Development of (17)O NMR approach for fast imaging of cerebral metabolic rate of oxygen in rat brain at high field

A comprehensive technique was developed for using three-dimensional (17)O magnetic resonance spectroscopic imaging at 9.4T for rapidly imaging the cerebral metabolic rate of oxygen consumption (CMRO(2)) in the rat brain during a two-min inhalation of (17)O(2). The CMRO(2) value (2.19 +/- 0.14 micromol/g/min, n = 7) was determined in the rat anesthetized with alpha-chloralose by independent and concurrent (17)O NMR measurements of cerebral H(2)17O content, arterial input function, and cerebral perfusion. CMRO(2) values obtained were consistent with the literature results for similar conditions. Our results reveal that, because of its superior sensitivity at ultra-high fields, the (17)O magnetic resonance spectroscopic imaging approach is capable of detecting small dynamic changes of metabolic H(2)17O during a short inhalation of (17)O(2) gas, and ultimately, for imaging CMRO(2) in the small rat brain. This study provides a crucial step toward the goal of developing a robust and noninvasive (17)O NMR approach for imaging CMRO(2) in animal and human brains that can be used for studying the central role of oxidative metabolism in brain function under normal and diseased conditions, as well as for understanding the mechanisms underlying functional MRI.     

The Tie-2 ligand angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies

The angiopoietins Ang-1 and Ang-2 have been identified as ligands with opposing functions of the receptor tyrosine kinase Tie-2 regulating endothelial cell survival and vascular maturation. Ang-1 acts in a paracrine agonistic manner, whereas Ang-2 appears to act primarily as an autocrine antagonistic regulator. To shed further light on the complexity of autocrine/paracrine agonistic/antagonistic functions of the angiopoietin/Tie-2 system, we have studied Ang-2 synthesis and secretion in different populations of wild-type and retrovirally Ang-2-transduced endothelial cells. Endogenous and overexpressed endothelial cell Ang-2 is expressed in a characteristic granular pattern indicative of a cytoplasmic storage granule. Light and electron microscopic double staining revealed Ang-2 colocalization with von Willebrand factor, identifying Ang-2 as a Weibel-Palade body molecule. Costaining with P-selectin showed that storage of Ang-2 and P-selectin in Weibel-Palade bodies is mutually exclusive. Stored Ang-2 has a long half-life of more than 18 hours and can be secreted within minutes of stimulation (eg, by phorbol 12-myristate 13-acetate [PMA], thrombin, and histamine). Collectively, the identification of Ang-2 as a stored, rapidly available molecule in endothelial cells strongly suggests functions of the angiopoietin/Tie-2 system beyond the established roles during angiogenesis likely to be involved in rapid vascular homeostatic reactions such as inflammation and coagulation.     

Arterial embolism to the upper extremity in a patient with factor V Leiden mutation (APC resistance)--a case report and review of the literature

Factor V Leiden mutation has emerged as one of the leading abnormalities in inherited blood coagulation disorders, resulting in a markedly increased risk for deep leg vein thrombosis. A 24-year-old woman presented with acute onset of critical ischemia of her left thumb and index finger. Intraarterial angiography revealed an embolus in the distal radial artery and a thrombotic occlusion of the digital artery of the thumb and index finger. Immediate therapy encompassed a selective surgical embolectomy of the distal radial artery followed by a local intraarterial lysis that was continued for 3 days. Additionally, therapeutic anticoagulation and vasodilating drugs (prostaglandin E) were administered. Within 2 days, capillary refill reappeared and the initial loss of sensory function at the tip of the thumb and index finger diminished. A screening test for thrombophilic disorders led to the diagnosis of a heterozygous mutation of factor V (Leiden mutation). Arterial thromboembolic events of factor V Leiden mutation are rare and have to date been described only in the supraaortic and coronary circulation. Therefore, the arterial embolism to the left hand presented in this report constitutes a rarity that could be successfully salvaged by the combined use of a vascular surgical procedure and intensified medical management.     

Anatomically corrected malposition of the great arteries, inflow ventricular septal defect, and subaortic stenosis: diagnostic and operative implications

Intracardiac correction of the combination of rare congenital heart lesion of anatomically corrected malposition of the great arteries, severe muscular as well as fibrous subaortic stenosis, and ventricular septal defects in the inflow and the trabecular septum in a 2 1/2 years old boy is reported. There was an additional large secundum atrial septal defect. The operation consisted of resection of the subaortic stenosis by a combined transatrial and transaortic approach, thereby regaining a wide outflow tract of the systemic ventricle. Both the ventricular and atrial septal defects were closed with Dacron patches. Reinvestigation 10 days after the operation revealed a residual pressure gradient across the outflow of the systemic ventricle of less than 10 mmHg. A secondary ventricular septal defect was detected in the anterior muscular septum at the same time and closed with a patch in a reintervention. At discharge from the hospital two weeks later the child was in excellent condition and had a stable sinus rhythm. The preoperative differential diagnosis of a double outlet right ventricle with L-malposition of the aorta as well as possible surgical approaches are discussed.     

Limb trauma with arterial injury: long-term performance of venous interposition grafts

BACKGROUND: In contrast to arterial occlusive disease, data on long-term outcomes after vein grafts in limb trauma with arterial injury are sparse. PATIENTS: From 1991 through 2001, 22 trauma victims received 23 interposition vein grafts performed by an interdisciplinary team of trauma and vascular surgeons. Indications included both blunt and penetrating injuries with critical limb ischemia in the majority of cases. RESULTS: Operative treatment of the injured vessels (brachial n = 5, radial/ulnar n = 7, popliteal n = 6, tibial n = 3, pedal n = 2) encompassed venous interposition graft of either saphenous (n = 15) or cephalic vein (n = 8). All patients survived the operative procedure. 4 graft occlusions were noted and 3 major amputations had to be performed (one despite patent graft). 13 patients (76%) were available for duplex ultrasound examination after a mean follow-up of 59 months where patent grafts could be detected in all cases. CONCLUSION: A multidisciplinary approach ensures optimal treatment strategy of arterial injury in extremity trauma. Interposition vein grafts provide durable long-term results and should be attempted even in single-vessel injuries of forearm and lower leg.     

Determination of cardiac output by an angle and diameter independent dual beam Doppler technique in critically ill infants.

OBJECTIVE--To compare cardiac output measurements in critically ill infants by the dual beam Doppler and thermodilution techniques. DESIGN--Prospective direct comparison of the two techniques. For statistical evaluation one randomly assigned paired measurement of every patient was used. SETTING--Paediatric intensive care unit in a university hospital. PATIENTS--18 infants after open heart surgery aged 4-25 months (weight 4-10 kg). INTERVENTIONS--Cardiac output measurements by dual beam Doppler and thermodilution techniques were performed within 10 minutes of each other and without knowledge of the results of the other methods. Multiple measurements were performed on some patients with a pharmacological or electrophysiological intervention or with a minimum of six hours between each pair of measurements. MEASUREMENTS AND MAIN RESULTS--Three patients were excluded because of an inadequate Doppler signal or a significant residual shunt. Cardiac output measurements ranged from 0.4 to 2.2 l/min for the thermodilution technique and from 0.5 to 2.1 l/min for the dual beam Doppler technique. Agreement between both methods was acceptable. The mean difference between the two methods was 0.026 l/min with two standard deviations ranging from -0.20 to 0.26 l/min. CONCLUSION--The dual beam Doppler technique was shown to have promise for the non-invasive determination of cardiac output in critically ill infants.     

Sensitivity of MRI resonance frequency to the orientation of brain tissue microstructure

Recent advances in high-field (≥7 T) MRI have made it possible to study the fine structure of the human brain at the level of fiber bundles and cortical layers. In particular, techniques aimed at detecting MRI resonance frequency shifts originating from local variation in magnetic susceptibility and other sources have greatly improved the visualization of these structures. A recent theoretical study [He X, Yablonskiy DA (2009) Proc Natl Acad Sci USA 106:13558–13563] suggests that MRI resonance frequency may report not only on tissue composition, but also on microscopic compartmentalization of susceptibility inclusions and their orientation relative to the magnetic field. The proposed sensitivity to tissue structure may greatly expand the information available with conventional MRI techniques. To investigate this possibility, we studied postmortem tissue samples from human corpus callosum with an experimental design that allowed separation of microstructural effects from confounding macrostructural effects. The results show that MRI resonance frequency does depend on microstructural orientation. Furthermore, the spatial distribution of the resonance frequency shift suggests an origin related to anisotropic susceptibility effects rather than microscopic compartmentalization. This anisotropy, which has been shown to depend on molecular ordering, may provide valuable information about tissue molecular structure.     

The BTB-Kelch protein KLEIP controls endothelial migration and sprouting angiogenesis

Sprouting and invasive migration of endothelial cells are important steps of the angiogenic cascade. Vascular endothelial growth factor (VEGF) induces angiogenesis by activating intracellular signal transduction cascades, which regulate endothelial cell morphology and function. BTB-kelch proteins are intracellular proteins that control cellular architecture and cellular functions. The BTB-kelch protein KLEIP has been characterized as an actin-binding protein that interacts with the nucleotide exchange factor ECT2. We report that KLEIP is preferentially expressed in endothelial cells, suggesting that it may play a critical role in controlling the functions of migrating, proliferating, and invading endothelial cells during angiogenesis. KLEIP mRNA level in endothelial cells is strongly regulated by hypoxia which is controlled by hypoxia-inducible factor-1alpha. Functional analysis of KLEIP in endothelial cells revealed that it acts as an essential downstream regulator of VEGF- and basic fibroblast growth factor-induced migration and in-gel sprouting angiogenesis. Yet, it is not involved in controlling VEGF- or basic fibroblast growth factor-mediated proliferative responses. The depletion of KLEIP in endothelial cells blunted the VEGF-induced activation of the monomeric GTPase RhoA but did not alter the VEGF-stimulated activation of extracellular signal-regulated kinase 1/2. Moreover, VEGF induced a physical association of KLEIP with the guanine nucleotide-exchange factor ECT2, the depletion of which also blunted VEGF-induced sprouting. We conclude that the BTB-kelch protein KLEIP is a novel regulator of endothelial function during angiogenesis that controls the VEGF-induced activation of Rho GTPases.