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Conventional type 1 dendritic cells (cDC1s) are critical for innate sensing of cancer, yet they are scarce in the tumor microenvironment (TME). Here, we present a protocol to identify and isolate cDC1 subsets from murine implantable tumors for subsequent transcriptomic profiling using a flow sorting-based strategy. We describe steps for cell culture of mouse tumors, tumoral growth, dissociation and isolation of tumoral cells, extracellular staining, and cell sorting. We then detail procedures for RNA isolation, mRNA library preparation, and sequencing. For complete details on the use and execution of this protocol, please refer to Papadas et al.1.

We describe the survey design and science goals for One-hundred-deg2 DECam Imaging in Narrowbands (ODIN), a NOIRLab survey using the Dark Energy Camera (DECam) to obtain deep (AB ∼ 25.7) narrowband images over an unprecedented area of sky. The three custom-built narrowband filters, N419, N501, and N673, have central wavelengths of 419, 501, and 673 nm and respective FWHM of 7.5, 7.6, and 10.0 nm, corresponding to Lyα at z = 2.4, 3.1, and 4.5 and cosmic times of 2.8, 2.1, and 1.4 Gyr, respectively. When combined with even deeper, public broadband data from the Hyper Suprime-Cam, DECam, and in the future, the Legacy Survey of Space and Time, the ODIN narrowband images will enable the selection of over 100,000 Lyα-emitting (LAE) galaxies at these epochs. ODIN-selected LAEs will identify protoclusters as galaxy overdensities, and the deep narrowband images enable detection of highly extended Lyα blobs (LABs). Primary science goals include measuring the clustering strength and dark matter halo connection of LAEs, LABs, and protoclusters, and their respective relationship to filaments in the cosmic web. The three epochs allow for the redshift evolution of these properties to be determined during the period known as Cosmic Noon, where star formation was at its peak. The narrowband filter wavelengths are designed to enable interloper rejection and further scientific studies by revealing [O ii] and [O iii] at z = 0.34, Lyα and He ii 1640 at z = 3.1, and Lyman continuum plus Lyα at z = 4.5. Ancillary science includes similar studies of the lower-redshift emission-line galaxy samples and investigations of nearby star-forming galaxies resolved into numerous [O iii] and [S ii] emitting regions.

Advances in immunotherapy in the last decade have revolutionized treatment paradigms across multiple cancer diagnoses. However, only a minority of patients derive durable benefit and progress with traditional approaches, such as cancer vaccines, remains unsatisfactory. A key to overcoming these barriers resides with a deeper understanding of tumor antigen presentation and the complex and dynamic heterogeneity of tumor-infiltrating antigen-presenting cells (APCs). Reminiscent of the 'second touch' hypothesis proposed by Klaus Ley for CD4+ T cell differentiation, the acquisition of full effector potential by lymph node- primed CD8+ T cells requires a second round of co-stimulation at the site where the antigen originated, i.e. the tumor bed. The tumor stroma holds a prime role in this process by hosting specialized APC niches, apparently distinct from tertiary lymphoid structures, that support second antigenic touch encounters and CD8+ T cell effector proliferation and differentiation. We propose that APC within second-touch niches become licensed for co-stimulation through stromal-derived instructive signals emulating embryonic or wound-healing provisional matrix remodeling. These immunostimulatory roles of stroma contrast with its widely accepted view as a physical and functional 'immune barrier'. Stromal control of antigen presentation makes evolutionary sense as the host stroma-tumor interface constitutes the prime line of homeostatic 'defense' against the emerging tumor. In this review, we outline how stroma-derived signals and cells regulate tumor antigen presentation and T-cell effector differentiation in the tumor bed. The re-definition of tumor stroma as immune rheostat rather than as inflexible immune barrier harbors significant untapped therapeutic opportunity.

BACKGROUND: Golgi apparatus (GA) is assembled as a crescent-like ribbon in mammalian cells under immunofluorescence microscope without knowing the shaping mechanisms. It is estimated that roughly 1/5 of the genes encoding kinases or phosphatases in human genome participate in the assembly of Golgi ribbon, reflecting protein modifications play major roles in building Golgi ribbon. METHODS: To explore how Golgi ribbon is shaped as a crescent-like structure under the guidance of protein modifications, we identified a protein complex containing the scaffold proteins Ajuba, two known GA regulators including the protein kinase Aurora-A and the protein arginine methyltransferase PRMT5, and the common substrate of Aurora-A and PRMT5, HURP. Mutual modifications and activation of PRMT5 and Aurora-A in the complex leads to methylation and in turn phosphorylation of HURP, thereby producing HURP p725. The HURP p725 localizes to GA vicinity and its distribution pattern looks like GA morphology. Correlation study of the HURP p725 statuses and GA structure, site-directed mutagenesis and knockdown-rescue experiments were employed to identify the modified HURP as a key regulator assembling GA as a crescent ribbon. RESULTS: The cells containing no or extended distribution of HURP p725 have dispersed GA membranes or longer GA. Knockdown of HURP fragmentized GA and HURP wild type could, while its phosphorylation deficiency mutant 725A could not, restore crescent Golgi ribbon in HURP depleted cells, collectively indicating a crescent GA-constructing activity of HURP p725. HURP p725 is transported, by GA membrane-associated ARF1, Dynein and its cargo adaptor Golgin-160, to cell center where HURP p725 forms crescent fibers, binds and stabilizes Golgi assembly factors (GAFs) including TRIP11, GRASP65 and GM130, thereby dictating the formation of crescent Golgi ribbon at nuclear periphery. CONCLUSIONS: The Ajuba/PRMT5/Aurora-A complex integrates the signals of protein methylation and phosphorylation to HURP, and the HURP p725 organizes GA by stabilizing and recruiting GAFs to its crescent-like structure, therefore shaping GA as a crescent ribbon. Therefore, the HURP p725 fiber serves a template to construct GA according to its shape. Video Abstract.

En este estudio se tratan los aspectos culturales, en concreto, los que están relacionados con la cultura española y la cultura china. En primer lugar, tratamos aspectos culturales comunes o diferenciadores entre España y Taiwán según el Modelo de las seis dimensiones elaborado por Hofstede. En segundo lugar, analizamos las aportaciones realizadas por Hall respecto del Modelo contextual. Los resultados de este estudio tienen importantes implicaciones didácticas que podrían servir de guía para la enseñanza de ELE, y también, esperamos que la aplicación de las citadas teorías a diversos campos sirva de ayuda a los interesados.

Background: Skill integration is vital in students' mastery development and is especially prominent in developing code tracing skills which are foundational to programming, an increasingly important area in the current STEM education. However, instructional design to support skill integration in learning technologies has been limited. Objectives: The current work presents the development and empirical evaluation of instructional design targeting students' difficulties in code tracing particularly in integrating component skills in the Trace Table Tutor (T3), an intelligent tutoring system. Methods: Beyond the instructional features of active learning, step-level support, and individualized problem selection of intelligent tutoring systems (ITS), the instructional design of T3 (e.g., hints, problem types, problem selection) was optimized to target skill integration based on a domain model where integrative skills were represented as combinations of component skills. We conducted an experimental study in a university-level introductory Python programming course and obtained three findings. Results and Conclusions: First, the instructional features of the ITS technology support effective learning of code tracing, as evidenced by significant learning gains (medium-to-large effect sizes). Second, performance data supports the existence of integrative skills beyond component skills. Third, an instructional design focused on integrative skills yields learning benefits beyond a design without such focus, such as improving performance efficiency (medium-to-large effect sizes). Major Takeaways: Our work demonstrates the value of designing for skill integration in learning technologies and the effectiveness of the ITS technology for computing education, as well as provides general implications for designing learning technologies to foster robust learning.