Nerve Growth Factor Sensitisation of TRPA1 on Sensory Neurones

Background: Sensory neurones from the trigeminal nerve innervate the oro-facial region and teeth. Transient receptor potential channels (TRPs) expressed by these neurones are responsible for relaying sensory information such as changes in ambient temperature, mechanical sensations and pain. Study of TRP channel expression and regulation in human sensory neurones therefore merits investigation to improve our understanding of allodynia and hyperalgesia. Objective: The objective of this study was to differentiate human dental pulp stem cells (hDPSCs) towards a neuronal phenotype (peripheral neuronal equivalents; PNEs) and employ this model to study TRP channel sensitisation. Method: hDPSCs were enriched by preferential adhesion to fibronectin, plated on coverslips (thickness 0) coated with poly-l-ornithine and laminin and then differentiated for 7 days in neurobasal A medium with additional supplementation. A whole cell patch clamp technique was used to investigate whether TRP channels on PNE membranes were modulated in the presence of nerve growth factor (NGF). PNEs were treated with NGF for 20 minutes immediately before experimentation and then stimulated for TRPA1 activity using cinnamaldehyde. Peak currents were read at 80 mV and -80 mV and compared to peak currents recorded in untreated PNEs. Data were analysed and plotted using Clampfit9 software (Molecular Devices, Sunnyvale, California, USA). Result: Results showed for the first time that pre-treatment of PNEs by NGF produced significantly larger inward and outward currents demonstrating that TRPA1 channels on PNE membranes were capable of becoming sensitised following treatment with NGF. Conclusion: Sensitisation of TRPA1 by NGF provides evidence of a mechanism for rapid neuronal sensitisation that is independent of TRPA1 gene expression

An in vitro model of nociceptors in human trigeminal nerves

Background: The oro-facial region is densely innervated by the trigeminal nerve, which when stimulated can induce noxious pain sensation and contribute to neurogenic inflammation in local tissues. Recent research on the expression of specialised ion channels on the trigeminal nerve has highlighted the need to undertake more extensive studies on ion channel expression/functionality with the aim of elucidating their role in pain sensations. A major family of such ion channels is the transient receptor potential (TRP) channels which are activated by a wide variety of thermal, mechanical or chemical stimuli and merit investigation as possible druggable targets for future analgesics.
Objective: Study of TRP channel expression and regulation in oro-facial tissues is hindered by the fact that the cell bodies of neurons innervating these tissues are located in the trigeminal ganglion. Using dental pulp stem cells differentiated towards peripheral neuronal equivalents (PNEs), we sought to determine TRP channel expression, functionality and potential modulation by cytokines in this novel model.
Method: Dental pulp stem cells (DPSCs) were grown on substrate-coated tissue culture plates and differentiated towards a neuronal phenotype using neuronal induction media. Quantitative polymerase chain reaction (qPCR) was performed on PNEs +/-cytokine treatment. Ion channel functionality was investigated using whole cell patch clamping.
Result: qPCR analysis showed that PNEs expressed the TRP channels TRPA1, TRPV1, TRPV4 and TRPM8. TRPA1 was the most abundantly expressed TRP channel studied whereas TRPM8 was lowly expressed. TRP channel expression was shown to be regulated by treatment with inflammatory cytokines. Patch clamp studies using specific agonists and antagonists for TRPA1 and TRPV1 showed these channels were functional.
Conclusion: PNEs differentiated from DPSCs provide a suitable model for TRP channel expression, regulation, and sensistisation in oro-facial tissues. This human neuronal model has potential for use in pre-clinical studies of novel analgesics.

Effect of the Presence of MEA on the CO2 Capture Ability of Superbase Ionic Liquids

The miscibility of monoethanolamine (MEA) in five superbase ionic liquids (ILs), namely the trihexyl-tetradecylphosphonium benzotriazolide ([P66614][Bentriz]), trihexyl-tetradecylphosphonium benzimidazolide ([P66614][Benzim]), trihexyl-tetradecylphosphonium 1,2,3-triazolide ([P66614][123Triz]), trihexyl-tetradecylphosphonium 1,2,4-triazolide ([P66614][124Triz]), and trihexyl-tetradecylphosphonium imidazolide ([P66614][Im]) was determined at 295.15 K using 1H NMR spectroscopy. The solubility of carbon dioxide (CO2) in equimolar (IL + MEA) mixtures was then studied experimentally using a gravimetric technique at 295.15 K and 0.1 MPa. The effect of MEA on the CO2 capture ability of these ILs was investigated together with the viscosity of these systems in the presence or absence of CO2 to evaluate their practical application in CO2 capture processes. The effect of the presence of MEA on the rate of CO2 uptake was also studied. The study showed that the MEA can enhance CO2 absorption over the ideal values in the case of [P66614][123Triz] and [P66614][Bentriz] while in the other systems the mixtures behave ideally. A comparison of the effect of MEA addition with the addition of water to these superbase ILs showed that similar trends were observed in each case for the individual ILs studied.

Use of emerging technologies to assess differences in outdoor physical activity in St. Louis, Missouri.

Introduction: Abundant evidence shows that regular physical activity (PA) is an effective strategy for preventing obesity in people of diverse socioeconomic status (SES) and racial groups. The proportion of PA performed in parks and how this differs by proximate neighborhood SES has not been thoroughly investigated. The present project analyzes online public web data feeds to assess differences in outdoor PA by neighborhood SES in St. Louis, MO, USA.
Methods: First, running and walking routes submitted by users of the website MapMyRun.com were downloaded. The website enables participants to plan, map, record, and share their exercise routes and outdoor activities like runs, walks, and hikes in an online database. Next, the routes were visually illustrated using geographic information systems. Thereafter, using park data and 2010 Missouri census poverty data, the odds of running and walking routes traversing a low-SES neighborhood, and traversing a park in a low-SES neighborhood were examined in comparison to the odds of routes traversing higher-SES neighborhoods and higher-SES parks.
Results: Results show that a majority of running and walking routes occur in or at least traverse through a park. However, this finding does not hold when comparing low-SES neighborhoods to higher-SES neighborhoods in St. Louis. The odds of running in a park in a low-SES neighborhood were 54% lower than running in a park in a higher-SES neighborhood (OR = 0.46, CI = 0.17-1.23). The odds of walking in a park in a low-SES neighborhood were 17% lower than walking in a park in a higher-SES neighborhood (OR = 0.83, CI = 0.26-2.61).
Conclusion: The novel methods of this study include the use of inexpensive, unobtrusive, and publicly available web data feeds to examine PA in parks and differences by neighborhood SES. Emerging technologies like MapMyRun.com present significant advantages to enhance tracking of user-defined PA across large geographic and temporal settings.

Learning from Outdoor Webcams: Surveillance of Physical Activity Across Environments

Abstract
Publicly available, outdoor webcams continuously view the world and share images. These cameras include traffic cams, campus cams, ski-resort cams, etc. The Archive of Many Outdoor Scenes (AMOS) is a project aiming to geolocate, annotate, archive, and visualize these cameras and images to serve as a resource for a wide variety of scientific applications. The AMOS dataset has archived over 750 million images of outdoor environments from 27,000 webcams since 2006. Our goal is to utilize the AMOS image dataset and crowdsourcing to develop reliable and valid tools to improve physical activity assessment via online, outdoor webcam capture of global physical activity patterns and urban built environment characteristics.
This project’s grand scale-up of capturing physical activity patterns and built environments is a methodological step forward in advancing a real-time, non-labor intensive assessment using webcams, crowdsourcing, and eventually machine learning. The combined use of webcams capturing outdoor scenes every 30 min and crowdsources providing the labor of annotating the scenes allows for accelerated public health surveillance related to physical activity across numerous built environments. The ultimate goal of this public health and computer vision collaboration is to develop machine learning algorithms that will automatically identify and calculate physical activity patterns.

A high-value, low-cost bubble continuous positive airway pressure system for low-resource settings:technical assessment and initial case reports

Acute respiratory infections are the leading cause of global child mortality. In the developing world, nasal oxygen therapy is often the only treatment option for babies who are suffering from respiratory distress. Without the added pressure of bubble Continuous Positive Airway Pressure (bCPAP) which helps maintain alveoli open, babies struggle to breathe and can suffer serious complications, and frequently death. A stand-alone bCPAP device can cost $6,000, too expensive for most developing world hospitals. Here, we describe the design and technical evaluation of a new, rugged bCPAP system that can be made in small volume for a cost-of-goods of approximately $350. Moreover, because of its simple design--consumer-grade pumps, medical tubing, and regulators--it requires only the simple replacement of a <$1 diaphragm approximately every 2 years for maintenance. The low-cost bCPAP device delivers pressure and flow equivalent to those of a reference bCPAP system used in the developed world. We describe the initial clinical cases of a child with bronchiolitis and a neonate with respiratory distress who were treated successfully with the new bCPAP device.

Microneedle arrays as transdermal and intradermal drug delivery systems: Materials science, manufacture and commercial development

Transdermal drug delivery offers a number of advantages for the patient, due not only its non-invasive and convenient nature, but also factors such as avoidance of first pass metabolism and prevention of gastrointestinal degradation. It has been demonstrated that microneedle arrays can increase the number of compounds amenable to transdermal delivery by penetrating the skin's protective barrier, the stratum corneum, and creating a pathway for drug permeation to the dermal tissue below. Microneedles have been extensively investigated in recent decades for drug and vaccine delivery as well as minimally invasive patient monitoring/diagnosis. This review focuses on a range of critically important aspects of microneedle technology, namely their material composition, manufacturing techniques, methods of evaluation and commercial translation to the clinic for patient benefit and economic return. Microneedle research and development is finally now at the stage where commercialisation is a realistic possibility. However, progress is still required in the areas of scaled-up manufacture and regulatory approval.

Seasons, Family and Nation in American Horror Story

Arguably, the title of American Horror Story sets out an agenda for the program: this is not just a horror story, but it is a particularly American one. This chapter examines the way that the program uses seasonal celebrations as a way of expressing that national identity, with special emphasis on the importance of family to those celebrations. The particular seasonal celebrations focused on are those of Halloween and Christmas, each of which has associations with the supernatural. However, the use of the supernatural at those seasons is one which is particularly associated with the US, presenting Halloween as a time of supernatural incursion and horror, and of disruption to society and the normal order of things, while Christmas is presented more as a time of unity for the family. Where the supernatural emerges in American Christmas television, it is typically as a force to encourage togetherness and reconciliation, rather than as a dark reminder of the past. While these interpretations of these festivals have been broadcast abroad by American cultural products, not least American television, they have different associations and implications elsewhere, as will be shown. So the particular uses of these festivals is part of what marks American Horror Story out as American, as is the way that the program's narratives have been structured to fit in with US television scheduling. This chapter, then, argues that the structures of the narratives combines with their use of the festivals of Halloween and Christmas in order to enhance the sense of this series as a particularly American horror story.

Beneficial Effects of Berberine on Oxidized LDL-induced Cytotoxicity in Human Retinal Müller Cells

PURPOSE. Limited mechanistic understanding of diabetic retinopathy (DR) has hindered therapeutic advances. Berberine, an isoquinolone alkaloid, has shown favorable effects on glucose and lipid metabolism in animal and human studies, but effects on DR are unknown. We previously demonstrated intraretinal extravasation and modification of LDL in human diabetes, and toxicity of modified LDL to human retinal M¨uller cells. We now explore pathogenic effects of modified LDL on M¨uller cells, and the efficacy of berberine in mitigating this cytotoxicity. METHODS. Confluent human M¨uller cells were exposed to in vitro–modified ‘highly oxidized, glycated (HOG-) LDL versus native-LDL (N-LDL; 200 mg protein/L) for 6 or 24 hours, with/ without pretreatment with berberine (5 lM, 1 hour) and/or the adenosine monophosphate (AMP)-activated protein kinase (AMPK) inhibitor, Compound C (5 lM, 1 hour). Using techniques including Western blots, reactive oxygen species (ROS) detection assay, and quantitative real-time PCR, the following outcomes were assessed: cell viability (CCK-8 assay), autophagy (LC3, Beclin-1, ATG-5), apoptosis (cleaved caspase 3, cleaved poly-ADP ribose polymerase), oxidative stress (ROS, nuclear factor erythroid 2-related factor 2, glutathione peroxidase 1, NADPH oxidase 4), angiogenesis (VEGF, pigment epithelium-derived factor), inflammation (inducible nitric oxide synthase, intercellular adhesion molecule 1, IL-6, IL-8, TNF-a), and glial cell activation (glial fibrillary acidic protein). RESULTS. Native-LDL had no effect on cultured human M¨uller cells, but HOG-LDL exhibited marked toxicity, significantly decreasing viability and inducing autophagy, apoptosis, oxidative stress, expression of angiogenic factors, inflammation, and glial cell activation. Berberine attenuated all the effects of HOG-LDL (all P < 0.05), and its effects were mitigated by AMPK inhibition (P < 0.05). CONCLUSIONS. Berberine inhibits modified LDL-induced M¨uller cell injury by activating the AMPK pathway, and merits further study as an agent for preventing and/or treating DR.

Techno-Economic Feasibility of Selective CO2 Capture Processes from Biogas Streams using Ionic Liquids as Physical Absorbents

Biogas from anaerobic digestion of sewage sludge is a renewable resource with high energy content, which is formed mainly of CH4 (40-75 vol.%) and CO2 (15-60 vol.%) Other components such as water (H2O, 5-10 vol.%) and trace amounts of hydrogen sulfide and siloxanes can also be present. A CH4-rich stream can be produced by removing the CO2 and other impurities so that the upgraded bio-methane can be injected into the natural gas grid or used as a vehicle fuel. The main objective of this paper is to develop a new modeling methodology to assess the technical and economic performance of biogas upgrading processes using ionic liquids which physically absorb CO2. Three different ionic liquids, namely the 1-ethyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]imide, 1-hexyl-3-methylimidazoliumbis[(trifluoromethyl)sulfonyl]imide and trihexyl(tetradecyl)phosphonium bis[(trifluoromethyl)sulfonyl]imide, are considered for CO2 capture in a pressure-swing regenerative absorption process. The simulation software Aspen Plus and Aspen Process Economic Analyzer is used to account for mass and energy balances as well as equipment cost. In all cases, the biogas upgrading plant consists of a multistage compressor for biogas compression, a packed absorption column for CO2 absorption, a flash evaporator for solvent regeneration, a centrifugal pump for solvent recirculation, a pre-absorber solvent cooler and a gas turbine for electricity recovery. The evaluated processes are compared in terms of energy efficiency, capital investment and bio-methane production costs. The overall plant efficiency ranges from 71-86 % whereas the bio-methane production cost ranges from £6.26-7.76 per GJ (LHV). A sensitivity analysis is also performed to determine how several technical and economic parameters affect the bio-methane production costs. The results of this study show that the simulation methodology developed can predict plant efficiencies and production costs of large scale CO2 capture processes using ionic liquids without having to rely on gas solubility experimental data.