Localizing Protein in 3D Neural Stem Cell Culture: a Hybrid Visualization Methodology

The importance of 3-dimensional (3D) topography in influencing neural stem and progenitor cell (NPC) phenotype is widely acknowledged yet challenging to study. When dissociated from embryonic or post-natal brain, single NPCs will proliferate in suspension to form neurospheres. Daughter cells within these cultures spontaneously adopt distinct developmental lineages (neurons, oligodendrocytes, and astrocytes) over the course of expansion despite being exposed to the same extracellular milieu. This progression recapitulates many of the stages observed over the course of neurogenesis and gliogenesis in post-natal brain and is often used to study basic NPC biology within a controlled environment. Assessing the full impact of 3D topography and cellular positioning within these cultures on NPC fate is, however, difficult. To localize target proteins and identify NPC lineages by immunocytochemistry, free-floating neurospheres must be plated on a substrate or serially sectioned. This processing is required to ensure equivalent cell permeabilization and antibody access throughout the sphere. As a result, 2D epifluorescent images of cryosections or confocal reconstructions of 3D Z-stacks can only provide spatial information about cell position within discrete physical or digital 3D slices and do not visualize cellular position in the intact sphere. Here, to reiterate the topography of the neurosphere culture and permit spatial analysis of protein expression throughout the entire culture, we present a protocol for isolation, expansion, and serial sectioning of post-natal hippocampal neurospheres suitable for epifluorescent or confocal immunodetection of target proteins. Connexin29 (Cx29) is analyzed as an example. Next, using a hybrid of graphic editing and 3D modelling softwares rigorously applied to maintain biological detail, we describe how to re-assemble the 3D structural positioning of these images and digitally map labelled cells within the complete neurosphere. This methodology enables visualization and analysis of the cellular position of target proteins and cells throughout the entire 3D culture topography and will facilitate a more detailed analysis of the spatial relationships between cells over the course of neurogenesis and gliogenesis in vitro

Localizing Protein in 3D Neural Stem Cell Culture: a Hybrid Visualization Methodology

The importance of 3-dimensional (3D) topography in influencing neural stem and progenitor cell (NPC) phenotype is widely acknowledged yet challenging to study. When dissociated from embryonic or post-natal brain, single NPCs will proliferate in suspension to form neurospheres. Daughter cells within these cultures spontaneously adopt distinct developmental lineages (neurons, oligodendrocytes, and astrocytes) over the course of expansion despite being exposed to the same extracellular milieu. This progression recapitulates many of the stages observed over the course of neurogenesis and gliogenesis in post-natal brain and is often used to study basic NPC biology within a controlled environment. Assessing the full impact of 3D topography and cellular positioning within these cultures on NPC fate is, however, difficult. To localize target proteins and identify NPC lineages by immunocytochemistry, free-floating neurospheres must be plated on a substrate or serially sectioned. This processing is required to ensure equivalent cell permeabilization and antibody access throughout the sphere. As a result, 2D epifluorescent images of cryosections or confocal reconstructions of 3D Z-stacks can only provide spatial information about cell position within discrete physical or digital 3D slices and do not visualize cellular position in the intact sphere. Here, to reiterate the topography of the neurosphere culture and permit spatial analysis of protein expression throughout the entire culture, we present a protocol for isolation, expansion, and serial sectioning of post-natal hippocampal neurospheres suitable for epifluorescent or confocal immunodetection of target proteins. Connexin29 (Cx29) is analyzed as an example. Next, using a hybrid of graphic editing and 3D modelling softwares rigorously applied to maintain biological detail, we describe how to re-assemble the 3D structural positioning of these images and digitally map labelled cells within the complete neurosphere. This methodology enables visualization and analysis of the cellular position of target proteins and cells throughout the entire 3D culture topography and will facilitate a more detailed analysis of the spatial relationships between cells over the course of neurogenesis and gliogenesis in vitro

Factors associated with uptake of influenza vaccine in people aged 50 to 64 years in Hong Kong: a case-control study.

BACKGROUND: In Hong Kong, people aged 50-64 years were added as a recommended priority group (recommended group) for influenza vaccination by the Department of Health (DH) starting from 2011/12 onwards. The coverage rate of influenza vaccination for this age group was suboptimal at 8.5 % in 2012/13. This study investigates the factors associated with the uptake of influenza vaccination among adults in Hong Kong aged 50-64 years. METHODS: A case-control study was conducted in communities by street intercept interviews from 17 July to 15 August 2013. Cases were adults aged 50-64 years who had received the influenza vaccine in 2011/12 or 2012/13, while controls were the same as the cases, except they had not received the influenza vaccine in 2011/12 or 2012/13. Multiple logistic regression analysis was performed on the data to explore the associations between vaccination status and the variables. RESULTS: Six hundred and four respondents in total were interviewed and included in the analysis. There were 193 cases (vaccinated) and 411 controls (non-vaccinated), with a case-to-control ratio of 1:2.1. The following were strongly associated with vaccination compared to other factors: 'eligible for free government vaccine' (OR6.38, 95 % CI, 3.43-11.87, p 80 %) were willing to be vaccinated if it was free. CONCLUSIONS: Factors related to free and convenient vaccination, the perception of the severity of symptoms when contracting influenza had a comparatively strong association with influenza vaccination uptake amongst 50-64 year olds, compared to other factors

Virtual Heritage Learning Environments

. The change and restrictions in how we react with cultural heritage because of the COVID-19 pandemic has created an urgency in advancing remote and digital access to objects and sites. This paper outlines the process for developing Virtual Learning Environments (VLEs) using digital recording and modelling of architectural heritage and archaeology. Virtual Reality (VR) software, game engine platforms and WEB platforms are outlined which can be applied to represent heritage sites in addition to emerging screen based technological learning systems. The application Historic Building Information Modelling (HBIM) and Game Engine Platforms for creating Virtual Learning Environments (VLEs) is also examined. The design-theory based on Virtual Learning Objects for cultural heritage is explored. Two case studies are explored for their potential to create Virtual Heritage Learning Environments. Finally, a design framework is proposed for developing Virtual Heritage Learning Environments

Reduction of Glucocorticoid Receptor Function in Chronic Fatigue Syndrome

Glucocorticoid receptor (GR) function may have aetiopathogenic significance in chronic fatigue syndrome (CFS), via its essential role in mediating inflammatory responses as well as in hypothalamic-pituitary-adrenal axis regulation. GR function can be estimated ex vivo by measuring dexamethasone (dex) modulation of cytokine response to lipopolysaccharide (LPS), and in vivo using the impact of dex on cortisol levels. This study aimed to compare the GR function between CFS (n = 48), primary Sjögren’s syndrome (a disease group control) (n = 27), and sedentary healthy controls (HCs) (n = 20), and to investigate its relationship with clinical measures. In the GR ex vivo response assay, whole blood was diluted and incubated with LPS (to stimulate cytokine production), with or without 10 or 100 nanomolar concentrations of dex. Cytometric bead array (CBA) and flow cytometry enabled quantification of cytokine levels (TNFα, interleukin- (IL-) 6, and IL-10) in the supernatants. In the in vivo response assay, five plasma samples were taken for determination of total cortisol concentration using ELISA at half-hourly intervals on two consecutive mornings separated by ingestion of 0.5 mg of dex at 11 pm. The association of the data from the in vivo and ex vivo analyses with reported childhood adversity was also examined. CFS patients had reduced LPS-induced IL-6 and TNFα production compared to both control groups and reduced suppression of TNFα by the higher dose of dex compared to HCs. Cortisol levels, before or after dex, did not differ between CFS and HCs. Cortisol levels were more variable in CFS than HCs. In the combined group (CFS plus HC), cortisol concentrations positively and ex vivo GR function (determined by dex-mediated suppression of IL-10) negatively correlated with childhood adversity score. The results do not support the hypothesis that GR dysregulation is aetiopathogenic in CFS and suggest that current and future endocrine cross-sectional studies in CFS may be vulnerable to the confounding influence of childhood trauma which is likely increased by comorbid depression

Augmented Reality Markerless Multi-Image Outdoor Tracking System for the Historical Buildings on Parliament Hill

[EN] Augmented Reality (AR) applications have experienced extraordinary growth recently, evolving into a well-established method for the dissemination and communication of content related to cultural heritage¿including education. AR applications have been used in museums and gallery exhibitions and virtual reconstructions of historic interiors. However, the circumstances of an outdoor environment can be problematic. This paper presents a methodology to develop immersive AR applications based on the recognition of outdoor buildings. To demonstrate this methodology, a case study focused on the Parliament Buildings National Historic Site in Ottawa, Canada has been conducted. The site is currently undergoing a multiyear rehabilitation program that will make access to parts of this national monument inaccessible to the public. AR experiences, including simulated photo merging of historic and present content, are proposed as one tool that can enrich the Parliament Hill visit during the rehabilitation. Outdoor AR experiences are limited by factors, such as variable lighting (and shadows) conditions, caused by changes in the environment (objects height and orientation, obstructions, occlusions), the weather, and the time of day. This paper proposes a workflow to solve some of these issues from a multi-image tracking approach.This work has been developed under the framework of the New Paradigms/New Tools for Heritage Conservation in Canada, a project funded through the Social Sciences and Humanities Research Council of Canada (SSHRC).Blanco-Pons, S.; Carrión-Ruiz, B.; Duong, M.; Chartrand, J.; Fai, S.; Lerma, JL. (2019). Augmented Reality Markerless Multi-Image Outdoor Tracking System for the Historical Buildings on Parliament Hill. Sustainability. 11(16):1-15. https://doi.org/10.3390/su11164268S1151116Bekele, M. K., Pierdicca, R., Frontoni, E., Malinverni, E. S., & Gain, J. (2018). A Survey of Augmented, Virtual, and Mixed Reality for Cultural Heritage. Journal on Computing and Cultural Heritage, 11(2), 1-36. doi:10.1145/3145534Gimeno, J., Portalés, C., Coma, I., Fernández, M., & Martínez, B. (2017). Combining traditional and indirect augmented reality for indoor crowded environments. A case study on the Casa Batlló museum. Computers & Graphics, 69, 92-103. doi:10.1016/j.cag.2017.09.001Kolivand, H., El Rhalibi, A., Shahrizal Sunar, M., & Saba, T. (2018). ReVitAge: Realistic virtual heritage taking shadows and sky illumination into account. Journal of Cultural Heritage, 32, 166-175. doi:10.1016/j.culher.2018.01.020Amakawa, J., & Westin, J. (2017). New Philadelphia: using augmented reality to interpret slavery and reconstruction era historical sites. International Journal of Heritage Studies, 24(3), 315-331. doi:10.1080/13527258.2017.1378909Kim, J.-B., & Park, C. (2011). Development of Mobile AR Tour Application for the National Palace Museum of Korea. Lecture Notes in Computer Science, 55-60. doi:10.1007/978-3-642-22021-0_7Barrile, V., Fotia, A., Bilotta, G., & De Carlo, D. (2019). Integration of geomatics methodologies and creation of a cultural heritage app using augmented reality. Virtual Archaeology Review, 10(20), 40. doi:10.4995/var.2019.10361Analysis of Tracking Accuracy for Single-Camera Square-Marker-Based Tracking. In Third Workshop on Virtual and Augmented Reality of the GI-Fachgruppe VR/AR, Koblenz, Germany, 2006http://campar.in.tum.de/Chair/PublicationDetail?pub=pentenrieder2006giCirulis, A., & Brigmanis, K. B. (2013). 3D Outdoor Augmented Reality for Architecture and Urban Planning. Procedia Computer Science, 25, 71-79. doi:10.1016/j.procs.2013.11.009You, S., Neumann, U., & Azuma, R. (1999). Orientation tracking for outdoor augmented reality registration. IEEE Computer Graphics and Applications, 19(6), 36-42. doi:10.1109/38.799738Wither, J., Tsai, Y.-T., & Azuma, R. (2011). Indirect augmented reality. Computers & Graphics, 35(4), 810-822. doi:10.1016/j.cag.2011.04.010Radkowski, R., & Oliver, J. (2013). Natural Feature Tracking Augmented Reality for On-Site Assembly Assistance Systems. Lecture Notes in Computer Science, 281-290. doi:10.1007/978-3-642-39420-1_30Rao, J., Qiao, Y., Ren, F., Wang, J., & Du, Q. (2017). A Mobile Outdoor Augmented Reality Method Combining Deep Learning Object Detection and Spatial Relationships for Geovisualization. Sensors, 17(9), 1951. doi:10.3390/s17091951Hoppe, H., DeRose, T., Duchamp, T., McDonald, J., & Stuetzle, W. (1993). Mesh optimization. Proceedings of the 20th annual conference on Computer graphics and interactive techniques - SIGGRAPH ’93. doi:10.1145/166117.166119Rossignac, J., & Borrel, P. (1993). Multi-resolution 3D approximations for rendering complex scenes. Modeling in Computer Graphics, 455-465. doi:10.1007/978-3-642-78114-8_29Gross, M. H., Staadt, O. G., & Gatti, R. (1996). Efficient triangular surface approximations using wavelets and quadtree data structures. IEEE Transactions on Visualization and Computer Graphics, 2(2), 130-143. doi:10.1109/2945.506225Botsch, M., Pauly, M., Rossl, C., Bischoff, S., & Kobbelt, L. (2006). Geometric modeling based on triangle meshes. ACM SIGGRAPH 2006 Courses on - SIGGRAPH ’06. doi:10.1145/1185657.1185839Pietroni, N., Tarini, M., & Cignoni, P. (2010). Almost Isometric Mesh Parameterization through Abstract Domains. IEEE Transactions on Visualization and Computer Graphics, 16(4), 621-635. doi:10.1109/tvcg.2009.96Khan, D., Yan, D.-M., Ding, F., Zhuang, Y., & Zhang, X. (2018). Surface remeshing with robust user-guided segmentation. Computational Visual Media, 4(2), 113-122. doi:10.1007/s41095-018-0107-yGuidi, G., Russo, M., Ercoli, S., Remondino, F., Rizzi, A., & Menna, F. (2009). A Multi-Resolution Methodology for the 3D Modeling of Large and Complex Archeological Areas. International Journal of Architectural Computing, 7(1), 39-55. doi:10.1260/147807709788549439Remondino, F., & El-Hakim, S. (2006). Image-based 3D Modelling: A Review. The Photogrammetric Record, 21(115), 269-291. doi:10.1111/j.1477-9730.2006.00383.xBruno, F., Bruno, S., De Sensi, G., Luchi, M.-L., Mancuso, S., & Muzzupappa, M. (2010). From 3D reconstruction to virtual reality: A complete methodology for digital archaeological exhibition. Journal of Cultural Heritage, 11(1), 42-49. doi:10.1016/j.culher.2009.02.006Unity, The Photogrammetry Workflowhttps://unity.com/solutions/photogrammetry.Blanco, S., Carrión, B., & Lerma, J. L. (2016). REVIEW OF AUGMENTED REALITY AND VIRTUAL REALITY TECHNIQUES IN ROCK ART. Proceedings of the ARQUEOLÓGICA 2.0 8th International Congress on Archaeology, Computer Graphics, Cultural Heritage and Innovation. doi:10.4995/arqueologica8.2016.3561Behzadan, A. H., & Kamat, V. R. (2010). Scalable Algorithm for Resolving Incorrect Occlusion in Dynamic Augmented Reality Engineering Environments. Computer-Aided Civil and Infrastructure Engineering, 25(1), 3-19. doi:10.1111/j.1467-8667.2009.00601.xTian, Y., Long, Y., Xia, D., Yao, H., & Zhang, J. (2015). Handling occlusions in augmented reality based on 3D reconstruction method. Neurocomputing, 156, 96-104. doi:10.1016/j.neucom.2014.12.081Tian, Y., Guan, T., & Wang, C. (2010). Real-Time Occlusion Handling in Augmented Reality Based on an Object Tracking Approach. Sensors, 10(4), 2885-2900. doi:10.3390/s10040288

A consensus on the use of daylight photodynamic therapy in the UK

Background: Actinic keratoses (AKs) are a consequence of chronic exposure to ultraviolet radiation. Treatment of chronically photo-damaged skin and AKs is driven by risk of progression to squamous cell carcinoma, as well as for symptomatic relief. Conventional photodynamic therapy (c-PDT) is indicated when AKs are multiple or confluent and if patients respond poorly or are unable to tolerate other therapies. c-PDT is limited by the field size that can be treated in single sessions and can cause significant discomfort.Objective: Recent studies investigated daylight illumination to activate protoporphyrin IX and daylight-PDT (d-PDT) is now licensed in the UK for face and scalp AKs. A group of experts met to discuss application of d-PDT with methyl aminolevulinate (MAL) and develop a UK consensus statement, specific to UK weather conditions.Methods: The UK consensus recommendations were reached among eight experts, who reviewed recent studies on d-PDT, assessed UK meteorological data and discussed personal experiences of d-PDT for AKs.Results: Recommendations from these discussions provide guidance on d-PDT use, specifically regarding patient selection, therapeutic indications, when to treat, skin preparation, MAL application and daylight exposure for patients with AKs.Conclusions: This UK expert consensus provides practical guidance for UK application of d-PDT

Vertical Diabatic Heating Structure of the MJO: Intercomparison between Recent Reanalyses and TRMM Estimates

Capitalizing on recently released reanalysis datasets and diabatic heating estimates based on Tropical Rainfall Measuring Mission (TRMM), the authors have conducted a composite analysis of vertical anomalous heating structures associated with the Madden–Julian oscillation (MJO). Because diabatic heating lies at the heart of prevailing MJO theories, the intention of this effort is to provide new insights into the fundamental physics of the MJO. However, some discrepancies in the composite vertical MJO heating profiles are noted among the datasets, particularly between three reanalyses and three TRMM estimates. A westward tilting with altitude in the vertical heating structure of the MJO is clearly evident during its eastward propagation based on three reanalysis datasets, which is particularly pronounced when the MJO migrates from the equatorial eastern Indian Ocean (EEIO) to the western Pacific (WP). In contrast, this vertical tilt in heating structure is not readily seen in the three TRMM products. Moreover, a transition from a shallow to deep heating structure associated with the MJO is clearly evident in a pressure–time plot over both the EEIO and WP in three reanalysis datasets. Although this vertical heating structure transition is detectable over the WP in two TRMM products, it is weakly defined in another dataset over the WP and in all three TRMM datasets over the EEIO. The vertical structures of radiative heating QR associated with the MJO are also analyzed based on TRMM and two reanalysis datasets. A westward vertical tilt in QR is apparent in all these datasets: that is, the low-level QR is largely in phase of convection, whereas QR in the upper troposphere lags the maximum convection. The results also suggest a potentially important role of radiative heating for the MJO, particularly over the Indian Ocean. Caveats in heating estimates based on both the reanalysis datasets and TRMM are briefly discussed

Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification

A rapid means of assessing reproductive status in rodents is useful not only in the study of reproductive dysfunction but is also required for the production of new mouse models of disease and investigations into the hormonal regulation of tissue degeneration (or regeneration) following pathological challenge. The murine reproductive (or estrous) cycle is divided into 4 stages: proestrus, estrus, metestrus, and diestrus. Defined fluctuations in circulating levels of the ovarian steroids 17-β-estradiol and progesterone, the gonadotropins luteinizing and follicle stimulating hormones, and the luteotropic hormone prolactin signal transition through these reproductive stages. Changes in cell typology within the murine vaginal canal reflect these underlying endocrine events. Daily assessment of the relative ratio of nucleated epithelial cells, cornified squamous epithelial cells, and leukocytes present in vaginal smears can be used to identify murine estrous stages. The degree of invasiveness, however, employed in collecting these samples can alter reproductive status and elicit an inflammatory response that can confound cytological assessment of smears. Here, we describe a simple, non-invasive protocol that can be used to determine the stage of the estrous cycle of a female mouse without altering her reproductive cycle. We detail how to differentiate between the four stages of the estrous cycle by collection and analysis of predominant cell typology in vaginal smears and we show how these changes can be interpreted with respect to endocrine status