Factor XIII improves platelet adhesion to fibrinogen by protein disulfide isomerase-mediated activity

BACKGROUND Factor XIII (FXIII), a plasma pro-transglutaminase, consists of two A subunits and two B subunits (FXIIIA2B2). Following activation by thrombin, it cross-links fibrin chains at the final step of coagulation. We previously reported that FXIII subunit A (FXIIIA) serves as a protein disulfide isomerase (PDI), and that PDI promotes platelet adhesion and aggregation. OBJECTIVE This study sought to examine possible mechanistic effect of FXIII on platelet adhesion to fibrinogen; specifically, the role of its PDI activity. METHODS Ex vivo experiments: Blood platelets derived from five patients with hereditary FXIIIA deficiency before and after treatment with Fibrogammin-P (FXIIIA2B2 concentrate) were washed and incubated on immobilized fibrinogen. Bound platelets were stained and counted by microscopy. In vitro experiments: Platelets derived from patients before treatment and five healthy controls were washed and analyzed for adhesion in the presence or absence of Fibrogammin-P or recombinant FXIII (FXIIIA2 concentrate). RESULTS In ex vivo experiments, one hour after Fibrogammin-P treatment, mean (±SEM) platelet adhesion to fibrinogen increased by 27±2.32% (p<0.001). In in vitro experiments, treatment with Fibrogammin-P or recombinant FXIII (10IU/mL each) enhanced platelet adhesion to fibrinogen (in patients, by 29.95±6.7% and 29.05±5.3%, respectively; in controls, by 26.06±3.24% and 26.91±4.72, respectively; p<0.04 for all). Iodoacetamide-treated FXIII (I-FXIII), where transglutaminase activity is blocked, showed similar enhanced adhesion as untreated FXIII. By contrast, addition of an antibody that specifically blocks FXIIIA-PDI activity inhibited FXIII-mediated platelet adhesion to fibrinogen by 65%. CONCLUSION These findings indicate that FXIII-induced enhancement of platelet adhesion is mediated by FXIII-PDI activity

Chronic in vitro shear stress stimulates endothelial cell retention on prosthetic vascular grafts and reduces subsequent in vivo neointimal thickness

AbstractObjective: The absence of endothelial cells at the luminal surface of a prosthetic vascular graft potentiates thrombosis and neointimal hyperplasia, which are common causes of graft failure in humans. This study tested the hypothesis that pretreatment with chronic in vitro shear stress enhances subsequent endothelial cell retention on vascular grafts implanted in vivo. Methods: Cultured endothelial cells derived from Fischer 344 rat aorta were seeded onto the luminal surface of 1.5-mm internal diameter polyurethane vascular grafts. The seeded grafts were treated for 3 days with 1 dyne/cm2 shear stress and then for an additional 3 days with 1 or 25 dyne/cm2 shear stress in vitro. The grafts then were implanted as aortic interposition grafts into syngeneic rats in vivo. Grafts that were similarly seeded with endothelial cells but not treated with shear stress and grafts that were not seeded with endothelial cells served as controls. The surgical hemostasis time was monitored. Endothelial cell identity, density, and graft patency rate were evaluated 24 hours after implantation. Endothelial cell identity in vivo was confirmed with cells transduced in vitro with β-galactosidase complementary DNA in a replication-deficient adenoviral vector. Histologic, scanning electron microscopic, and immunohistochemical analyses were performed 1 week and 3 months after implantation to establish cell identity and to measure neointimal thickness. Results: The pretreatment with 25 dyne/cm2—but not with 0 or 1 dyne/cm2—shear stress resulted in the retention of fully confluent endothelial cell monolayers on the grafts 24 hours after implantation in vivo. Retention of seeded endothelial cells was confirmed by the observation that β-galactosidase transduced cells were retained as a monolayer 24 hours after implantation in vivo. In the grafts with adherent endothelial cells that were pretreated with shear stress, immediate graft thrombosis was inhibited and surgical hemostasis time was significantly prolonged. Confluent intimal endothelial cell monolayers also were present 1 week and 3 months after implantation. However, 1 week after implantation, macrophage infiltration was observed beneath the luminal cell monolayer. Three months after the implantation in vivo, subendothelial neointimal cells that contained α–smooth muscle actin were present. The thickness of this neointima averaged 41 ± 12 μm and 60 ± 23 μm in endothelial cell–seeded grafts that were pretreated with 25 dyne/cm2 shear stress and 1 dyne/cm2 shear stress, respectively, and 158 ± 46 μm in grafts that were not seeded with endothelial cells. Conclusion: The effect of chronic shear stress on the enhancement of endothelial cell retention in vitro can be exploited to fully endothelialize synthetic vascular grafts, which reduces immediate in vivo graft thrombosis and subsequent neointimal thickness. (J Vasc Surg 1999;29:157-67.

Design and Analysis of a Logless Dynamic Reconfiguration Protocol

Distributed replication systems based on the replicated state machine model have become ubiquitous as the foundation of modern database systems. To ensure availability in the presence of faults, these systems must be able to dynamically replace failed nodes with healthy ones via dynamic reconfiguration. MongoDB is a document oriented database with a distributed replication mechanism derived from the Raft protocol. In this paper, we present MongoRaftReconfig, a novel dynamic reconfiguration protocol for the MongoDB replication system. MongoRaftReconfig utilizes a logless approach to managing configuration state and decouples the processing of configuration changes from the main database operation log. The protocol's design was influenced by engineering constraints faced when attempting to redesign an unsafe, legacy reconfiguration mechanism that existed previously in MongoDB. We provide a safety proof of MongoRaftReconfig, along with a formal specification in TLA+. To our knowledge, this is the first published safety proof and formal specification of a reconfiguration protocol for a Raft-based system. We also present results from model checking its safety properties on finite protocol instances. Finally, we discuss the conceptual novelties of MongoRaftReconfig, how it can be understood as an optimized and generalized version of the single server reconfiguration algorithm of Raft, and present an experimental evaluation of how its optimizations can provide performance benefits for reconfigurations.Comment: 35 pages, 2 figure

Absence of the brachial artery: Report of a rare human variation and review of upper extremity arterial anomalies

AbstractVariations in the arterial anatomy of the upper extremities, although uncommon, occur in up to one in five patients. Most of these variants occur in either the radial or ulnar artery; brachial artery variations are less common. The case we report is a rare anomaly consisting of brachial artery agenesis or regression. The brachial artery was absent from its origin but reconstituted as a normal-appearing vessel 3 cm above the antecubital fossa. The profunda brachii artery and the superior and inferior ulnar collateral arteries were also absent in this patient. The axillary artery served as the main collateral to the forearm. This constellation of anomalies has not been previously described or explained by developmental models in humans and other primates. We speculate that failure of development or arrest of specific vascular anlage in the upper extremity occurs at particular embryological stages because of unknown factors. (J Vasc Surg 2001;33:191-4.

Endovascular management of hepatic artery pseudoaneurysm hemorrhage complicating pancreaticoduodenectomy

Gastrointestinal bleeding is a morbid complication of pancreaticoduodenectomy. Determining its etiology is often a daunting challenge in that both common and unusual mechanisms may be operative. Visceral artery pseudoaneurysms, although rare, must be considered in that minimally invasive means are available for effective therapy. Our recent experience with two cases highlights the importance for both general and vascular surgeons to be aware of the diagnostic and therapeutic role for early angiography and deployment of endovascular techniques to achieve a successful outcome

Safe Environmental Envelopes of Discrete Systems

A safety verification task involves verifying a system against a desired safety property under certain assumptions about the environment. However, these environmental assumptions may occasionally be violated due to modeling errors or faults. Ideally, the system guarantees its critical properties even under some of these violations, i.e., the system is \emph{robust} against environmental deviations. This paper proposes a notion of \emph{robustness} as an explicit, first-class property of a transition system that captures how robust it is against possible \emph{deviations} in the environment. We modeled deviations as a set of \emph{transitions} that may be added to the original environment. Our robustness notion then describes the safety envelope of this system, i.e., it captures all sets of extra environment transitions for which the system still guarantees a desired property. We show that being able to explicitly reason about robustness enables new types of system analysis and design tasks beyond the common verification problem stated above. We demonstrate the application of our framework on case studies involving a radiation therapy interface, an electronic voting machine, a fare collection protocol, and a medical pump device.Comment: Full version of CAV23 pape