Generation of Single-Cell Suspensions from Mouse Neural Tissue

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a repertoire of cell surface molecules, or molecular determinants, through which it can be identified and characterized. Currently, robust cell identification and separation technologies require single-cell preparations to be generated while simultaneously limiting cell death and destruction of characteristic surface protein. The gentleMACS Dissociator, when used in combination with trypsin or papain-based dissociation kits, can effectively and gently dissociate brain tissue while preserving antigen epitopes and limiting cell loss. Standardized preparation of single-cell suspensions is achieved using C Tubes and optimized, preset gentleMACS Programs. Once generated, single-cell suspensions can be treated with monoclonal conjugates like Anti-Prominin-1 MicroBeads, which identify neural progenitors, or purified further using Myelin Removal Beads

The Social and Cultural Context of Coping with Sickle Cell Disease: III. Stress, Coping Tasks, Family Functioning, and Children’s Adjustment

Conceptions of individual and family coping with sickle cell disease (SCD) must incorporate several disease and sociocultural factors. This article proposes an integrative model and tests the relative contribution of model parameters to the prediction of social, academic, and psychological adjustment of children with SCD. The individual coping and family functioning variables most highly predictive of the child’s psychological outcomes (anxiety, depression, and positive mood) include parental psychological functioning, maturity demands made of the ill child, and the quality of relations with parents and siblings. Academic adjustment was significantly predicted by parental academic expectations and by the child’s rejection of a restrictive sick role. Competent social functioning also was predicted by the extent to which the ill child rejected the role of being sick.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/67288/2/10.1177_0095798499025003006.pd

Deliverable 1.1 review document on the management of marine areas with particular regard on concepts, objectives, frameworks and tools to implement, monitor, and evaluate spatially managed areas

The main objectives if this document were to review the existing information on spatial management of marine areas, identifying the relevant policy objectives, to identify parameters linked to the success or failure of the various Spatially Managed marine Areas (SMAs) regimes, to report on methods and tools used in monitoring and evaluation of the state of SMAs, and to identify gaps and weaknesses in the existing frameworks in relation to the implementation, monitoring, evaluation and management of SMAs. The document is naturally divided in two sections: Section 1 reviews the concepts, objectives, drivers, policy and management framework, and extraneous factors related to the design, implementation and evaluation of SMAs; Section 2 reviews the tools and methods to monitor and evaluate seabed habitats and marine populations.peer-reviewe

Human matrix metalloproteinases: An ubiquitarian class of enzymes involved in several pathological processes

Human matrix metalloproteinases (MMPs) belong to the M10 family of the MA clan of endopeptidases. They are ubiquitarian enzymes, structurally characterized by an active site where a Zn(2+) atom, coordinated by three histidines, plays the catalytic role, assisted by a glutamic acid as a general base. Various MMPs display different domain composition, which is very important for macromolecular substrates recognition. Substrate specificity is very different among MMPs, being often associated to their cellular compartmentalization and/or cellular type where they are expressed. An extensive review of the different MMPs structural and functional features is integrated with their pathological role in several types of diseases, spanning from cancer to cardiovascular diseases and to neurodegeneration. It emerges a very complex and crucial role played by these enzymes in many physiological and pathological processes

Anti-ACSA-2 defines a novel monoclonal antibody for prospective isolation of living neonatal and adult astrocytes

International audienceAstrocytes are the most abundant cell type of the central nervous system and cover a broad range of functionalities. We report here the generation of a novel monoclonal antibody, anti-astrocyte cell surface antigen-2 (Anti-ACSA-2). Flow cytometry, immunohistochemistry and immunocytochemistry revealed that Anti-ACSA-2 reacted specifically with a not yet identified glycosylated surface molecule of murine astrocytes at all developmental stages. It did not show any labeling of non-astroglial cells such as neurons, oligodendrocytes, NG2+ cells, microglia, endothelial cells, leukocytes, or erythrocytes. Co-labeling studies of GLAST and ACSA-2 showed largely overlapping expression. However, there were also notable differences in protein expression levels and frequencies of single-positive subpopulations of cells in some regions of the CNS such as cerebellum, most prominently at early postnatal stages. In the neurogenic niches, the dentate gyrus of the hippocampus and the subventricular zone (SVZ), again a general overlap with slight differences in expression levels were observed. ACSA-2 was unlike GLAST not sensitive to papain-based tissue dissociation and allowed for a highly effective, acute, specific, and prospective purification of viable astrocytes based on a new rapid sorting procedure using Anti-ACSA-2 directly coupled to superparamagnetic MicroBeads. In conclusion, ACSA-2 appears to be a new surface marker for astrocytes, radial glia, neural stem cells and bipotent glial progenitor cells which opens up the possibility of further dissecting the characteristics of astroglial subpopulations and lineages

Ecosystem-based marine spatial management: Review of concepts, policies, tools, and critical issues

Conventional sectoral management and piecemeal governance are considered less and less appropriate in pursuit of sustainable development. Ecosystem based marine spatial management (EB-MSM) is an approach that recognizes the full array of interactions within an ecosystem, including human uses, rather than considering single issues, species, or ecosystem services in isolation. Marine spatial planning and ocean zoning are emerging concepts that can support EB-MSM. EB-MSM is driven by high-level goals that managers aim to achieve through the implementation of measures. High-level goals and objectives need to be translated into more operational objectives before specific targets, limits and measures can be elaborated. Monitoring, evaluation and adaptation are necessary to ensure that marine management measures are both effective and efficient. Solid monitoring frameworks are the foundation of adaptive management, as they provide the necessary information to evaluate performance and the effectiveness of management actions. Marine protected areas (MPAs) - possibly set up in networks - constitute a key component in EB-MSM policies and practises and have been applied as a cornerstone in conservation of marine biodiversity, management of fish populations, development of coastal tourism, etc. Moreover, MPA experiences have provided methods and concepts (such as zoning) to a wider EB-MSM context. The assignment of values to biophysical features of the marine environment allows the direct assessment of related management choices and may assist EB-MSM. A range of monetary valuation techniques have been proposed to reduce attributes of goods and services to a single metric. However, in the marine environment such an approach is often over simplistic, and thus less reductive techniques may be necessary. Rather than producing a single metric, the results of non-monetary assessments guide policy allowing weight to be given as necessary to potential areas of conflict and consensus. Strategies to take into account climate change effects and geohazard risks in EB-MSM have been applied or proposed worldwide. EB-MSM regimes must be alert to such risks and flexible to account for changes