To develop a dual hormone artificial pancreas (DHAP) a liquid stable glucagon component is needed. Currently available glucagon products have limited stability after reconstitution and are therefore not suited for DHAP usage. Dasiglucagon is a novel liquid stable glucagon analog, which does not require mixing prior to use. In this randomized double-blind trial 17 patients with T1DM received four single subcutaneous doses (0.03, 0.08, 0.2 and 0.6 mg) of dasiglucagon (4 mg/ml) under hypoglycemic (plasma glucose (PG) 56-66 mg/dL) or euglycemic (PG 100 mg/dL) conditions. For comparison, identical doses of Lilly GlucagonTM (GLUCA) were investigated at euglycemia. In the euglycemic setting dasiglucagon showed rapid and dose-dependent PG-increases of mean 39, 56, 72 and 80 mg/dL at 30 minutes post-dosing (30, 49 and 71 mg/dl for GLUCA) (Figure). During hypoglycemia, dasiglucagon provided similar PG-increases of 24, 64, 87 and 94 mg/dL 30 minutes post dose and rapidly corrected PG levels to normoglycemia (PG ?70 mg/dL) with a median time of 14, 10, 6 and 6 minutes. In conclusion, low doses of dasiglucagon rapidly and effectively increase PG from euglycemia and hypoglycemia in a titratable manner. Therefore, dasiglucagon should be a good candidate for use in dual-hormone closed-loop AP systems.Figure. Different Doses of Dasiglucagon and GLUCA, PG Levels (Mean ±SEM). Disclosure U. Hovelmann: None. M. Braendholt Olsen: Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Self; Zealand Pharma A/S. U. Mouritzen: Employee; Self; Zealand Pharma A/S. Stock/Shareholder; Self; Novo Nordisk A/S. D. Lamers: None. B. Kronshage: None. T. Heise: Research Support; Self; ADOCIA, Boehringer Ingelheim GmbH, Dance Biopharm, Eli Lilly and Company, Janssen Research & Development, MedImmune, Merck Sharp & Dohme Corp., Mylan, Nordic Bioscience, Novo Nordisk A/S, Poxel SA, Roche Diagnostics Corporation, Saniona, Sanofi, Senseonics, Zealand Pharma A/S. Advisory Panel; Self; Novo Nordisk A/S, Mylan. Speaker's Bureau; Self; Dexcom, Inc., Eli Lilly and Company, Novo Nordisk A/S, Sanofi.

Fragestellung: PEDF gehört zur Familie der nicht-inhibierenden Serpine und ist ein multi-funktionales Protein mit neurotrophen und anti-angiogenen Eigenschaften. Seit kurzem ist bekannt, dass Typ 2 Diabetiker erhöhte PEDF Konzentrationen im Serum aufweisen. In Mäusen konnte gezeigt werden, dass die PEDF Expression im Fettgewebe positiv mit Adipositas und Insulinresistenz korreliert. Durch die Sekretionsanalyse von in vitro differenzierten humanen Adipozyten mittels 2D-PAGE und MALDI-MS konnten wir zeigen, dass PEDF eines der abundantesten Proteine im Sekretom der humanen Fettzelle ist. Ziel dieser Studie war es, die Regulation und autokrine Funktion von PEDF in humanen Adipozyten zu untersuchen und die parakrinen Effekte auf humane Skelettmuskelzellen (hSkMC) und glatte Muskelzellen (hSMC) zu bestimmen.

Objective: Pigment epithelium-derived factor (PEDF) is a multifunctional protein with neurotrophic and anti-angiogenic properties. More recently it became evident that PEDF is upregulated in patients with type 2 diabetes and also contributes to insulin resistance in mice. During characterization of the secretome of in vitro differentiated human adipocytes by two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-MS, we found that PEDF is one of the most abundant proteins released by adipocytes. The aim of this study was to investigate the regulation and autocrine function of PEDF in human adipocytes and to determine its paracrine effects on human skeletal muscle cells (hSkMC) and human smooth muscle cells (hSMC). Methods and results: Human primary adipocytes secrete 130 ng ml −1 PEDF over 24 h from 1 million cells, which is extremely high as compared with adiponectin, interleukin-6 (IL-6) or IL-8. This release of PEDF is significantly higher than from other primary cells, such as adipose-tissue located macrophages (50-times), hSkMC and hSMC (5-times). PEDF protein expression significantly increases during adipogenesis, which is paralleled by increased PEDF secretion. Furthermore, tumor necrosis factor-α and hypoxia significantly downregulate PEDF protein levels. PEDF secretion was significantly reduced by troglitazone and hypoxia and significantly increased by insulin. Treatment of adipocytes and hSkMC with PEDF induced insulin resistance in adipocytes, skeletal and smooth muscle cells at the level of insulin-stimulated Akt phosphorylation, which was dose dependent and more prominent in adipocytes. Furthermore, inflammatory nuclear factor-κB (NF-κB) signaling was induced by PEDF. In hSMC, PEDF induced proliferation (1.7-fold) and acutely activated proliferative and inflammatory signaling pathways (NF-κB, p38 mitogen-activated protein kinase and mammalian target of rapamycin). Conclusion: PEDF is one of the most abundant adipokines and its secretion is inversely regulated by insulin and hypoxia. PEDF induces insulin resistance in adipocytes and hSkMC and leads to inflammatory signaling in hSMC. Because of these diverse actions, PEDF is a key adipokine, which could have an important role in diabetes and obesity-related disorders.

Fragestellung: Als endokrines Organ setzt das Fettgewebe eine große Anzahl von Peptiden und Proteinen frei. Diese sogenannten Adipokine sind an der Entwicklung verschiedener Krankheitsbilder wie dem Metabolischen Syndrom oder dem Typ 2 Diabetes beteiligt, wobei deren genaue Zusammensetzung zum großen Teil noch unbekannt ist. Die detaillierte Charakterisierung des Sekretoms der Fettzelle kann somit einen entscheidenden Beitrag zur Identifizierung neuer potentieller, krankheitsassoziierter Biomarker leisten.

Fragestellung: Die gesteigerte Fettgewebsmasse bei der Adipositas stellt einen großen Risikofaktor für die Entstehung der Atherosklerose dar. Perivaskuläres Fett ist im Kontext der Adipositas und dem kardiometabolischen Syndrom als Auslöser für Inflammation und Proliferation in der Gefäßwand beschrieben. Adiponectin, ein bekanntes anti-inflammatorisches Adipokin, kann autokrin das Sekretom der Fettzellen verändern, und die Produktion von inflammatorischen Adipokinen reduzieren. Ziel dieser Studie ist es, durch Adiponectin regulierte Fettzellprodukte mittels 2-D-DIGE Multiplexanalyse zu charakterisieren, und seine anti-atherogene Wirkung auf Ebene von koronaren glatten Muskelzellen (SMC) zu bestimmen.

Fragestellung: Bei der Entwicklung von Atherosklerose spielt die abnorme Proliferation und Migration der glatten Muskelzellen (SMC) eine wichtige Rolle. Ein unabhängiger Risikofaktor für Atherosklerose ist Adipositas. Das Fettgewebe ist ein endokrines Organ, welches verschiedene pro-inflammatorische Adipokine produziert und ebenso freie Fettsäuren abgibt, wie zum Beispiel Ölsäure (OA), welche bekannt dafür ist den „synthetischen“ Typ der SMC in der Atherosklerose zu begünstigen. Wir haben in dieser Studie untersucht, wie Adipokine, OA und die Kombination von beiden die Proliferation der SMC beeinflussen und welche Signalwege beteiligt sind.

Fragestellung: Das Fettgewebe ist als endokrines Organ durch die Produktion einer Vielzahl von inflammatorischen Adipokinen, wie IL-6, TNF alpha, PAI-1 oder Angiotensin gekennzeichnet. Die gesteigerte Freisetzung dieser Adipokine kann negativ auf die Gefäßwand einwirken und spielt möglicherweise eine entscheidende Rolle bei der Entstehung der Atherosklerose. Adiponectin, ein bekanntes anti-inflammatorisches Adipokin, kann autokrin das Sekretom der Fettzelle verändern und die Produktion anti-inflammatorischer Adipokine reduzieren. Ziel dieser Studie war es, die durch Adiponectin regulierten Fettzellprodukte durch vergleichende Proteinpattern-Analyse zu charakterisieren und die direkte anti-atherogene Wirkung auf Gefäßzellen (koronare glatte Muskelzellen, SMC) zu bestimmen.

A very small proportion of newborns fail to establish a normal respiratory pattern without some assistance at birth and newborns requiring resuscitative measures at birth should have a skilled practitioner able to provide it. In this small hospital midwives felt unskilled in newborn resuscitation and paediatricians were not always immediately available. A stakeholder group gathered to discuss the problem. A training program was implemented that improved the skills, confidence and support of the attending midwife to resuscitate the newborn effectively. The aim was to improve the outcome for the infant whilst maintaining the mother's choice of birthing in a small local hospital close to family and friends.

TRPV5 and TRPV6 constitute the Ca2+ influx pathway in a variety of epithelial cells. Here, we identified S100A10 as the first auxiliary protein of these epithelial Ca2+ channels using yeast two‐hybrid and GST pull‐down assays. This S100 protein forms a heterotetrameric complex with annexin 2 and associates specifically with the conserved sequence VATTV located in the C‐terminal tail of TRPV5 and TRPV6. Of these five amino acids, the first threonine plays a crucial role since the corresponding mutants (TRPV5 T599A and TRPV6 T600A) exhibited a diminished capacity to bind S100A10, were redistributed to a subplasma membrane area and did not display channel activity. Using GST pull‐down and co‐immunoprecipitation assays we demonstrated that annexin 2 is part of the TRPV5–S100A10 complex. Furthermore, the S100A10–annexin 2 pair colocalizes with the Ca2+ channels in TRPV5‐expressing renal tubules and TRPV6‐expressing duodenal cells. Importantly, downregulation of annexin 2 using annexin 2‐specific small interfering RNA inhibited TRPV5 and TRPV6‐mediated currents in transfected HEK293 cells. In conclusion, the S100A10–annexin 2 complex plays a crucial role in routing of TRPV5 and TRPV6 to plasma membrane.

Städt. Klinik Dortmund, Medizinische Klinik, Beurhausstrasse 40, 4600 Dortmund, Federal Republic of Germany