Single-cell spatial landscapes of the lung tumour immune microenvironment.

Single-cell technologies have revealed the complexity of the tumour immune microenvironment with unparalleled resolution1-9. Most clinical strategies rely on histopathological stratification of tumour subtypes, yet the spatial context of single-cell phenotypes within these stratified subgroups is poorly understood. Here we apply imaging mass cytometry to characterize the tumour and immunological landscape of samples from 416 patients with lung adenocarcinoma across five histological patterns. We resolve more than 1.6 million cells, enabling spatial analysis of immune lineages and activation states with distinct clinical correlates, including survival. Using deep learning, we can predict with high accuracy those patients who will progress after surgery using a single 1-mm2 tumour core, which could be informative for clinical management following surgical resection. Our dataset represents a valuable resource for the non-small cell lung cancer research community and exemplifies the utility of spatial resolution within single-cell analyses. This study also highlights how artificial intelligence can improve our understanding of microenvironmental features that underlie cancer progression and may influence future clinical practice.

Microglia are involved in phagocytosis and extracellular digestion during Zika virus encephalitis in young adult immunodeficient mice.

BACKGROUND: Zika virus (ZIKV) has been associated with several neurological complications in adult patients. METHODS: We used a mouse model deficient in TRIF and IPS-1 adaptor proteins, which are involved in type I interferon production, to study the role of microglia during brain infection by ZIKV. Young adult mice were infected intravenously with the contemporary ZIKV strain PRVABC59 (1 × 105 PFUs/100 µL). RESULTS: Infected mice did not present overt clinical signs of the disease nor body weight loss compared with noninfected animals. However, mice exhibited a viremia and a brain viral load that were maximal (1.3 × 105 genome copies/mL and 9.8 × 107 genome copies/g of brain) on days 3 and 7 post-infection (p.i.), respectively. Immunohistochemistry analysis showed that ZIKV antigens were distributed in several regions of the brain, especially the dorsal hippocampus. The number of Iba1+/TMEM119+ microglia remained similar in infected versus noninfected mice, but their cell body and arborization areas significantly increased in the stratum radiatum and stratum lacunosum-moleculare layers of the dorsal hippocampus cornu ammoni (CA)1, indicating a reactive state. Ultrastructural analyses also revealed that microglia displayed increased phagocytic activities and extracellular digestion of degraded elements during infection. Mice pharmacologically depleted in microglia with PLX5622 presented a higher brain viral load compared to untreated group (2.8 × 1010 versus 8.5 × 108 genome copies/g of brain on day 10 p.i.) as well as an increased number of ZIKV antigens labeled with immunogold in the cytoplasm and endoplasmic reticulum of neurons and astrocytes indicating an enhanced viral replication. Furthermore, endosomes of astrocytes contained nanogold particles together with digested materials, suggesting a compensatory phagocytic activity upon microglial depletion. CONCLUSIONS: These results indicate that microglia are involved in the control of ZIKV replication and/or its elimination in the brain. After depletion of microglia, the removal of ZIKV-infected cells by phagocytosis could be partly compensated by astrocytes.

Droplet Digital PCR and Immunohistochemistry Techniques to Detect Zika Virus in the Central Nervous System of Mice.

Detection of Zika virus (ZIKV) in the central nervous system (CNS) is a critical step when studying the pathogenesis of the infection in animal models. Both viral load determination and immunohistochemistry (IHC) staining are useful methods to quantitatively and qualitatively characterize viral infections in target tissues. Here, we describe viral RNA load determination by droplet digital PCR as well as protein detection by polymer-based IHC as effective techniques to quantify and localize ZIKV in the CNS of mice.

Impact of a patient safety curriculum for nurse anesthesia students.

Patient safety has become an important aspect of national health care initiatives. The purpose of this evaluation was to measure the impact of a patient safety education series for students enrolled in a nurse anesthesia program. Baseline surveys that measured patient safety competencies across three domains, attitudes, skills and knowledge, were administered to the students. A patient safety education series was delivered to the cohort and the survey was then readministered. Mean scores were compared using independent samples t tests. Attitude scores did not change from baseline to posttest. Participants scored higher on posttest means for both the patient safety skills and knowledge domains. Incorporating patient safety content into the nurse anesthesia master's degree curriculum may enhance clinicians' skills and knowledge related to patient safety, and the addition of a patient safety curriculum is important during the formative education process.

Comprehensive Assessment of PD-L1 Staining Heterogeneity in Pulmonary Adenocarcinomas Using Tissue Microarrays: Impact of the Architecture Pattern and the Number of Cores.

Checkpoint inhibitors directed against programmed death receptor 1 (PD-1) and its ligand (PD-L1) changed the treatment of advanced lung non-small cell carcinomas. The decision to treat patients is influenced by PD-L1 expression by tumor cells, but evidence indicates that this staining is heterogenous within a tumor. As PD-L1 staining is tested mostly on biopsies, false negative results can occur due to sampling issues. The clinical impact of this heterogeneity has not been established. We selected 241 patients who underwent pulmonary resection for adenocarcinoma. Tissue microarrays were constructed with five 1 mm cores representative of the histologic patterns observed in each tumor and stained for PD-L1. For each core, the histologic pattern and the percentage of PD-L1 positive tumor cells were noted. Staining heterogeneity was defined as cases with both positive and negative cores at positivity thresholds of 1%, 10%, and 50% of tumor cells. At the 50% cut-off, 37.8% of patients were PD-L1 positive, whereas 22.4% showed staining heterogeneity. Among patients with 1 negative core, 26.5% also had a positive core and could have been misclassified based on 1 biopsy. Mean staining of PD-L1 was higher in solid (47.9%) and micropapillary (24.2%) patterns and was lower in acinar (14.1%), papillary (3.4%), and lepidic (6.4%) architectures. A significant proportion of patients presented a heterogenous staining for PD-L1. A total of 26.5% of patients negative on 1 core turned out to be positive on another core, which raises the consideration of rebiopsy, in particular when lepidic, acinar, or papillary patterns are observed on a biopsy.

Refocusing research priorities in schools of nursing.

It is critical for schools of nursing to periodically reassess their scholarly programs to ensure that their conceptual framework and approaches address current challenges and enhance productivity. This article describes the process undertaken at Columbia University School of Nursing to evaluate scholarly enterprise so that it remains relevant and responsive to changing trends and to revise our research conceptual model to be reflective of the foci of our clinicians and researchers. As part of a larger strategic initiative, a two-phase Research Excellence Planning and Implementation Workgroup was convened, consisting of a broad representation of faculty and administrative staff, with an overall goal of expanding scholarly capacity. During Phase I, members developed measurable outcomes and tactics and revised the school's conceptual research model. In Phase II, the workgroup implemented and monitored tactics and presented final recommendations to the dean. To measure progress, faculty members completed a survey to establish baseline scholarship and collaboration with results indicating room for growth in interdisciplinary and inter-institutional collaboration. Ongoing assessment of outcomes includes Web-based tracking of scholarly activities and follow-up surveys to monitor expansion of faculty collaboration. We recommend this process to other schools committed to sustainable, increasingly relevant scholarship.