Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride

The final stage of in vitro osteogenic differentiation is characterized by the production of mineral deposits containing calcium cations and inorganic phosphates, which populate the extracellular matrix surrounding the cell monolayer. Conventional histological techniques for the assessment of mineralization, such as Von Kossa and Alizarin Red S staining, are end-point techniques requiring cell fixation. Moreover, in both cases staining quantitation requires dye extraction which irreversibly alters the ECM conformation and structure, therefore preventing the use of the sample for further analysis. In this study, the use of Tetracycline hydrochloride (TC) is proposed for the non-destructive staining, quantitation and imaging of mineralizing bone-like nodules in live cultures of human bone marrow mesenchymal stem cells (MSCs) cultured under osteogenic conditions. Overnight administration of TC to living cells was shown not to alter the metabolic activity or the progression of cell differentiation. When applied to differentiating cultures, cell exposure to serial doses of TC was found to produce quantifiable fluorescence emission specifically in osteogenic cultures. Incubation with TC enabled fluorescence imaging of mineralised areas in live cultures and the combination with other fluorophores using appropriate filters. These results demonstrate that serial TC administration over the differentiation time course provides a qualitative and quantitative tool for the monitoring and evaluation of the differentiation process in live cells

A New Spin on Research Translation: The Boston Consensus Conference on Human Biomonitoring

BACKGROUND. Translating research to make it more understandable and effective (research translation) has been declared a priority in environmental health but does not always include communication to the public or residents of communities affected by environmental hazards. Their unique perspectives are also commonly missing from discussions about science and technology policy. The consensus conference process, developed in Denmark, offers a way to address this gap. OBJECTIVES. The Boston Consensus Conference on Human Biomonitoring, held in Boston, Massachusetts, in the fall of 2006, was designed to educate and elicit input from 15 Boston-area residents on the scientifically complex topic of human biomonitoring for environmental chemicals. This lay panel considered the many ethical, legal, and scientific issues surrounding biomonitoring and prepared a report expressing their views. DISCUSSION. The lay panel's findings provide a distinct and important voice on the expanding use of biomonitoring. In some cases, such as a call for opt-in reporting of biomonitoring results to study participants, they mirror recommendations raised elsewhere. Other conclusions have not been heard previously, including the recommendation that an individual's results should be statutorily exempted from the medical record unless permission is granted, and the opportunity to use biomonitoring data to stimulate green chemistry. CONCLUSION. The consensus conference model addresses both aspects of a broader conception of research translation: engaging the public in scientific questions, and bringing their unique perspectives to bear on public health research, practice, and policy. In this specific application, a lay panel's recommendations on biomonitoring surveillance, communication, and ethics have practical implications for the conduct of biomonitoring studies and surveillance programs.National Institute of Environmental Health Sciences (R25 ES12084, P42ES007381

What Does It Take to Make it to the Polling Station? The Effects of Campaign Activities on Electoral Participation

This study explores the extent to which campaign visibility facilitates electoral participation, using data from first- and second-order elections in Britain. Our contribution to the existing literature is threefold. First, we assess whether the effects of campaign effort are conditioned by marginality, finding that campaign mobilisation gets out the vote regardless of the competitiveness of the race. Second, we look at the relative ability of different campaign activities to stimulate turnout, detecting significant differences. Third, we show that the effects of campaign effort on electoral participation are rather similar in first- and second-order elections. These findings suggest that a greater level of electoral information provided by campaign activities does reduce the costs of voting. Local campaigns play a key role in bringing voters to the polls in marginal and non-marginal races, and at general elections as much as at second-order elections

Development of a Coherent Doppler Lidar for Precision Maneuvering and Landing of Space Vehicles

A coherent Doppler lidar has been developed to address NASAs need for a high-performance, compact, and cost-effective velocity and altitude sensor onboard its landing vehicles. Future robotic and manned missions to planetary bodies require precise ground-relative velocity vector and altitude data to execute complex descent maneuvers and safe, soft landing at a pre-designated site. This lidar sensor, referred to as a Navigation Doppler Lidar, meets the required performance of landing missions while complying with vehicle size, mass, and power constraints. Operating from over five kilometers altitude, the lidar obtains velocity and range precision measurements with 2 cm/sec and 2 meters, respectively, dominated by the vehicle motion. After a series of flight tests onboard helicopters and rocket-powered free-flyer vehicles, the Navigation Doppler Lidar is now being ruggedized for future missions to various destinations in the solar system

Porous calcium phosphate glass microspheres for orthobiologic applications

Orthobiologics is a rapidly advancing field utilising cell-based therapies and biomaterials to enable the body to repair and regenerate musculoskeletal tissues. This paper reports on a cost-effective flame spheroidisation process for production of novel porous glass microspheres from calcium phosphate-based glasses to encapsulate and deliver stem cells. Careful selection of the glass and pore-forming agent, along with a manufacturing method with the required processing window enabled the production of porous glass microspheres via a single-stage manufacturing process. The morphological and physical characterisation revealed porous microspheres with tailored surface and interconnected porosity (up to 76 ± 5%) with average pore size of 55 ± 8 µm and surface areas ranging from 0.34 to 0.9 m 2 g −1 . Furthermore, simple alteration of the processing parameters produced microspheres with alternate unique morphologies, such as with solid cores and surface porosity only. The tuneable porosity enabled control over their surface area, degradation profiles and hence ion release rates. Furthermore, cytocompatibility of the microspheres was assessed using human mesenchymal stem cells via direct cell culture experiments and analysis confirmed that they had migrated to within the centre of the microspheres. The novel microspheres developed have huge potential for tissue engineering and regenerative medicine applications. Statement of Significance: This manuscript highlights a simple cost-effective one-step process for manufacturing porous calcium phosphate-based glass microspheres with varying control over surface pores and fully interconnected porosity via a flame spheroidisation process. Moreover, a simple alteration of the processing parameters can produce microspheres which have a solid core with surface pores only. The tuneable porosity enabled control over their surface area, degradation profiles and hence ion release rates. The paper also shows that stem cells not only attach and proliferate but more importantly migrate to within the core of the porous microspheres, highlighting applications for bone tissue engineering and regenerative medicine

Reassessing the Role of Histaminergic Tuberomammillary Neurons in Arousal Control

The histaminergic neurons of the tuberomammillary nucleus (TMN(HDC)) of the posterior hypothalamus have long been implicated in promoting arousal. More recently, a role for GABAergic signaling by the TMN(HDC) neurons in arousal control has been proposed. Here, we investigated the effects of selective chronic disruption of GABA synthesis (via genetic deletion of the GABA synthesis enzyme, glutamic acid decarboxylase 67) or GABAergic transmission (via genetic deletion of the vesicular GABA transporter (VGAT)) in the TMN(HDC) neurons on sleep-wake in male mice. We also examined the effects of acute chemogenetic activation and optogenetic inhibition of TMN(HDC) neurons upon arousal in male mice. Unexpectedly, we found that neither disruption of GABA synthesis nor GABAergic transmission altered hourly sleep-wake quantities, perhaps because very few TMN(HDC) neurons coexpressed VGAT. Acute chemogenetic activation of TMN(HDC) neurons did not increase arousal levels above baseline but did enhance vigilance when the mice were exposed to a behavioral cage change challenge. Similarly, acute optogenetic inhibition had little effect upon baseline levels of arousal. In conclusion, we could not identify a role for GABA release by TMN(HDC) neurons in arousal control. Further, if TMN(HDC) neurons do release GABA, the mechanism by which they do so remains unclear. Our findings support the view that TMN(HDC) neurons may be important for enhancing arousal under certain conditions, such as exposure to a novel environment, but play only a minor role in behavioral and EEG arousal under baseline conditions.SIGNIFICANCE STATEMENT The histaminergic neurons of the tuberomammillary nucleus of the hypothalamus (TMN(HDC)) have long been thought to promote arousal. Additionally, TMN(HDC) neurons may counter-regulate the wake-promoting effects of histamine through co-release of the inhibitory neurotransmitter, GABA. Here, we show that impairing GABA signaling from TMN(HDC) neurons does not impact sleep-wake amounts and that few TMN(HDC) neurons contain the vesicular GABA transporter, which is presumably required to release GABA. We further show that acute activation or inhibition of TMN(HDC) neurons has limited effects upon baseline arousal levels and that activation enhances vigilance during a behavioral challenge. Counter to general belief, our findings support the view that TMN(HDC) neurons are neither necessary nor sufficient for the initiation and maintenance of arousal under baseline conditions

Participatory politics, environmental journalism and newspaper campaigns

This is an Author's Accepted Manuscript of an article published in Journalism Studies, 13(2), 210 - 225, 2012, copyright Taylor & Francis, available online at: http://www.tandfonline.com/10.1080/1461670X.2011.646398.This article explores the extent to which approaches to participatory politics might offer a more useful alternative to understanding the role of environmental journalism in a society where the old certainties have collapsed, only to be replaced by acute uncertainty. This uncertainty not only generates acute public anxiety about risks, it has also undermined confidence in the validity of long-standing premises about the ideal role of the media in society and journalistic professionalism. The consequence, this article argues, is that aspirations of objective reportage are outdated and ill-equipped to deal with many of the new risk stories environmental journalism covers. It is not a redrawing of boundaries that is needed but a wholesale relocation of our frameworks into approaches better suited to the socio-political conditions and uncertainties of late modernity. The exploration of participatory approaches is an attempt to suggest one way this might be done

Intra-articular Injection Administration in UK Ex-professional Footballers During Their Playing Careers and the Association with Post-career Knee Osteoarthritis

© 2020, The Author(s). Background: The long-term risk from knee intra-articular (KIA) injections in professional athletes such as ex-footballers remains unknown. The use of KIA injections is controversial and remains anecdotally prolific as it is perceived as being safe/beneficial. The aim of this study was to determine the number, type and frequency KIA injections administered to retired professional footballers during their playing careers and the associations with post-career knee osteoarthritis (KOA). Methods: This is a cross-sectional study involving a postal questionnaire (n = 1207) and subsequent knee radiographs in a random sample of questionnaire responders (n = 470). Footballers self-reported in the questionnaire whether they had received KIA injections and the estimated total number over the course of their playing career. Participant characteristics and football career-related details were also recorded. KOA was measured as self-reported knee pain (KP), total knee replacement (TKR) and radiographic KOA (RKOA). Results: 44.5% of footballers had received at least one KIA injection (mean: 7.5; SD ± 11.2) during their professional career. 71% of knee injections were cortisone/corticosteroid based. Multivariate logistic regression, adjusting for age, body mass index (BMI) and significant knee injury identified that footballers with injections weretwo times more likely to have KP (OR 1.81, 95% CI 1.40–2.34) and TKR (OR 2.21, 95% CI 1.43–3.42) than those without injections. However, there was no association with RKOA (OR 1.30, 95% CI 0.85–2.01). Given, the association with KP and TKR, we found a significant dose–response relationship as the more injections a player received (by dose–response groups), the greater the risk of KP and TKR outcomes after adjustment for knee injury and other confounders (p for trend < 0.01). Conclusion: On average, 8 KIA injections were given to the ex-footballers during their professional career. The most commonly administered injections were cortisone based. These injections associated with KP and TKR after they retired. The associations are independent of knee injuries and are dose dependent. The study suggests that there may have been excessive use of KIA injections to expedite return to play and this contributed to detrimental long-term outcomes such as KP and TKR post-retirement from professional football