Targeted subendothelial matrix oxidation by myeloperoxidase triggers myosin II-dependent de-adhesion and alters signaling in endothelial cells

During inflammation, myeloperoxidase (MPO) released by circulating leukocytes accumulates within the subendothelial matrix by binding to and transcytosing the endothelium. Oxidative reactions catalyzed by subendothelial-localized MPO are implicated as a key cause of endothelial dysfunction in inflammatory vascular diseases. Whilst the subendothelial matrix is a reactive target for MPO-derived oxidants in disease, the functional implications of oxidative matrix modification for the endothelium are largely unknown. Here we show that hypochlorous acid (HOCl) produced by endothelial-transcytosed MPO oxidizes the subendothelial matrix, involving covalent crosslinking of the adhesive matrix protein fibronectin. Real-time biosensor and live cell imaging studies showed that HOCl-mediated matrix oxidation triggers rapid membrane retraction from the substratum and adjacent cells (de-adhesion). This de-adhesion was linked with the alteration of Tyr-118 phosphorylation of paxillin, a key focal adhesion-dependent signaling process, as well as Rho kinase-dependent myosin light chain-2 phosphorylation. De-adhesion dynamics were dependent on the contractile state of cells, with myosin II inhibition with blebbistatin markedly attenuating the rate of membrane retraction. Rho kinase inhibition with Y-27632 also conferred protection, but not during the initial phase of membrane retraction, which was driven by pre-existing actomyosin tensile stress. Notably, diversion of MPO from HOCl production by thiocyanate and nitrite attenuated de-adhesion and associated signaling responses, despite the latter substrate supporting MPO-catalyzed fibronectin nitration. This study indicates that HOCl-mediated matrix oxidation by subendothelial MPO deposits may play an important and previously unrecognized role in altering endothelial adhesion, signaling and integrity during inflammatory vascular disorders

Self-Generated Magnetic Fields in Galactic Cooling Flows

Interstellar magnetic fields in elliptical galaxies are assumed to have their origin in stellar fields that accompany normal mass loss from an evolving population of old stars. The seed fields are amplified by interstellar turbulence driven by stellar mass loss and supernova events. These disordered fields are further amplified by time-dependent compression in the inward moving galactic cooling flow and are expected to dominate near the galactic core. Under favorable circumstances, fields similar in strength to those observed $B \sim 1-10~(r/10~kpc)^{-1.2}\mu$G can be generated solely from these natural galactic processes. In general the interstellar field throughout elliptical galaxies is determined by the outermost regions in the interstellar gas where the turbulent dynamo process can occur. Because of the long hydrodynamic flow times in galactic cooling flows, currently observed magnetic fields may result from periods of intense turbulent field amplification that occurred in the outer galaxy in the distant past. Particularly strong fields in ellipticals may result from ancient galactic mergers or shear turbulence introduced at the boundary between the interstellar gas and ambient cluster gas.Comment: 21 pages in AASTEX LaTeX with 2 figures; accepted by Astrophysical Journa

Loss of monomorphic and polymorphic HLA antigens in metastatic breast and colon carcinoma.

MHC class I antigens are intimately involved in intercellular communication, and recognition by cytotoxic T cells. Thus tumour cells that fail to express them may be at a growth or metastatic advantage. A series of ten colorectal and ten breast carcinomas, and their respective lymph node metastases, were examined immunohistologically using monoclonal antibodies (mAb) against both monomorphic and A2 polymorphic determinants, and beta-2-microglobulin (beta 2m). Four colon polypoid adenomas stained positively throughout, but 6/10 primary tumours had partial or complete loss of expression of monomorphic determinants using mAb W6/32: two node and the liver metastasis showed less, four more expression. Similar results were seen for beta 2m. HLA-A2 expression was absent or reduced in 4/4 colon tumours and all their metastases. Among the breast tumours, W6/32 staining was absent or reduced in 2/10, and node deposits showed two with less reactivity than their primary. Beta 2m staining was reduced or absent in 8/10 primaries and all the node metastases; in every case in which beta 2m was detected in the primary tumour their corresponding lymph node metastasis showed a decreased expression. HLA-A2 expression was absent or reduced in 3/4 primary breast carcinomas, and all their metastases. These results show that individual human colon and breast carcinomas often have a reduced HLA class I antigen expression, which apparently confers a metastatic advantage

Evidence for Interstitial Carbon in Nitrogenase FeMo Cofactor

The identity of the interstitial light atom in the center of the FeMo cofactor of nitrogenase has been enigmatic since its discovery. Atomic-resolution x-ray diffraction data and an electron spin echo envelope modulation (ESEEM) analysis now provide direct evidence that the ligand is a carbon species

Women with Polycystic Ovary Syndrome have an increased risk of major cardiovascular events: a population study

Context The effects of Polycystic Ovary Syndrome (PCOS) on cardiovascular morbidity and mortality are unclear. Objective To establish the relative risk of myocardial infarction (MI), stroke, angina, revascularization and cardiovascular mortality for women with PCOS. Design Data were extracted from the Clinical Practice Research Datalink Aurum database. Patients with PCOS were matched to controls (1:1) by age, body mass index (BMI) category and primary care practice. The primary outcome was the time to major adverse cardiovascular event (MACE); a composite endpoint incorporating MI, stroke, angina, revascularization and cardiovascular mortality. Secondary outcomes were the individual MACE endpoints. Results Of 219,034 with a diagnosis of PCOS, 174,660 (79.7%) met the eligibility criteria and were matched. Crude rates of the composite endpoint, MI, stroke, angina, revascularization and cardiovascular mortality were respectively 82.7, 22.7, 27.4, 32.8, 10.5 and 6.97 per 100,000 patient-years for cases, and 64.3, 15.9, 25.7, 19.8, 7.13 and 7.75 per 100,000 patient-years for controls. In adjusted cox proportional hazard models (CPHM), the hazard ratios [HR] were 1.26 (95% confidence interval=1.13-1.41), 1.38 (1.11-1.72), 1.60 (1.32-1.94) and 1.50 (1.08-2.07) for the composite outcome, MI, angina and revascularization, respectively. In a time-dependent CPHM, weight gain (HR 1.01 [1.00-1.01]), prior type 2 diabetes (T2DM) (HR 2.40 [1.76-3.30]) and social deprivation (HR 1.53 [1.11-2.11]) increased risk of progression to the composite endpoint. Conclusions The risk of incident MI, angina and revascularization is increased in young women with PCOS. Weight and T2DM are potentially modifiable risk factors amenable to intervention

Nitrogenase FeMoco investigated by spatially resolved anomalous dispersion refinement

The [Mo:7Fe:9S:C] iron-molybdenum cofactor (FeMoco) of nitrogenase is the largest known metal cluster and catalyses the 6-electron reduction of dinitrogen to ammonium in biological nitrogen fixation. Only recently its atomic structure was clarified, while its reactivity and electronic structure remain under debate. Here we show that for its resting S=3/2 state the common iron oxidation state assignments must be reconsidered. By a spatially resolved refinement of the anomalous scattering contributions of the 7 Fe atoms of FeMoco, we conclude that three irons (Fe1/3/7) are more reduced than the other four (Fe2/4/5/6). Our data are in agreement with the recently revised oxidation state assignment for the molybdenum ion, providing the first spatially resolved picture of the resting-state electron distribution within FeMoco. This might provide the long-sought experimental basis for a generally accepted theoretical description of the cluster that is in line with available spectroscopic and functional data

Mechanism, localization and cure of atrial arrhythmias occurring after a new intraoperative endocardial radiofrequency ablation procedure for atrial fibrillation

AbstractOBJECTIVESThe purpose of this study was to test a new pattern of radiofrequency ablation for atrial fibrillation (AFib) intended to optimize atrial activation, and to demonstrate the usefulness of catheter techniques for mapping and ablation of postoperative atrial arrhythmias.BACKGROUNDLinear radiofrequency lesions have been used to cure AFib, but the optimal pattern of lesions is unknown and postoperative tachyarrhythmias are common.METHODSA radial pattern of linear radiofrequency lesions (Star) was made using an endocardial open surgical approach in 25 patients. Postoperative arrhythmias were induced and characterized during electrophysiological studies in 15 patients.RESULTSThe AFib was abolished in most patients (91%), but atrial flutter (AFlut) occurred in 96% of patients postoperatively. At postoperative electrophysiological studies, 37 flutter morphologies were studied in 15 patients (46% spontaneous, cycle length [CL] 223 ± 25 ms). Seven mechanisms (lesions discontinuity, n = 6; focal mechanism, n = 1) of AFlut were characterized in six patients. In these cases, flutter was abolished using further catheter radiofrequency ablation. In the remaining cases, flutter was usually localized to an area involving the interatrial septum, but no critical isthmus was identified for ablation. After 16 ± 10 months, 15 patients (65%) were asymptomatic with (n = 3) or without (n = 12) antiarrhythmic medications. Eight (35%) patients had persistent arrhythmias. Postoperative atrial electrical activation was near physiological.CONCLUSIONSThe AFib may be abolished using a radial pattern of linear endocardial radiofrequency lesions, but postoperative AFlut is common even when lesions are made under optimal conditions. Endocardial mapping techniques can be used to characterize the flutter mechanisms, thus enabling subsequent successful catheter ablation