ADAM10-Mediated Cleavage of ICAM-1 Is Involved in Neutrophil Transendothelial Migration

To efficiently cross the endothelial barrier during inflammation, neutrophils first firmly adhere to the endothelial surface using the endothelial adhesion molecule ICAM-1. Upon actual transmigration, the release from ICAM-1 is required. While Integrin LFA1/Mac1 de-activation is one described mechanism that leads to this, direct cleavage of ICAM-1 from the endothelium represents a second option. We found that a disintegrin and metalloprotease 10 (ADAM10) cleaves the extracellular domain of ICAM-1 from the endothelial surface. Silencing or inhibiting endothelial ADAM10 impaired the efficiency of neutrophils to cross the endothelium, suggesting that neutrophils use endothelial ADAM10 to dissociate from ICAM-1. Indeed, when measuring transmigration kinetics, neutrophils took almost twice as much time to finish the diapedesis step when ADAM10 was silenced. Importantly, we found increased levels of ICAM-1 on the transmigrating neutrophils when crossing an endothelial monolayer where such increased levels were not detected when neutrophils crossed bare filters. Using ICAM-1-GFP-expressing endothelial cells, we show that ICAM-1 presence on the neutrophils can also occur by membrane transfer from the endothelium to the neutrophil. Based on these findings, we conclude that endothelial ADAM10 contributes in part to neutrophil transendothelial migration by cleaving ICAM-1, thereby supporting the release of neutrophils from the endothelium during the final diapedesis step.

Effects of mesenchymal stem cell and amnion membrane transfer on prevention of pericardial adhesions

BackgroundTo investigate and compare the antiadhesive/antifibrotic effects of mesenchymal stem cells (MSC) and amnion membrane transfer (AMT) in a rat model.Material and methodsThree experimental and sham groups were formed using 30 Wistar-Albino rats. AMT and MSC were applied to the related groups. The control group was not treated. After 12 weeks follow-up, intracardiac blood and cardiac-pericardiac tissue samples were taken. The severity of adhesions and fibrosis were scored macroscopically and microscopically with Hematoxylin/Eosin and Masson’s trichrome staining. TNF-α, TGF-β, IL-1, PDGF, FGF, VEGF and Caspase-3 levels were measured with the ELISA method.ResultsSevere adhesions were observed in the AMT and control groups, but no adhesion was present in the MSC group. Pericardial thickness, increased vascularity, fibrosis, and collagen accumulation were similar between control and AMT groups, but were less in Sham and MSC groups. Between MSC and AMT groups, only Caspase-3 level was different, which is an apoptosis marker.ConclusionThe positive effects of MSC on adhesion, which we achieved in our study, suggest that it may prevent adhesion. AMT did not provide a positive effect. The correlation of Caspase-3 with postoperative adhesion/fibrosis should be examined in more detail.

IRS1/PI3K/AKT pathway signal involved in the regulation of glycolipid metabolic abnormalities by Mulberry (Morus alba L.) leaf extracts in 3T3-L1 adipocytes

Background Mulberry (Morus alba L.) leaf tea benefits the control of diabetes in Asian nations. This study was aim to investigate if the flavonoids, which extracts from mulberry leaves, could regulate the metabolism of glycolipid, and to investigate if flavonoids could regulate IRS1/PI3K/AKT pathway signal to affect the expression of FAS and membrane transfer capacity GLUT4 in 3T3-L1 adipocytes. Results Results revealed that flavonoids decreased the levels of free fatty acid and increased the glucose consumption and the levels of adiponectin and leptin in a dose-dependent manner, and remarkably increased the protein expression levels of p-IRS1, p-PI3K, p-Akt, total GLUT4, and membrane GLUT4, and decreased the protein expression levels of PTEN and FAS in 3T3-L1 adipocytes IR model. On the other hand, wortmannin (2 nM), a selective and irreversible PI3K inhibitor, significantly decreased the glucose consumption and the adiponectin and leptin levels, and increased the free fatty acid level in flavonoids treated 3T3-L1 adipocytes IR model. Furthermore, wortmannin (2 nM) partly eliminated the activation of PI3K/AKT signaling, the suppression of FAS, and the up-regulated membrane transfer capacity of GLUT4 in flavonoids treated 3T3-L1 adipocytes IR model. Conclusion In conclusion, our results illustrated that mulberry leaf extracts flavonoids alleviated the glycolipid metabolic abnormalities in 3T3-L1 adipocytes IR model, and the effect was associated with the activation of IRS1/PI3K/AKT pathway, the suppression of FAS, and the up-regulation of membrane transfer capacity of GLUT4.

Megakaryocyte emperipolesis mediates membrane transfer from intracytoplasmic neutrophils to platelets

Bone marrow megakaryocytes engulf neutrophils in a phenomenon termed emperipolesis. We show here that emperipolesis is a dynamic process mediated actively by both lineages, in part through the β2-integrin/ICAM-1/ezrin pathway. Tethered neutrophils enter in membrane-bound vesicles before penetrating into the megakaryocyte cytoplasm. Intracytoplasmic neutrophils develop membrane contiguity with the demarcation membrane system, thereby transferring membrane to the megakaryocyte and to daughter platelets. This phenomenon occurs in otherwise unmanipulated murine marrow in vivo, resulting in circulating platelets that bear membrane from non-megakaryocytic hematopoietic donors. Transit through megakaryocytes can be completed as rapidly as minutes, after which neutrophils egress intact. Emperipolesis is amplified in models of murine inflammation associated with platelet overproduction, contributing to platelet production in vitro and in vivo. These findings identify emperipolesis as a new cell-in-cell interaction that enables neutrophils and potentially other cells passing through the megakaryocyte cytoplasm to modulate the production and membrane content of platelets.

Megakaryocyte emperipolesis mediates membrane transfer from intracytoplasmic neutrophils to platelets

SummaryBone marrow megakaryocytes engulf neutrophils in a phenomenon termed emperipolesis. We show here that emperipolesis is a dynamic process mediated actively by both lineages, in part through the β2-integrin/ICAM-1/ezrin pathway. Tethered neutrophils enter in membrane-bound vesicles before penetrating into the megakaryocyte cytoplasm. Intracytoplasmic neutrophils develop membrane contiguity with the demarcation membrane system, thereby transferring membrane to the megakaryocyte and to daughter platelets. This phenomenon occurs in otherwise unmanipulated marrowin vivo, resulting in circulating platelets that bear membrane from non-megakaryocytic hematopoietic donors. Transit through megakaryocytes can be completed as rapidly as minutes, after which neutrophils egress intact. Emperipolesis is amplified in models of inflammation associated with platelet overproduction, contributing to platelet productionin vitroandin vivo.These findings identify emperipolesis as a new cell-in-cell interaction that enables neutrophils and potentially other cells passing through the megakaryocyte cytoplasm to modulate the production and membrane content of platelets.

Treatment of Pharmaceutical Process Wastewater with Hybrid Separation Method: Distillation and Hydrophilic Pervaporation

The work is motivated by an industrial problem, which is alcohol removal from pharmaceutical process wastewater. The aim of the study was to develop a complete hybrid operation is investigated. Ethanol dehydration, in combination with distillation and hydrophilic pervaporation, is used to investigate about the extent of separation of the ethanol-water mixture. The aim of this research is to rigorously model and optimize this hybrid operation in professional flowsheet simulator environment. The number of minimal theoretical plates of distillation column and minimal effective membrane transfer area are determined. Cost estimation is also examined according to Douglas methodology. Considering our results it can be concluded that, the distillation and hydrophilic pervaporation processes are suitable for separation ethanol and water in 99.5 weight percent purity