easySTORM: a robust, lower-cost approach to localisation and TIRF microscopy

TIRF and STORM microscopy are super-resolving fluorescence imaging modalities for which current implementations on standard microscopes can present significant complexity and cost. We present a straightforward and low-cost approach to implement STORM and TIRF taking advantage of multimode optical fibres and multimode diode lasers to provide the required excitation light. Combined with open source software and relatively simple protocols to prepare samples for STORM, including the use of Vectashield for non-TIRF imaging, this approach enables TIRF and STORM imaging of cells labelled with appropriate dyes or expressing suitable fluorescent proteins to become widely accessible at low cost

Bourgeois Modernity Versus the Historical Aristocracy in Christoph Meiners’s Political Thought

Christoph Meiners (1747–1810), a major historian and philosopher of the German late Enlightenment, has received increasing recognition as a significant thinker in the emergence of nineteenth-century racial theories. The scholarly focus on Meiners's hierarchical view of race and its legacy has led to the classification of his broader oeuvre as conservative, or even reactionary. By examining his Geschichte der Ungleichheit der Stände unter den vornehmsten europäischen Völkern (1792), written in response to the French Revolution and the contemporary circumstances of the Holy Roman Empire, this article sheds new light on his work, as well as on an under-researched line of thought in the 1790s. Rather than a conservative or reactionary work, this text is a radical critique of the German aristocracy that ultimately recommends the abolition of most significant aristocratic privileges and the overhaul of its membership in favour of the bourgeoisie. This article presents not only a more complex understanding of Christoph Meiners's ideas, but also calls for a reappraisal of the categories applied to late eighteenth- and early nineteenth-century intellectuals both in Germany and in Europe more broadly

Investing in New Technology in Pulmonary Medicine–Navigating the Tortuous Path to Success.

The introduction of new technologies offer the promise to advance medicine. This occurs alongside improved efforts to control costs of healthcare by hospital administrators, Center for Medicare and Medicaid Services’ (CMS) pivot to value programs, and commercial payers’ efforts to reduce reimbursement. These trends present a challenge for the pulmonologist, among others, who must navigate increasingly complex and highly scrutinized evaluation processes used to secure new technologies. Healthcare providers are turning toward value assessments, while simultaneously tasked with a mission of offering state-of-the art technologies and services. Pulmonologists desiring new technologies are thus faced with increased scrutiny in their evaluation of costs and clinical data to support investments. Consideration of this scrutiny and further evidence to temper the evaluation will improve the likelihood of adoption and patient access to clinically-impactful technology. The identification of this evidence may provide – to both administrators and pulmonary clinicians – a comprehensive view of the clinical and economic benefits of such technologies. It is imperative that all parties involved in the decision process work collaboratively to deploy value-added and clinically-impactful technologies. While a physician group might invest in such new technologies, the capital required often leads such decisions to a larger organization such as a hospital, healthcare system, or privately owned entity. This article aims to provide a framework for pulmonary clinicians to better understand the processes that purchasers use to evaluate new technologies, the pressures that influence their consideration, and what resources may be leveraged towards success

Cytomegalovirus ulceration of the oropharynx

Cytomegalovirus (CMV) is a common opportunistic infection in both iatrogenic and HIV-induced immunosuppression. The usual sites of involvement are the gastro-intestinal tract, retina and lung. We present three cases of CMV ulceration of the oropharynx. All three patients presented with symptoms localized to the oropharynx and in each case the diagnosis was only made on histological examination of ulcer biopsy specimens. The patients all responded well to ganciclovir treatment and at writing none have required maintenance therapy (7-11 months post diagnosis)

The prevalence and risk of immune restoration disease in HIV-infected patients treated with highly active antiretroviral therapy

Background It is becoming increasingly clear that, during successful highly active antiretroviral therapy (HAART), a proportion of treated patients develop opportunistic infections (OIs), referred to in this setting as immune restoration disease (IRD). We examined the risk of developing IRD in HAART-treated HIV-infected patients. Methods A retrospective study of a cohort including all 389 patients treated with HAART between I January 1998 and 31 May 2004 in our HIV unit was performed to evaluate the occurrence of and risk factors for IRD during HAART. Baseline and follow-up values of CD4 T-cell counts and plasma viral loads (pVLs) were compared to assess the success of HAART. Results During successful HAART (significant increase in CD4 T-cell counts and decrease in pVL), at least one IRD episode occurred in 65 patients (16.7%). The median time to IRD was 4.6 months (range 212 months). IRDs included dermatomal herpes zoster (26 patients), pulmonary tuberculosis (four patients), tuberculous exudative pericarditis (two patients), tuberculous lymphadenitis (two patients), cerebral toxoplasmosis (one patient), progressive multifocal leucoencephalopathy (PML) (one patient), inflamed molluscum (one patient), inflamed Candida albicans angular cheilitis (three patients), genital herpes simplex (two patients), tinea corporis (two patients), cytomegalovirus (CMV) retinitis (two patients), CMV vitritis (one patient) and hepatitis B (three patients) or C (fifteen patients). A baseline CD4 T-cell count below 100 cells/mu L was shown to be the single predictor [odds ratio (OR) 2.5, 95% confidence interval (CI) 0.9-6.4] of IRD, while a CD4 T-cell count increase to gt 400 cells/mu L, but not undetectable pVL, was a negative predictor of IRD (OR 0.3, 95% CI 0.1-0.8). Conclusions To avoid IRD in advanced patients, HAART should be initiated before the CD4 T-cell count falls below 100 cells/mu L

Harmony and discord:Development of political parties and social fragmentation in Hong Kong, 1980-2017

This paper seeks to examine why political parties in Hong Kong are fragmented and how the development of political parties in Hong Kong leads to social discord. Political parties started to emerge in Hong Kong in the 1980s. They had a golden opportunity to develop in the 1990s due to political reform, but why are political parties in Hong Kong still small, weak, with poor reputations and weak support? The author points out five factors that lead to the malfunction of political parties in Hong Kong. Although some factors are caused by the political parties themselves, the author argues that they are, all in all, constitutional or institutional factors, as they are long-term restraints directly set by the government since the colonial era. Due to the failure of party development in Hong Kong caused by constitutional and institutional restraints, the author will also discuss how this failure has lead to the political and social discord in the past two decades since the handover and the future

Age-Related Gene Expression in the Frontal Cortex Suggests Synaptic Function Changes in Specific Inhibitory Neuron Subtypes

Genome-wide expression profiling of the human brain has revealed genes that are differentially expressed across the lifespan. Characterizing these genes adds to our understanding of both normal functions and pathological conditions. Additionally, the specific cell-types that contribute to the motor, sensory and cognitive declines during aging are unclear. Here we test if age-related genes show higher expression in specific neural cell types. Our study leverages data from two sources of murine single-cell expression data and two sources of age-associations from large gene expression studies of postmortem human brain. We used nonparametric gene set analysis to test for age-related enrichment of genes associated with specific cell-types; we also restricted our analyses to specific gene ontology groups. Our analyses focused on a primary pair of single-cell expression data from the mouse visual cortex and age-related human post-mortem gene expression information from the orbitofrontal cortex. Additional pairings that used data from the hippocampus, prefrontal cortex, somatosensory cortex and blood were used to validate and test specificity of our findings. We found robust age-related up-regulation of genes that are highly expressed in oligodendrocytes and astrocytes, while genes highly expressed in layer 2/3 glutamatergic neurons were down-regulated across age. Genes not specific to any neural cell type were also down-regulated, possibly due to the bulk tissue source of the age-related genes. A gene ontology-driven dissection of the cell-type enriched genes highlighted the strong down-regulation of genes involved in synaptic transmission and cell-cell signaling in the Somatostatin (Sst) neuron subtype that expresses the cyclin dependent kinase 6 (Cdk6) and in the vasoactive intestinal peptide (Vip) neuron subtype expressing myosin binding protein C, slow type (Mybpc1). These findings provide new insights into cell specific susceptibility to normal aging, and suggest age-related synaptic changes in specific inhibitory neuron subtypes

The Tetrodotoxin Receptor of Voltage-Gated Sodium Channels—Perspectives from Interactions with μ-Conotoxins

Neurotoxin receptor site 1, in the outer vestibule of the conducting pore of voltage-gated sodium channels (VGSCs), was first functionally defined by its ability to bind the guanidinium-containing agents, tetrodotoxin (TTX) and saxitoxin (STX). Subsequent studies showed that peptide μ-conotoxins competed for binding at site 1. All of these natural inhibitors block single sodium channels in an all-or-none manner on binding. With the discovery of an increasing variety of μ-conotoxins, and the synthesis of numerous derivatives, observed interactions between the channel and these different ligands have become more complex. Certain μ-conotoxin derivatives block single-channel currents partially, rather than completely, thus enabling the demonstration of interactions between the bound toxin and the channel’s voltage sensor. Most recently, the relatively small μ-conotoxin KIIIA (16 amino acids) and its variants have been shown to bind simultaneously with TTX and exhibit both synergistic and antagonistic interactions with TTX. These interactions raise new pharmacological possibilities and place new constraints on the possible structures of the bound complexes of VGSCs with these toxins