Cell type-specific transcriptomics of esophageal adenocarcinoma as a scalable alternative for single cell transcriptomics

Single-cell transcriptomics have revolutionized our understanding of the cell composition of tumors and allowed us to identify new subtypes of cells. Despite rapid technological advancements, single-cell analysis remains resource-intense hampering the scalability that is required to profile a sufficient number of samples for clinical associations. Therefore, more scalable approaches are needed to understand the contribution of individual cell types to the development and treatment response of solid tumors such as esophageal adenocarcinoma where comprehensive genomic studies have only led to a small number of targeted therapies. Due to the limited treatment options and late diagnosis, esophageal adenocarcinoma has a poor prognosis. Understanding the interaction between and dysfunction of individual cell populations provides an opportunity for the development of new interventions. In an attempt to address the technological and clinical needs, we developed a protocol for the separation of esophageal carcinoma tissue into leukocytes (CD45+), epithelial cells (EpCAM+), and fibroblasts (two out of PDGFRα, CD90, anti-fibroblast) by fluorescence-activated cell sorting and subsequent RNA sequencing. We confirm successful separation of the three cell populations by mapping their transcriptomic profiles to reference cell lineage expression data. Gene-level analysis further supports the isolation of individual cell populations with high expression of CD3, CD4, CD8, CD19, and CD20 for leukocytes, CDH1 and MUC1 for epithelial cells, and FAP, SMA, COL1A1, and COL3A1 for fibroblasts. As a proof of concept, we profiled tumor samples of nine patients and explored expression differences in the three cell populations between tumor and normal tissue. Interestingly, we found that angiogenesis-related genes were upregulated in fibroblasts isolated from tumors compared with normal tissue. Overall, we suggest our protocol as a complementary and more scalable approach compared with single-cell RNA sequencing to investigate associations between clinical parameters and transcriptomic alterations of specific cell populations in esophageal adenocarcinoma

Observation of a first ντ\nu_\tau candidate in the OPERA experiment in the CNGS beam

The OPERA neutrino detector in the underground Gran Sasso Laboratory (LNGS) has been designed to perform the first detection of neutrino oscillations in direct appearance mode through the study of the $\nu_\mu\rightarrow\nu_\tau$ channel. The hybrid apparatus consists of an emulsion/lead target complemented by electronic detectors and it is placed in the high energy long-baseline CERN to LNGS beam (CNGS) 730 km away from the neutrino source. Runs with CNGS neutrinos were successfully carried out in 2008 and 2009. After a brief description of the beam, the experimental setup and the procedures used for the analysis of the neutrino events, we describe the topology and kinematics of a first candidate $\nu_\tau$ charged-current event satisfying the kinematical selection criteria. The background calculations and their cross-check are explained in detail and the significance of the event is assessed.Comment: 19 pages, 3 figure

A Comparison of Effects of Ambient Pressure on the Atomization Performance of Soybean Oil Methyl Ester and Dimethyl Ether Sprays

The purpose of this study is the experimental investigation of Soybean oil Methyl Ester (SME) and DiMethyl Ether (DME) spray characteristics injected through the common-rail injection system under various ambient pressures. A high pressure chamber that can be pressurized up to 4 MPa was utilized for a change of ambient pressure. In order to compare the spray development and atomization characteristics, the images of SME and DME were obtained by using a high speed camera with two metal halide lamps under various ambient pressures in the spray chamber. From these spray images, the spray characteristics such as the spray penetration from the nozzle tip, maximum radial distance, and spray diameter were measured and analyzed. In addition, the Sauter Mean Diameter (SMD) of two fuels under ambient pressure was analyzed using the droplet measuring system. It was revealed that the axial distance of spray from the nozzle tip of the SME spray is longer than that of DME spray under same injection condition. The axial penetration, maximum radial distance, and spray diameter decreased when the ambient pressure in the chamber increased. As the ambient pressure increased, the SMD decreased and the DME spray showed a superior atomization performance compared to the SME spray

Multifunctional porous microspheres encapsulating oncolytic bacterial spores and their potential for cancer immunotherapy

Clostridium novyi-NT (C. novyi-NT) is an anaerobic bacterium that can be used for targeted cancer therapy because it germinates selectively in the hypoxic regions of tumor tissues. However, systemic administration of C. novyi-NT spores cannot effectively treat tumors because of the limited intratumoral delivery of active spores. In this study, we demonstrated that multifunctional porous microspheres (MPMs) containing C. novyi-NT spores have the potential for image-guided local tumor therapy. The MPMs can be repositioned under an external magnetic field, enabling precise tumor targeting and retention. Polylactic acid-based MPMs were prepared using the oil-in-water emulsion technique and then coated with a cationic polyethyleneimine polymer prior to loading with negatively charged C. novyi-NT spores. The C. novyi-NT spores delivered by MPMs were released and germinated in a simulated tumor microenvironment, effectively secreting proteins cytotoxic to tumor cells. In addition, the germinated C. novyi-NT induced immunogenic death of the tumor cells and M1 polarization of macrophages. These results indicate that MPMs encapsulated with C. novyi-NT spores have great potential for image-guided cancer immunotherapy. © 2023 The Royal Society of Chemistry.11Nsciescopu