Cilia Proteins are Biomarkers of Altered Flow in the Vasculature

Cilia, microtubule-based organelles that project from the apical luminal surface of endothelial cells (ECs), are widely regarded as low-flow sensors. Previous reports suggest that upon high shear stress, cilia on the EC surface are lost, and more recent evidence suggests that deciliation—the physical removal of cilia from the cell surface—is a predominant mechanism for cilia loss in mammalian cells. Thus, we hypothesized that EC deciliation facilitated by changes in shear stress would manifest in increased abundance of cilia-related proteins in circulation. To test this hypothesis, we performed shear stress experiments that mimicked flow conditions from low to high shear stress in human primary cells and a zebrafish model system. In the primary cells, we showed that upon shear stress induction, indeed, ciliary fragments were observed in the effluent in vitro, and effluents contained ciliary proteins normally expressed in both endothelial and epithelial cells. In zebrafish, upon shear stress induction, fewer cilia-expressing ECs were observed. To test the translational relevance of these findings, we investigated our hypothesis using patient blood samples from sickle cell disease and found that plasma levels of ciliary proteins were elevated compared with healthy controls. Further, sickled red blood cells demonstrated high levels of ciliary protein (ARL13b) on their surface after adhesion to brain ECs. Brain ECs postinteraction with sickle RBCs showed high reactive oxygen species (ROS) levels. Attenuating ROS levels in brain ECs decreased cilia protein levels on RBCs and rescued ciliary protein levels in brain ECs. Collectively, these data suggest that cilia and ciliary proteins in circulation are detectable under various altered-flow conditions, which could serve as a surrogate biomarker of the damaged endothelium

Cilia proteins are biomarkers of altered flow in the vasculature

Cilia, microtubule-based organelles that project from the apical luminal surface of endothelial cells (ECs), are widely regarded as low-flow sensors. Previous reports suggest that upon high shear stress, cilia on the EC surface are lost, and more recent evidence suggests that deciliation- the physical removal of cilia from the cell surface-is a predominant mechanism for cilia loss in mammalian cells. Thus, we hypothesized that EC deciliation facilitated by changes in shear stress would manifest in increased abundance of cilia-related proteins in circulation. To test this hypothesis, we performed shear stress experiments that mimicked flow conditions from low to high shear stress in human primary cells and a zebrafish model system. In the primary cells, we showed that upon shear stress induction, indeed, ciliary fragments were observed in the effluent in vitro, and effluents contained ciliary proteins normally expressed in both endothelial and epithelial cells. In zebrafish, upon shear stress induction, fewer cilia-expressing ECs were observed. To test the translational relevance of these findings, we investigated our hypothesis using patient blood samples from sickle cell disease and found that plasma levels of ciliary proteins were elevated compared with healthy controls. Further, sickled red blood cells demonstrated high levels of ciliary protein (ARL13b) on their surface after adhesion to brain ECs. Brain ECs postinteraction with sickle RBCs showed high reactive oxygen species (ROS) levels. Attenuating ROS levels in brain ECs decreased cilia protein levels on RBCs and rescued ciliary protein levels in brain ECs. Collectively, these data suggest that cilia and ciliary proteins in circulation are detectable under various altered-flow conditions, which could serve as a surrogate biomarker of the damaged endothelium.This work was funded by Qatar National Research Fund, National Priority Research Program (NPRP 10-0123-170222 to HCY). ADS is supported by funds from the Department of Pediatrics, Herma Heart Institute, the National Center for Research Resources, and the National Center for Advancing Translational Sciences, NIH (UL1TR001436)

A framework linking ecosystem services and human well‐being: Saltmarsh as a case study

1. The ecosystem services approach is based on the interdependencies between nature and human well‐being. However, while the ecosystem services aspect of this approach is well‐developed, the human well‐being aspect remains unstructured and vaguely defined. 2. An integrated conceptual framework was developed by adapting and linking the UK National Ecosystem Assessment‐Follow On framework with human well‐being domains. 3. As well as benefits, the notion of disbenefits was incorporated to recognise the potentially detrimental effects from interacting with nature. Benefits and disbenefits occur at the social–ecological interface and are classified by the seven domains of human well‐being they affect. 4. The framework is applied to saltmarsh habitat as a case study, highlighting knowledge gaps and the potential applicability and usefulness of the framework. In saltmarsh, benefits mainly accrue at larger scales with a greater impact affecting local to global individuals, while disbenefits tend to occur at a smaller scale and impact in‐situ individuals. 5. The framework provides in‐depth insight into links, trade‐offs and dichotomies between benefits and disbenefits and human well‐being, and improves accessibility to the complex research area of human well‐being. 6. This research can be a useful tool to guide environmental and health policy and management, as well as stakeholder engagement

Salve Regina Arboretum Ten Year Plan to Reach Level III Accreditation

The Salve Regina University Arboretum, located in Newport, Rhode Island is currently registered as a Level II arboretum and is intertwined with the city of Newport Arboretum. The university now has intentions to reach Level III status, as part of a ten-year plan. This plan was developed by the students of the Spring 2018 BIO 255: Conservation Biology course, instructed by Dr. Jameson Chace, Associate Professor of biology at Salve Regina University. As part of a curriculum geared towards civic engagement, the class focused on creating and optimizing strategies that can be applied to the ten-year plan. These strategies were applied to the plan categorically: a team to inventory the current tree collection; a team to develop formal educational programming; a team for informal educational programming; a team to establish goals for conservation initiative related to the arboretum; a team dedicated to research related to arboreta; and a team to develop a list of species of special interest to add to the arboretum in the coming years. In the following document, each team’s strategies for the ten-year plan are outlined. Each of the components of this plan incorporate means to fulfill the conditions to meet Level III arboretum status so that the arboretum can apply for official registration. The aforementioned teams were tasked with designing a foundation on which to work up from. This includes formal educational programming to be applied to classroom settings and informal educational programming which can be applied to community outreach-based settings. The teams that worked to strengthen the arboretum’s mission of conservation focused on researching trees that can fit into the current landscape while providing some sort of benefit to the surrounding flora/fauna. Further, many of the species of interest, such as the chestnut, hold historical value to the greater Rhode Island region. In all, the Salve Regina Arboretum must achieve a total of 500 unique species of trees and woody plants as part of its efforts to apply for Level III status. In addition to the programming and research performed so far by the student teams, the arboretum must also hire a curator to manage the programming and to oversee the arboretum as a whole. Additionally, the arboretum must continue to actively collaborate with other arboreta and should encourage scientific research. It is important to recognize that the Salve Regina University Arboretum has already been utilized in the field of microbiology and has gained some attention at the university as a resource for further research and investigation. This ten year plan, along with resources within in it, is designed to provide a list of potential guidelines and ideas that can be applied for the arboretum’s benefit and growth. The Salve Regina University arboretum is a continually growing and developing part of the greater Newport, Rhode Island community, and will continue to strengthen its mission and that of the university which oversees its success.https://digitalcommons.salve.edu/bio255_arboretum/1000/thumbnail.jp

Genomic Analysis of the Hydrocarbon-Producing, Cellulolytic, Endophytic Fungus Ascocoryne sarcoides

The microbial conversion of solid cellulosic biomass to liquid biofuels may provide a renewable energy source for transportation fuels. Endophytes represent a promising group of organisms, as they are a mostly untapped reservoir of metabolic diversity. They are often able to degrade cellulose, and they can produce an extraordinary diversity of metabolites. The filamentous fungal endophyte Ascocoryne sarcoides was shown to produce potential-biofuel metabolites when grown on a cellulose-based medium; however, the genetic pathways needed for this production are unknown and the lack of genetic tools makes traditional reverse genetics difficult. We present the genomic characterization of A. sarcoides and use transcriptomic and metabolomic data to describe the genes involved in cellulose degradation and to provide hypotheses for the biofuel production pathways. In total, almost 80 biosynthetic clusters were identified, including several previously found only in plants. Additionally, many transcriptionally active regions outside of genes showed condition-specific expression, offering more evidence for the role of long non-coding RNA in gene regulation. This is one of the highest quality fungal genomes and, to our knowledge, the only thoroughly annotated and transcriptionally profiled fungal endophyte genome currently available. The analyses and datasets contribute to the study of cellulose degradation and biofuel production and provide the genomic foundation for the study of a model endophyte system

Heterozygosity for Pten Promotes Tumorigenesis in a Mouse Model of Medulloblastoma

BACKGROUND: Recent publications have described an important role for cross talk between PI-3 kinase and sonic hedgehog signaling pathways in the pathogenesis of medulloblastoma. METHODOLOGY/PRINCIPAL FINDINGS: We crossed mice with constitutive activation of Smoothened, SmoA1, with Pten deficient mice. Both constitutive and conditional Pten deficiency doubled the incidence of mice with symptoms of medulloblastoma and resulted in decreased survival. Analysis revealed a clear separation of gene signatures, with up-regulation of genes in the PI-3 kinase signaling pathway, including downstream activation of angiogenesis in SmoA1+/-; Pten +/- medulloblastomas. Western blotting and immunohistochemistry confirmed reduced or absent Pten, Akt activation, and increased angiogenesis in Pten deficient tumors. Down-regulated genes included genes in the sonic hedgehog pathway and tumor suppressor genes. SmoA1+/-; Pten +/+ medulloblastomas appeared classic in histology with increased proliferation and diffuse staining for apoptosis. In contrast, Pten deficient tumors exhibited extensive nodularity with neuronal differentiation separated by focal areas of intense staining for proliferation and virtually absent apoptosis. Examination of human medulloblastomas revealed low to absent PTEN expression in over half of the tumors. Kaplan-Meier analysis confirmed worse overall survival in patients whose tumor exhibited low to absent PTEN expression. CONCLUSIONS/SIGNIFICANCE: This suggests that PTEN expression is a marker of favorable prognosis and mouse models with activation of PI-3 kinase pathways may be important tools for preclinical evaluation of promising agents for the treatment of medulloblastoma

Demographic, clinical, and service-use characteristics related to the clinician’s recommendation to transition from child to adult mental health services

Purpose: The service configuration with distinct child and adolescent mental health services (CAMHS) and adult mental health services (AMHS) may be a barrier to continuity of care. Because of a lack of transition policy, CAMHS clinicians have to decide whether and when a young person should transition to AMHS. This study describes which characteristics are associated with the clinicians’ advice to continue treatment at AMHS. Methods: Demographic, family, clinical, treatment, and service-use characteristics of the MILESTONE cohort of 763 young people from 39 CAMHS in Europe were assessed using multi-informant and standardized assessment tools. Logistic mixed models were fitted to assess the relationship between these characteristics and clinicians’ transition recommendations. Results: Young people with higher clinician-rated severity of psychopathology scores, with self- and parent-reported need for ongoing treatment, with lower everyday functional skills and without self-reported psychotic experiences were more likely to be recommended to continue treatment. Among those who had been recommended to continue treatment, young people who used psychotropic medication, who had been in CAMHS for more than a year, and for whom appropriate AMHS were available were more likely to be recommended to continue treatment at AMHS. Young people whose parents indicated a need for ongoing treatment were more likely to be recommended to stay in CAMHS. Conclusion: Although the decision regarding continuity of treatment was mostly determined by a small set of clinical characteristics, the recommendation to continue treatment at AMHS was mostly affected by service-use related characteristics, such as the availability of appropriate services

Germline TET2 loss of function causes childhood immunodeficiency and lymphoma

Molecular dissection of inborn errors of immunity can help to elucidate the nonredundant functions of individual genes. We studied 3 children with an immune dysregulation syndrome of susceptibility to infection, lymphadenopathy, hepatosplenomegaly, developmental delay, autoimmunity, and lymphoma of B-cell (n = 2) or T-cell (n = 1) origin. All 3 showed early autologous T-cell reconstitution following allogeneic hematopoietic stem cell transplantation. By whole-exome sequencing, we identified rare homozygous germline missense or nonsense variants in a known epigenetic regulator of gene expression: ten-eleven translocation methylcytosine dioxygenase 2 (TET2). Mutated TET2 protein was absent or enzymatically defective for 5-hydroxymethylating activity, resulting in whole-blood DNA hypermethylation. Circulating T cells showed an abnormal immunophenotype including expanded double-negative, but depleted follicular helper, T-cell compartments and impaired Fas-dependent apoptosis in 2 of 3 patients. Moreover, TET2-deficient B cells showed defective class-switch recombination. The hematopoietic potential of patient-derived induced pluripotent stem cells was skewed toward the myeloid lineage. These are the first reported cases of autosomal-recessive germline TET2 deficiency in humans, causing clinically significant immunodeficiency and an autoimmune lymphoproliferative syndrome with marked predisposition to lymphoma. This disease phenotype demonstrates the broad role of TET2 within the human immune system