Toll-like receptor-induced cytokine secretion by human monocytes in healthy donors and septic patients assessed at the single cell level

Following the ligation of Toll-like receptors (TLRs) by bacterial-derived signature molecules, innate immune cells such as monocytes, macrophages and neutrophils initiate the inflammatory process by secreting chemokines and cytokines with a wide range of immunomodulatory effects. Under normal conditions, these mediators work to compartmentalize the inflammatory process and eradicate the intruder, while simultaneously recruiting more immune cells to the site of infection. In sepsis, however, the virtues of this containment are lost as the response against a disseminated bacterial infection becomes systemic with an excessive production of cytokines that can lead to tissue injury, organ dysfunction and ultimately death. Here we have investigated TLR-induced cytokine secretion in vitro by monocytes and granulocytes from healthy donors and septic patients using the ELISpot and/or FluoroSpot assays. By looking at the secretion of cytokines directly ex vivo, the ELISpot assay offered the potential of being able to better define the immunological status of septic patients. We investigated the lipopolysaccharide (LPS)-induced cytokine secretion by peripheral blood mononuclear cells (PBMC) and granulocytes from healthy donors using the ELISpot assay. Cytokines (IL-1β, IL-6, IL-10, IL-12p40, TNF-α, GM-CSF) and chemokines (IL-8, MIP-1β) important in inflammatory processes were assessed. Granulocytes were found to selectively secrete IL-8 and MIP-1β. Also TNF-α was secreted but by considerably fewer cells. In contrast, PBMCs secreted all evaluated cytokines with CD14+ monocytes being the main source of production. Next, we analyzed the cytokine secretion by enriched monocytes from healthy donors in response to LPS and lipoteichoic acid (LTA). The FluoroSpot technique allowed for the simultaneous analysis of two cytokines from the same population of isolated cells. With this approach, a recurring pattern of cytokine co-secretion was observed, identifying several distinct cytokine-secreting profiles for the TNF-α-, GM-CSF-, IL-10-, IL-12p40- secreting monocytes and those secreting IL-6 or IL-1β. Finally, the spontaneous and LPS-induced cytokine secretion by total leukocytes isolated from septic patients (n=32) and healthy controls (n=30) was evaluated using the ELISpot assay. Surprisingly, we found no increase in the number of constitutively cytokine-secreting cells from any of the septic patients despite significantly increased levels of cytokines (IL-6, IL-1β, TNF- α, GM-CSF, IL-10, IL-12p40) in their plasma. Simultaneously, the LPS-induced in vitro capacity revealed a maintained (IL-6, TNF-α) as well as reduced (IL-1β, GM- CSF, IL-10, IL-12p40) number of cytokine-secreting monocytes in the patients compared to normal donors. This selective reduction for some of the cytokines could be correlated with disease severity. In conclusion, our data indicate that circulating monocytes are not the major source of increased cytokine levels in patients with sepsis

Intiaanit ovat edelleen keskuudessamme

Rani-Henrik Andersson ja Markku Henriksson: Intiaanit – Pohjois- Amerikan alkuperäiskansojen historia. Gaudeamus 2010

Between Unity and Diversity : The Application of the Charter of Fundamental Rights in the Light of the Principle of Subsidiarity

The aim of this study is to examine the application of the Charter of Fundamental Rights of the European Union in the light of the principle of subsidiarity in EU law. The Charter constitutes a point of intersection of subsidiarity and fundamental rights, brought together by their common purpose of guiding the exercise of public powers and reconciling a diversity of Member State values with the need for European unity. With a particular focus on the case law of the Court of Justice of the European Union, the study embraces the principle of subsidiarity for the purpose of interpreting the legal nature of the Charter and systemising its application. The formal link between subsidiarity and fundamental rights under the Charter is found in Article 51(1). In order to clarify the legal nature of the Charter and the context in which it operates, the study looks into how fundamental rights and subsidiarity have developed in EU law as well as how they are interrelated. The subsidiary nature of EU fundamental rights law is demonstrated by the restrictive personal and material scope of the Charter. Similarly, the principle of subsidiarity is used to analyse the Charter’s minimum and maximum level of protection. The Court’s application of the Charter is analysed by examining the expressions that the principle of subsidiarity takes when the Charter is balanced with economic freedoms, applied in conjunction with other norms, used to invalidate EU norms or called to set aside national legal acts. In addition to this, the analysis considers the role of subsidiarity in cases where the Charter was not applied, either because the Court lacked jurisdiction or remained silent on its applicability. The study demonstrates how the legal nature of the Charter to a great extent can be explained through the principle of subsidiarity. However, the EU legal order has its proper understanding of subsidiarity in relation to fundamental rights. Fundamental rights protection is only one of several EU objectives that need to be balanced when determining the appropriate level of action. Ultimately, the application of the Charter is triggered by the need to preserve the primacy, unity and effectiveness of EU law. Only in exceptional circumstances, these requirements may be overridden with reference to the principle of legal certainty, as recognised by the Charter. Conversely, subsidiarity as an interpretative principle of EU fundamental rights law may also contribute to enhanced legal certainty regarding the application of the Charter.Syftet med denna avhandling är att undersöka tillämpningen av Europeiska unionens stadga om de grundläggande rättigheterna i ljuset av den EU-rättsliga subsidiaritetsprincipen. Stadgan utgör en skärningspunkt för subsidiaritet och grundläggande rättigheter, vilka förenas av en gemensam strävan att reglera utövningen av offentlig makt och sammanjämka medlemsstaternas mångfacetterade värderingar med behovet av europeisk gemenskap. Med särskilt fokus på rättspraxis från Europeiska unionens domstol antar undersökningen subsidiaritetsprincipen som ett led i uttolkningen av stadgans rättsliga status och systematiseringen av dess tillämpning. Den formella kopplingen mellan subsidiaritet och grundläggande rättigheter återfinns i artikel 51(1) i stadgan. Avhandlingen undersöker hur de grundläggande rättigheterna och subsidiaritetsprincipen har utvecklats inom EU-rätten samt deras inbördes förhållande, i syfte att klargöra stadgans rättsliga status och den kontext i vilken den verkar. Den subsidiära karaktär som EU-rättens grundläggande rättigheter besitter framgår av stadgans begränsade personkrets och tillämpningsområde. Subsidiaritetsprincipen kan även tillämpas på analysen av minimi- och maximinivån på det skydd som stadgan erbjuder. Domstolens tillämpning av stadgan analyseras med utgångspunkt i de uttryck som subsidiaritetsprincipen tar när stadgan vägs mot de ekonomiska friheterna, tillämpas i kombination med andra rättsnormer, används för att ogiltigförklara EU-rättsliga normer eller åberopas för att åsidosätta nationell lagstiftning. Därtill beaktas betydelsen av subsidiaritetsprincipen i rättsfall där stadgan inte tillämpats, antingen eftersom domstolen saknat behörighet eller låtit bli att uttala sig om dess tillämplighet. Avhandlingen visar hur stadgans rättsliga status i stor utsträckning kan förklaras med hjälp av subsidiaritetsprincipen. EU-rätten har emellertid en särpräglad tolkning av subsidiaritetsprincipen i förhållande till grundläggande rättigheter. Skyddet för de grundläggande rättigheterna är endast en av EU:s många målsättningar som måste vägas mot varandra när domstolen fastställer på vilken nivå en åtgärd ska anses ändamålsenlig. Särskilt behovet att bevara EU-rättens företräde, enhetlighet och verkan kan ses som utlösande faktorer för tillämpningen av stadgan. Endast i undantagsfall kan dessa förutsättningar åsidosättas med hänvisning till individens rättssäkerhet som erkänns i stadgan. I egenskap av tolkningsprincip för EU:s grundläggande rättigheter kan subsidiaritetsprincipen samtidigt också anses bidra till ökad förutsebarhet vad gäller stadgans tillämplighet

Feasibility of using smartphones by village health workers for pregnancy registration and effectiveness of mobile phone text messages on reduction of homebirths in rural Uganda

INTRODUCTION: Homebirths are common in low and middle income countries and are associated with poor child survival. We assessed the feasibility of using smartphones by village health workers for pregnancy registration and the effectiveness of health text messages (SMS) sent to pregnant women through village health workers in reducing homebirths in rural Uganda. METHODS: A non-randomised intervention study was undertaken in 26 villages. In the intervention arm, village health workers registered pregnant women (n = 262) in 13 villages using a smartphone app (doForm) and paper forms and gestation age-timed SMS were sent through village health workers to the pregnant women. In 13 control villages, (n = 263) pregnant women were registered on paper forms only and no SMS was sent. The main outcome was place of birth measured through a self-report. Logistic regression with generalised estimating equations was used to explore the effect of the intervention. RESULTS: Comparing 795 corresponding data fields on phone and paper revealed that numeric variable fields were 86%-95% similar while text fields were 38%-48% similar. Of the 525 pregnant women followed, 83 (15.8%) delivered at home. In the adjusted analysis, the intervention was associated with lower odds of homebirths [AOR = 0.38, 95%CI (0.15-0.97)]. Muslim religion [AOR = 4.0, 95%CI (1.72-9.34)], primary or no maternal education [AOR = 2.51, 95%CI (1.00-6.35)] and health facility distance ≥ 2 km [AOR = 2.26, 95%CI (0.95-5.40)] were independently associated with homebirths. CONCLUSION: Village health workers can register pregnant women at home using phones and relay gestation age specific SMS to them to effectively reduce homebirths

Adverse pregnancy outcomes in rural Uganda (1996-2013): trends and associated factors from serial cross sectional surveys.

OBJECTIVE: Community based evidence on pregnancy outcomes in rural Africa is lacking yet it is needed to guide maternal and child health interventions. We estimated and compared adverse pregnancy outcomes and associated factors in rural south-western Uganda using two survey methods. METHODS: Within a general population cohort, between 1996 and 2013, women aged 15-49 years were interviewed on their pregnancy outcome in the past 12 months (method 1). During 2012-13, women in the same cohort were interviewed on their lifetime experience of pregnancy outcomes (method 2). Adverse pregnancy outcome was defined as abortions or stillbirths. We used random effects logistic regression for method 1 and negative binomial regression with robust clustered standard errors for method 2 to explore factors associated with adverse outcome. RESULTS: One third of women reported an adverse pregnancy outcome; 10.8% (abortion = 8.4%, stillbirth = 2.4%) by method 1 and 8.5% (abortion = 7.2%, stillbirth = 1.3%) by method 2. Abortion rates were similar (10.8 vs 10.5) per 1000 women and stillbirth rates differed (26.2 vs 13.8) per 1000 births by methods 1 and 2 respectively. Abortion risk increased with age of mother, non-attendance of antenatal care and proximity to the road. Lifetime stillbirth risk increased with age. Abortion and stillbirth risk reduced with increasing parity. DISCUSSION: Both methods had a high level of agreement in estimating abortion rate but were markedly below national estimates. Stillbirth rate estimated by method 1 was double that estimated by method 2 but method 1 estimate was more consistent with the national estimates. CONCLUSION: Strategies to improve prospective community level data collection to reduce reporting biases are needed to guide maternal health interventions