FluoroSpot Analysis of TLR-Activated Monocytes Reveals Several Distinct Cytokine-Secreting Subpopulations

Monocytes have long been considered a heterogeneous group of cells both in terms of morphology and function. In humans, three distinct subsets have been described based on their differential expression of the cell surface markers CD14 and CD16. However, the relationship between these subsets and the production of cytokines has for the most part been based on ELISA measurements, making it difficult to draw conclusions as to their functional profile on the cellular level. In this study, we have investigated lipoteichoic acid (LTA)- and lipopolysaccharide (LPS)-induced cytokine secretion by monocytes using the FluoroSpot technique. This method measures the number of cytokine-secreting cells on the single-cell level and uses fluorescent detection, allowing for the simultaneous analysis of two cytokines from the same population of isolated cells. By this approach, human monocytes from healthy volunteers could be divided into several subgroups as IL-1β, IL-6, TNF-α and MIP-1β were secreted by larger populations of responding cells (25.9–39.2%) compared with the smaller populations of GM-CSF (9.1%), IL-10 (1.3%) and IL-12p40 (1.2%). Furthermore, when studying co-secretion in FluoroSpot, an intricate relationship between the monocytes secreting IL-1β and/or IL-6 and those secreting TNF-α, MIP-1β, GM-CSF, IL-10 and IL-12p40 was revealed. In this way, dissecting the secretion pattern of the monocytes in response to TLR-2 or TLR-4 stimulation, several subpopulations with distinct cytokine-secreting profiles could be identified

Effects of resuscitation with crystalloid fluids on cardiac function in patients with severe sepsis

<p>Abstract</p> <p>Background</p> <p>The use of hypertonic crystalloid solutions, including sodium chloride and bicarbonate, for treating severe sepsis has been much debated in previous investigations. We have investigated the effects of three crystalloid solutions on fluid resuscitation in severe sepsis patients with hypotension.</p> <p>Methods</p> <p>Ninety-four severe sepsis patients with hypotension were randomly assigned to three groups. The patients received the following injections within 15 min at initial treatment: Ns group (n = 32), 5 ml/kg normal saline; Hs group (n = 30), with 5 ml/kg 3.5% sodium chloride; and Sb group (n = 32), 5 ml/kg 5% sodium bicarbonate. Cardiac output (CO), systolic blood pressure, mean arterial pressure (MAP), body temperature, heart rate, respiratory rate and blood gases were measured.</p> <p>Results</p> <p>There were no differences among the three groups in CO, MAP, heart rate or respiratory rate during the 120 min trial or the 8 hour follow-up, and no significant differences in observed mortality rate after 28 days. However, improvement of MAP and CO started earlier in the Sb group than in the Ns and Hs groups. Sodium bicarbonate increased the base excess but did not alter blood pH, lactic acid or [HCO₃]^-values; and neither 3.5% hypertonic saline nor 5% sodium bicarbonate altered the Na⁺, K⁺, Ca²⁺or Cl^-levels.</p> <p>Conclusion</p> <p>All three crystalloid solutions may be used for initial volume loading in severe sepsis, and sodium bicarbonate confers a limited benefit on humans with severe sepsis.</p> <p>Trial registration</p> <p>ISRCTN36748319.</p

Endothelin receptor antagonism and hypertonic solutions in experimental endotoxin shock

Sepsis is a common and serious condition among patients in the intensive care unit. It is characterized by a systemic inflammatory response, resulting in cardiovascular instability and impaired organ tissue oxygenation due to hypoperfusion and inflammatory changes. The supportive treatment includes fluid resuscitation and other measures to promote organ perfusion. Hypertonic saline/dextran (HSD) resuscitation has shown improved haemodynamics in haemorrhage. In addition, immuno-modulatory effects have been reported in experimental studies. These effects would in theory also be of benefit in septic shock. The vasoconstrictive peptide endothelin (ET) is involved in several of the sepsis manifestations. Experimental results of ET antagonism in endotoxemia have demonstrated beneficial effects on cardiac performance and regional perfusion. Hypotension due to vasodilation may however be of concern in septic conditions, requiring measures to secure perfusion pressure. The overall aim of this thesis was to evaluate two different interventions and their ability to affect vital organ perfusion through effects on systemic and regional haemodynamics in porcine endotoxin shock. Specifically, the cardiovascular effects of HSD resuscitation were further evaluated, with focus on regional perfusion. In addition, effects of the new parenteral ET antagonist, tezosentan, on systemic and regional haemodynamics were investigated. Furthermore, tezosentan was combined with HSD with the purpose of preventing ET antagonism-related hypotension, and possibly augment positive cardiovascular effects. In addition, early pro-inflammatory cytokine reponse was studied with both treatments to further evaluate their immunomodulating potential. A model of endotoxin shock in anaesthetized pigs was used in two series of experiments. Haemodynamic responses were assessed using a pulmonary artery catheter, invasive arterial blood pressure and ultrasonic flow probes positioned around the portal vein and the renal and carotid arteries. Plasma TNF-á and IL-6 levels were monitored. Endotoxemia resulted in a hypodynamic shock including reduced regional blood flow and development of metabolic acidosis. An inflammatory response with increases in cytokine and endothelin-1 levels was elicited. HSD resuscitation transiently improved cardiac index and mean arterial pressure. Regional blood flow increased in the renal and carotid, but not portal circulation. The progression of acidosis was delayed and the macro- and microcirculatory improvements resulted in improved survival rates. Tezosentan treatment resulted in vasodilation, which improved cardiac index, reversed pulmonary hypertension and increased portal and carotid, but not renal blood flow. Addition of HSD to tezosentan did not increase mean arterial pressure, but markedly increased portal and carotid perfusion. None of the interventions affected TNF-á and IL-6 response. In conclusion, transient beneficial effects of HSD on systemic and renal haemodynamics were confirmed, and improved early survival indicated. From a clinical perspective, HSD resuscitation may gain time for other treatments to have their effects. The immunomodulatory potential of HSD in endotoxemia was not supported by the results from this study. However, ET antagonism as a means to promote organ perfusion was supported by the results. The improved regional flow demonstrated with combined tezosentan/HSD emphasizes the importance of additional measures to secure perfusion pressure during ET antagonism

Attachment in Preschool : Educational Approaches for Children with Previous Insecure Attachment

Detta examensarbete handlar om förskollärares erfarenheter av arbetet med barn som har en otrygg anknytning. Syftet med arbetet är att undersöka förskollärares beskrivningar av arbetet med barn med otrygg anknytning i kontext till ett utsatt område. För att kunna besvara våra frågeställningar har vi valt att använda följande frågor: Vilka metoder med avsikt att främja en trygg förskolemiljö beskrivs? Vilka utmaningar uppmärksammas? I vårt examensarbete har vi använt oss av en kvalitativ metod där vi använt semistrukturerade intervjuer för att samla in den data vi behöver. Studiens respondenter är förskollärare som arbetar på olika förskolor i utsatta områden. I vårt resultat kan vi se att förskollärarna använder olika strategier och metoder för att identifiera barn som har en otrygg anknytning, men även för att stödja dessa barn på bästa sätt. Vi kan även se i vårt resultat att förskollärarna stöter på utmaningar i arbetet med dessa barn. De utmaningar som lyfts fram är språk och kommunikation, stora barngrupper och brist på förskollärare med rätt utbildning

Attachment in Preschool : Educational Approaches for Children with Previous Insecure Attachment

Detta examensarbete handlar om förskollärares erfarenheter av arbetet med barn som har en otrygg anknytning. Syftet med arbetet är att undersöka förskollärares beskrivningar av arbetet med barn med otrygg anknytning i kontext till ett utsatt område. För att kunna besvara våra frågeställningar har vi valt att använda följande frågor: Vilka metoder med avsikt att främja en trygg förskolemiljö beskrivs? Vilka utmaningar uppmärksammas? I vårt examensarbete har vi använt oss av en kvalitativ metod där vi använt semistrukturerade intervjuer för att samla in den data vi behöver. Studiens respondenter är förskollärare som arbetar på olika förskolor i utsatta områden. I vårt resultat kan vi se att förskollärarna använder olika strategier och metoder för att identifiera barn som har en otrygg anknytning, men även för att stödja dessa barn på bästa sätt. Vi kan även se i vårt resultat att förskollärarna stöter på utmaningar i arbetet med dessa barn. De utmaningar som lyfts fram är språk och kommunikation, stora barngrupper och brist på förskollärare med rätt utbildning

#Bodygoals : En kvalitativ studie om hur Instagram påverkar unga tjejers kroppsuppfattning

Instagram är en fotodelningsapplikation som i stor utsträckning används av den yngre befolkningen. Det har visat sig att unga tjejer är den målgrupp som i störst utsträckning påverkas av det innehållet som publiceras på Instagram, framförallt när det kommer till aspekter gällande utseende och kroppsideal. Syftet med studien var att undersöka hur unga tjejer förhåller sig till Instagram samt vilka erfarenheter de har av det kroppsrelaterade innehåll som framställs på applikationen och hur det i sin tur kan komma att påverka deras kroppsuppfattning. Studien bygger på en kvalitativ ansats och empirin har samlats in genom fokusgrupper, i vilka sex tjejer i åldrarna 18 till 19 år deltog, samt genom netnografi. Genom en kvalitativ innehållsanalys har fem teman växt fram, som sedan legat till grund för hur resultatet presenterats. Av resultatet framgår att unga tjejer påverkas såväl medvetet som omedvetet av de kroppsrelaterade innehåll som publiceras på Instagram, vilket leder till både positiva och negativa känslor om den egna kroppen.2018-06-07</p