Essentials
Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease.
Fluorescence‐based flow cytometric analysis is suitable to detect low abundant EV subsets.
Particles ...of non‐interest can induce false‐positive light scatter and fluorescent signals.
Interference of particles of non‐interest can be monitored by analyzing serial dilutions.
Summary
Background
Extracellular vesicles (EVs) in plasma are increasingly being recognized as potential biomarkers. EV analysis for diagnostic purposes should be robust and should allow analysis of EV subsets with a wide range of abundance and in a large number of patient samples. Flow cytometry offers possibilities to meet these criteria, as it allows multiparameter analysis of individual EVs. However, analysis of plasma EVs is challenging, because of their size and heterogeneity, and the presence of other submicrometer‐sized particles in plasma that could interfere with EV analysis.
Objectives
To explore whether fluorescence‐based flow cytometric analysis of EV subsets is suitable when the EVs of interest are present in low abundance in a background of non‐labeled or differently labeled EVs and particles.
Methods
Fluorescently labeled EVs of interest were spiked at different ratios in full plasma, purified plasma components, or (non‐)fluorescent polystyrene beads, and subsequently analyzed by flow cytometry with fluorescence threshold triggering.
Results
We found that light scatter detection of low‐abundance or rare EV subsets during fluorescence threshold triggering was severely affected by particles of non‐interest, owing to coincidence and swarming. Importantly, we show that interfering particles labeled with different fluorophores induced false‐positive fluorescent signals on the particles of interest. These unwanted effects could only be discerned and controlled by performing serial dilutions and analyzing light scatter and fluorescence parameters.
Conclusions
We demonstrate how particles of non‐interest in plasma can impact on the light scatter and fluorescence detection of low‐abundance EVs of interest during fluorescence‐based flow cytometric analysis, and provide a means to prevent erroneous data interpretation.
Objectives To assess the prevalence of the 19 neuropsychiatric (NP) syndromes in systemic lupus erythematosus (SLE) patients, as defined by the American College of Rheumatology (ACR) in 1999, and ...better understand the reasons for interstudy variability of prevalence estimates, by performing a meta-analysis of relevant publications. Methods A literature search from April 1999 to May 2008 was performed to identify studies investigating NP syndromes in patients with definite SLE, applying the 1999 ACR case definitions and having a sample size of at least 30 patients. Excluded were studies that did not relate to all 19 NPSLE syndromes, presented duplicate data, or were irrelevant. Results Seventeen of 112 identified studies matched the inclusion criteria, reporting on a total of 5057 SLE patients, including 1439 NPSLE patients, with 2709 NPSLE syndromes. In a subanalysis of the 10 higher quality prospective and elicited studies (2049 patients) using the random-effects model, the prevalence of NP syndromes in SLE patients was estimated to be 56.3% (95% CI 42.5%-74.7%), and the most frequent NP syndromes were headache 28.3% (18.2%-44.1%), mood disorders 20.7% (11.5%-37.4%), cognitive dysfunction 19.7% (10.7%-36%), seizures 9.9% (4.8%-20.5%), and cerebrovascular disease 8.0% (4.5%-14.3%), although significant between-study heterogeneity was present ( P < 0.05). Autonomic disorder and Guillain-Barré syndrome carried a prevalence of less than 0.1%. No case of plexopathy was reported. Conclusions NP syndromes were estimated to exist in more than half of SLE patients. The most prevalent manifestations were headache, mood disorders, and cognitive dysfunction. A major limitation of the study was the significant heterogeneity of prevalence estimates between studies.
Abstract Objective To assess whether and how the rankings of the world's health systems based on disability adjusted life expectancy as done in the 2000 World Health Report change when using the ...narrower concept of mortality amenable to health care, an outcome more closely linked to health system performance. Design Analysis of mortality amenable to health care (including and excluding ischaemic heart disease). Main outcome measure Age standardised mortality from causes amenable to health care Setting 19 countries belonging to the Organisation for Economic Cooperation and Development. Results Rankings based on mortality amenable to health care (excluding ischaemic heart disease) differed substantially from rankings of health attainment given in the 2000 World Health Report. No country retained the same position. Rankings for southern European countries and Japan, which had performed well in the report, fell sharply, whereas those of the Nordic countries improved. Some middle ranking countries (United Kingdom, Netherlands) also fell considerably; New Zealand improved its position. Rankings changed when ischaemic heart disease was included as amenable to health care. Conclusion The 2000 World Health Report has been cited widely to support claims for the merits of otherwise different health systems. High levels of health attainment in well performing countries may be a consequence of good fortune in geography, and thus dietary habits, and success in the health effects of policies in other sectors. When assessed in terms of achievements that are more explicitly linked to health care, their performance may not be as good.
Multimorbidity, the simultaneous presence of multiple health conditions in an individual, is an increasingly common phenomenon globally. The systematic assessment of the quality of care delivered to ...people with multimorbidity will be key to informing the organization of services for meeting their complex needs. Yet, current assessments tend to focus on single conditions and do not capture the complex processes that are required for providing care for people with multimorbidity. We conducted a scoping review on quality of care and multimorbidity in selected databases in June 2018 and identified 87 documents as eligible for review, predominantly original research and reviews from North America, Europe and Australasia and mostly frequently related to primary care settings. We synthesized data qualitatively in terms of perceived challenges, evidence and proposed metrics. Findings reveal that the association between quality of care and multimorbidity is complex and depends on the conditions involved (quality appears to be higher for those with concordant conditions, and lower in the presence of discordant conditions) and the approach used for measuring quality (quality appears to be higher in people with multimorbidity when measured using condition/drug‐specific process or intermediate outcome indicators, and worse when using patient‐centred reports of experiences of care). People with discordant multimorbidity may be disadvantaged by current approaches to quality assessment, particularly when they are linked to financial incentives. A better understanding of models of care that best meet the needs of this group is needed for developing appropriate quality assessment frameworks. Capturing patient preferences and values and incorporate patients’ voices in the form of patient‐reported experiences and outcomes of care will be critical towards the achievement of high‐performing health systems that are responsive to the needs of people with multimorbidity.
Content List – Read more articles from the symposium: “Multimorbidity research at the cross‐roads: developing the evidence for clinical practice and health policy”.
The role of microRNA in nutritional control Nolte‐’t Hoen, E. N. M.; Van Rooij, E.; Bushell, M. ...
Journal of internal medicine,
August 2015, Letnik:
278, Številka:
2
Journal Article
Recenzirano
Odprti dostop
MicroRNAs (miRNAs) are one of a growing class of noncoding RNAs that are involved in the regulation of a wide range of metabolic processes including cellular differentiation, cell proliferation and ...apoptosis. The generation of miRNA is regulated in complex ways, for example by small interfering RNAs (small nucleolar and nuclear RNAs) and various other metabolites. This complexity of control is likely to explain how a relatively small part of the DNA that codes for proteins has enabled the evolution of such complex organisms as mammals. Non‐protein‐coding DNA is therefore thought to carry the memory of early evolutionary steps that led to progressively complex metabolic controls. Clinically, miRNAs are becoming increasingly important following the recognition that some congenital abnormalities can be traced to defects in miRNA processing. The potential for manipulating metabolism and affecting disease processes by the pharmaceutical or biological targeting of specific miRNA pathways is now being tested. miRNAs are also released into the extracellular milieu after packaging by cells into nano‐sized extracellular vesicles. Such vesicles can be taken up by adjacent and possibly more distant cells, thereby allowing coordinated intercellular communication in specific tissues. Extracellular miRNAs found in the blood stream may also serve as novel biomarkers for both diagnosing specific forms of cancer and assessing the likelihood of metastasis, and as powerful prognostic indices for various cancers. Here, we discuss the role of intracellular and extracellular miRNAs in nutritional control of various (patho)physiological processes. In this review, we provide an update of the presentations from the 25th Marabou Symposium (Stockholm, 14–16 June 2013) entitled ‘Role of miRNA in health and nutrition’, attended by 50 international experts.
FOR A COMPILATION OF THE GENERAL DISCUSSION AT THE MARABOU SYMPOSIUM, click here http://www.marabousymposium.org/
Introduction
Tissue factor pathway inhibitor (TFPI) is an endogenous inhibitor of the extrinsic pathway that negatively regulates thrombin production during coagulation. Under haemophilic conditions, ...where the intrinsic coagulation pathway is impaired, inhibition of TFPI may improve clotting.
Aim
We investigated the ex vivo effects of a human TFPI neutralizing antibody, marstacimab (previously PF‐06741086), in coagulation assays including rotational thromboelastometry (ROTEM), thrombin generation assay (TGA) and the dilute prothrombin time (dPT) assay, performed in haemophilic whole blood and plasmas. We compared the effects of marstacimab to the effects of recombinant coagulation factors and investigated the reproducibility of marstacimab in restoring haemostasis by comparing its effect in whole blood collected from the same study participants on differing days.
Methods
Citrated whole blood and plasmas obtained from haemophilia participants were supplemented ex vivo with vehicle, marstacimab, recombinant FVIII (rFVIII) or recombinant factor IX (rFIX) and analysed in ROTEM, TGA and the dPT assay using low tissue factor concentrations to trigger coagulation.
Results
Marstacimab induced pro‐coagulant responses in ROTEM parameters including reduction in clotting times and increases in angle. Similarly, participant plasmas supplemented with marstacimab exhibited improvements in TGA parameters, including reduced lag times, increased peak thrombin concentrations and reductions in dPT clotting time. Concentrations of marstacimab tested showed activity comparable to addition of rFVIII or rFIX and were reproducible.
Conclusions
These studies show the ex vivo potency of marstacimab in restoring haemostasis in whole blood and plasmas from haemophilia participants and comparability to ex vivo reconstitution with recombination coagulation factors.
Did the global financial crisis and its aftermath impact upon the performance of health systems in Europe? We investigated trends in amenable and other mortality in the EU since 2000 across 28 EU ...countries.
We use WHO detailed mortality files from 28 EU countries to calculate age-standardized deaths rates from amenable and other causes. We then use joinpoint regression to analyse trends in mortality before and after the onset of the economic crisis in Europe in 2008.
Amenable and other mortality have declined in the EU since 2000, albeit faster for amenable mortality. We observed increases in amenable mortality following the global financial crisis for females in Estonia from -4.53 annual percentage change (APC) in 2005-12 to 0.03 APC in 2012-14 and Slovenia (from -4.22 APC in 2000-13 to 0.73 in 2013-15) as well as males and females in Greece(males: from -2.93 APC in 2000-10 to 0.01 APC in 2010-13; females: from -3.48 APC in 2000-10 to 0.06 APC in 2010-13). Other mortality continued to decline for these populations. Increases in deaths from infectious diseases before and after the crisis played a substantial part in reversals in Estonia, Slovenia and Greece.
There is evidence that amenable mortality rose in Greece and, among females in Estonia and Slovenia. However, in most countries, trends in amenable mortality rates appeared to be unaffected by the crisis.
Summary
The scorecard summarises key indicators of the burden of osteoporosis and its management in each of the member states of the European Union. The resulting scorecard elements were then ...assembled on a single sheet to provide a unique overview of osteoporosis in Europe.
Introduction
The scorecard for osteoporosis in Europe (SCOPE) is an independent project that seeks to raise awareness of osteoporosis care in Europe. The aim of this project was to develop a scorecard and background documents to draw attention to gaps and inequalities in the provision of primary and secondary prevention of fractures due to osteoporosis.
Methods
The SCOPE panel reviewed the information available on osteoporosis and the resulting fractures for each of the 27 countries of the European Union (EU27). The information researched covered four domains: background information (e.g. the burden of osteoporosis and fractures), policy framework, service provision and service uptake e.g. the proportion of men and women at high risk that do not receive treatment (the treatment gap).
Results
There was a marked difference in fracture risk among the EU27. Of concern was the marked heterogeneity in the policy framework, service provision and service uptake for osteoporotic fracture that bore little relation to the fracture burden. For example, despite the wide availability of treatments to prevent fractures, in the majority of the EU27, only a minority of patients at high risk receive treatment for osteoporosis even after their first fracture. The elements of each domain in each country were scored and coded using a traffic light system (red, orange, green) and used to synthesise a scorecard. The resulting scorecard elements were then assembled on a single sheet to provide a unique overview of osteoporosis in Europe.
Conclusions
The scorecard will enable healthcare professionals and policy makers to assess their country’s general approach to the disease and provide indicators to inform future provision of healthcare.
We have determined the production yields for radionuclides in Al
2O
3, SiO
2, S, Ar, K
2SO
4, CaCO
3, Fe, Ni and Cu targets, which were irradiated with slow negative muons at the Paul Scherrer ...Institute in Villigen (Switzerland). The fluences of the stopped negative muons were determined by measuring the muonic X-rays. The concentrations of the long-lived and short-lived radionuclides were measured with accelerator mass spectrometry (AMS) and γ-spectroscopy, respectively. Special emphasis was put on the radionuclides
10Be,
14C and
26Al produced in quartz targets,
26Al in Al
2O
3 and S targets,
36Cl in K
2SO
4 and CaCO
3 targets, and
53Mn in Fe
2O
3 targets. These targets were selected because they are also the naturally occurring target minerals for cosmic ray interactions in typical rocks. We also present results of calculations for depth-dependent production rates of radionuclides produced after cosmic ray μ
− capture, as well as cosmic ray-induced production rates of geologically relevant radionuclides produced by the nucleonic component, by μ
− capture, by fast muons and by neutron capture.
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