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 (ECM) 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 nondestructive staining, quantitation, and imaging of mineralizing bone-like nodules in live cultures of human bone marrow mesenchymal stem cells 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 mineralized 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

Bioresorbable composite bone fracture repair plates: manufacture and characterisation

This study reports on Bioresorbable composites manufactured using PLA as matrix and phosphate-based glass fibres as reinforcement. Composites were manufactured with varying volume fraction (from 25% - 45%) and mechanically tested

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

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

Alpha-defensin and the Synovasure lateral flow device for the diagnosis of prosthetic joint infection: a systematic review and meta-analysis

AimsThe aim of this review is to evaluate the available literature and to calculate a pooled diagnostic sensitivity and specificity for the different alpha-defensin test systems to diagnose peri-prosthetic infection.Materials and MethodsStudies using alpha-defensin or Synovasure to diagnose periprosthetic joint infection were identified from systematic searches of electronic databases. Study quality was evaluated using the QUADAS tool. Meta-analysis was completed using a bivariate model.ResultsEleven eligible studies were included. Median QUADAS score was 13 [I.Q.R. 13-13] out of 14. Significant conflicts of interest were identified in five studies. Pooled sensitivity for the laboratory alpha-defensin test was 0.95 (95% CI 0.91-0.98) and specificity 0.97 (95% CI 0.95-0.98) for four studies with a threshold level of 5.2mgl-a. Pooled sensitivity for the lateral flow cassette test was 0.85 (95% CI 0.74-0.92) and specificity was 0.90 (95% CI 0.91-0.98). There was a statistically significant difference in sensitivity but not specificity.ConclusionLaboratory based alpha-defensin testing remains a promising tool for diagnosing periprosthetic joint infection. The lateral flow cassette has a significantly lower performance and pooled results are comparable to the leucocyte esterase test. Further studies are essential before the widespread adoption of the lateral flow cassette alpha-defensin test. Take home messageAlpha defensin testing for periprosthetic joint infection has excellent sensitivity and specificity when performed in a laboratory. The pooled sensitivity and specificity is much lower when results are combined for the lateral flow cassette (Synovasure devise

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

Branding of Government Services: Benefits and Challenges

Till recently, brand management was perceived as a function exclusive to commercial organizations. However, the experiences of several nations have shown that branding can be an effective tool for managing government services also. This paper discusses the benefits of the current practices of branding in governance. A coherent branding strategy assists the departments in the formulation and delivery of services by having clarity of purpose and consistency in communication. The citizen benefits in various ways from a streamlined execution of the strategy. The paper also identifies the challenges for brand management to grow in the governance sphere

Functional outcome and complications following surgery for Dupuytren’s disease: a multi-centre cross-sectional study

Variables associated with recurrent Dupuytren’s disease, or a ‘diathesis’, have been investigated, but those associated with functional outcome and complications are less well studied. Outcomes 1 or 5 years after an aponeurotomy, fasciectomy or dermofasciectomy were assessed by patient interview and examination at five UK centres. A total of 432 procedures were studied. The reoperation rate did not differ at 1 year (p = 0.396, Chi-square test with Monte Carlo simulation), but was higher after aponeurotomy in the 5-year group (30%, versus 6% after fasciectomy and 0% after dermofasciectomy, p = 0.003, Chi square test with Monte Carlo simulation). Loss of function (DASH>15) did not differ between procedures at 5 years, even when reoperation and other variables were controlled. Diabetes, female gender and previous ipsilateral surgery were associated with poorer function in logistic regression analysis. The variables associated with poor function after treatments differ from diathesis variables. Aponeurotomy had lower complication rates than fasciectomy and dermofasciectomy. This may counterbalance the former’s higher recurrence rate and explain why aponeurotomy demonstrated similar long-term functional outcome compared with excisional surgery in this study