Call for a reference model of chronic hind limb ischemia to investigate therapeutic angiogenesis

A large number of studies utilize animal models to investigate therapeutic angiogenesis. However, the lack of a standardized experimental model leaves the comparison of different studies problematic. To establish a reference model of prolonged moderate tissue ischemia, we created unilateral hind limb ischemia in athymic rnu-rats by surgical excision of the femoral vessels. Blood flow of the limb was monitored for 60 days by laser Doppler imaging. Following a short postoperative period of substantially depressed perfusion, the animals showed a status of moderate hind limb ischemia from day 14 onwards. Thereafter, the perfusion remained at a constant level (55.5% of normal value) until the end of the observation period. Histopathological assessment of the ischemic musculature on postoperative days 28 and 60 showed essentially no inflammatory cell infiltrate or fibrosis. However, the mitochondrial activity and capillary-to-fiber ratio of the muscular tissue was reduced to 52.7% of normal, presenting with a significant weakness of the ischemic limb evidenced by a progressive decline in performance. Intramuscular injection of culture-expanded human endothelial progenitor cells (EPC) resulted in a significant increase in blood flow (82.0+/-3.5% of normal), capillary density (1.60+/-0.08/muscle fiber) and smooth muscle covered arterioles (8.0+/-0.6/high power field) in the ischemic hind limb as compared to controls (55.0+/-3.1%; 0.99+/-0.03; 5.0+/-0.2). In conclusion, chronic, moderate hind limb ischemia with consistently reduced perfusion levels persisting over a prolonged period can be established reliably in rnu athymic nude rats and is responsive to pro-angiogenic treatments such as EPC transplantation. This study provides a detailed protocol of a highly reproducible reference model to test novel therapeutic options for limb ischemia.

Novel cell-free strategy for therapeutic angiogenesis: in vitro generated conditioned medium can replace progenitor cell transplantation

BACKGROUND: Current evidence suggests that endothelial progenitor cells (EPC) contribute to ischemic tissue repair by both secretion of paracrine factors and incorporation into developing vessels. We tested the hypothesis that cell-free administration of paracrine factors secreted by cultured EPC may achieve an angiogenic effect equivalent to cell therapy. METHODOLOGY/PRINCIPAL FINDINGS: EPC-derived conditioned medium (EPC-CM) was obtained from culture expanded EPC subjected to 72 hours of hypoxia. In vitro, EPC-CM significantly inhibited apoptosis of mature endothelial cells and promoted angiogenesis in a rat aortic ring assay. The therapeutic potential of EPC-CM as compared to EPC transplantation was evaluated in a rat model of chronic hindlimb ischemia. Serial intramuscular injections of EPC-CM and EPC both significantly increased hindlimb blood flow assessed by laser Doppler (81.2+/-2.9% and 83.7+/-3.0% vs. 53.5+/-2.4% of normal, P<0.01) and improved muscle performance. A significantly increased capillary density (1.62+/-0.03 and 1.68+/-0.05/muscle fiber, P<0.05), enhanced vascular maturation (8.6+/-0.3 and 8.1+/-0.4/HPF, P<0.05) and muscle viability corroborated the findings of improved hindlimb perfusion and muscle function. Furthermore, EPC-CM transplantation stimulated the mobilization of bone marrow (BM)-derived EPC compared to control (678.7+/-44.1 vs. 340.0+/-29.1 CD34(+)/CD45(-) cells/1x10(5) mononuclear cells, P<0.05) and their recruitment to the ischemic muscles (5.9+/-0.7 vs. 2.6+/-0.4 CD34(+) cells/HPF, P<0.001) 3 days after the last injection. CONCLUSIONS/SIGNIFICANCE: Intramuscular injection of EPC-CM is as effective as cell transplantation for promoting tissue revascularization and functional recovery. Owing to the technical and practical limitations of cell therapy, cell free conditioned media may represent a potent alternative for therapeutic angiogenesis in ischemic cardiovascular diseases.

Cell-based therapy facilitates venous thrombus resolution

Endothelial progenitor cells (EPC) are involved in many healing processes in cardiovascular diseases and can be found in spontaneously resolving venous thrombi. The purpose of the present study was to investigate whether the therapeutic administration of EPC might enhance the resolution of venous thrombi. For this purpose, venous thrombosis was induced in the infrarenal inferior vena cava (IVC) in 28 athymic nude rats. Culture expanded EPC derived from human peripheral blood mononuclear cells were injected intravenously two and four days after thrombus induction. Recanalisation of the IVC and thrombus organisation were assessed by laser Doppler measurements of the blood flow and immunohistochemical detection of endothelialised luminal structures in the thrombus. EPC transplantation resulted in significantly enhanced thrombus neovascularisation (capillary density: 186.6 +/- 26.7/HPF vs. 78 +/- 12.3/HPF, p<0.01; area covered by capillaries: 8.9 +/- 1.7 microm(2) vs. 2.5 +/- 1.3 microm(2), p<0.01) and was accompanied by a substantial increase in intra-thrombus blood flow (perfusion ratio: 0.7 +/- 0.07 vs. 0.3 +/- 0.08, p<0.02). These results were paralleled by augmented macrophage recruitment into resolving thrombi in the animals treated with EPC (39.4 +/- 4.7/HPF vs. 11.6 +/- 1.9/HPF, p<0.01). Our data suggest that EPC transplantation might be of clinical value to facilitate venous thrombus resolution in cases where current therapeutic options have limited success.

Integrating Pricing and Inventory Control: Is it Worth the Effort?

In this paper we first show that the gains achievable by integrating pricing and inventory control are usually small for classical demand functions. We then introduce reference price models and demonstrate that for this class of demand functions the benefits of integration with inventory control are substantially increased due to the price dynamics. We also provide some analytical results for this more complex model. We thus conclude that integrated pricing/inventory models could repeat the success of revenue management in practice if reference price effects are included in the demand model and the properties of this new model are better understood.

Realtime Logistics – Ein Algorithmus zur parallelisierbaren Bestimmung von Transportkollisionen in Materialflusssystemen

In diesem Beitrag zur Komplexitätsanalyse von Materialflusssystemen unter Zeitrestriktionen wird ein Algorithmus vorgestellt, der die Bestimmung von Transportkollisionen als parallelisierbares Problem betrachtet und dessen Datenstrukturen auf die Analyse der Wechselbeziehungen von Lastobjekten ausgerichtet ist. Am Beispiel eines Deadlockszenarios wird die Funktionsweise des Algorithmus dargestellt und gezeigt, dass die explizite Betrachtung von zeitlichen und räumlichen Abhängigkeiten unter Lastobjekten eine Deadlockerkennung möglich macht. Der Algorithmus bildet die Grundlage für weitere Anwendungen in der Analyse der Echtzeitfähigkeit von Materialflusssystemen.

Entwicklung von energieautarken, intelligenten Ladehilfsmitteln am Beispiel des inBin

Das autonome, intelligente Ladehilfsmittel verkörpert die Idee des Internets der Dinge in der Intralogistik in Reinform. Am Beispiel des inBin wird das Energy-Harvesting in der Intralogistik betrachtet und gezeigt, dass ein Behälter mit komplexen logistischen Funktionen unter realistischer Umgebungsbeleuchtung durch Solarzellen betrieben werden kann.

Kollaborative Planung dezentral gesteuerter Materialflusssysteme in der Intralogistik

Der Beitrag stellt eine Kollaborationssoftware vor, die im Rahmen des AiF-Forschungsprojektes „KoDeMat“ entwickelt wurde. Der Fokus wird auf die Problemfelder der fehlenden Standardisierung und Anpassbarkeit im Bereich von fördertechnischen Anlagen gerichtet. Ziel ist, unter Zuhilfenahme von standardisierten, kollaborativen Engineeringprozessen, eine unternehmensübergreifende Planung, Realisierung und einen Umbau von komplexen dezentral gesteuerten Intralogistiksystemen sowie deren Betrieb effizient zu ermöglichen.

Planung und Berechnung der systemischen Leistungsverfügbarkeit komplexer Logistiksysteme

Das Verständnis von Leistungsverfügbarkeit, wie sie in der VDI-Richtlinie 4486 definiert ist, reicht für die Planung komplexer, dynamischer und teil-autonomer Systeme nicht aus. Die Definition in der VDI 4486 setzt den Fokus ausschließlich auf den Erfüllungsgrad vereinbarter Prozesse bei der Inbetriebnahme lo-gistischer Anlagen und regelt die Messungen und Be-rechnungen der Leistungsverfügbarkeit zu diesem Zeitpunkt. Es bleibt die Frage, wie ein Materialflusssystem für eine spezifizierte Leistungsverfügbarkeit geplant werden kann. Dazu werden die Wirkzusammenhänge zwischen dem logistischen System und seinen Sub-Systemen z.B. vertikale Integration von Wirkzusammenhängen der Instandhaltung, von Echtzeiteffekten der Kommunikationsprozesse oder Effekten der Ma-schinensteuerung, betrachtet.

Entwicklung eines Versuchsfelds für große Systeme intelligenter Behälter

Im Rahmen dieses Artikels werden aktuelle Forschungsarbeiten zum Einsatz von Energy-Harvesting und Ultra-Low-Power-Geräten in Materialflusssystemen beschrieben. Ein besonderes Augenmerk wird auf die inBin-Plattform, das Energy-Harvesting und deren Auswirkungen auf die Leistungsverfügbarkeit gelegt. Dazu werden die Hardwareplattform und Architektur der inBin-Plattform sowie der Aufbau eines Versuchsfelds detailliert erläutert. Des Weiteren wird ein Ansatz zur Modellierung und Simulation von Systemen mit einer großen Anzahl von inBin-Plattformen vorgestellt. Darüber hinaus werden die Ergebnisse zweier simulierter Szenarien und mögliche Folgen für die Planung zukünftiger Materialflusssysteme betrachtet.

Mitochondrial Outer Membrane Proteome of Trypanosoma brucei Reveals Novel Factors Required to Maintain Mitochondrial Morphology

Trypanosoma brucei is a unicellular parasite that causes devastating diseases in humans and animals. It diverged from most other eukaryotes very early in evolution and, as a consequence, has an unusual mitochondrial biology. Moreover, mitochondrial functions and morphology are highly regulated throughout the life cycle of the parasite. The outer mitochondrial membrane defines the boundary of the organelle. Its properties are therefore key for understanding how the cytosol and mitochondria communicate and how the organelle is integrated into the metabolism of the whole cell. We have purified the mitochondrial outer membrane of T. brucei and characterized its proteome using label-free quantitative mass spectrometry for protein abundance profiling in combination with statistical analysis. Our results show that the trypanosomal outer membrane proteome consists of 82 proteins, two-thirds of which have never been associated with mitochondria before. 40 proteins share homology with proteins of known functions. The function of 42 proteins, 33 of which are specific to trypanosomatids, remains unknown. 11 proteins are essential for the disease-causing bloodstream form of T. brucei and therefore may be exploited as novel drug targets. A comparison with the outer membrane proteome of yeast defines a set of 17 common proteins that are likely present in the mitochondrial outer membrane of all eukaryotes. Known factors involved in the regulation of mitochondrial morphology are virtually absent in T. brucei. Interestingly, RNAi-mediated ablation of three outer membrane proteins of unknown function resulted in a collapse of the network-like mitochondrion of procyclic cells and for the first time identified factors that control mitochondrial shape in T. brucei.