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My essay is a close reading of the initial poem of the third book of Statius’ Silvae. Its primary topic is the dedication by Pollius Felix of his new temple to Hercules. As the narrative unfolds, we trace how the shrine came into being and the accompanying metamorphoses of the landscape and of the character of the god himself. Pollius and his wife, Polla, play major roles in the proceedings. Statius’s own virtuosity is on constant display not only in his skill as a wordsmith but in his suggestive bows to a series of genres and in his rich allusiveness, especially to Virgil.

This chapter offers a close reading of Poem 2 from the fourth book of the Silvae of Statius. Statius’ brilliant word usage in describing a banquet at Domitian’s palace and the architectural magnificence of the emperor’s along with, more generally, the poet’s use of intertexts evoke Virgil’s poetry, as well as Horace’s. Here Statius pushes language to the limits of praise. And yet the recurrent theme in Silvae 4.2 of Dido’s welcome banquet for Aeneas, along with echoes of Virgil’s exiled goatherd of Eclogue 1, Meliboeus, provide a tragic reminder of the precarity of human life and happiness as well as of the limits of poetic power.

This chapter introduces readers to the volume on the Silvae. It showcases Statius’ complex, rich intertextuality on display in his exploration of villa culture, of patronage and friendship, and of life on the Bay of Naples rather than at court in Rome. The importance of Virgil’s poetry, both epic and pastoral, above all stands out as a major influence upon the Silvae. The introduction also offers summaries of Putnam’s individual essays on the Silvae.

This chapter offers a close reading of Poem 3 from the second book of the Silvae of Statius. A genethliacon and, in conclusion, eulogy for the writer’s friend Atedius Melior, it directs us to the latter’s garden and centres on an expansive aition, of the poet’s invention, devoted to the god Pan’s attraction for the nymph Pholoe. The chapter’s primary focus rests on Statius’ brilliant word usage and, more generally, on the intellectual background on which he draws. It refers regularly to the poetry of Horace, Tibullus, and especially Virgil in my search to elucidate Statius’ original genius.

This chapter offers a close reading of Poem 4 from the second book of the Silvae of Statius. In a tribute to Melior’s recently deceased parrot, Statius’ brilliant word usage and, more generally, the poet’s use of intertexts evoke the poetry of Lucretius, Virgil, Horace, and especially Ovid in an original composition that brings the ‘dead pet poem’ into the imperial age by emphasising the power of words and their confrontation with silence.

This chapter is the longest in the collection and the most ambitious; it makes a fitting conclusion to the volume. It offers a close examination of Statius’ Silvae 3.2, a propempticon for Maecius Celer. Using earlier Latin farewell poems as intellectual background, in particular Horace’s third ode, Putnam illustrates the poet’s originality. In particular he points out that Silvae 3.2 is a highly complex poem that employs a variety of genres, such as lyric and epic, and especially the Aeneid, as Statius crafts a meditation on a series of curricula vitae that illuminate Celer’s own voyage. Various themes, moreover, such as the dangers of seafaring, the violence of Roman imperialism, Egyptian tourism, the neutralization of a bloodied historic, and literary past are skillfully woven together in a double, courageous journey, the literal journey of Celer to his military post in Syria, and the metaphorical journey of Statius to the challenging conclusion of his epic poem.

This chapter presents a detailed literary analysis of Statius’s Silvae 1.3 which honours Manilius Vopiscus and his Tiburtine villa. My reading follows the progress of the poem, as it first outlines the quality of the domain as a whole, then examines aspects of what a visitor might encounter. I survey Statius’ varied use of ekphrasis and trace his patterns of allusion, especially to the work of Virgil. Eroticism is a salient theme throughout as is the importance of a series of bodies of water, especially the river Anio, to the poem’s evolution to a celebration of beauty harmonised in both nature and art.

Mesenchymal stem cells (MSCs) have broad-based therapeutic potential in regenerative medicine. However, a major barrier to their clinical utility is that MSCs from different tissues are highly variable in their regenerative properties. In this study, we defined the molecular and phenotypic identities of different MSC populations from different osseous tissue sites of different patients and, additionally, determined their respective regenerative properties. MSCs from 6 patients were isolated from either bone marrow of the iliac crest (BMSCs) or alveolar bone tissue (aBMSCs), and flow cytometry revealed that regardless of the tissue source, MSC immunotypes had the same expression of MSC markers CD73, CD90, and CD105. However, transcriptomic analyses revealed 589 genes differentially expressed (DE) between BMSCs and aBMSCs, including eightfold higher levels of bone morphogenetic protein 4 (BMP-4) in aBMSCs. In striking contrast, gene expression of MSCs derived from the same tissue, but between different patients (i.e., BMSCs to BMSCs, aBMSCs to aBMSCs), showed only 38 DE BMSC genes and 51 DE aBMSC genes. A protein array showed that aBMSC and BMSC produced equivalent levels of angiogenic cytokines; however, when placed in angiogenesis model systems, aBMSCs induced significantly more capillaries in vitro and in vivo. Finally, cell transplantation of MSCS into osseous defects showed that the bone regenerative capacity of aBMSCs was significantly greater than that of BMSCs. This study is the first to link the molecular, phenotypic, and regenerative properties of different MSCs from different patients and provides novel insights toward MSC differences based on the osseous tissue origin.

Hydrogels are often used to study the impact of biomechanical and topographical cues on cell behavior. Conventional hydrogels are designed a priori, with characteristics that cannot be dynamically changed in an externally controlled, user-defined manner. We developed a composite hydrogel, termed an acoustically-responsive scaffold (ARS), that enables non-invasive, spatiotemporally controlled modulation of mechanical and morphological properties using focused ultrasound. An ARS consists of a phase-shift emulsion distributed in a fibrin matrix. Ultrasound non-thermally vaporizes the emulsion into bubbles, which induces localized, radial compaction and stiffening of the fibrin matrix. In this in vitro study, we investigate how this mechanism can control the differentiation of fibroblasts into myofibroblasts, a transition correlated with substrate stiffness on 2D substrates. Matrix compaction and stiffening was shown to be highly localized using confocal and atomic force microscopies, respectively. Myofibroblast phenotype, evaluated by α-smooth muscle actin (α-SMA) immunocytochemistry, significantly increased in matrix regions proximal to bubbles compared to distal regions, irrespective of the addition of exogenous transforming growth factor-β1 (TGF-β1). Introduction of the TGF-β1 receptor inhibitor SB431542 abrogated the proximal enhancement. This approach providing spatiotemporal control over biophysical signals and resulting cell behavior could aid in better understanding fibrotic disease progression and the development of therapeutic interventions for chronic wounds. STATEMENT OF SIGNIFICANCE: Hydrogels are used in cell culture to recapitulate both biochemical and biophysical aspects of the native extracellular matrix. Biophysical cues like stiffness can impact cell behavior. However, with conventional hydrogels, there is a limited ability to actively modulate stiffness after polymerization. We have developed an ultrasound-based method of spatiotemporally-controlling mechanical and morphological properties within a composite hydrogel, termed an acoustically-responsive scaffold (ARS). Upon exposure to ultrasound, bubbles are non-thermally generated within the fibrin matrix of an ARS, thereby locally compacting and stiffening the matrix. We demonstrate how ARSs control the differentiation of fibroblasts into myofibroblasts in 2D. This approach could assist with the study of fibrosis and the development of therapies for chronic wounds.

Pro-angiogenic growth factors have been studied as potential therapeutics for cardiovascular diseases like critical limb ischemia (CLI). However, the translation of these factors has remained a challenge, in part, due to problems associated with safe and effective delivery. Here, we describe a hydrogel-based delivery system for growth factors where release is modulated by focused ultrasound (FUS), specifically a mechanism termed acoustic droplet vaporization. With these fibrin-based, acoustically-responsive scaffolds (ARSs), release of a growth factor is non-invasively and spatiotemporally-controlled in an on-demand manner using non-thermal FUS. In vitro studies demonstrated sustained release of basic fibroblast growth factor (bFGF) from the ARSs using repeated applications of FUS. In in vivo studies, ARSs containing bFGF were implanted in mice following induction of hind limb ischemia, a preclinical model of CLI. During the 4-week study, mice in the ARS + FUS group longitudinally exhibited significantly more perfusion and less visible necrosis compared to other experimental groups. Additionally, significantly greater angiogenesis and less fibrosis were observed for the ARS + FUS group. Overall, these results highlight a promising, FUS-based method of delivering a pro-angiogenic growth factor for stimulating angiogenesis and reperfusion in a cardiovascular disease model. More broadly, these results could be used to personalize the delivery of therapeutics in different regenerative applications by actively controlling the release of a growth factor.

Vascularization is a critical step following implantation of an engineered tissue construct in order to maintain its viability. The ability to spatially pattern or direct vascularization could be therapeutically beneficial for anastomosis and vessel in-growth. However, acellular and cell-based strategies to stimulate vascularization typically do not afford this control. We have developed an ultrasound-based method of spatially- controlling regenerative processes using acellular, composite hydrogels termed acoustically-responsive scaffolds (ARSs). An ARS consists of a fibrin matrix doped with a phase-shift double emulsion (PSDE). A therapeutic payload, which is initially contained within the PSDE, is released by an ultrasound-mediated process called acoustic droplet vaporization (ADV). During ADV, the perfluorocarbon (PFC) phase within the PSDE is vaporized into a gas bubble. In this study, we generated ex situ four different spatial patterns of ADV within ARSs containing basic fibroblast growth factor (bFGF), which were subcutaneously implanted in mice. The PFC species within the PSDE significantly affected the morphology of the ARS, based on the stability of the gas bubble generated by ADV, which impacted host cell migration. Irrespective of PFC, significantly greater cell proliferation (i.e., up to 2.9-fold) and angiogenesis (i.e., up to 3.7-fold) were observed adjacent to +ADV regions of the ARSs compared to -ADV regions. The morphology of the PSDE, macrophage infiltration, and perfusion in the implant region were also quantified. These results demonstrate that spatially-defined patterns of ADV within an ARS can elicit spatially-defined patterns of angiogenesis. Overall, these finding can be applied to improve strategies for spatially-controlling vascularization. Statement of significance Vascularization is a critical step following implantation of an engineered tissue. The ability to spatially pattern or direct vascularization could be therapeutically beneficial for inosculation and vessel in-growth. However, acellular and cell-based strategies to stimulate vascularization typically do not afford this control. We have developed an ultrasound-based method of spatially-controlling angiogenesis using acellular, composite hydrogels termed acoustically-responsive scaffolds (ARSs). An ARS consists of a fibrin matrix doped with a phase-shift double emulsion (PSDE). An ultrasound-mediated process called acoustic droplet vaporization (ADV) was used to release basic fibroblast growth factor (bFGF), which was initially contained within the PSDE. We demonstrate that spatially-defined patterns of ADV within an ARS can elicit spatially-defined patterns of angiogenesis in vivo. Overall, these finding can improve strategies for spatially-controlling vascularization.

This article summarizes research and risk reduction that will inform acquisition decisions regarding NOAA’s future national operational weather radar network. A key alternative being evaluated is polarimetric phased-array radar (PAR). Research indicates PAR can plausibly achieve fast, adaptive volumetric scanning, with associated benefits for severe-weather warning performance. We assess these benefits using storm observations and analyses, observing system simulation experiments, and real radar-data assimilation studies. Changes in the number and/or locations of radars in the future network could improve coverage at low altitude. Analysis of benefits that might be so realized indicates the possibility for additional improvement in severe-weather and flash-flood warning performance, with associated reduction in casualties. Simulations are used to evaluate techniques for rapid volumetric scanning and assess data quality characteristics of PAR. Finally, we describe progress in developing methods to compensate for polarimetric variable estimate biases introduced by electronic beam-steering. A research-to-operations (R2O) strategy for the PAR alternative for the WSR-88D replacement network is presented.

Supportive stromal cells of mesenchymal origins regulate vascular morphogenesis in developmental, pathological, and regenerative contexts, contributing to vessel formation, maturation, and long-term stability, in part via the secretion of bioactive molecules. In this work, we adapted a microfluidic lab-on-a-chip system that enables the formation and perfusion of microvascular capillary beds with connections to arteriole-scale endothelialized channels to explore how stromal cell (SC) identity influences endothelial cell (EC) morphogenesis. We compared and contrasted lung fibroblasts (LFs), dermal fibroblasts (DFs), and bone marrow-derived mesenchymal stem cells (MSCs) for their abilities to support endothelial morphogenesis and subsequent perfusion of microvascular networks formed in fibrin hydrogels within the microfluidic device. We demonstrated that while all 3 SC types supported EC morphogenesis, LFs in particular resulted in microvascular morphologies with the highest total network length, vessel diameter, and vessel interconnectivity across a range of SC-EC ratio and density conditions. Not only did LFs support robust vascular morphology, but also, they were the only SC type to support functional perfusion of the resultant capillary beds. Lastly, we identified heightened traction stress produced by LFs as a possible mechanism by which LFs enhance endothelial morphogenesis in 3D compared to other SC types examined. This study provides a unique comparison of three different SC types and their role in supporting the formation of microvasculature that could provide insights for the choice of cells for vascular cell-based therapies and the regulation of tissue-specific vasculature.

Revascularization of ischemic tissues is a major barrier to restoring tissue function in many pathologies. Delivery of pro-angiogenic factors has shown some benefit, but it is difficult to recapitulate the complex set of factors required to form stable vasculature. Cell-based therapies and pre-vascularized tissues have shown promise, but the former require time for vascular assembly in situ while the latter require invasive surgery to implant vascularized scaffolds. Here, we developed cell-laden fibrin microbeads that can be pre-cultured to form primitive vascular networks within the modular structures. These microbeads can be delivered in a minimally invasive manner and form functional microvasculature in vivo. Microbeads containing endothelial cells and stromal fibroblasts were pre-cultured for 3 days in vitro and then injected within a fibrin matrix into subcutaneous pockets on the dorsal flanks of SCID mice. Vessels deployed from these pre-cultured microbeads formed functional connections to host vasculature within 3 days and exhibited extensive, mature vessel coverage after 7 days in vivo. Cellular microbeads showed vascularization potential comparable to bulk cellular hydrogels in this pilot study. Furthermore, our findings highlight some potentially advantageous characteristics of pre-cultured microbeads, such as volume preservation and vascular network distribution, which may be beneficial for treating ischemic diseases.

Hydrogels are commonly used in regenerative medicine for the delivery of growth factors (GFs). The spatial and temporal presentations of GFs are critical for directing regenerative processes, yet conventional hydrogels do not enable such control. We have developed a composite hydrogel, termed an acoustically-responsive scaffold (ARS), where release of a GF is non-invasively and spatiotemporally-controlled using focused ultrasound. The ARS consists of a fibrin matrix doped with a GF-loaded, phase-shift emulsion. The GF is released when the ARS is exposed to suprathreshold ultrasound via a mechanism termed acoustic droplet vaporization. In this study, we investigate how different spatial patterns of suprathreshold ultrasound can impact the biological response upon in vivo implantation of an ARS containing basic fibroblast growth factor (bFGF). ARSs were fabricated with either perfluorohexane (bFGF-C6-ARS) or perflurooctane (bFGF-C8-ARS) within the phase-shift emulsion. Ultrasound generated stable bubbles in bFGF-C6-ARS, which inhibited matrix compaction, whereas transiently stable bubbles were generated in bFGF-C8-ARS, which decreased in height by 44% within one day of implantation. The rate of bFGF release and distance of host cell migration were up to 6.8-fold and 8.1-fold greater, respectively, in bFGF-C8-ARS versus bFGF-C6-ARS. Ultrasound increased the formation of macropores within the fibrin matrix of bFGF-C8-ARS by 2.7-fold. These results demonstrate that spatially patterning suprathreshold ultrasound within bFGF-C8-ARS can be used to elicit a spatially-directed response from the host. Overall, these findings can be used in developing strategies to spatially pattern regenerative processes. STATEMENT OF SIGNIFICANCE: Hydrogels are commonly used in regenerative medicine for the delivery of growth factors (GFs). The spatial and temporal presentations of GFs are critical for directing regenerative processes, yet conventional hydrogels do not enable such control. We have developed a composite hydrogel, termed an acoustically-responsive scaffold (ARS), where GF release is non-invasively and spatiotemporally-controlled using focused ultrasound. The ARS consists of a fibrin matrix doped with a phase-shift emulsion loaded with GF, which is released when the ARS is exposed to ultrasound. In this in vivo study, we demonstrate that spatially patterning ultrasound within an ARS containing basic fibroblast growth factor (bFGF) can elicit a spatially-directed response from the host. Overall, these findings can be used in developing strategies to spatially pattern regenerative processes.

Acoustically-responsive scaffolds (ARSs) are composite hydrogels that respond to ultrasound in an on-demand, spatiotemporally-controlled manner due to the presence of a phase-shift emulsion. When exposed to ultrasound, a gas bubble is formed within each emulsion droplet via a mechanism termed acoustic droplet vaporization (ADV). In previous in vitro and in vivo studies, we demonstrated that ADV can control regenerative processes by releasing growth factors and/or modulating micromechanics in ARSs. Precise, spatial patterning of emulsion within an ARS could be beneficial for ADV-induced modulation of biochemical and biophysical cues. However, precise patterning is limited using conventional bulk polymerization techniques. Here, we developed an extrusion-based method for bioprinting ARSs with micropatterned structures. Emulsions were loaded within bioink formulations containing fibrin, hyaluronic acid and/or alginate. Experimental as well as theoretical studies elucidated the interrelations between printing parameters, needle geometry, rheological properties of the bioink, and the process-induced mechanical stresses during bioprinting. The shear thinning properties of the bioinks enabled use of lower extrusion pressures resulting in decreased shear stresses and shorter residence times, thereby facilitating high viability for cell-loaded bioinks. Bioprinting yielded greater alignment of fibrin fibers in ARSs compared to conventionally polymerized ARSs. Bioprinted ARSs also enabled generation of ADV at high spatial resolutions, which were otherwise not achievable in conventional ARSs, and acoustically-driven collapse of ADV-induced bubbles. Overall, bioprinting could aid in optimizing ARSs for therapeutic applications.

A potential replacement candidate for the aging operational WSR-88D infrastructure currently in place is the phased array radar (PAR) system. The current WSR-88Ds take ~5 min to produce a full volumetric scan of the atmosphere, whereas PAR technology allows for full volumetric scanning of the same atmosphere every ~1 min. How this increase in temporal frequency of radar observations might affect the National Severe Storms Laboratory’s (NSSL) Warn-on-Forecast system (WoFS), which is a storm-scale ensemble data assimilation and forecast system for severe convective weather, is unclear. Since radar data assimilation is critical for the WoFS, this study explores the optimal temporal frequency of PAR observations for storm-scale data assimilation using the 31 May 2013 El Reno, Oklahoma, tornadic supercell event. The National Severe Storms Laboratory’s National Weather Radar Testbed PAR in Norman, Oklahoma, began scanning this event more than an hour before the first (and strongest) tornado developed near El Reno, and scanned most of the tornadic supercell’s evolution. Several experiments using various cycling and data frequencies to synchronously and asynchronously assimilate these PAR observations are conducted to produce analyses and very short-term forecasts of the El Reno supercell. Forecasts of low-level reflectivity and midlevel updraft helicity are subjectively evaluated and objectively verified using spatial and object-based techniques. Results indicate that assimilating more frequent PAR observations can lead to more accurate analyses and probabilistic forecasts of the El Reno supercell at longer lead times. Hence, PAR is a promising radar platform for WoFS.

Hydrogels are commonly used for the delivery of bioactive molecules, especially growth factors and cytokines capable of stimulating tissue regeneration. Regenerative processes are regulated by the concentrations and spatiotemporal presentations of these molecules. With conventional hydrogels, these critical delivery parameters cannot be actively modulated following implantation. We have developed composite hydrogel scaffolds where payload release is non-invasively modulated, in an on-demand manner, using ultrasound. These acoustically-responsive scaffolds (ARSs) consist of a fibrin matrix doped with a payload-carrying, perfluorocarbon double emulsion. Previously, acoustic droplet vaporization (ADV) was used to trigger release of a pro-angiogenic growth factor, encapsulated in the ARS, which stimulated blood vessel formation in vivo. In the present study, we assess how characteristics of the monodispersed emulsion, fibrin matrix, and ultrasound impact ADV thresholds and the release efficiency of a dextran payload. ADV thresholds increased with the molecular weight of the PFC in the emulsion and inversely with the volume fraction of emulsion in the ARS. Payload release from ARSs with perfluoroheptane (C7) or perfluorooctane (C8) emulsions was dependent on the number of z-planes of US used to generate ADV and inversely dependent on the lateral spacing. Conversely, release from ARSs with perfluoropentane (C5) or perfluorohexane (C6) emulsions was less dependent on these US exposure parameters. Following ADV, payload diffusion decreased significantly in ARSs with C5 or C6 emulsions compared to ARSs with C7 or C8 emulsions. The expansion of the ARS following ADV decreased with the molecular weight of the PFC. Non-selective release increased with the molecular weight of the PFC and thrombin concentration. Overall, these findings can be used for optimization of ARS properties and US parameters in future therapeutic applications.

Real polarimetric radar observations are directly assimilated for the first time using the ensemble Kalman filter (EnKF) for a supercell case from 20 May 2013 in Oklahoma. A double-moment microphysics scheme and advanced polarimetric radar observation operators are used together to estimate the model states. Lookup tables for the observation operators are developed based on T-matrix scattering amplitudes for all hydrometeor categories, which improve upon previous curved-fitted approximations of T-matrix scattering amplitudes or the Rayleigh approximation. Two experiments are conducted: one assimilates reflectivity (Z) and radial velocity (Vr) (EXPZ), and one assimilates in addition differential reflectivity (ZDR) below the observed melting level at ~2-km height (EXPZZDR). In the EnKF analyses, EXPZZDR exhibits a ZDR arc that better matches observations than EXPZ. EXPZZDR also has higher ZDR above 2 km, consistent with the observed ZDR column. Additionally, EXPZZDR has an improved estimate of the model microphysical states. Specifically, the rain mean mass diameter (Dnr) in EXPZZDR is higher in the ZDR arc region and the total rain number concentration (Ntr) is lower downshear in the forward flank than EXPZ when compared to values retrieved from the polarimetric observations. Finally, a negative gradient of hail mean mass diameter (Dnh) is found in the right-forward flank of the EXPZZDR analysis, which supports previous findings indicating that size sorting of hail, as opposed to rain, has a more significant impact on low-level polarimetric signatures. This paper represents a proof-of-concept study demonstrating the value of assimilating polarimetric radar data in improving the analysis of features and states related to microphysics in supercell storms.

After decades of research and development, the WSR-88D (NEXRAD) network in the United States was upgraded with dual-polarization capability, providing polarimetric radar data (PRD) that have the potential to improve weather observations, quantification, forecasting, and warnings. The weather radar networks in China and other countries are also being upgraded with dual-polarization capability. Now, with radar polarimetry technology having matured, and PRD available both nationally and globally, it is important to understand the current status and future challenges and opportunities. The potential impact of PRD has been limited by their oftentimes subjective and empirical use. More importantly, the community has not begun to regularly derive from PRD the state parameters, such as water mixing ratios and number concentrations, used in numerical weather prediction (NWP) models. In this review, we summarize the current status of weather radar polarimetry, discuss the issues and limitations of PRD usage, and explore potential approaches to more efficiently use PRD for quantitative precipitation estimation and forecasting based on statistical retrieval with physical constraints where prior information is used and observation error is included. This approach aligns the observation-based retrievals favored by the radar meteorology community with the model-based analysis of the NWP community. We also examine the challenges and opportunities of polarimetric phased array radar research and development for future weather observation.

Background The novel SARS‐CoV‐2 virus inflicts far‐reaching health decrements, both directly and through secondary inflammatory stimulation. To date, there is little information regarding the effects of COVID‐19 on the heart after infection, especially among young healthy adults. Purpose We sought to determine whether contracting SARS‐CoV‐2 affects cardiac function in young, otherwise healthy adults and whether these alterations recede after testing positive for SARS‐CoV‐2. Methods Transthoracic echocardiography was performed on 7 subjects with SARS‐CoV‐2 (3F/4M, 21±1y, 24±2 kgᐧm‐1) who tested positive 3‐4 weeks prior to baseline testing followed by 1‐month and 2‐month follow‐up testing. A parasternal long‐axis view was utilized to measure septal and left ventricular posterior wall thickness, left ventricular volume, and left ventricular outflow tract dimensions. A four‐chamber view was utilized to measure mitral and tricuspid valve, left ventricular, and pulmonary vein function. Data are mean±SD. Results Stroke volume (Baseline: 44±11ml, 1‐month: 42±13ml, 2‐month: 54±12ml) and stroke index (Baseline: 24±6ml/m^2, 1‐month: 22±7ml/m^2, 2‐month: 29±6ml/m^2) were elevated at 2‐month follow‐up (p

Assimilation of dual-polarization (dual-pol) observations provides more accurate storm-scale analyses to initialize forecasts of severe convective thunderstorms. This study investigates the impact assimilating experimental sector-scan dual-pol observations has on storm-scale ensemble forecasts and how this impact changes over different data assimilation (DA) windows using the ensemble Kalman filter (EnKF). Ensemble forecasts are initialized after 30, 45, and 60 minutes of DA for two sets of experiments that assimilate either reflectivity and radial velocity only (EXPZ) or reflectivity and radial velocity plus differential reflectivity (EXPZZDR). This study uses the 31 May 2013 Oklahoma event which included multiple storms that produced tornadoes and severe hail, with focus placed on two storms that impacted El Reno and Stillwater during the event.The earliest initialized forecast of EXPZZDR better predicts the evolution of the El Reno storm than EXPZ, but the two sets of experiments become similar at subsequent forecast times. However, the later EXPZZDR forecasts of the Stillwater storm, which organized towards the end of the DA window, produce improved results compared to EXPZ, in which the storm is less intense and weakens. Evaluation of forecast products for supercell mesocyclones (updraft helicity [UH]) and hail show similar results with earlier EXPZZDR forecasts better predicting the UH swaths of the El Reno storm and later forecasts producing improved UH and hail swaths for the Stillwater storm. The results indicate that the assimilation of ZDR over fewer DA cycles can produce improved forecasts when DA windows sufficiently cover storms during their initial development and organization.

Current treatment options for critical limb ischemia (CLI), an advanced type of peripheral artery disease (PAD), are grossly insufficient. Clinical trials have evaluated exogenous, pro-angiogenic growth factors to stimulate angiogenesis in PAD/CLI. However, conventional administration routes are ineffective for growth factors, which have contributed to translational challenges. We have developed a composite hydrogel scaffold, termed an acoustically-responsive scaffold (ARS), where growth factor delivery is controlled via acoustic droplet vaporization (ADV). The ARS consists of a phase-shift emulsion contained within a fibrin matrix. We demonstrate in a mouse model of CLI that ADV can be used to enhance angiogenesis and perfusion via the release of basic fibroblast growth factor (bFGF) from implanted ARSs.

Acoustically-responsive scaffolds (ARSs), which are composite fibrin hydrogels, have been used to deliver regenerative molecules. ARSs respond to ultrasound in an on-demand, spatiotemporally-controlled manner via a mechanism termed acoustic droplet vaporization (ADV). Here, we study the ADV-induced, time-dependent micromechanical and microstructural changes to the fibrin matrix in ARSs using confocal fluorescence microscopy as well as atomic force microscopy. ARSs, containing phase-shift double emulsion (PSDE, mean diameter: 6.3 μm), were exposed to focused ultrasound to generate ADV - the phase transitioning of the PSDE into gas bubbles. As a result of ADV-induced mechanical strain, localized restructuring of fibrin occurred at the bubble-fibrin interface, leading to formation of locally denser regions. ADV-generated bubbles significantly reduced fibrin pore size and quantity within the ARS. Two types of ADV-generated bubble responses were observed in ARSs: super-shelled spherical bubbles, with a growth rate of 31 μm per day in diameter, as well as fluid-filled macropores, possibly as a result of acoustically-driven microjetting. Due to the strain stiffening behavior of fibrin, ADV induced a 4-fold increase in stiffness in regions of the ARS proximal to the ADV-generated bubble versus distal regions. These results highlight that the mechanical and structural microenvironment within an ARS can be spatiotemporally modulated using ultrasound, which could be used to control cellular processes and further the understanding of ADV-triggered drug delivery for regenerative applications.

Extracellular matrix (ECM) remodeling is essential for the process of capillary morphogenesis. Here we employed synthetic poly(ethylene glycol) (PEG) hydrogels engineered with proteolytic specificity to either matrix metalloproteinases (MMPs), plasmin, or both to investigate the relative contributions of MMP- and plasmin-mediated ECM remodeling to vessel formation in a 3D-model of capillary self-assembly analogous to vasculogenesis. We first demonstrated a role for both MMP- and plasmin-mediated mechanisms of ECM remodeling in an endothelial-fibroblast co-culture model of vasculogenesis in fibrin hydrogels using inhibitors of MMPs and plasmin. When this co-culture model was employed in engineered PEG hydrogels with selective protease sensitivity, we observed robust capillary morphogenesis only in MMP-sensitive matrices. Fibroblast spreading in plasmin-selective hydrogels confirmed this difference was due to protease preference by endothelial cells, not due to limitations of the matrix itself. In hydrogels engineered with crosslinks that were dually susceptible to MMPs and plasmin, capillary morphogenesis was unchanged. These findings highlight the critical importance of MMP-mediated degradation during vasculogenesis and provide strong evidence to justify the preferential selection of MMP-degradable peptide crosslinkers in synthetic hydrogels used to study vascular morphogenesis and promote vascularization. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B:2507-2516, 2019.

Forming functional blood vessel networks is a major clinical challenge in the fields of tissue engineering and therapeutic angiogenesis. Cell-based strategies to promote neovascularization have been widely explored, but cell sourcing remains a significant limitation. Induced-pluripotent stem cell-derived endothelial cells (iPSC-ECs) are a promising, potentially autologous, alternative cell source. However, it is unclear whether iPSC-ECs form the same robust microvasculature in vivo documented for other EC sources. In this study, we utilized a well-established in vivo model, in which ECs (iPSC-EC or human umbilical vein endothelial cells [HUVEC]) were coinjected with normal human lung fibroblasts (NHLFs) and a fibrin matrix into the dorsal flank of severe combined immunodeficiency mice to assess their ability to form functional microvasculature. Qualitatively, iPSC-ECs were capable of vessel formation and perfusion and demonstrated similar vessel morphologies to HUVECs. However, quantitatively, iPSC-ECs exhibited a two-fold reduction in vessel density and a three-fold reduction in the number of perfused vessels compared with HUVECs. Further analysis revealed the presence of collagen-IV and α-smooth muscle actin were significantly lower around iPSC-EC/NHLF vasculature than in HUVEC/NHLF implants, suggesting reduced vessel maturity. Collectively, these results demonstrate the need for increased iPSC-EC maturation for clinical translation to be realized.

During normal development, the extracellular matrix (ECM) regulates cell fate mechanically and biochemically. However, the ECM's influence on lineage reprogramming, a process by which a cell's developmental cycle is reversed to attain a progenitor-like cell state followed by subsequent differentiation into a desired cell phenotype, is unknown. Using a material mimetic of the ECM, here we show that ligand identity, ligand density, and substrate modulus modulate indirect cardiac reprogramming efficiency, but were not individually correlated with phenotypic outcomes in a predictive manner. Alternatively, we developed a data-driven model using partial least squares regression to relate short-term cell states, defined by quantitative mechanosensitive responses to different material environments, with long-term changes in phenotype. This model was validated by accurately predicting the reprogramming outcomes on a different material platform. Collectively, these findings suggest a means to rapidly screen candidate biomaterials that support reprogramming with high efficiency, without subjecting cells to the entire reprogramming process.

A major translational challenge in the fields of therapeutic angiogenesis and regenerative medicine is the need to create functional microvasculature. The purpose of this study was to assess whether a potentially autologous endothelial cell (EC) source derived from human induced pluripotent stem cells (iPSC-ECs) can form the same robust, stable microvasculature as previously documented for other sources of ECs. We utilized a well-established in vitro assay, in which endothelial cell-coated (iPSC-EC or HUVEC) beads were co-embedded with fibroblasts in a 3D fibrin matrix to assess their ability to form stable microvessels. iPSC-ECs exhibited a five-fold reduction in capillary network formation compared to HUVECs. Increasing matrix density reduced sprouting, although this effect was attenuated by distributing the NHLFs throughout the matrix. Inhibition of both MMP- and plasmin-mediated fibrinolysis was required to completely block sprouting of both HUVECs and iPSC-ECs. Further analysis revealed MMP-9 expression and activity were significantly lower in iPSC-EC/NHLF co-cultures than in HUVEC/NHLF co-cultures at later time points, which may account for the observed deficiencies in angiogenic sprouting of the iPSC-ECs. Collectively, these findings suggest fundamental differences in EC phenotypes must be better understood to enable the promise and potential of iPSC-ECs for clinical translation to be realized.