Heparinase selectively sheds heparan sulphate from the endothelial glycocalyx

A healthy vascular endothelium is coated by the endothelial glycocalyx. Its main constituents are transmembrane syndecans and bound heparan sulphates. This structure maintains the physiological endothelial permeability barrier and prevents leukocyte and platelet adhesion, thereby mitigating inflammation and tissue oedema. Heparinase, a bacteria] analogue to heparanase, is known to attack the glycocalyx. However, the exact extent and specificity of degradation is unresolved. We show by electron microscopy, immunohistological staining and quantitative measurements of the constituent parts, that heparinase selectively sheds heparan sulphate from the glycocalyx, but not the synclecans

The Influence of Matter-Antimatter Domains on Big Bang Nucleosynthesis

In der vorliegenden Arbeit habe ich mich mit den Auswirkungen eventuell im fruehen Universum vorhandener Antimaterieregionen auf die Haeufigkeiten der leichten Elemente beschaeftigt. Praktisch das gesamte Deuterium und der ueberwiegende Teil der Helium-4 Kerne, die wir heute im Universum beobachten, wurden in einem fuehen Evolutionsstadium des Kosmos — nur wenige Minuten nach dem Urknall — gebildet. In der Theorie der sogenannten Primordialen Nukleosynthese — oder auch Big Bang Nukleosynthese (BBN) — werden die relativen Haeufigkeiten der einzelnen Kerne abhangig von den genauen physikalischen Bedingungen im jungen Universum vorhergesagt. Die im Rahmen des Standardmodells der Kosmologie vorhergesagten Elementhaeufigkeiten stimmen im Allgemeinen gut mit aus Beobachtungen abgeleiteten Werten ueberein. Dies begr uendet den großen Erfolg dieser Theorie und macht sie zu einem der Grundpfeiler des kosmologischen Standardmodells. Denkbare Erweiterungen des Standardmodells koennen jedoch potentiell Auswirkungen auf den Ablauf der Kernsynthese haben. Da aber jedes glaubwuerdige Szenario ebenso wie die Standardtheorie die aus den Beobachtungen abgeleiteten Haeufigkeiten vorhersagen muss, duerfen die H¨aufigkeiten nur minimal beeinflusst werden. Diese Ueberlegungen gestatten es uns, die Kernsynthese als ”Werkzeug“ zur Untersuchung der physikalischen Bedingungen im jungen Universum zu verwenden. Dies ist bereits in der Vergangenheit vielfach praktiziert worden. Eine haeufig untersuchte Variante ist die sogenannte inhomogene Nukleosynthese. In einem solchen Modell wird eine Grundannahme des kosmologischen Standardmodells, die Homogenitaet der Verteilung der baryonischen Materie im jungen Universum, fallengelassen. Das von mir untersuchte Szenario geht noch einen Schritt weiter und laeßt auch Fluktuationen in der Baryonendichte mit negativem Vorzeichen zu. In einem solchen Modell besteht das junge Universum aus getrennten Materie- und Antimaterieregionen. Diese Art spezieller Anfangsbedingungen wird in einigen Modellen der elektroschwachen Baryogenese vorhergesagt. Solche Materie- und Antimaterieregionen werden sich gegenseitig annihilieren, sobald der Transport von Baryonen ueber die Grenzen der Regionen moeglich ist. Nach der vollst¨andigen Annihilation aller Antimaterieregionen bleibt nur der im Zuge der Baryogenese gebildete Ueberschuß an Materie uebrig. Zur numerischen Behandlung dieses Problems habe ich ein Computerprogramm entwickelt. In diesem Programm werden sowohl die nuklearen Reaktionen, die zum Aufbau der leichten Elemente fuehren, als auch Annihilationen beruecksichtigt. Da die Kernsynthese und die Annihilation der Antimaterieregionen im expandierenden Universum ablaufen, und die genauenWerte der einzelnen thermodynamischen Variablen, wie Druck, Dichte und Temperatur der beteiligten Teilchen, von entscheidender Wichtigkeit sind, muss das Programm auf dem Hintergrund der Expansion des Kosmos gerechnet werden. Weiterhin musste neben den Reaktionen, die zwischen den einzelnen Nukleonen ablaufen koennen, auch der Transport von Nukleonen und Antinukleonen in die jeweilige Anti-Region behandelt werden. Diese Transportprozesse werden zu fruehen Zeiten durch Diffusion von Baryonen beschrieben, zu spaeten Zeiten hingegen durch hydrodynamische Expansion von Regionen mit hoeherer Dichte gegen solche mit niedrigerer Dichte. Abhaengig vom Zeitpunkt der Annihilation k¨onnen die Haeufigkeiten der leichten Elemente durch zwei Haupteffekte beeinflusst werden. Im Zuge der Heliumsynthese, die bei einer kosmischen Temperatur von etwa 80 keV ablaeuft, werden praktisch alle freien Neutronen in Helium-4 Kerne eingebaut. Die primordiale Helium-4 Haeufigkeit haengt also stark von der Anzahl verfuegbarer Neutronen ab. Zu Zeiten vor der Heliumsynthese laeuft der Transport von Baryonzahl ueber die Domaenengrenzen durch Neutronendiffusion ab, Protonen koennen auf Grund ihrer elektrischen Ladung nur ueber wesentlich kuerzere Distanzen diffundieren. Fruehe Annihilation wird also bevorzugt auf Neutronen stattfinden und fuehrt so zu einer Reduzierung der Neutronendichte, und damit unmittelbar auch zu einer geringeren Menge an primordial produziertem Helium-4. Sind die Antimaterieregionen groeßer als die Diffusionslaenge von Neutronen zur Zeit der Heliumsynthese, ist ein nennenswerter Transport von Baryonzahl erst zu wesentlich sp¨ateren Zeiten moeglich. Antiprotonen, die nun in die Materieregion eindringen, koennen sowohl auf Protonen als auch auf die bereits gebildeten Helium-4 Kerne annihilieren. Weiterhin koennen die Helium-4 Kerne auch durch die im Annihilationprozess entstehenden Gammaquanten photodisintegriert werden. Beide Prozesse fuehren zur Bildung energetischer Sekundaerkerne, in erster Linie Helium-3. Diese energetischen Kerne koennen in einem weiteren Schritt durch nicht-thermische Reaktionen mit Helium-4 Kernen Lithium-6 Kerne bilden. Sp¨ate Annihilation wird also zu einer erhoehten Helium-3 und Lithium-6 Haeufigkeit im Vergleich zum Standardszenario fuehren. Als ein wichtiges Ergebnis meiner Arbeit habe ich auf Grund dieser Effekte Schranken sowohl an den maximal erlaubten Antimateriegehalt im jungen Universum, als auch an den Zeitpunkt der Annihilation, bestimmt durch die Groeße der Antimaterieregionen, hergeleitet. Diese neuen und rigiden Schranken decken einen weiten Annihilationszeitraum ab, von der Epoche des Ausfrierens der schwachen Wechselwirkungen bei einer Temperatur von etwa 1 MeV bis hinunter zur Epoche der Rekombination bei einer kosmischen Temperatur von etwa 10 nisse wesentlich restriktiver. Der relative Antimateriegehalt in Regionen die unmittelbar nach dem Ende der Kernsynthese annihilieren kann beispielsweise nicht hoeher als wenige Prozent der gesamten baryonischen Materie sein, fuer spaetere Annihilation sinkt dieser Wert um mehr als zwei Groeßenordnungen. In einem zweiten Hauptaspekt meiner Arbeit habe ich gezeigt, dass die durchaus im Detail vorhandenen Diskrepanzen zwischen den im Standardszenario der Big Bang Nukleosynthese vorhergesagten Elementh aeufigkeiten und den aus Beobachtungen abgeleiteten Werten durch die Praesenz einer gewissen Menge Antimaterie in einem bestimmten Laengenskalenbereich beseitigt werden koennen. Weiterhin habe ich untersucht, ob die im Standardszenario g ueltige obere Grenze fuer die Baryonendichte im Universum in einem Szenario mit Antimateriedom aenen ebenso gueltig ist. Auf Grund meiner Ergebnisse erscheint es sehr unwahrscheinlich, dass die Baryonendichte in einem Materie-Antimaterie Szenario wesentlich gr¨oßer sein kann, als im Standardszenario vorhergesagt

Perspectives in Microvascular Fluid Handling: Does the Distribution of Coagulation Factors in Human Myocardium Comply with Plasma Extravasation in Venular Coronary Segments?

Background: Heterogeneity of vascular permeability has been suggested for the coronary system. Whereas arteriolar and capillary segments are tight, plasma proteins pass readily into the interstitial space at venular sites. Fittingly, lymphatic fluid is able to coagulate. However, heart tissue contains high concentrations of tissue factor, presumably enabling bleeding to be stopped immediately in this vital organ. The distribution of pro- and anti-coagulatively active factors in human heart tissue has now been determined in relation to the types of microvessels. Methods and Results: Samples of healthy explanted hearts and dilated cardiomyopathic hearts were immunohistochemically stained. Albumin was found throughout the interstitial space. Tissue factor was packed tightly around arterioles and capillaries, whereas the tissue surrounding venules and small veins was practically free of this starter of coagulation. Thrombomodulin was present at the luminal surface of all vessel segments and especially at venular endothelial cell junctions. Its product, the anticoagulant protein C, appeared only at discrete extravascular sites, mainly next to capillaries. These distribution patterns were basically identical in the healthy and diseased hearts, suggesting a general principle. Conclusions: Venular extravasation of plasma proteins probably would not bring prothrombin into intimate contact with tissue factor, avoiding interstitial coagulation in the absence of injury. Generation of activated protein C via thrombomodulin is favored in the vicinity of venular gaps, should thrombin occur inside coronary vessels. This regionalization of distribution supports the proposed physiological heterogeneity of the vascular barrier and complies with the passage of plasma proteins into the lymphatic system of the heart. Copyright (C) 2010 S. Karger AG, Base

Small-volume resuscitation with hyperoncotic albumin: a systematic review of randomized clinical trials

Background Small-volume resuscitation can rapidly correct hypovolemia. Hyperoncotic albumin solutions, long in clinical use, are suitable for small-volume resuscitation; however, their clinical benefits remain uncertain. Methods Randomized clinical trials comparing hyperoncotic albumin with a control regimen for volume expansion were sought by multiple methods, including computer searches of bibliographic databases, perusal of reference lists, and manual searching. Major findings were qualitatively summarized. In addition, a quantitative meta-analysis was performed on available survival data. Results In all, 25 randomized clinical trials with a total of 1,485 patients were included. In surgery, hyperoncotic albumin preserved renal function and reduced intestinal edema compared with control fluids. In trauma and sepsis, cardiac index and oxygenation were higher after administration of hydroxyethyl starch than hyperoncotic albumin. Improved treatment response and renal function, shorter hospital stay and lower costs of care were reported in patients with liver disease receiving hyperoncotic albumin. Edema and morbidity were decreased in high-risk neonates after hyperoncotic albumin administration. Disability was reduced by therapy with hyperoncotic albumin in brain injury. There was no evidence of deleterious effects attributable to hyperoncotic albumin. Survival was unaffected by hyperoncotic albumin (pooled relative risk, 0.95; 95% confidence interval 0.78 to 1.17). Conclusion In some clinical indications, randomized trial evidence has suggested certain benefits of hyperoncotic albumin such as reductions in morbidity, renal impairment and edema. However, further clinical trials are needed, particularly in surgery, trauma and sepsis

Exogenous nitric oxide requires an endothelial glycocalyx to prevent postischemic coronary vascular leak in guinea pig hearts

Introduction Postischemic injury to the coronary vascular endothelium, in particular to the endothelial glycocalyx, may provoke fluid extravasation. Shedding of the glycocalyx is triggered by redox stress encountered during reperfusion and should be alleviated by the radical scavenger nitric oxide (NO). The objective of this study was to investigate the effect of exogenous administration of NO during reperfusion on both coronary endothelial glycocalyx and vascular integrity. Methods Isolated guinea pig hearts were subjected to 15 minutes of warm global ischemia followed by 20 minutes of reperfusion in the absence (Control group) and presence (NO group) of 4 mu M NO. In further experiments, the endothelial glycocalyx was enzymatically degraded by means of heparinase followed by reperfusion without (HEP group) and with NO (HEP+NO group). Results Ischemia and reperfusion severely damaged the endothelial glycocalyx. Shedding of heparan sulfate and damage assessed by electron microscopy were less in the presence of NO. Compared with baseline, coronary fluid extravasation increased after ischemia in the Control, HEP, and HEP+NO groups but remained almost unchanged in the NO group. Tissue edema was significantly attenuated in this group. Coronary vascular resistance rose by 25% to 30% during reperfusion, but not when NO was applied, irrespective of the state of the glycocalyx. Acute postischemic myocardial release of lactate was comparable in the four groups, whereas release of adenine nucleotide catabolites was reduced 42% by NO. The coronary venous level of uric acid, a potent antioxidant and scavenger of peroxynitrite, paradoxically decreased during postischemic infusion of NO. Conclusion The cardioprotective effect of NO in postischemic reperfusion includes prevention of coronary vascular leak and interstitial edema and a tendency to forestall both no-reflow and degradation of the endothelial glycocalyx

Comparison of 6 % hydroxyethyl starch and 5 % albumin for volume replacement therapy in patients undergoing cystectomy (CHART): study protocol for a randomized controlled trial

Background The use of artificial colloids is currently controversial, especially in Central Europe Several studies demonstrated a worse outcome in intensive care unit patients with the use of hydroxyethyl starch. This recently even led to a drug warning about use of hydroxyethyl starch products in patients admitted to the intensive care unit. The data on hydroxyethyl starch in non–critically ill patients are insufficient to support perioperative use. Methods/Design We are conducting a single-center, open-label, randomized, comparative trial with two parallel patient groups to compare human albumin 5 % (test drug) with hydroxyethyl starch 6 % 130/0.4 (comparator). The primary endpoint is cystatin C ratio, calculated as the ratio of the cystatin value at day 90 after surgery relative to the preoperative value. Secondary objectives are inter alia the evaluation of the influence of human albumin and hydroxyethyl starch on further laboratory chemical and clinical parameters, glycocalyx shedding, intensive care unit and hospital stay and acute kidney injury as defined by RIFLE criteria (risk of renal dysfunction, injury to the kidney, failure of kidney function, loss of kidney function, and end-stage kidney disease) criteria. Discussion There is a general lack of evidence on the relative safety and effects of hydroxyethyl starch compared with human albumin for volume replacement in a perioperative setting. Previously conducted studies of surgical patients in which researchers have compared different hydroxyethyl starch products included too few patients to properly evaluate clinical important outcomes such as renal function. In the present study in a high-risk patient population undergoing a major surgical intervention, we will determine if perioperative fluid replacement with human albumin 5 % will have a long-term advantage over a third-generation hydroxyethyl starch 130/0.4 on the progression of renal dysfunction until 90 days after surgery

The endothelial glycocalyx prefers albumin for evoking shear stress-induced, nitric oxide-mediated coronary dilatation

Background: Shear stress induces coronary dilatation via production of nitric oxide ( NO). This should involve the endothelial glycocalyx ( EG). A greater effect was expected of albumin versus hydroxyethyl starch ( HES) perfusion, because albumin seals coronary leaks more effectively than HES in an EG-dependent way. Methods: Isolated hearts ( guinea pigs) were perfused at constant pressure with Krebs-Henseleit buffer augmented with 1/3 volume 5% human albumin or 6% HES ( 200/0.5 or 450/0.7). Coronary flow was also determined after EG digestion ( heparinase) and with nitro-L-arginine ( NO-L-Ag). Results: Coronary flow ( 9.50 +/- 1.09, 5.10 +/- 0.49, 4.87 +/- 1.19 and 4.15 +/- 0.09 ml/ min/ g for `albumin', `HES 200', `HES 450' and `control', respectively, n = 5-6) did not correlate with perfusate viscosity ( 0.83, 1.02, 1.24 and 0.77 cP, respectively). NO-L-Ag and heparinase diminished dilatation by albumin, but not additively. Alone NO-L-Ag suppressed coronary flow during infusion of HES 450. Electron microscopy revealed a coronary EG of 300 nm, reduced to 20 nm after heparinase. Cultured endothelial cells possessed an EG of 20 nm to begin with. Conclusions: Albumin induces greater endothelial shear stress than HES, despite lower viscosity, provided the EG contains negative groups. HES 450 causes some NO-mediated dilatation via even a rudimentary EG. Cultured endothelial cells express only a rudimentary glycocalyx, limiting their usefulness as a model system. Copyright (c) 2007 S. Karger AG, Basel

Causes of metabolic acidosis in canine hemorrhagic shock: role of unmeasured ions

Introduction: Metabolic acidosis during hemorrhagic shock is common and conventionally considered to be due to hyperlactatemia. There is increasing awareness, however, that other nonlactate, unmeasured anions contribute to this type of acidosis. Methods: Eleven anesthetized dogs were hemorrhaged to a mean arterial pressure of 45 mm Hg and were kept at this level until a metabolic oxygen debt of 120 mLO2/kg body weight had evolved. Blood pH, partial pressure of carbon dioxide, and concentrations of sodium, potassium, magnesium, calcium, chloride, lactate, albumin, and phosphate were measured at baseline, in shock, and during 3 hours post-therapy. Strong ion difference and the amount of weak plasma acid were calculated. To detect the presence of unmeasured anions, anion gap and strong ion gap were determined. Capillary electrophoresis was used to identify potential contributors to unmeasured anions. Results: During induction of shock, pH decreased significantly from 7.41 to 7.19. The transient increase in lactate concentration from 1.5 to 5.5 mEq/L during shock was not sufficient to explain the transient increases in anion gap (+11.0 mEq/L) and strong ion gap (+7.1 mEq/L), suggesting that substantial amounts of unmeasured anions must have been generated. Capillary electrophoresis revealed increases in serum concentration of acetate (2.2 mEq/L), citrate (2.2 mEq/L), alpha-ketoglutarate (35.3 microEq/L), fumarate (6.2 microEq/L), sulfate (0.1 mEq/L), and urate (55.9 microEq/L) after shock induction. Conclusion: Large amounts of unmeasured anions were generated after hemorrhage in this highly standardized model of hemorrhagic shock. Capillary electrophoresis suggested that the hitherto unmeasured anions citrate and acetate, but not sulfate, contributed significantly to the changes in strong ion gap associated with induction of shock

Ferrocenyl-coupled n-heterocyclic carbene complexes of gold(i): a successful approach to multinuclear anticancer drugs

Four gold(I) carbene complexes featuring 4-ferrocenyl substituted imidazol-2-ylidene ligands were investigated for antiproliferative and antivascular properties. They were active against a panel of seven cancer cell lines, including multidrug-resistant ones, with low micromolar or nanomolar IC50 (72 h) values, according to their lipophilicity and cellular uptake. The delocalised lipophilic cationic complexes 8 and 10 acted by increasing the reactive oxygen species in two ways: via a genuine ferrocene effect and by inhibiting the thioredoxin reductase. Both complexes gave rise to a reorganization of the F-actin cytoskeleton in endothelial and melanoma cells, associated with a G1 phase cell cycle arrest and a retarded cell migration. They proved antiangiogenic in tube formation assays with endothelial cells and vascular-disruptive on real blood vessels in the chorioallantoic membrane of chicken eggs. Biscarbene complex 10 was also tolerated well by mice where it led to a volume reduction of xenograft tumors by up to 80%