detonation transition at ≈0.5 M _⊙ seen opposite to the line of sight of the observer (−30°). The inner electron capture core is dominated by energy deposition from γ -rays, whereas a broader region is dominated by positron deposition, placing SN 2021aefx at +415 days in the transitional phase of the evolution to the positron-dominated regime. The formerly “flat-tilted” profile at 9 μ m now has a significant contribution from [Ni iv ], [Fe ii ], and [Fe iii ] and less from [Ar iii ], which alters the shape of the feature as positrons mostly excite the low-velocity Ar. Overall, the strength of the stable Ni features in the spectrum is dominated by positron transport rather than the Ni mass. Based on multidimensional models, our analysis is consistent with a single-spot, close-to-central ignition with an indication of a preexisting turbulent velocity field and excludes a multiple-spot, off-center ignition.

We present a JWST/MIRI low-resolution mid-infrared (MIR) spectroscopic observation of the normal Type Ia supernova (SN Ia) SN 2021aefx at +323 days past rest-frame B -band maximum light. The spectrum ranges from 4 to 14 μ m and shows many unique qualities, including a flat-topped [Ar iii ] 8.991 μ m profile, a strongly tilted [Co iii ] 11.888 μ m feature, and multiple stable Ni lines. These features provide critical information about the physics of the explosion. The observations are compared to synthetic spectra from detailed non–local thermodynamic equilibrium multidimensional models. The results of the best-fitting model are used to identify the components of the spectral blends and provide a quantitative comparison to the explosion physics. Emission line profiles and the presence of electron capture elements are used to constrain the mass of the exploding white dwarf (WD) and the chemical asymmetries in the ejecta. We show that the observations of SN 2021aefx are consistent with an off-center delayed detonation explosion of a near–Chandrasekhar mass ( M _Ch ) WD at a viewing angle of −30° relative to the point of the deflagration to detonation transition. From the strengths of the stable Ni lines, we determine that there is little to no mixing in the central regions of the ejecta. Based on both the presence of stable Ni and the Ar velocity distributions, we obtain a strict lower limit of 1.2 M _⊙ for the initial WD, implying that most sub- M _Ch explosions models are not viable models for SN 2021aefx. The analysis here shows the crucial importance of MIR spectra in distinguishing between explosion scenarios for SNe Ia.

Summary: Photoreceptors (PRs) are the primary visual sensory cells, and their loss leads to blindness that is currently incurable. Although cell replacement therapy holds promise, success is hindered by our limited understanding of PR axon growth during development and regeneration. Here, we generate retinal organoids from human pluripotent stem cells to study the mechanisms of PR process extension. We find that early-born PRs exhibit autonomous axon extension from dynamic terminals. However, as PRs age from 40 to 80 days of differentiation, they lose dynamic terminals on 2D substrata and in 3D retinal organoids. Interestingly, PRs without motile terminals are still capable of extending axons but only by process stretching via attachment to motile non-PR cells. Immobile PR terminals of late-born PRs have fewer and less organized actin filaments but more synaptic proteins compared with early-born PR terminals. These findings may help inform the development of PR transplantation therapies.

We present multiwavelength time-series spectroscopy of SN 2013aa and SN 2017cbv, two Type Ia supernovae (SNe Ia) on the outskirts of the same host galaxy, NGC 5643. This work utilizes new nebular-phase near-infrared (NIR) spectra obtained by the Carnegie Supernova Project-II, in addition to previously published optical and NIR spectra. Using nebular-phase [Fe ii ] lines in the optical and NIR, we examine the explosion kinematics and test the efficacy of several common emission-line-fitting techniques. The NIR [Fe ii ] 1.644 μ m line provides the most robust velocity measurements against variations due to the choice of the fit method and line blending. The resulting effects on velocity measurements due to choosing different fit methods, initial fit parameters, continuum and line profile functions, and fit region boundaries were also investigated. The NIR [Fe ii ] velocities yield the same radial shift direction as velocities measured using the optical [Fe ii ] λ 7155 line, but the sizes of the shifts are consistently and substantially lower, pointing to a potential issue in optical studies. The NIR [Fe ii ] 1.644 μ m emission profile shows a lack of significant asymmetry in both SNe, and the observed low velocities elevate the importance for correcting for any velocity contribution from the host galaxy’s rotation. The low [Fe ii ] velocities measured in the NIR at nebular phases disfavor progenitor scenarios in close double-degenerate systems for both SN 2013aa and SN 2017cbv. The time evolution of the NIR [Fe ii ] 1.644 μ m line also indicates moderately high progenitor white dwarf central density and potentially high magnetic fields.

Objectives Explore international consensus on nomenclatures of suicidal behaviours and analyse differences in terminology between high-income countries (HICs) and low/middle-income countries (LMICs).Design An online survey of members of the International Organisation for Suicide Prevention (IASP) used multiple-choice questions and vignettes to assess the four dimensions of the definition of suicidal behaviour: outcome, intent, knowledge and agency.Setting International.Participants Respondents included 126 individuals, 37 from 30 LMICs and 89 from 33 HICs. They included 40 IASP national representatives (65% response rate), IASP regular members (20% response rate) and six respondents from six additional countries identified by other organisations.Outcome measures Definitions of English-language terms for suicidal behaviours.Results The recommended definition of ‘suicide’ describes a fatal act initiated and carried out by the actors themselves. The definition of ‘suicide attempt’ was restricted to non-fatal acts with intent to die, whereas definition of ‘self-harm’ more broadly referred to acts with varying motives, including the wish to die. Almost all respondents agreed about the definitions of ‘suicidal ideation’, ‘death wishes’ and ‘suicide plan’. ‘Aborted suicide attempt’ and ‘interrupted suicide attempt’ were not considered components of ‘preparatory suicidal behaviour’. There were several differences between representatives from HICs and LMICs.Conclusion This international opinion survey provided the basis for developing a transcultural nomenclature of suicidal behaviour. Future developments of this nomenclature should be tested in larger samples of professionals, including LMICs may be a challenge.

Bone marrow derived human Mesenchymal Stem Cells (hMSCs) are an attractive candidate for regenerative medicine. However, their harvest can be invasive, painful, and expensive, making it difficult to supply the enormous amount of pure hMSCs needed for future allogeneic therapies. Because of this, a robust method of scaled bioreactor culture must be designed to supply the need for high purity, high density hMSC yields. Here we test a scaled down model of a novel bioreactor consisting of an unsubmerged 3D printed Polylactic Acid (PLA) lattice matrix wetted by culture media. The growth matrix is uniform, replicable, and biocompatible, enabling homogenous cell culture in three dimensions. The goal of this study was to prove that hMSCs would culture well in this novel bioreactor design. The system tested resulted in comparable stem cell yields to other cell culture systems using bone marrow derived hMSCs, while maintaining viability (96.54% ±2.82), high purity (>98% expression of combined positive markers), and differentiation potential.

Replacement heifer rearing is critical for the future of dairy operations, to improve genetic merit and maintain herd size. A myriad of options exist on how to manage, feed, and ultimately raise replacement heifers. Pasture is perceived to offer optimal welfare and an economical housing system for replacement animals, but confinement systems are gaining popularity. This study investigates the costs associated with replacement heifer management decisions from birth to calving, considering the factors of housing systems, labor, feed, and health. The objective of this study was to develop an economic model to determine the cost of raising a replacement heifer managed in confinement, dry-lot, and pasture-based scenarios post-weaning. We accounted for variation in feed, labor, and health inputs and quantified the impact of these individual management decisions. An economic simulation with 10,000 iterations were completed for each situation using @Risk and PrecisionTree add-ons (Palisade Corporation, Ithaca, NY) where health incidence, commodity prices, and management variables were made stochastic. Published literature or sample farm data created parameters used in Pert distributions. Costs and biological responses were reflective of published surveys, literature, and market conditions. Management decision inputs had 3 main factors: housing type, ration composition, and labor utilization. Housing systems were calculated separately for confinement, dry-lot, and pasture scenarios. The mean total cost (min, max) to raise a replacement heifer from birth to calving, assuming the same pre-weaning strategy of group housing with an automatic calf feeder, was found to be $1,919.02 ($1,777.25, $2,100.57), $1,593.57 ($1,490.30, $1,737.26), and $1,335.84 ($1,266.69, $1,423.94) for confinement, dry-lot, and pasture, respectively. Total housing cost per replacement heifer was $423.05, $117.96, and $207.96 for confinement, dry-lot, and pasture management systems, respectively. When compared to total cost, housing contributed 21% for confinement, 7% for dry-lot, and 15% for pasture. Upon analysis of all scenarios, utilizing pasture to raise heifers resulted in a lower overall cost when compared to confinement housing options. Percentage breakdowns of feed, labor, housing, and fixed and variable costs provided more information on efficiency rather than total cost, which makes each situation different in relation to on-farm cost. This cost analysis is critical to assisting farms in making decisions in the utilization of their resources for replacement dairy heifers.

Summary: Retinal ganglion cells (RGCs) are the projection neurons of the retina and transmit visual information to postsynaptic targets in the brain. While this function is shared among nearly all RGCs, this class of cell is remarkably diverse, comprised of multiple subtypes. Previous efforts have identified numerous RGC subtypes in animal models, but less attention has been paid to human RGCs. Thus, efforts of this study examined the diversity of RGCs differentiated from human pluripotent stem cells (hPSCs) and characterized defined subtypes through the expression of subtype-specific markers. Further investigation of these subtypes was achieved using single-cell transcriptomics, confirming the combinatorial expression of molecular markers associated with these subtypes, and also provided insight into more subtype-specific markers. Thus, the results of this study describe the derivation of RGC subtypes from hPSCs and will support the future exploration of phenotypic and functional diversity within human RGCs. : In this article, Langer and colleagues present extensive characterization of RGC subtypes derived from human pluripotent stem cells, with multiple subtypes identified by subtype-specific molecular markers. Their results present a more detailed analysis of RGC diversity in human cells and yield the use of different markers to identify RGC subtypes. Keywords: iPSC, retina, retinal ganglion cell, RGC subtype, stem cell, ipRGC, alpha RGC, direction selective RGC, RNA-seq

Cell culture systems have greatly expanded our understanding of how bacterial pathogens target signaling pathways to manipulate the host and cause infection. Advances in genetic engineering have allowed for the creation of fluorescent protein readouts within signaling pathways, but these techniques have been underutilized in pathogen biology. Here, we genetically engineered a lung cell line with fluorescent reporters for extracellular signal-related kinase (ERK) and the downstream transcription factor FOS-related antigen 1 (Fra1) and evaluated signaling after inoculation with pathogenic and non-pathogenic bacteria. Cells were inoculated with 100 colony-forming units of Acinetobacter baylyi, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus agalactiae, or Staphylococcus epidermidis and imaged in a multi-mode reader. The alamarBlue cell viability assay was used as a reference test and showed that pathogenic P. aeruginosa induced significant (p < 0.05) cell death after 8 h in both wild-type and engineered cell lines compared to non-pathogenic S. epidermidis. In engineered cells, we found that Fra1 signaling was disrupted in as little as 4 h after inoculation with bacterial pathogens compared to delayed disruption in signaling by non-pathogenic S. epidermidis. Overall, we demonstrate that low levels of pathogenic versus non-pathogenic bacteria can be rapidly and sensitively screened based on ERK-Fra1 signaling.

Abstract Reporter lines generated in human pluripotent stem cells can be highly useful for the analysis of specific cell types and lineages in live cultures. We created the first human rod reporter line using CRISPR/Cas9 genome editing to replace one allele of the Neural Retina Leucine zipper (NRL) gene with an eGFP transgene in the WA09 human embryonic stem cell (hESC) line. After confirming successful targeting, three-dimensional optic vesicle structures were produced to examine reporter specificity and to track rod differentiation in culture. The NRL +/eGFP hESC line robustly and exclusively labeled the entirety of rods throughout differentiation, eventually revealing highly mature structural features. This line provides a valuable tool for studying human rod development and disease and testing therapeutic strategies for retinitis pigmentosa.