Azithromycin treatment modifies airway and blood gene expression networks in neutrophilic COPD.

Long-term, low-dose azithromycin reduces exacerbation frequency in chronic obstructive pulmonary disease (COPD), yet the mechanism remains unclear. This study characterised genome-wide gene expression changes in patients with neutrophilic COPD following long-term, low-dose azithromycin treatment. Patients with neutrophilic COPD (>61% or >162×104 cells per mL sputum neutrophils) were randomised to receive either azithromycin or placebo for 12 weeks. Sputum and blood were obtained before and after 12 weeks of treatment. Gene expression was defined using microarrays. Networks were analysed using the Search Tool for the Retrieval of Interacting Gene database. In sputum, 403 genes were differentially expressed following azithromycin treatment (171 downregulated and 232 upregulated), and three following placebo treatment (one downregulated and two upregulated) compared to baseline (adjusted p1.5). In blood, 138 genes were differentially expressed with azithromycin (121 downregulated and 17 upregulated), and zero with placebo compared to baseline (adjusted p1.3). Network analysis revealed one key network in both sputum (14 genes) and blood (46 genes), involving interferon-stimulated genes, human leukocyte antigens and genes regulating T-cell responses. Long-term, low-dose azithromycin is associated with downregulation of genes regulating antigen presentation, interferon and T-cell responses, and numerous inflammatory pathways in the airways and blood of neutrophilic COPD patients

The effect of azithromycin in adults with stable neutrophilic COPD: A double blind randomised, placebo controlled trial

Background: Chronic Obstructive Pulmonary Disease (COPD) is a progressive airway disease characterised by neutrophilic airway inflammation or bronchitis. Neutrophilic bronchitis is associated with both bacterial colonisation and lung function decline and is common in exacerbations of COPD. Despite current available therapies to control inflammation, neutrophilic bronchitis remains common. This study tested the hypothesis that azithromycin treatment, as an add-on to standard medication, would significantly reduce airway neutrophil and neutrophils chemokine (CXCL8) levels, as well as bacterial load. We conducted a randomised, double-blind, placebo-controlled study in COPD participants with stable neutrophilic bronchitis. Methods: Eligible participants (n = 30) were randomised to azithromycin 250 mg daily or placebo for 12 weeks in addition to their standard respiratory medications. Sputum was induced at screening, randomisation and monthly for a 12 week treatment period and processed for differential cell counts, CXCL8 and neutrophil elastase assessment. Quantitative bacteriology was assessed in sputum samples at randomisation and the end of treatment visit. Severe exacerbations where symptoms increased requiring unscheduled treatment were recorded during the 12 week treatment period and for 14 weeks following treatment. A sub-group of participants underwent chest computed tomography scans (n = 15). Results: Nine participants with neutrophilic bronchitis had a potentially pathogenic bacteria isolated and the median total bacterial load of all participants was 5.22×107 cfu/mL. Azithromycin treatment resulted in a non-significant reduction in sputum neutrophil proportion, CXCL8 levels and bacterial load. The mean severe exacerbation rate was 0.33 per person per 26 weeks in the azithromycin group compared to 0.93 exacerbations per person in the placebo group (incidence rate ratio (95%CI): 0.37 (0.11,1.21), p = 0.062). For participants who underwent chest CT scans, no alterations were observed. Conclusions: In stable COPD with neutrophilic bronchitis, add-on azithromycin therapy showed a trend to reduced severe exacerbations sputum neutrophils, CXCL8 levels and bacterial load. Future studies with a larger sample size are warranted. Trial Registration: Australian New Zealand Clinical Trials Registry ACTRN12609000259246. © 2014 Simpson et al

Mutations and altered expression of SERPINF1 in patients with familial otosclerosis

Otosclerosis is a relatively common heterogenous condition, characterized by abnormal bone remodelling in the otic capsule leading to fixation of the stapedial footplate and an associated conductive hearing loss. Although familial linkage and candidate gene association studies have been performed in recent years, little progress has been made in identifying disease-causing genes. Here, we used whole-exome sequencing in four families exhibiting dominantly inherited otosclerosis to identify 23 candidate variants (reduced to 9 after segregation analysis) for further investigation in a secondary cohort of 84 familial cases. Multiple mutations were found in the SERPINF1 (Serpin Peptidase Inhibitor, Clade F) gene which encodes PEDF (pigment epithelium-derived factor), a potent inhibitor of angiogenesis and known regulator of bone density. Six rare heterozygous SERPINF1 variants were found in seven patients in our familial otosclerosis cohort; three are missense mutations predicted to be deleterious to protein function. The other three variants are all located in the 5'-untranslated region (UTR) of an alternative spliced transcript SERPINF1-012 RNA-seq analysis demonstrated that this is the major SERPINF1 transcript in human stapes bone. Analysis of stapes from two patients with the 5'-UTR mutations showed that they had reduced expression of SERPINF1-012 All three 5'-UTR mutations are predicted to occur within transcription factor binding sites and reporter gene assays confirmed that they affect gene expression levels. Furthermore, RT-qPCR analysis of stapes bone cDNA showed that SERPINF1-012 expression is reduced in otosclerosis patients with and without SERPINF1 mutations, suggesting that it may be a common pathogenic pathway in the disease

Partially linear censored quantile regression

Censored regression quantile (CRQ) methods provide a powerful and flexible approach to the analysis of censored survival data when standard linear models are felt to be appropriate. In many cases however, greater flexibility is desired to go beyond the usual multiple regression paradigm. One area of common interest is that of partially linear models: one (or more) of the explanatory covariates are assumed to act on the response through a non-linear function. Here the CRQ approach of Portnoy (J Am Stat Assoc 98:1001–1012, 2003) is extended to this partially linear setting. Basic consistency results are presented. A simulation experiment and unemployment example justify the value of the partially linear approach over methods based on the Cox proportional hazards model and on methods not permitting nonlinearity

Mixed material wear particle isolation from periprosthetic tissue surrounding total joint replacements

Submicron-sized wear particles are generally accepted as a potential cause of aseptic loosening when produced in sufficient volumes. With the accelerating use of increasingly wear-resistant biomaterials, identifying such particles and evaluating their biological response is becoming more challenging. Highly sensitive wear particle isolation methods have been developed but these methods cannot isolate the complete spectrum of particle types present in individual tissue samples. Two established techniques were modified to create one novel method to isolate both high- and low-density materials from periprosthetic tissue samples. Ten total hip replacement and eight total knee replacement tissue samples were processed. All particle types were characterized using high resolution scanning electron microscopy. UHMWPE and a range of high-density materials were isolated from all tissue samples, including: polymethylmethacrylate, zirconium dioxide, titanium alloy, cobalt chromium alloy and stainless steel. This feasibility study demonstrates the coexistence of mixed particle types in periprosthetic tissues and provides researchers with high-resolution images of clinically relevant wear particles that could be used as a reference for future in vitro biological response studies

Isolation and characterisation of wear debris surrounding failed total ankle replacements

Aseptic loosening and osteolysis continue to be a short- to mid-term problem for total ankle replacement (TAR) devices. The production of wear particles may contribute to poor performance, but their characteristics are not well understood. This study aimed to determine the chemical composition, size and morphology of wear particles surrounding failed TARs. A recently developed wear particle isolation method capable of isolating both high- and low-density materials was applied to 20 retrieved periprosthetic tissue samples from 15 failed TARs of three different brands. Isolated particles were imaged using ultra-high-resolution imaging and characterised manually to determine their chemical composition, size, and morphology. Six different materials were identified, which included: UHMWPE, calcium phosphate (CaP), cobalt chromium alloy (CoCr), commercially pure titanium, titanium alloy and stainless steel. Eighteen of the 20 samples contained three or more different wear particle material types. In addition to sub-micron UHMWPE particles, which were present in all samples, elongated micron-sized shards of CaP and flakes of CoCr were commonly isolated from tissues surrounding AES TARs. The mixed particles identified in this study demonstrate the existence of a complex periprosthetic environment surrounding TAR devices. The presence of such particles suggests that early failure of devices may be due in part to the multifaceted biological cascade that ensues after particle release. This study could be used to support the validation of clinically-relevant wear simulator testing, pre-clinical assessment of fixation wear and biological response studies to improve the performance of next generation ankle replacement devices

Deconstructing, Addressing, and Eliminating Racial and Ethnic Inequities in Prostate Cancer Care

Context Men of African ancestry have demonstrated markedly higher rates of prostate cancer mortality than men of other races and ethnicities around the world. In fact, the highest rates of prostate cancer mortality worldwide are found in the Caribbean and Sub-Saharan West Africa, and among men of African descent in the USA. Addressing this inequity in prostate cancer care and outcomes requires a focused research approach that creates durable solutions to address the structural, social, environmental, and health factors that create racial disparities in care and outcomes. Objective To introduce a conceptual model for evaluating racial inequities in prostate cancer care to facilitate the development of translational research studies and interventions. Evidence acquisition A collaborative review of literature relevant to racial inequities in prostate cancer care and outcomes was performed. Existing literature was used to highlight various components of the conceptual model to inform future research and interventions toward equitable care and outcomes. Evidence synthesis Racial inequities in prostate cancer outcomes are driven by a series of structural and social determinants of health that impact exposures, mediators, and outcomes. Social determinants of equity, such as laws/policies, economic systems, and structural racism, affect the inequitable access to environmental and neighborhood exposures, in addition to health care access. Although the incidence disparity remains problematic, various studies have demonstrated parity in outcomes when social and health factors, such as access to equitable care, are normalized. Few studies have tested interventions to reduce inequities in prostate cancer among Black men. Conclusions Worldwide, men of African ancestry demonstrate worse outcomes in prostate cancer, a phenomenon driven largely by social factors that inform biologic, environmental, and health care risks. A conceptual model was presented that organizes the many factors that influence prostate cancer incidence and mortality. Within that framework, we must understand the current state of inequities in clinical prostate cancer practice, the optimal state of what equitable practice would be, and how achieving equity in prostate cancer care balances costs, benefits, and harms. More robust characterization of the sources of prostate cancer inequities should inform testing of ambitious and innovative interventions as we work toward equity in care and outcomes. Patient summary Men of African ancestry demonstrate the highest rates of prostate cancer mortality, which may be reduced through social interventions. We present a framework for formalizing the identification of the drivers of prostate cancer inequities to facilitate the development of interventions and trials to eradicate them

A functional Magnetic Resonance Imaging study of patients with Polar Type II/III complex shoulder instability

The pathophysiology of Stanmore Classification Polar type II/III shoulder instability is not well understood. Functional Magnetic Resonance Imaging was used to measure brain activity in response to forward flexion and abduction in 16 patients with Polar Type II/III shoulder instability and 16 age-matched controls. When a cluster level correction was applied patients showed significantly greater brain activity than controls in primary motor cortex (BA4), supramarginal gyrus (BA40), inferior frontal gyrus (BA44), precentral gyrus (BA6) and middle frontal gyrus (BA6): the latter region is considered premotor cortex. Using voxel level correction within these five regions a unique activation was found in the primary motor cortex (BA4) at MNI coordinates -38 -26 56. Activation was greater in controls compared to patients in the parahippocampal gyrus (BA27) and perirhinal cortex (BA36). These findings show, for the first time, neural differences in patients with complex shoulder instability, and suggest that patients are in some sense working harder or differently to maintain shoulder stability, with brain activity similar to early stage motor sequence learning. It will help to understand the condition, design better therapies and improve treatment of this group; avoiding the common clinical misconception that their recurrent shoulder dislocations are a form of attention-seeking

Prenatal hypoxia induces increased cardiac contractility on a background of decreased capillary density.

Background: Chronic hypoxia in utero (CHU) is one of the most common insults to fetal development and may be associated with poor cardiac recovery from ischaemia-reperfusion injury,yet the effects on normal cardiac mechanical performance are poorly understood. Methods: Pregnant female wistar rats were exposed to hypoxia (12% oxygen, balance nitrogen)for days 10–20 of pregnancy. Pups were born into normal room air and weaned normally. At 10 weeks of age, hearts were excised under anaesthesia and underwent retrograde 'Langendorff' perfusion. Mechanical performance was measured at constant filling pressure (100 cm H2O) with intraventricular balloon. Left ventricular free wall was dissected away and capillary density estimated following alkaline phosphatase staining. Expression of SERCA2a and Nitric Oxide Synthases (NOS) proteins were estimated by immunoblotting. Results: CHU significantly increased body mass (P < 0.001) compared with age-matched control rats but was without effect on relative cardiac mass. For incremental increases in left ventricular balloon volume, diastolic pressure was preserved. However, systolic pressure was significantly greater following CHU for balloon volume = 50 μl (P < 0.01) and up to 200 μl (P < 0.05). For higher balloon volumes systolic pressure was not significantly different from control. Developed pressures were correspondingly increased relative to controls for balloon volumes up to 250 μl (P < 0.05).Left ventricular free wall capillary density was significantly decreased in both epicardium (18%; P <0.05) and endocardium (11%; P < 0.05) despite preserved coronary flow. Western blot analysis revealed no change to the expression of SERCA2a or nNOS but immuno-detectable eNOS protein was significantly decreased (P < 0.001) in cardiac tissue following chronic hypoxia in utero. Conclusion: These data offer potential mechanisms for poor recovery following ischaemia, including decreased coronary flow reserve and impaired angiogenesis with subsequent detrimental effects of post-natal cardiac performance