A nephrogenic progenitor cell (NP) with cancer stem cell characteristics driving Wilms tumor (WT) using spatial transcriptomics, bulk and single cell RNA sequencing, and complementary in vitro and transplantation experiments is identified and characterized. NP from WT samples with NP from the developing human kidney is compared. Cells expressing SIX2 and CITED1 fulfill cancer stem cell criteria by reliably recapitulating WT in transplantation studies. It is shown that self-renewal versus differentiation in SIX2+CITED1+ cells is regulated by the interplay between integrins ITGβ1 and ITGβ4. The spatial transcriptomic analysis defines gene expression maps of SIX2+CITED1+ cells in WT samples and identifies the interactive gene networks involved in WT development. These studies define SIX2+CITED1+ cells as the nephrogenic-like cancer stem cells of WT and points to the renal developmental transcriptome changes as a possible driver in regulating WT formation and progression.

•Choroid and choriocapillaris are potential important targets of therapy early in AMD.•Macular vasculature pathology, like choriocapillaris flow deficits, can be measured using OCT-A.•Subretinal drusenoid deposits result from biogenesis pathways involved in lipid homeostasis.•How to target (over- or under-express) the HtrA1 gene associated with AMD is controversial.•Optimal gene therapies for AMD depend on retinal delivery site and specificity to retinal cell.

You have accessJournal of UrologyCME1 May 2022PD01-11 CHARACTERIZATION OF WILMS TUMOR AND HUMAN FETAL KIDNEY USING SPATIAL TRANSCRIPTOMICS Astgik Petrosyan, Valentina Villani, Paola Aguiari, Matthew Thornton, Zhou Shengmei, Brendan Grubbs, Marie Csete, Roger De Filippo, Kevin Lemley, Sargis Sedrakyan, Stefano Da Sacco, and Laura Perin Astgik PetrosyanAstgik Petrosyan More articles by this author , Valentina VillaniValentina Villani More articles by this author , Paola AguiariPaola Aguiari More articles by this author , Matthew ThorntonMatthew Thornton More articles by this author , Zhou ShengmeiZhou Shengmei More articles by this author , Brendan GrubbsBrendan Grubbs More articles by this author , Marie CseteMarie Csete More articles by this author , Roger De FilippoRoger De Filippo More articles by this author , Kevin LemleyKevin Lemley More articles by this author , Sargis SedrakyanSargis Sedrakyan More articles by this author , Stefano Da SaccoStefano Da Sacco More articles by this author , and Laura PerinLaura Perin More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002516.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Growing evidence links Wilms tumor (WT) to aberrant nephrogenesis. Studies highlighted the genetic complexity of WT, but little is known about the molecular mechanisms that regulate WT development. Using Spatial transcriptomics (ST), which allows analysis of the gene expression based on morphological context, we showed important differences between WT subtypes and defined the interactive gene networks involved in WT development using human fetal kidney (hFK) as reference. METHODS: Using Visium 10x Genomics, we generated spatial maps of gene expression in human fetal kidney (hFK) andfavorable (stage III) and unfavorable (stage I) WTs. Data were analyzed using Space Ranger software v1.0.0, Seurat v3.2, Panther V14, and Loupe Cell Browser and further analyzed against our previously generated bulk/sc-RNA seq data on the same samples. RESULTS: ST identified specific clusters in hFK that closely recapitulated the developmental stages of normal nephrogenesis (nephrogenic zone, glomeruli, tubules, and stroma). Unfavorable WT and favorable WT clusters showed heterogeneity of the tumor landscape (blastema, epithelium, and stroma and non-renal phenotypes). Blastema in WT favorable vs. WT unfavorable, though histologically identical, presented different transcriptomics profiles. WTs also showed gene expression typical of muscle tissue (or other non-renal phenotypes) rather than mature kidney structures, which correlated with the histologic absence of mature tubules and glomeruli. Comparative RNA-seq analysis identified cells expressing SIX2 and CITED1 as the root cells of the origin of the WT. Unfavorable WT expressed a higher level of CITED1 in blastema foci and higher expression of uncommitted genes and modulation of inductive nephrogenic signals like WNT and FGF. We also identified genes expressed specifically in WT subtypes and performed a preliminary characterization of the immune milieu of WT. CONCLUSIONS: The spatiotemporal mapping combined with different transcriptomic data highlighted the heterogeneity of the WT subtypes confirming uncommitted nephron progenitors as driving the development of WT. We identified genes that may allow for better stratification of WT and potential therapeutic targets for distinct WT subtypes. Source of Funding: GOFARR Fund © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e33 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Astgik Petrosyan More articles by this author Valentina Villani More articles by this author Paola Aguiari More articles by this author Matthew Thornton More articles by this author Zhou Shengmei More articles by this author Brendan Grubbs More articles by this author Marie Csete More articles by this author Roger De Filippo More articles by this author Kevin Lemley More articles by this author Sargis Sedrakyan More articles by this author Stefano Da Sacco More articles by this author Laura Perin More articles by this author Expand All Advertisement PDF downloadLoading ...

Aging is associated with loss of function across organ systems, contributing to systemic frailty. Loss of skeletal muscle mass and function, in particular, is a major source of frailty in older adults, severely impacting quality of life. Some loss of muscle mass and strength with aging is inevitable, and sarcopenia, the severe loss of muscle mass with aging, is common. Sarcopenia is determined in part by genetics but can be modified by lifestyle choices. The pathophysiologic underpinnings of sarcopenia are complex and multifactorial. In this review, the causes of sarcopenia are surveyed at the systems, cell, subcellular, and molecular levels with emphasis on the interplay between these various causes of this degenerative disease process.

Dramatic variation in SARS-CoV-2 virulence and transmission between hosts has driven the COVID-19 pandemic. The complexity and dynamics of the immune response present a challenge to understanding variation in SARS-CoV-2 infections. To address this challenge, we apply control theory, a framework used to study complex feedback systems, to establish rigorous mathematical bounds on immune responses. Two mechanisms of SARS-CoV-2 biology are sufficient to create extreme variation between hosts: (1) a sparsely expressed host receptor and (2) potent, but not unique, suppression of interferon. The resulting model unifies disparate and unexplained features of the SARS-CoV-2 pandemic, predicts features of future viruses that threaten to cause pandemics, and identifies potential interventions.

ABSTRACTBACKGROUND AND PURPOSEIn hepatic encephalopathy (HE), osmotic stressors promoting brain edema result in a compensatory drop in the astrocyte metabolite myo‐inositol (mI). Identifying differences between nonalcoholic steatohepatitis (NASH) with and without HE and healthy controls using proton magnetic resonance spectroscopy (MRS) and evaluating hypoalbuminemia and hyperammonemia as osmotic stressors that predict the reduction of mI allow further understanding of mechanisms that promote brain edema in HE. The aim of this study was to assess brain edema in HE using characteristic MRS markers and serum albumin.METHODSWe evaluated between group differences among 19 NASH cirrhosis without HE (Crhs‐HE) (age = 63 ± 8.7), 9 NASH cirrhosis with HE (Crhs+HE) (age = 63 ± 9.2), and 16 controls (age = 57.8 ± 11.7) using 1H MRS. Glutamine (Gln/tCr) and serum albumin were evaluated as predictors of myo‐inositol (mI/tCr) using linear regression. Statistical significance was set at P < .05 with adjustment for multiple comparisons.RESULTSBrain mI/tCr was decreased, and Gln/tCr increased in Crhs+HE compared to Crhs‐HE and controls in both brain regions (P < .001 for all). Evaluated together as joint predictors, serum albumin but not Gln/tCr significantly predicted mI/tCr in GM (P = .02 and P = .2, respectively) and PWM (P = .01 and P = .1, respectively).CONCLUSIONLow mI/tCr and increased Gln/tCr were characteristics of Crhs+HE. Low serum albumin was the strongest predictor of brain osmotic stress indicated by reduced mI/tCr, with no residual independent association seen for brain Gln/tCr concentration. This suggests that hypoalbuminemia in chronic liver disease may promote brain edema in HE.

Significant progress has been made to advance stem cell products as potential therapies for kidney diseases: various kinds of stem cells can restore renal function in preclinical models of acute and chronic kidney injury. Nonetheless this literature contains contradictory results, and for this reason, we focus this review on reasons for apparent discrepancies in the literature, because they contribute to difficulty in translating renal regenerative therapies. Differences in methodologies used to derive and culture stem cells, even those from the same source, in addition to the lack of standardized renal disease animal models (both acute and chronic), are important considerations underlying contradictory results in the literature. We propose that harmonized rigorous protocols for characterization, handling, and delivery of stem cells in vivo could significantly advance the field, and present details of some suggested approaches to foster translation in the field of renal regeneration. Our goal is to encourage coordination of methodologies (standardization) and long-lasting collaborations to improve protocols and models to lead to reproducible, interpretable, high-quality preclinical data. This approach will certainly increase our chance to 1 day offer stem cell therapeutic options for patients with all-too-common renal diseases. Stem Cells Translational Medicine 2019;8:82-92

Correction for 'The value of universally available raw NMR data for transparency, reproducibility, and integrity in natural product research' by James B. McAlpine et al., Nat. Prod. Rep., 2018, DOI: .

Human parthenogenetic stem cells (hpSC) are pluripotent stem cells that can be derived from a blastocyst inner cell mass. But, unlike traditional blastocyst sources of human embryonic stem cell (hESC) lines, the blastocyst source of hpSC is created by stimulating an unfertilized egg to divide. So, an embryo is not destroyed in the process of generating hpSC, and the hpSC diploid genome is completely maternal. In fundamental ways, hpSC are similar to hESC: under the right conditions both divide indefinitely. Both differentiate into cells of all three germ layers. Importantly, the hpSC genome can be homozygous at HLA loci, making them immunologically less complex to match in potential therapeutic applications. This unique feature may be advantageous in applying cell therapies and holds the key to hpSC use in future clinical applications. Genetic and epigenetic instability of hpSC is a potential problem in hpSC clinical application, but may also be a problem for cell therapies from hESC and induced pluripotent stem cells (iPSC). Similarly, the relative thresholds of individual hpSC (and hESC and hiPSC) lines to differentiate into particular cell types differ from line to line and require further study. Significantly less work has been done on hpSC than hESC or iPSC so that clinical application of hpSC will require many more years of research.

We examined data and patterns in clinical islet transplant studies registered on ClinicalTrials.gov (CTgov) for treatment of type 1 diabetes (T1D), with a goal of extracting insights to apply in the design of a pluripotent stem cell-derived islet therapy. Clinical islet transplantation, as a cell therapy (rather than solid organ transplant) is a unique precedent for stem cell-based islet therapies. Registration activity shows that the field is not growing significantly, and newer registrations suggest that the reasons for stagnation include need for a more optimal site of infusion/transplantation, and especially a need for better immune protective strategies to advance a more effective and durable therapy for T1D. Stem Cells Translational Medicine 2019;8:209&214.

Significant progress has been made to advance stem cell products as potential therapies for kidney diseases: various kinds of stem cells can restore renal function in preclinical models of acute and chronic kidney injury. Nonetheless this literature contains contradictory results, and for this reason, we focus this review on reasons for apparent discrepancies in the literature, because they contribute to difficulty in translating renal regenerative therapies. Differences in methodologies used to derive and culture stem cells, even those from the same source, in addition to the lack of standardized renal disease animal models (both acute and chronic), are important considerations underlying contradictory results in the literature. We propose that harmonized rigorous protocols for characterization, handling, and delivery of stem cells in vivo could significantly advance the field, and present details of some suggested approaches to foster translation in the field of renal regeneration. Our goal is to encourage coordination of methodologies (standardization) and long‐lasting collaborations to improve protocols and models to lead to reproducible, interpretable, high‐quality preclinical data. This approach will certainly increase our chance to 1 day offer stem cell therapeutic options for patients with all‐too‐common renal diseases. Stem Cells Translational Medicine 2019;8:82–92

Objective: The study is based on previously reported mathematical analysis of arterial waveform that extracts hidden oscillations in the waveform that we called intrinsic frequencies. The goal of this clinical study was to compare the accuracy of left ventricular ejection fraction derived from intrinsic frequencies noninvasively versus left ventricular ejection fraction obtained with cardiac MRI, the most accurate method for left ventricular ejection fraction measurement. Design: After informed consent, in one visit, subjects underwent cardiac MRI examination and noninvasive capture of a carotid waveform using an iPhone camera (The waveform is captured using a custom app that constructs the waveform from skin displacement images during the cardiac cycle.). The waveform was analyzed using intrinsic frequency algorithm. Setting: Outpatient MRI facility. Subjects: Adults able to undergo MRI were referred by local physicians or self-referred in response to local advertisement and included patients with heart failure with reduced ejection fraction diagnosed by a cardiologist. Interventions: Standard cardiac MRI sequences were used, with periodic breath holding for image stabilization. To minimize motion artifact, the iPhone camera was held in a cradle over the carotid artery during iPhone measurements. Measurements and Main Results: Regardless of neck morphology, carotid waveforms were captured in all subjects, within seconds to minutes. Seventy-two patients were studied, ranging in age from 20 to 92 years old. The main endpoint of analysis was left ventricular ejection fraction; overall, the correlation between ejection fraction–iPhone and ejection fraction–MRI was 0.74 (r = 0.74; p < 0.0001; ejection fraction–MRI = 0.93 × [ejection fraction–iPhone] + 1.9). Conclusions: Analysis of carotid waveforms using intrinsic frequency methods can be used to document left ventricular ejection fraction with accuracy comparable with that of MRI. The measurements require no training to perform or interpret, no calibration, and can be repeated at the bedside to generate almost continuous analysis of left ventricular ejection fraction without arterial cannulation.

A considerable number of studies have focused on solving the inverse problem of ECG using various modeling techniques. Successful modeling uses mostly surface to surface mapping, considering the heart as a surface source or even a point source for the purpose of eliminating complexity of problem. However, many cardiovascular abnormalities cannot be fully understood or diagnosed without more understanding of cross-layer electrophysiological (EP) activities in the ventricle including the repolarization ECG waveform for diagnosis. Here, we propose a systematic approach to build a numeric model for ECG to explore the heterogeneous EP properties in ventricles.

Early coronary artery reperfusion improves outcomes for patients with ST-segment elevation myocardial infarction (STEMI), but morbidity and mortality after STEMI remain unacceptably high. The primary deficits seen in these patients include inadequate pump function, owing to rapid infarction of muscle in the first few hours of treatment, and adverse remodelling of the heart in the months that follow. Given that attempts to further reduce myocardial infarct size beyond early reperfusion in clinical trials have so far been disappointing, effective therapies are still needed to protect the reperfused myocardium. In this Review, we discuss several approaches to preserving the reperfused heart, such as therapies that target the mechanisms involved in mitochondrial bioenergetics, pyroptosis, and autophagy, as well as treatments that harness the cardioprotective properties of inhaled anaesthetic agents. We also discuss potential therapies focused on correcting the no-reflow phenomenon and its effect on healing and adverse left ventricular remodelling.

Objectives: To assess the knowledge and use of the Assessment, prevention, and management of pain; spontaneous awakening and breathing trials; Choice of analgesia and sedation; Delirium assessment; Early mobility and exercise; and Family engagement and empowerment (ABCDEF) bundle to implement the Pain, Agitation, Delirium guidelines. Design: Worldwide online survey. Setting: Intensive care. Intervention: A cross-sectional online survey using the Delphi method was administered to intensivists worldwide, to assess the knowledge and use of all aspects of the ABCDEF bundle. Measurement and Main Results: There were 1,521 respondents from 47 countries, 57% had implemented the ABCDEF bundle, with varying degrees of compliance across continents. Most of the respondents (83%) used a scale to evaluate pain. Spontaneous awakening trials and spontaneous breathing trials are performed in 66% and 67% of the responder ICUs, respectively. Sedation scale was used in 89% of ICUs. Delirium monitoring was implemented in 70% of ICUs, but only 42% used a validated delirium tool. Likewise, early mobilization was "prescribed" by most, but 69% had no mobility team and 79% used no formal mobility scale. Only 36% of the respondents assessed ICU-acquired weakness. Family members were actively involved in 67% of ICUs; however, only 33% used dedicated staff to support families and only 35% reported that their unit was open 24 hr/d for family visits. Conclusions: The current implementation of the ABCDEF bundle varies across individual components and regions. We identified specific targets for quality improvement and adoption of the ABCDEF bundle. Our data reflect a significant but incomplete shift toward patient-andfamily-centered ICU care in accordance with the Pain, Agitation, Delirium guidelines.

Background: Hemorrhagic shock is a leading cause of preventable death in young adults, both military and civilian. The volatile anesthetic isoflurane can protect (precondition) against cardiac repe...