Spatial atlas of the mouse central nervous system at molecular resolution.

Spatially charting molecular cell types at single-cell resolution across the 3D volume is critical for illustrating the molecular basis of brain anatomy and functions. Single-cell RNA sequencing has profiled molecular cell types in the mouse brain1,2, but cannot capture their spatial organization. Here we used an in situ sequencing method, STARmap PLUS3,4, to profile 1,022 genes in 3D at a voxel size of 194 × 194 × 345 nm3, mapping 1.09 million high-quality cells across the adult mouse brain and spinal cord. We developed computational pipelines to segment, cluster and annotate 230 molecular cell types by single-cell gene expression and 106 molecular tissue regions by spatial niche gene expression. Joint analysis of molecular cell types and molecular tissue regions enabled a systematic molecular spatial cell-type nomenclature and identification of tissue architectures that were undefined in established brain anatomy. To create a transcriptome-wide spatial atlas, we integrated STARmap PLUS measurements with a published single-cell RNA-sequencing atlas1, imputing single-cell expression profiles of 11,844 genes. Finally, we delineated viral tropisms of a brain-wide transgene delivery tool, AAV-PHP.eB5,6. Together, this annotated dataset provides a single-cell resource that integrates the molecular spatial atlas, brain anatomy and the accessibility to genetic manipulation of the mammalian central nervous system.

Identifying the Physical and Mental Healthcare Needs of Opioid Treatment Program Clients.

Background: Individuals with opioid use disorder (OUD) often have significant medical and behavioral health needs that are unaddressed. Opioid treatment programs (OTP) are uniquely positioned to provide integrated services for OUD, physical and mental health but are underutilized for this purpose. This study aims to describe the physical and mental healthcare needs of OTP clients in order to inform integrated care implementation in OTPs. Method: OTP clients (n = 1261) in an integrated care program in the Bronx borough of New York City were assessed for mental health symptoms (e.g., anxiety, depression), chronic disease indicators (e.g., blood pressure, cholesterol), and general functioning (e.g., capability of managing healthcare needs). Results: Symptoms of anxiety, post-traumatic stress, and depression were common. Self-reported health status and level of functioning were generally poor. Heavy smoking and obesity were the most frequent physical health risks. Other chronic disease indicators (e.g., blood pressure) showed 25-46% may be at risk. Sixty percent had multiple mental health risks and 85% had multiple physical health risks. Older clients had a higher rate of hypertension and diabetes risk than younger clients. Conclusions: Integrated care programs in OTPs must be prepared to address and coordinate care for chronic mental and physical health conditions in addition to OUD.

Revised guidelines for the endodontic education of dentistry students in Australia and New Zealand (FEBRUARY 2021).

These revised guidelines for contemporary endodontic education in Australia and New Zealand (version 2021) propose the minimum criteria for the training of dentistry students. The document contains a definition of endodontics and a description of the scope of endodontics. It proposes a general outline for education programmes in endodontics as part of general dental practice.

Revised guidelines for educational requirements for specialisation in endodontics in Australia and New Zealand (July 2020).

These revised guidelines developed by the Australian Society of Endodontology Inc. and the Australian and New Zealand Academy of Endodontists for educational requirements for specialisation in endodontics in Australia and New Zealand (version 2020) propose minimum criteria for training specialists in our field. The document contains a definition of endodontics and a description of the scope of endodontics. It proposes criteria for selection of the students and describes the proposed main features and a general outline of the education programme.

Behavior of mineral trioxide aggregate apical plugs and root fillings under cyclic loading.

AIM: The aim of the present study was to develop a methodology for constant monitoring of the resistance to fluid flow during functional loading and to then compare the efficacy of 4-mm mineral trioxide aggregate (MTA) apical plugs and full-length MTA root fillings. METHODS: The root canals of 24 single-rooted teeth were instrumented and assigned into two groups: group 1 (filled with a 4-mm MTA apical plug and gutta percha) and group 2 (filled with MTA to cementoenamel junction level). All teeth were subjected to stepwise cyclic loading with a constant simulated interstitial pressure of 25 cm H2 O applied apically. A Flodec machine (De Marco Engineering, Geneva, Switzerland) constantly monitored the rate and direction of fluid flow. Student's t-test, Kruskal-Wallis test, Fisher's exact test, and Newcombe-Wilson test were used for statistical analysis, with P < 0.05. RESULTS: Crown fractures occurred in 21 teeth with no change in the rate of fluid flow. Three teeth (filled with full-length MTA) showed sudden change in the rate of fluid flow through the root canal after cyclic loading. CONCLUSIONS: The methodology reliably monitored fluid flow during cyclic loading. There was no difference between a 4-mm MTA apical plug and full-length MTA root filling after cyclic loading using a dynamic fluid-flow monitoring technique.

Characterization of Wnt-1 and Wnt-2 induced growth alterations and signaling pathways in NIH3T3 fibroblasts.

Members of the Wnt family induce mouse mammary tumors and partially transform mammary epithelial cells in culture. However, their mechanism of transformation remains to be elucidated. In NIH3T3 mouse embryo fibroblasts, a standard transformation model, Wnt-1 and Wnt-2 were shown to induce altered properties including increased saturation density and growth in soft agar. Such cells also exhibited increased cell-cell adhesiveness. However, unlike oncogenes such as PDGFB or ras, Wnt-1 and -2 failed to induce detectable transformed foci following transfection, and stable NIH3T3 transfectants lacked tumor forming capacity. Wnt-1 and -2 transfectants exhibited increased uncomplexed, cytosolic beta-catenin, which was not observed with PDGFB, ras or erbB2 transfectants. In transient transfection, Wnt-1 and -2 induced a rapid increase in cytosolic beta-catenin but no detectable increase in the phosphorylated activated forms of MAP kinase. In contrast, ras was a potent activator of MAP kinase but had no effect on free beta-catenin levels. These findings establish that both Wnt signaling and pattern of growth alterations differ from those of oncogenes which activate proliferative signaling pathways in NIH3T3 cells.