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Microdissection of the Majority Transcriptome in Single-Cell Quality Shows Scientific Importance and Myeloid Cellular material Heterogeneity in Lung Adenocarcinoma.
Explore the longitudinal CT-based radiomics to demonstrate the changing trend of radiotherapy response and to determine at which point after the onset of treatment radiomics exhibit the greatest change for stage III NSCLC patients.

Ten stage III NSCLC patients in line with inclusion criteria were enrolled retrospectively, each of whom received radiotherapy or concurrent chemo-radiotherapy and performed eight series of follow-up CT imaging. Longitudinal radiomics were extracted on region of interest from the eight registered images, then two steps were conducted to select significant features as indicators of tumor change 1) stable features were selected by Kendall rank correlation; 2) texture feature types with a steadily changing trend were retained and intensity features with stable change trends were selected to represent the large number of them. Next, the trend and rate of tumor change were analyzed using the Delta method and Curve-fitting method. Finally, the statistics in the distribution of stableerapy.
The COVID-19 pandemic caused a rapid pivot from in-person to virtual residency interviews across the United States. We present a survey we conducted about the attitudes and opinions of anesthesiology program directors with regard to the 2021 virtual interview process.

This was a 13-question online survey disseminated to 142 anesthesiology residency program directors in February 2021, asking them to compare the most recent interview cycle to their experience with prior cycles.

There were 46 (37%) respondents. Generally, respondents saw an increase in applicants and reported perceiving worse interpersonal relationships with applicants, significantly so in programs with small resident classes.

Past research has focused on the benefits of virtual interviews for the applicant, but these should be evaluated in tandem with increased difficulties for the interviewers.
Past research has focused on the benefits of virtual interviews for the applicant, but these should be evaluated in tandem with increased difficulties for the interviewers.Recent work argues that changes in people's responses to the same question over time should be thought of as reflecting a fixed baseline subject to temporary local influences, rather than durable changes in response to new information. Distinguishing between these two individual-level process-a settled dispositions model and an active updating model-is important because these individual-level processes underlie different theories of population-level social change. This article introduces an alternative method for adjudicating between these two models based on structural equation modeling. This model provides a close fit to the theoretical models outlined in previous work. Applying this method to more than 500 questions in the General Social Survey's three-wave panels, we find even stronger evidence than previous work that most survey responses reflect settled dispositions developed prior to adulthood.Oxygen plays a critical role in the photopolymerization process resulting in the formation of solid structures from liquid resins during three-dimensional (3D) printing it acts as a polymerization inhibitor. Upon exposure to light, oxygen is depleted. As a result, the polymerization process becomes activated. Electron paramagnetic resonance (EPR) imaging is described as a tool to visualize changes in oxygen distribution caused by light exposure. This nondestructive method uses radio waves and, therefore, is not constrained by optical opacity offering greater penetrating depth. Three proof-of-principle imaging experiments were demonstrated (1) spatial propagation of the photopolymerization process; (2) oxygen depletion as a result of postcuring; and (3) oxygen visualization in a 3D printed spiral model. Commercial stereolithography (SLA) resin was used in these experiments. Lithium octa-n-butoxynaphthalocyanine (LiNc-BuO) probe was mixed with the resin to permit oxygen imaging. Li-naphthalocyanine probes are routinely used in various EPR applications because of their long-term stability and high functional sensitivity to oxygen. In this study, we demonstrate that EPR imaging has the potential to become a powerful visualization tool in the development of 3D printing technology, including bioprinting and tissue engineering.Recent clinical successes have intensified interest in using adeno-associated virus (AAV) vectors for therapeutic gene delivery. The liver is a key clinical target, given its critical physiological functions and involvement in a wide range of genetic diseases. Here, we report the bioengineering of a set of next-generation AAV vectors, named AAV-SYDs (where "SYD" stands for Sydney, Australia), with increased human hepato-tropism in a liver xenograft mouse model repopulated with primary human hepatocytes. We followed a two-step process that staggered directed evolution and domain-swapping approaches. Using DNA-family shuffling, we first mapped key AAV capsid regions responsible for efficient human hepatocyte transduction in vivo. Focusing on these regions, we next applied domain-swapping strategies to identify and study key capsid residues that enhance primary human hepatocyte uptake and transgene expression. Our findings underscore the potential of AAV-SYDs as liver gene therapy vectors and provide insights into the mechanism responsible for their enhanced transduction profile.More than 50 lysosomal storage diseases (LSDs) are associated with lysosomal dysfunctions with the frequency of 15,000 live births. As a result of missing enzyme activity, the lysosome dysfunction accumulates undegraded or partially degraded molecules, affecting the entire body. Most of them are life-threatening diseases where patients could die within the first or second decade of life. Approximately 20 LSDs have the approved treatments, which do not provide the cure for the disorder. Therefore, the delivery of missing genes through gene therapy is a promising approach for LSDs. Over the years, ex vivo lentiviral-mediated gene therapy for LSDs has been approached using different strategies. Several clinical trials for LSDs are under investigation.Ex vivo lentiviral-mediated gene therapy needs optimization in dose, time of delivery, and promoter-driven expression. Choosing suitable promoters seems to be one of the important factors for the effective expression of the dysfunctional enzyme. This review summarizes the research on therapy for LSDs that has used different lentiviral vectors, emphasizing gene promoters.Understanding pulmonary diseases requires robust culture models that are reproducible, sustainable in long-term culture, physiologically relevant, and suitable for assessment of therapeutic interventions. Primary human lung cells are physiologically relevant but cannot be cultured in vitro long term and, although engineered organoids are an attractive choice, they do not phenotypically recapitulate the lung parenchyma; overall, these models do not allow for the generation of reliable disease models. Recently, we described a new cell culture platform based on H441 cells that are grown at the air-liquid interface to produce the SALI culture model, for studying and correcting the rare interstitial lung disease surfactant protein B (SPB) deficiency. Here, we report the characterization of the effects of SALI culture conditions on the transcriptional profile of the constituent H441 cells. Selleck ESI-09 We further analyze the transcriptomics of the model in the context of surfactant metabolism and the disease phenotype through SFTPB knockout SALI cultures. By comparing the gene expression profile of SALI cultures with that of human lung parenchyma obtained via single-cell RNA sequencing, we found that SALI cultures are remarkably similar to human alveolar type II cells, implying clinical relevance of the SALI culture platform as a non-diseased human lung alveolar cell model.OCT4 is a key mediator of induced pluripotent stem cell (iPSC) reprogramming, but the mechanistic insights into the role of exogenous OCT4 and timelines that initiate pluripotency remain to be resolved. Here, using measles reprogramming vectors, we present microRNA (miRNA) targeting of exogenous OCT4 to shut down its expression during the mesenchymal to the epithelial transition phase of reprogramming. We showed that exogenous OCT4 is required only for the initiation of reprogramming and is dispensable for the maturation stage. However, the continuous expression of SOX2, KLF4, and c-MYC is necessary for the maturation stage of the iPSC. Additionally, we demonstrate a novel application of miRNA targeting in a viral vector to contextually control the vector/transgene, ultimately leading to an improved reprogramming efficiency. This novel approach could be applied to other systems for improving the efficiency of vector-induced processes.Adeno-associated viruses (AAVs) represent important gene therapy vectors with several approved clinical applications and numerous more in clinical trials. Genome packaging is an essential step in the bioprocessing of AAVs and needs to be tightly monitored to ensure the proper delivery of transgenes and the production of effective drugs. Current methods to monitor genome packaging have limited sensitivity, a high demand on labor, and struggle to distinguish between packaging of the intended genome or unwanted side-products. Here we show that Orbitrap-based charge-detection mass spectrometry allows the very sensitive quantification of all these different AAV bioprocessing products. A protocol is presented that allows the quantification of genome-packed AAV preparations in under half an hour, requiring only micro-liter quantities of typical AAV preparations with ∼1013 viral capsids per milliliter. The method quickly assesses the integrity and amount of genome packed AAV particles to support AAV bioprocessing and characterization of this rapidly emerging class of advanced drug therapies.Over the past decade, numerous gene-editing platforms which alter host DNA in a highly specific and targeted fashion have been described. Two notable examples are zinc finger nucleases (ZFNs), the first gene-editing platform to be tested in clinical trials, and more recently, CRISPR/Cas9. Although CRISPR/Cas9 approaches have become arguably the most popular platform in the field, the therapeutic advantages and disadvantages of each strategy are only beginning to emerge. We have established a nonhuman primate (NHP) model that serves as a strong predictor of successful gene therapy and gene-editing approaches in humans; our recent work shows that ZFN-edited hematopoietic stem and progenitor cells (HSPCs) engraft at lower levels than CRISPR/Cas9-edited cells. Here, we investigate the mechanisms underlying this difference. We show that optimized culture conditions, including defined serum-free media, augment engraftment of gene-edited NHP HSPCs in a mouse xenograft model. Furthermore, we identify intracellular RNases as major barriers for mRNA-encoded nucleases relative to preformed enzymatically active CRISPR/Cas9 ribonucleoprotein (RNP) complexes. We conclude that CRISPR/Cas9 RNP gene editing is more stable and efficient than ZFN mRNA-based delivery and identify co-delivered RNase inhibitors as a strategy to enhance the expression of gene-editing proteins from mRNA intermediates.
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