Agarose Spot as a Comparative Method for in situ Analysis of Simultaneous Chemotactic Responses to Multiple Chemokines

yesWe describe a novel protocol to quantitatively and simultaneously compare the chemotactic responses of cells towards different chemokines. In this protocol, droplets of agarose gel containing different chemokines are applied onto the surface of a Petri dish, and then immersed under culture medium in which cells are suspended. As chemokine molecules diffuse away from the spot, a transient chemoattractant gradient is established across the spots. Cells expressing the corresponding cognate chemokine receptors migrate against this gradient by crawling under the agarose spots towards their centre. We show that this migration is chemokine-specific; meaning that only cells that express the cognate chemokine cell surface receptor, migrate under the spot containing its corresponding chemokine ligand. Furthermore, we show that migration under the agarose spot can be modulated by selective small molecule antagonists present in the cell culture medium

Long-term follow-up with Granulocyte and Monocyte Apheresis re-treatment in patients with chronically active inflammatory bowel disease

<p>Abstract</p> <p>Background</p> <p>Patients with IBD and chronic inflammation refractory to conventional therapy often demonstrate higher risk of serious complications. Combinations of immunosuppression and biological treatment as well as surgical intervention are often used in this patient group. Hence, there is need for additional treatment options. In this observational study, focused on re-treatment and long-term results, Granulocyte/Monocyte Adsorption (GMA, Adacolumn^®) treatment has been investigated to study efficacy, safety and quality of life in IBD-patients with chronic activity.</p> <p>Methods</p> <p>Fifteen patients with ulcerative colitis and 25 patients with Crohn's disease, both groups with chronically active inflammation refractory to conventional medication were included in this observational study. The patients received 5-10 GMA sessions, and the clinical activity was assessed at baseline, after each completed course, and at week 10 and 20 by disease activity index, endoscopy and quality of life evaluation. Relapsed patients were re-treated by GMA in this follow-up study up to 58 months.</p> <p>Results</p> <p>Clinical response was seen in 85% and complete remission in 65% of the patients. Ten patients in the UC-group (66%) and 16 patients in the CD-group (64%) maintained clinical and endoscopic remission for an average of 14 months. Fourteen patients who relapsed after showing initial remission were re-treated with GMA and 13 (93%) went into a second remission. Following further relapses, all of seven patients were successfully re-treated for the third time, all of three patients for the fourth time and one for a fifth time.</p> <p>Conclusions</p> <p>IBD-patients with chronic inflammation despite conventional therapy seem to benefit from GMA. Re-treatment of relapsing remission patients seems to be effective.</p

Pulsatility of insulin release – a clinically important phenomenon

The mechanisms and clinical importance of pulsatile insulin release are presented against the background of more than half a century of companionship with the islets of Langerhans. The insulin-secreting β-cells are oscillators with intrinsic variations of cytoplasmic ATP and Ca2+. Within the islets the β-cells are mutually entrained into a common rhythm by gap junctions and diffusible factors (ATP). Synchronization of the different islets in the pancreas is supposed to be due to adjustment of the oscillations to the same phase by neural output of acetylcholine and ATP. Studies of hormone secretion from the perfused pancreas of rats and mice revealed that glucose induces pulses of glucagon anti-synchronous with pulses of insulin and somatostatin. The anti-synchrony may result from a paracrine action of somatostatin on the glucagon-producing α-cells. Purinoceptors have a key function for pulsatile release of islet hormones. It was possible to remove the glucagon and somatostatin pulses with maintenance of those of insulin with an inhibitor of the P2Y1 receptors. Knock-out of the adenosine A1 receptor prolonged the pulses of glucagon and somatostatin without affecting the duration of the insulin pulses. Studies of isolated human islets indicate similar relations between pulses of insulin, glucagon, and somatostatin as found during perfusion of the rodent pancreas. The observation of reversed cycles of insulin and glucagon adds to the understanding how the islets regulate hepatic glucose production. Current protocols for pulsatile intravenous infusion therapy (PIVIT) should be modified to mimic the anti-synchrony between insulin and glucagon normally seen in the portal blood

P023 Prevalence of malnutrition, risk of malnutrition and quality of life among patients with Inflammatory Bowel Disease

Abstract Background The relapsing and remitting course of inflammatory bowel disease (IBD) with inflammation and gut symptoms may result in an elevated risk of malnutrition and impaired quality of life. Patients with pro-longed active IBD are at the highest risk of developing impaired nutritional status. The aim of this study was to determine the prevalence of malnutrition and risk of malnutrition in outpatients with inflammatory bowel disease. Furthermore, to study their body composition, identify factors associated with malnutrition and investigate the association between malnutrition and quality of life. Methods A total of 328 IBD outpatients, (154 [47 %] with Crohn’s disease (CD) and 174 [53%] with ulcerative colitis (UC)) at Linköping University hospital were included in this population-based cross-sectional study. Patients underwent malnutrition assessment using the recently published Global Leadership Initiative on Malnutrition (GLIM) criteria. Data about body mass index, weight loss and dietary problems were collected and used to evaluate risk of malnutrition. The Short Health Scale (SHS) was used to measure quality of life. Results Malnutrition was detected in 33 patients (10,1 %) and 162 patients (55,0 %) had an increased risk of malnutrition. Malnutrition was associated with lower quality of life in the entire patient cohort and there was no difference between diagnoses. The prevalence of malnutrition was significantly higher among women (73% vs 27 %). Malnourished patients and those at risk of impaired nutritional status were more likely to have had active disease during the preceding 12 months compared to patients without malnutrition or at risk of malnutrition (27 % vs 12% [p=0.015] and 15 % vs 7% [p=0.037] respectively). Surprisingly, a higher calprotectin value was detected among patients without risk of malnutrition when compared to those who were at risk (median 46 [IQR 17 – 106] vs 59 [IQR 25 – 288] respectively, p=0,009). Women with IBD had significantly lower fat-free mass index (FFMI) compared to men. Patients with CD had reduced quality of life compared to patients with UC. Conclusion The prevalence of malnutrition in IBD-outpatients was 10% and more than half of the participants were at risk of malnutrition. Malnutrition was associated with poor quality of life. Malnutrition screening might be important in this population as ongoing inflammation and active disease seem to be apparent even among patients without clear nutrition difficulties or weight problems, but might influence nutritional status and quality of life negatively in the long term. Patients with IBD should therefore be screened for malnutrition and risk of malnutrition on a regular basis. </jats:sec