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Inflammation and thrombosis are increasingly recognized as interrelated biologic processes. Endothelial cell expression of thrombomodulin (TM), a key component of the anticoagulant protein C pathway, is potently inhibited by inflammatory cytokines. Because the mechanism underlying this effect is largely unknown, we investigated a potential role for the inflammatory transcription factor nuclear factor-kappa B (NF-κB). Blocking NF-κB activation effectively prevented cytokine-induced down-regulation of TM, both in vitro and in a mouse model of tumor necrosis factor-α (TNF-α)–mediated lung injury. Although the TM promoter lacks a classic NF-κB consensus site, it does contain tandem Ets transcription factor binding sites previously shown to be important for both constitutive TM gene expression and cytokine-induced repression. Using electrophoretic mobility shift assay and chromatin immunoprecipitation, we found that multiple Ets species bind to the TNF-α response element within the TM promoter. Although cytokine exposure did not alter Ets factor binding, it did reduce binding of p300, a coactivator required by Ets for full transcriptional activity. Overexpression of p300 also prevented TM repression by cytokines. We conclude that NF-κB is a critical mediator of TM repression by cytokines. Further evidence suggests a mechanism involving competition by NF-κB for limited pools of the transcriptional coactivator p300 necessary for TM gene expression.

Introduction Superoxide anion may contribute to erectile dysfunction (ED) in diabetes mellitus by reducing cavernosal nitric oxide (NO) bioavailability. The purpose of this study was to determine if gene transfer of extracellular superoxide dismutase (EC‐SOD) can reduce superoxide anion formation and determine if this reactive oxygen species may contribute to diabetes‐related ED in an experimental model of diabetes. Methods Three groups of animals were utilized: (1) control; (2) streptozotocin (STZ)‐diabetic rats [60 mg/kg intraperitoneally (ip)] intracavernosally injected with AdCMVβgal (negative control); and (3) STZ‐rats intracavernosally injected with AdCMVEC‐SOD. Two months after ip injection of STZ, groups 2 and 3 were transfected with the adenoviruses and 2 days after transfection, all animals underwent cavernosal nerve stimulation (CNS) to assess erectile function. Confocal microscopy for superoxide anion and von Willebrand Factor (vWF) was performed in the STZ‐diabetic rat. Superoxide anion production, total SOD activity, and cyclic guanosine monophosphate (cGMP) levels were measured in each experimental group of rats. Results 248 Confocal microscopy demonstrated superoxide in smooth muscle and endothelial cells of the STZ‐rat cavernosum and colocalized with vWF in the endothelium. Higher superoxide anion levels and decreased cGMP levels were found in the penis of STZ‐rats at a time when erectile function was reduced. Two days after administration of AdCMVEC‐SOD, superoxide anion levels were significantly lower in the penis of STZ‐rats. Total SOD activity and cavernosal cGMP was increased in the penis of EC‐SOD‐transfected rats. STZ‐rats transfected with AdCMVEC‐SOD had a peak intracavernosal pressure (ICP) and total ICP to CNS that was similar to control rats. Conclusions These data demonstrate that in vivo adenoviral gene transfer of EC‐SOD can reduce corporal superoxide anion levels and raise cavernosal cGMP levels by increasing NO bioavailability thus restoring erectile function in the STZ‐diabetic rat.

The ca. 3.12 Ga Whundo Group forms an exotic terrane on the western margin of the old (≥3.4 Ga) nucleus of the Pilbara Craton in northwestern Australia. It is exceptional amongst Mesoarchaean, or older, volcanic sequences in that it preserves a range of geological and geochemical features that together provide unambiguous evidence of modern-style subduction processes. The group includes boninites, interlayered tholeiitic and calc-alkaline volcanics, Nb-enriched basalts and adakites. Low Th/La (0.07–0.14) and Ce/Yb (

background Reversible left ventricular dysfunction precipitated by emotional stress has been reported, but the mechanism remains unknown. methods We evaluated 19 patients who presented with left ventricular dysfunction after sudden emotional stress. All patients underwent coronary angiography and serial echocardiography; five underwent endomyocardial biopsy. Plasma catecholamine levels in 13 patients with stress-related myocardial dysfunction were compared with those in 7 patients with Killip class III myocardial infarction. results The median age of patients with stress-induced cardiomyopathy was 63 years, and 95 percent were women. Clinical presentations included chest pain, pulmonary edema, and cardiogenic shock. Diffuse T-wave inversion and a prolonged QT interval occurred in most patients. Seventeen patients had mildly elevated serum troponin I levels, but only 1 of 19 had angiographic evidence of clinically significant coronary disease. Severe left ventricular dysfunction was present on admission (median ejection fraction, 0.20; interquartile range, 0.15 to 0.30) and rapidly resolved in all patients (ejection fraction at two to four weeks, 0.60; interquartile range, 0.55 to 0.65; P

The molecular mechanism for priapism is not well characterized. Although the nitric oxide (NO) pathway is known to mediate penile erection under normal conditions, we hypothesized that the mechanism of priapism rests in aberrant downstream signaling of this pathway based on our previous findings that mice lacking the gene for endothelial nitric oxide synthase (eNOS —/— ) and mice lacking both neuronal NOS (nNOS) and eNOS (nNOS —/— , eNOS —/— ) have a tendency for priapic activity. We investigated the role of downstream guanylate cyclase and phosphodiesterase type 5 (PDE5A) expression and function in mediating these responses in eNOS —/— and nNOS —/— , eNOS —/— mice. Erectile responses to both cavernous nerve stimulation and intracavernosal injection of the NO donor diethylamine-NONOate were augmented in eNOS —/— and nNOS —/— , eNOS —/— mice but not in WT or nNOS —/— mice. PDE5A protein expression and activity and cGMP levels were significantly lower in eNOS —/— and nNOS —/— , eNOS —/— mice, and this effect was reproduced in WT corpus cavernosum exposed to NOS inhibitors. Moreover, cavernous nerve stimulation was associated with a marked augmentation of cavernosal cGMP levels, suggesting that, although lower at baseline, the production of cGMP is unchecked in eNOS —/— and nNOS —/— , eNOS —/— mice upon neurostimulation. Transfection of eNOS —/— mice with an adenovirus encoding eNOS resulted in a normalization of PDE5A protein and activity as well as a correction of priapic activity. Coupled with the observation that sickle cell disease mice (which show a priapism phenotype) evince dysregulated PDE5A expression/activity, these data suggest that PDE5A dysregulation is a fundamental mechanism for priapism.

Examination of an extensive adakite geochemical database identifies two distinct compositional groups. One consists of high-SiO2 adakites (HSA) which is considered to represent subducted basaltic slab-melts that have reacted with peridotite during ascent through mantle wedge. The second group consists of low-SiO2 adakites (LSA) which we interpret to have formed by melting of a peridotitic mantle wedge whose composition has been modified by reaction with felsic slab-melts. The chemical composition of less differentiated (primitive) Archaean tonalite–trondhjemite–granodiorite (TTG) magmas evolvedfrom4.0to2.5Ga.Mg#(molecularMg/(Mg+Fe 2+ ),Ni,andCrcontentsincreasedoverthisperiodoftimeandweinterpret thesechangesintermsofchangesinthedegreetowhichtheTTGmagmasinteractedwithmantleperidotite.Overthesame period, concentrations of (CaO+Na2O) and Sr also increased, as the amount of plagioclase, residual from basalt melting, decreased in responsetoincreasedpressuresatthesiteofslab-melting. IntheEarlyArchaean,itappearsthattheseinteractionswereveryrareor absentthus leading tothe conclusion that subduction was typically flat andlacked the developmentofa mantle wedge.In contrast, therelativelylowerheatproduction by~2.5Gameant thatslab-melting occurredatgreaterdepth,whereplagioclasewasnolonger stable, and where the development of a thick mantle wedge ensured interaction between the slab-melts and mantle peridotite. Close compositional similarities between HSA and Late Archaean TTG (Tb~3.0 Ga) strongly suggest a petrogenetic analogy. However, an analogy between the older Archaean TTG and HSA is not complete because evidence for mantle wedge interaction is missing in most Early Archaean TTGs. Late Archaean sanukitoids and the compositionally similar Closepet-type granites have compositions significantly different from TTG of all ages. However, they show some affinity with LSAwhich could be considered as their possible analogue. These magmas are all thought to result from melting of a mantle peridotite whose composition has been modified by reaction with slab-melts.

Deep seismic reflection data across the Archaean Eastern Goldfields Province, northeastern Yilgarn Craton, Western Australia, have provided information on its crustal architecture and on several of its highly mineralised belts. The seismic reflection data allow interpretation of several prominent crustal scale features, including an eastward thickening of the crust, subdivision of the crust into three broad layers, the presence of a prominent east dip to the majority of the reflections and the interpretation of three east-dipping crustal-penetrating shear zones. These east-dipping shear zones are major structures that subdivide the region into four terranes. Major orogenic gold deposits in the Eastern Goldfields Province are spatially associated with these major structures. The Laverton Tectonic Zone, for example, is a highly mineralised corridor that contains several world-class gold deposits plus many smaller deposits. Other non crustal-penetrating structures within the area do not appear to be as well endowed metallogenically as the Laverton structure. The seismic reflection data have also imaged a series of low-angle shear zones within and beneath the granite–greenstone terranes. Where the low-angle shear zones intersect the major crustal-penetrating structures, a wedge shaped geometry is formed. This geometry forms a suitable fluid focusing wedge in which upward to subhorizontal moving fluids are focused and then distributed into the nearby complexly deformed greenstones.

Rare Archaean light rare earth element (LREE)-enriched mafic rocks derived from a strongly refractory mantle source show a range of features in common with modern boninites. These Archaean second-stage melts are divided into at least two distinct groups—Whundo-type and Whitney-type. Whundo-type rocks are most like modern boninites in terms of their composition and association with tholeiitic to calc-alkaline mafic to intermediate volcanics. Small compositional differences compared to modern boninites, including higher Al2O3 and heavy REE (HREE), probably reflect secular changes in mantle temperatures and a more garnet-rich residual source. Whundo-type rocks are known from 3.12 and 2.8 Ga assemblages and are true Archaean analogues of modern boninites. Whitney-type rocks occur throughout the Archaean, as far back as ca. 3.8 Ga, and are closely associated with ultramafic magmatism including komatiites, in an affiliation unlike that of modern subduction zones. They are characterised by very high Al2O3 and HREE concentrations, and their extremely depleted compositions require a source which at some stage was more garnet-rich than the source for either modern boninites or Whundo-type second-stage melts. Low La/Yb and La/Gd ratios compared to Whundo-type rocks and modern boninites either reflect very weak subduction-related metasomatism of the mantle source or very limited crustal assimilation by a refractory-mantle derived melt. Regardless, the petrogenesis of the Whitney-type rocks appears either directly or indirectly related to plume magmatism. If Whitney-type rocks have a boninitic petrogenesis then a plume related model similar to that proposed for the modern Tongan high-Ca boninites might apply, but with uniquely Archaean source compositions and source enrichment processes. Second-stage melts from Barberton (S. Africa –3.5 Ga) and ca. 3.0 Ga rocks from the central Pilbara (Australia) have features in common with both Whundo- and Whitney-types, but appear more closely related to the Whitney-type. Subduction zone processes essentially the same as those that produce modern boninites have operated since at least ~3.12 Ga, while a uniquely Archaean boninite-forming process, involving more buoyant oceanic plates and very inefficient mantle-source enrichment, may have occurred before then.

Significant impairment in endothelial-derived nitric oxide is present in the diabetic corpus cavernosum. RhoA/Rho-kinase may suppress endothelial nitric oxide synthase (eNOS). Here, we tested the hypothesis that RhoA/Rho-kinase contributes to diabetes-related erectile dysfunction and down-regulation of eNOS in the streptozotocin (STZ)-diabetic rat penis. Colocalization of Rho-kinase and eNOS protein was present in the endothelium of the corpus cavernosum. RhoA/Rho-kinase protein abundance and MYPT-1 phosphorylation at Thr-696 were elevated in the STZ-diabetic rat penis. In addition, eNOS protein expression, cavernosal constitutive NOS activity, and cGMP levels were reduced in the STZ-diabetic penis. To assess the functional role of RhoA/Rho-kinase in the penis, we evaluated the effects of an adeno-associated virus encoding the dominant-negative RhoA mutant (AAVTCMV19NRhoA) on RhoA/Rho-kinase and eNOS and erectile function in vivo in the STZ-diabetic rat. STZ-diabetic rats transfected with AAVCMVT19NRhoA had a reduction in RhoA/Rho-kinase and MYPT-1 phosphorylation at a time when cavernosal eNOS protein, constitutive NOS activity, and cGMP levels were restored to levels found in the control rats. There was a significant decrease in erectile response to cavernosal nerve stimulation in the STZ-diabetic rat. AAVT19NRhoA gene transfer improved erectile responses in the STZ-diabetic rat to values similar to control. These data demonstrate a previously undescribed mechanism for the down-regulation of penile eNOS in diabetes mediated by activation of the RhoA/Rho-kinase pathway. Importantly, these data imply that inhibition of RhoA/Rho-kinase improves eNOS protein content and activity thus restoring erectile function in diabetes.

The Eastern Goldfields Province (EGP) is a late Archaean orogenic belt located on the eastern margin of the Yilgarn Craton. The EGP is characterised by a prominent NNW-trending tectonic grain developed largely during regional ∼E-W oriented shortening. A simplistic sequence of largely progressive compressional deformation events (D 1 to D 4+ ) has been used by previous workers to explain the development of the EGP. Regional ‘D 2 ’ was thought to be responsible for most of the finite strain, and therefore a reliable marker event for correlating deformation histories across the region. A re-examination of the deformation history has shown that ‘D 2 ’ (Kalgoorlie Orogen of Weinberg et al. [Precambrian Res. 120 (2003) 219–239]) was not a single progressive event. Rather, it was episodic and involved several stages of switching of the tectonic mode (compression–extension switching) between ∼2665 and ∼2655 Ma. The sequence of events during ‘D 2 ’ was an early stage of ∼E-W shortening (D 2a ), followed by extension with basin formation and normal fault movement (D 2E ), and finally a second stage of ∼E-W compression (D 2b ). We introduce this sequence of events as the Wangkathaa Orogeny, more complex in tectonics and broader in geographical scope than the Kalgoorlie Orogen. Features of the Wangkathaa Orogeny are well illustrated by the geometry and geological relationships that occur in the Kurnalpi Terrane (Welcome Well area) and the Kalgoorlie Terrane (Ora Banda, Kambalda, and the Boorara Domains) of the central and southern EGP. Available geochronology indicates that this sequence of events was likely to have been diachronous across the entire EGP and the eastern Southern Cross Province (of the Yilgarn Craton). Recognition of these separate (episodic) stages of ∼E-W oriented compression and extension is important in our understanding the tectonic evolution of the EGP. Switching tectonic mode between compression and extension will change the state of mean-stress in a rock mass (e.g. from net dilation to net constriction and vice versa), and is an effective method in transporting and trapping mineral-rich fluids. The paradox of the different timings established for ‘D 2 ’ across the province are explained by the Wangkathaa Orogeny, especially if it were diachronous, as different workers dated different phases of an episodic orogeny.

Nitric oxide (NO) functions principally as a diffusible paracrine effector. The exception is in cardiomyocytes where both NO synthases (NOS) and target proteins coexist, allowing NO to work in an autocrine/intracrine fashion. However, the most abundant myocyte isoform (NOS3) is far more expressed in vascular endothelium; thus, the in vivo contribution of myocyte-NOS3 remains less clear. The present study tested this role by transfecting whole hearts of NOS3-null (NOS3 −/− ) mice with adenovirus-expressing NOS3 coupled to a α-MHC promoter (AdV NOS3 ), comparing results to hearts transfected with marker-gene β-galactosidase (AdVβ gal ). Total myocardial NOS3 protein and activity were restored to near wild-type (WT) levels in NOS3 −/− +AdV NOS3 hearts, and NOS3 relocalized normally with caveolin-3. Ejection function by pressure-volume analysis was enhanced in NOS3 −/− +AdVβ gal over WT or NOS3 −/− +AdV NOS3 . More prominently, isoproterenol (ISO)-stimulated systolic and diastolic function in WT was amplified in NOS3 −/− +AdVβ gal , whereas NOS3 −/− +AdV NOS3 returned the response to control. ISO-activated systolic function was inhibited 85% by concomitant muscarinic stimulation (carbachol) in NOS3 −/− +AdV NOS3 but not NOS3 −/− +AdVβ gal hearts. Lastly, NOS3 −/− +AdVβ gal mice displayed enhanced inotropy and lusitropy over WT at slower heart rates but a blunted rate augmentation versus controls. A more positive rate response was restored in NOS3 −/− +AdV NOS3 ( P

Erectile dysfunction associated with diabetes mellitus is caused in part by disordered endothelial smooth muscle relaxation, neuropathy, and a decrease in cavernosal nitric oxide synthase (NOS) activity. The purpose of this study was to determine whether a combination of sildenafil and adenoviral gene transfer of endothelial NOS (eNOS) could enhance the erectile response in diabetic rats. Five groups of animals were utilized: (1) age-matched control rats, (2) streptozotocin (STZ)-induced diabetic rats (60 mg/kg i.p.), (3) STZ-rats + sildenafil (2 mg/kg i.v.), (4) STZ-rats transfected with AdCMVbetagal or AdCMVeNOS, and (5) STZ-rats transfected with AdCMVeNOS +sildenafil (2 mg/kg i.v.). At 2 months after i.p. injection of STZ, groups 4 and 5 were transfected with the adenoviruses and 1-2 days after transfection, all animals underwent cavernosal nerve stimulation (CNS) to assess erectile function. Cyclic 3',5'-guanosine monophosphate (cGMP) levels were assessed in the cavernosal tissue. STZ-diabetic rats had a significant decrease in erectile function as determined by the peak intracavernosal pressure (ICP) and total ICP (area under the erectile curve; AUC) after CNS when compared to control rats. STZ-diabetic rats+AdCMVeNOS had a peak ICP and AUC, which were similar to control animals. STZ-diabetic rats administered sildenafil demonstrated a significant increase in peak ICP at the 5 and 7.5 V settings, while the AUC was significantly increased at all voltage (V) settings. The increase in both ICP and AUC of STZ-diabetic rats transfected with AdCMVeNOS at all V settings was greater than STZ-diabetic rats transfected with AdCMVbetagal. STZ-diabetic rats transfected with AdCMVeNOS and administered sildenafil had a significant increase in total ICP that was greater than eNOS gene therapy alone. Cavernosal cGMP levels were significantly decreased in STZ-diabetic rats, but were increased after transfection with AdCMVeNOS to values greater than control animals. In conclusion, overexpression of eNOS and cGMP in combination with sildenafil significantly increased both the peak ICP and total ICP to CNS in the STZ-diabetic rat, which was similar to the response observed in control rats. Moreover, the total erectile response was greater in STZ-diabetic rats receiving eNOS gene therapy plus sildenafil than STZ-rats receiving sildenafil or eNOS gene therapy alone.

Background— Viral gene transfer to the whole heart in vivo has been achieved in several mammalian species but remained difficult to accomplish in murine hearts. We postulated that a key impediment derives from the use of proximal aortic occlusion during virus injection, because this eliminates coronary perfusion gradients in mice as aortic root and left ventricle pressures equalize. Methods and Results— Pressure-volume analysis confirmed these mechanics. In contrast, descending aortic occlusion with whole-body cooling (20°C) preserved transmyocardial perfusion gradients and allowed for sustained (>10-minute) dwell times in an upper-body perfusion circuit. This approach yielded robust cardiac transfection with adenovirus (AdV) and adeno-associated virus (AAV) injected into the left ventricle cavity or more simply via a central vein. Cardio-specific expression was achieved with a myocyte-specific promotor. Optimal AdV transfection required 9-minute aortic occlusion, versus 5-minute occlusion for AAV. Using this method, we examined the in vivo function of phospholamban (PLB) by stably transfecting PLB-null mice with AAV encoding PLB (AAV PLB ). AAV PLB restored PLB protein to near control levels that colocalized with SERCA2A in cardiomyocytes. At baseline, PLB-null hearts exhibited enhanced systolic and diastolic function, but frequency-dependent reserve was blunted versus wild-type controls. These properties, particularly the frequency response, returned toward control 3 months after AAV PLB transfection. Conclusions— The new simplified approach for murine whole-heart viral transfection should assist molecular physiology studies.

Erectile dysfunction (ED) is defined as the consistent inability to obtain or maintain an erection for satisfactory sexual intercourse. Basic science research on erectile physiology has been devoted to investigating the pathogenesis of ED and has led to the conclusion that ED is predominately a disease of vascular origin. The incidence of ED dramatically increases in men with diabetes mellitus, hypercholesterolemia, and cardiovascular disease. Loss of the functional integrity of the endothelium and subsequent endothelial dysfunction plays an integral role in the occurrence of ED in this cohort of men. This communication reviews the role of the vascular endothelium in erectile physiology and the influence of endothelial dysfunction in the pathogenesis of ED. Future pharmacological and gene therapy interventions to restore endothelial function may represent exciting new therapeutic strategies for the treatment of ED. Penile erection is a neurovascular phenomenon that depends upon neural integrity, a functional vascular system, and healthy cavernosal tissues (Giuliano et al, 1995). Normal erectile function involves 3 synergistic and simultaneous processes: 1) neurologically mediated increase in penile arterial inflow, 2) relaxation of cavernosal smooth muscle, and 3) restriction of venous outflow from the penis. The corpus cavernosum of the penis is composed of a meshwork of interconnected smooth muscle cells lined by vascular endothelium. Of note, endothelial cells and underlying smooth muscle also line the small resistance helicine arteries that supply blood to the corpus cavernosum during penile tumescence. Pathological alteration in the anatomy of the penile vasculature or impairment of any combination of neurovascular processes can result

Subduction of oceanic crust at an unusually low-angle has been proposed as a model for the growth of continental crust older than about 2.5 Ga. At modern zones of low-angle-, or flat-subduction, magmatic additions to new crust come from partial melting of both the subducting oceanic crust (slab) and the thin wedge of mantle above the slab. Evidence for both a slab and wedge source is preserved in most late Archaean (3.0–2.5 Ga) terrains, but we find little evidence that a mantle wedge contributed to crustal growth prior to ∼3.1 Ga. This lack of evidence in part reflects a dearth of exposed crust aged between 3.0 and 3.3 Ga, but also suggests that subduction enriched mantle source regions did not develop before ∼3.3 Ga and possibly not before 3.1 Ga. In contrast to most modern terrains and some late-Archaean terrains, early Archaean (>∼3.3 Ga) continental crust evolved through direct melting of thick mafic crust. We invoke a process of subduction that does not include the development of a mantle wedge, and call this process Archaean flat-subduction to distinguish it from modern low-angle subduction.

This study compares uncertainties of equivalent doses after internal contamination by 125 I, 129 I or 131 I. Uncertainties were calculated using reported distributions of physiological parameters and Monte Carlo simulation. In adults, uncertainties increase from 131 I to 125 I and 129 I with 1% of the population receiving 3.9, 4.0 and 7.2 times the median dose for the respective isotopes. In newboms, these values were 7.5, 12.3 and 19.0 for 131 I, 125 I and 129 I respectively. The ratio of the beta dose delivered to the epithelium versus a homogeneous distributed dose was estimated for different iodine concentrations in colloid, epithelium and interstitium. In adults, for 131 I, about 40% of the beta dose was delivered to the epithelial cells whereas this fraction varied depending on the concentration for 125 I and 129 I, i.e. 20-30% at a relative epithelial concentration of 20% and 7-14% at a concentration of 3%. Small variations were observed depending on age.

We determined whether adenoviral gene transfer of endothelial nitric oxide synthase (eNOS) to the penis of streptozotocin induced diabetic rats could improve the impaired erectile response.Two experimental groups of animals were transfected with adenoviruses, including streptozotocin (Sigma Chemical Company, St. Louis, Missouri) diabetic rats with AdCMVbetagal and streptozotocin diabetic rats with AdCMVeNOS. At 1 to 2 days after transfection these study animals underwent cavernous nerve stimulation to assess erectile function and their responses were compared with those of age matched control rats. In control and transfected streptozotocin diabetic rats eNOS and neuronal NOS (nNOS) were examined by Western blot analysis. Constitutive and inducible NOS activities were evaluated in the presence and absence of calcium by L-arginine to L-citrulline conversion and nitrate plus nitrite levels were measured. In control and streptozotocin diabetic penes beta-galactosidase activity and localization were determined.After transfection with AdCMVbetagal beta-galactosidase was localized to the endothelium and smooth muscle cells of the streptozotocin diabetic rat penis. Streptozotocin diabetic rats had a significant decrease in erectile function, as determined by peak and total intracavernous pressure (area under the curve) after cavernous nerve stimulation compared with control rats. Streptozotocin diabetic rats transfected with AdCMVeNOS had peak intracavernous pressure and area under the curve similar to those in control animals. This change in erectile function was a result of eNOS over expression with an increase in eNOS protein expression and constitutive NOS activity as well as an increase in nitric oxide biosynthesis, as reflected by an increase in cavernous nitrate plus nitrite formation. There was no change in nNOS protein expression or calcium independent conversion of NOS (inducible NOS activity).Adenoviral gene transfer of eNOS significantly increased peak and total intracavernous pressure to cavernous nerve stimulation in streptozotocin diabetic rats to a value similar to the response observed in control rats. Our results suggest that eNOS contributes significantly to the physiology of penile erection. These data demonstrate that in vivo adenoviral gene transfer of eNOS can physiologically improve erectile function in the streptozotocin diabetic rat.

Increased superoxide anion (O[Formula: see text]·) may contribute to vascular dysfunction in aging. In aged cavernosal tissue, lucigenin-enhanced chemiluminescence demonstrated a threefold increase in superoxide formation, and the oxidative fluorescent probe hydroethidine indicated higher superoxide levels throughout the aged penis. This increase in superoxide was associated with impaired cavernosal nerve-mediated and agonist-induced erectile responses, increased nitrotyrosine staining, and lower cGMP levels, but no compensatory change in cavernosal extracellular (EC)-superoxide dismutase (EC-SOD) mRNA or protein. In vivo adenoviral (Ad) gene transfer of EC-SOD to the penis resulted in higher expression of EC-SOD mRNA, protein, SOD activity, cGMP levels, and lower nitrotyrosine staining. Transfection with AdCMVEC-SOD resulted in a significant increase in erectile response to cavernosal nerve stimulation, ACh, and zaprinast to a magnitude similar to young rats. These data provide evidence in support of the hypothesis that erectile dysfunction associated with aging is related in part to an increase in cavernosal O[Formula: see text]· formation. Gene-transfer of EC-SOD reduces superoxide formation and restores age-associated erectile function and may represent a novel therapeutic target for the treatment of erectile dysfunction.

We report the case of a 51-year-old woman who presents with a 2-week history of episodes of pressure like chest pain. The initial electrocardiogram was not indicative of myocardial ischemia or infarction and the cardiac enzymes remained normal during the initial hospital day. However, the precordial T waves inverted and progressively deepened on the second hospital day and the patient underwent cardiac catheterization with percutaneous coronary angioplasty and stent placement of the left anterior descending coronary artery with good results. The postprocedure electrocardiogram showed complete resolution of the inverted precordial T waves. The development of new T-wave inversions in the precordial leads of patients presenting with unstable angina is predictive of significant stenosis of the left anterior descending coronary artery. This subgroup of patients has a poor prognosis if medical therapy alone is instituted. Early cardiac catheterization and revascularization is recommended for these patients. Evidence has shown that 75% patients with these electrocardiogram changes who are not revascularized developed extensive anterior wall infarction within a few weeks.

The etiologies of erectile dysfunction (ED) after nerve-sparing radical prostatectomy have not been clearly elucidated. The aim of this study was to evaluate the effects of cavernous nerve injury on cavernous fibrosis, and to consider measures to prevent irreversible damage to the cavernous tissues. Twenty male Sprague-Dawley rats constituted the study population. The animals were divided into 2 groups; group 1 consisted of sham-operated rats (n = 10), and group 2 consisted of rats that underwent incision of both cavernous nerves (n = 10). Three months later, all rats underwent intracavernous papaverine injection (300 and 600 mg), and intracorporal pressures were recorded. Transforming growth factor-beta(1) (TGF-beta(1)) messenger RNA (mRNA) expression from rat penile tissue was measured using reverse transcriptase-polymerase chain reaction. Hypoxia-inducible factor-1alpha (HIF-1alpha), TGF-beta(1), and collagen I and III protein expressions were determined by Western blot analysis and immunohistochemical staining. Erectile function as studied with intracavernosal papaverine injection and histological analysis of penile cross-sections at 3 months was similar in both groups. TGF-beta(1) mRNA expression, HIF-1alpha, TGF-beta(1), and collagen I and III protein expressions were significantly greater in the neurotomy group. Immunohistochemical staining for TGF-beta(1), HIF-1alpha, and collagen III were qualitatively more positive in the neurotomy group, whereas collagen I staining was similar. This study demonstrates an increase in TGF-beta(1), HIF-1alpha, and collagen III synthesis in rat cavernosal smooth musculature after cavernous neurotomies. In theory, cavernous fibrosis may be reduced by employing various vasoactive agents or interventions that increase oxygenation to the corporal tissues during the postoperative period.

Responses to human CGRP, adrenomedullin (ADM), and proadrenomedullin NH2-terminal 20 peptide (PAMP) were studied in small human thymic arteries. CGRP, ADM, and PAMP produced concentration-dependent vasodilator responses in arteries preconstricted with the thromboxane mimic U-46619. Responses to ADM and PAMP were attenuated, whereas responses to CGRP were not altered by endothelial denudation. Inhibitors of nitric oxide synthase and guanylyl cyclase attenuated responses to ADM and PAMP but not to CGRP. The CGRP1 receptor antagonist CGRP(8–37) attenuated responses to CGRP and ADM but not to PAMP. Responses to CGRP were reduced by SQ-22536 and Rp-cAMPS, inhibitors of adenylyl cyclase and PKA. These data suggest that responses to CGRP and ADM are mediated by CGRP(8–37)-sensitive receptors and that the endothelial ADM receptor induces vasodilation by a nitric oxide-guanylyl cyclase mechanism, whereas a smooth muscle CGRP receptor signals by a cAMP-dependent mechanism. A different endothelial receptor recognizes PAMP and signals by a nitric oxide-dependent mechanism.

Nitric oxide (NO) plays critical roles in the regulation of integrated cardiac and vascular function and homeostasis. An understanding of the physiologic role and relative contribution of the three NO synthase isoforms (neuronal--NOS1, inducible--NOS2, and endothelial--NOS3) is imperative to comprehend derangements of the NO signaling pathway in the failing cardiovascular system. Several theories of NO and its regulation have developed as explanations for the divergent observations from studies in health and disease states. Here we review the physiologic and pathophysiologic influence of NO on cardiac function, in a framework that considers several theories of altered NO signaling in heart failure. We discuss the notion of spatial compartmentalization of NO signaling within the myocyte in an effort to reconcile many controversies about derangements in the influences of NO in the heart and vasculature.

HIV-associated pulmonary arterial hypertension (HIV-PAH) is a serious complication that develops in approximately 1:200 of HIV-infected individuals, and an even higher prevalence of echocardiographic signs of pulmonary hypertension indicates that the disease may be more common than previously thought (1–3). Mortality is high, even in the era of antiretroviral therapy (ART) (4). Lower CD4 cell count has been associated with HIV-PAH, but successful treatment with ART has not reduced prevalence, suggesting that factors other than HIV may contribute to disease (4). Although HIV viral proteins have been suggested to play a role in the pathogenesis of HIV-PAH, not all patients with HIV develop the disease, leading to the investigation of other risk factors, or “second-hit” events. Intravenous drug use in HIV infection is one potential “second hit” that has been associated with HIV-PAH (5); however, the contribution of specific illicit drugs such as opioids or amphetamine derivatives, and the mechanism by which these drugs contribute to pulmonary vasculopathy in HIV-PAH, is not known. Figure 1. Multiple hits are likely involved in the pathogenesis of HIV-associated pulmonary arterial hypertension. In this issue of the Journal, Spikes and colleagues (pp. 1235–1243) study a nonhuman primate model to investigate the effect of morphine on simian immunodeficiency virus (SIV) infection and the development of pulmonary hypertension (6). They report that rhesus macaques pretreated with morphine for 26 weeks, followed by inoculation with macrophage-tropic SIVmacR71/17E virus, and subsequent treatment with intramuscular morphine for 31 weeks after inoculation, develop significant pulmonary vascular remodeling including plexiform lesions, whereas animals either infected with SIV alone or treated with morphine alone did not. Although there is mild to moderate inflammation in all animals, the authors report increased perivascular inflammation in the SIV/morphine group. In addition, increased macrophage lung infiltration and elevated plasma levels of monocyte chemotactic protein-1 and interleukin-8 are associated with vascular remodeling in the SIV-infected, morphine-treated macaques. The current article’s key strength lies in its in vitro studies exploring potential mechanistic underpinnings for the observations made in the SIV-infected, morphine-treated macaques. In the in vitro studies, the synergistic effects of HIV viral proteins and morphine on proliferation, apoptosis, and oxidative stress in human pulmonary microvascular endothelial cells suggest that SIV/HIV viral proteins and morphine may initially interact to increase apoptosis, with endothelial injury subsequently promoting the proliferation of apoptosis-resistant cells. This aberrant healing process is hypothesized to lead to angio-obliteration, potentially through increased production of reactive oxygen species and, in cases of chronic morphine and HIV trans-activator of transcription (Tat) protein exposure, increased phosphorylation and activation of vascular endothelial growth factor-2 receptor. These studies support the concept that HIV-PAH is a “multiple-hit” phenomenon and that opiate-induced endothelial cell injury may be one such insult. Such findings are consistent with previous reports in humans describing increased prevalence of injection drug use in the epidemiology of HIV-PAH (1). Other illicit drugs such as cocaine may also increase risk of HIV-PAH. Dhillon and colleagues previously described an in vitro model in which combined treatment with HIV Tat protein and cocaine increases pulmonary artery endothelial cell permeability and smooth muscle cell proliferation (7). The current findings are thought-provoking and lead to the following questions. First, what role does severity of SIV infection, as measured by CD4 T-cell levels and viral load, play in PAH pathogenesis? Second, it is interesting that all three HIV viral proteins studied had similar proapoptotic and proliferative effects on endothelial cells. Were there any HIV viral proteins such as gag, pol, or rev that did not produce this effect? Finally, the SIV and morphine-treated group developed Pneumocystis infection in 60% of the animals. Given prior findings by Swain and coworkers in which Pneumocystis exposure increased the likelihood of developing pulmonary hypertension in a rodent model, Pneumocystis colonization or infection could have also contributed to the development of disease (8). This work extends previous studies demonstrating that SIV-infected macaques develop pulmonary vascular lesions consistent with HIV-PAH (9, 10). Although it has been previously postulated that HIV viral proteins play a role in the pathogenesis of HIV-PAH, the mechanism by which this happens is not known. HIV transgenic rats that express seven of the nine HIV viral proteins develop increased right ventricular pressures, right ventricular hypertrophy, and pulmonary vascular remodeling consistent with pulmonary hypertension (11). Although direct infection of endothelial cells by HIV has never been demonstrated, it is possible that soluble HIV proteins affect host target cells and lead to HIV-PAH. For example, HIV-negative regulatory factor (Nef) protein has been identified within the endothelial cells of complex plexiform lesions in Simian/human chimeric immunodeficiency virus (SHIV)-nef–infected rhesus macaques and HIV-infected humans (12). Similar to humans with idiopathic pulmonary arterial hypertension, aberrant Golgi trafficking has been reported in SHIV-nef–infected macaques (13). Macaques infected with SIV or SHIV-envelope (env) protein also develop pathologic changes (10) as well as hemodynamic changes consistent with pulmonary hypertension (George, unpublished data). Both Env and Tat have been associated with increased oxidative stress in endothelial cells (14). Several other potential pathways have been described in the literature. In a murine Pneumocystis model of immune restoration, mice that had previously been infected with (and cleared) Pneumocystis developed physiologic and pathologic changes consistent with pulmonary hypertension (8). Autoimmune mechanisms have also been invoked in HIV-PAH, with an increased prevalence of the disease in individuals with specific HLA-DR alleles (15). The effect of immunologic aging and premature cellular senescence in HIV and PAH is also an avenue currently under exploration (16). The current studies by Spikes and colleagues are the first to investigate the potential pathogenic role of opioid drugs in the development of HIV-PAH. This study is a provocative initial look at the potentially synergistic interactions between morphine and the HIV viral proteins Tat, Nef, and Env in the pathogenesis of PAH. Future studies may further explore the impact of other drugs such as methamphetamines and other opioids. Identifying these risk factors as well as understanding the multiple hits in the pathogenesis of the HIV-PAH will help identify high-risk individuals and optimize treatments for this fatal disease.

In this issue of Circulation , Parikh and colleagues1 present an elegant study that lies at the intersection of 2 recent major developments in cardiovascular research—the recognition of the role of microRNAs in normal and abnormal cardiovascular biology, and the introduction of computational systems biology approaches to elucidate the mechanisms of cardiovascular disease. This work provides convincing evidence for the central role of microRNA-21 (miR-21) in the pathogenesis of pulmonary hypertension (PH) generally and pulmonary arterial hypertension (PAH) more specifically, and suggests a potential 2-hit rationale for incomplete penetrance of mutations in BMPR2 in heritable PAH. Article see p 1520 MicroRNAs are small noncoding RNA molecules 17 to 23 nucleotides long that bind to target mRNA molecules and repress their translation by cleavage, sequestration, degradation, or ribosomal inhibition. There are >1000 microRNAs in the genome, regulating ≈30% of gene expression.2 Although they were initially discovered in Caenorhabditis elegans in 1993, their role in mammalian cardiovascular biology has been recognized only in the last decade. Global deletion of functional microRNAs in smooth muscle cells in mice leads to impairment of vascular smooth muscle cell development, differentiation, and contraction, abrogated aortic development, extensive hemorrhage, and embryonic lethality.3 Similar deletion of microRNAs in endothelial cells is compatible with life, but leads to impaired angiogenesis.4 Thus, microRNAs are essential for vascular development and function. Further work, some of which is discussed below, has shown that dysregulation of microRNAs is associated with vascular disease. To establish the role of miR-21 in PH, Parikh and colleagues used an innovative approach integrating network analysis of existing genome-wide mRNA expression data, algorithmic prediction of microRNA targets, previously established concepts of biological pathways involved in PH, and in vitro and in vivo experimentation. First, they established a consolidated interactome of molecular interactions known to occur …

Calcitonin gene-related peptide (CGRP) is believed to play an important role in maintaining low pulmonary vascular resistance (PVR) and may be involved in modulating the pulmonary vascular response to chronic hypoxia. In the present study, an adenoviral vector encoding CGRP (AdRSVCGRP) was used to examine the effects of in vivo gene transfer of CGRP to the lung on increases in PVR, right ventricular mass, and pulmonary vascular remodeling that occurs in chronic hypoxia in the mouse. Following intratracheal administration of AdRSVCGRP or reporter gene mice were exposed to 16 days of chronic hypoxia (FIO 2 0.10). The increase in pulmonary arterial pressure (PAP), PVR, right ventricular mass, and pulmonary vascular remodeling in response to chronic hypoxia was attenuated in animals overexpressing CGRP, whereas systemic arterial pressure was not altered. Following exposure to hypoxia, a subgroup of mice were treated with capsaicin, which did not significantly alter CGRP expression in the mouse lung. These data show that in vivo transfer of the CGRP gene to the lung attenuates the increase in PVR, right ventricular mass, and pulmonary vascular remodeling in chronically hypoxic mice with little effect on the systemic circulation. Moreover, these data suggest that adenoviral gene transfer of CGRP to the lung results in expression of the gene product in non-neural tissue.

Peyronie's disease is an idiopathic, localized connective tissue disorder of the penis which involves the tunica albuginea of the corpus cavernosum and the adjacent areolar space. Peyronie's disease is characterized by local changes in the collagen and elastic fiber composition of the tunica albuginea. The formation of fibrotic plaques alters penile anatomy and can cause different degrees of bending and narrowing, as well as penile pain and erectile dysfunction. Though long recognized as an important clinical entity of the male genitalia, the etiology of this disease has remained poorly understood. Until recently there have been no studies to examine the role nitric oxide (NO) and nitric oxide synthase (NOS) isoforms may play in the onset and progression of Peyronie's disease. NO is a potent biological mediator with diverse physiological and pathophysiological roles. The purpose of this review is to describe each of the NOS isoforms and their potential roles in the pathophysiology of Peyronie's disease, with particular emphasis on the regulation of endothelial and inducible NOS isoforms.

The present study was undertaken to investigate the effects of treatment with the angiotensin-converting enzyme (ACE) inhibitor enalapril in a mouse model of pulmonary hypertension induced by bleomycin. Bleomycin-induced lung injury in mice is mediated by enhanced tumor necrosis factor-α (TNF) expression in the lung, which determines the murine strain sensitivity to bleomycin, and murine strains are sensitive (C57BL/6) or resistant (BALB/c). Bleomycin induced significant pulmonary hypertension in C57BL/6, but not in BALB/c, mice; average pulmonary arterial pressure (PAP) was 26.4 ± 2.5 mmHg ( P < 0.05) vs. 15.2 ± 3 mmHg, respectively. Bleomycin treatment induced activation of nuclear factor (NF)-κB and activator protein (AP)-1 and enhanced collagen and TNF mRNA expression in the lung of C57BL/6 but not in BALB/c mice. Double TNF receptor-deficient mice (in a C57BL/6 background) that do not activate NF-κB or AP-1 in response to bleomycin did not develop bleomycin-induced pulmonary hypertension (PAP 14 ± 3 mmHg). Treatment of C57BL/6 mice with enalapril significantly ( P < 0.05) inhibited the development of pulmonary hypertension after bleomycin exposure. Enalapril treatment inhibited NF-κB and AP-1 activation, the enhanced TNF and collagen mRNA expression, and the deposition of collagen in bleomycin-exposed C57BL/6 mice. These results suggest that ACE inhibitor treatment decreases lung injury and the development of pulmonary hypertension in bleomycin-treated mice.

Pulmonary hypertension (PH) is a chronic pulmonary complication of sickle cell disease (SCD) and a leading cause of death in adults. The exact molecular causes of SCD-associated PH are unknown although pathologic hemolysis related to chronic nitric oxide depletion and oxidative stress are recognized as contributing factors. We have reported that the protein thrombospondin-1 (TSP1) is elevated in the plasma of patients with SCD and by interacting with its receptor CD47 limits vasodilation of distal pulmonary arteries ex vivo . We hypothesized that the TSP1-CD47 interaction may promote PH in the BERK sickle mouse model. To test this hypothesis, we inquired whether TSP1 and CD47 are upregulated in the lungs of BERK mice and patients with SCD-associated PH. We found that lungs of age matched male 13-14 months old BERK sickle mice overexpressed TSP1 and CD47 as compared to both C57BL/6J and Hemi BERK mice and also confirmed our prior findings that sickle mice develop PH as they age. Immunohistochemistry of lung sections from 6 patients with SCD-associated PH revealed increased levels of CD47 as compared to sections from patients without PH or overt lung disease. We interrogated the TSP1-CD47 axis in vivo by generating chimeric animals with a sickle erythropoiesis on a CD47KO background by transplanting BERK sickle bone marrow into C57BL/6J (n=24) and CD47 knockout (CD47KO, n=27) mice. Fully engrafted chimeric mice underwent pulmonary hemodynamic assessment after 6 months. Right ventricular (RV) pressures were lower in Sickle-to-CD47KO chimeras as compared to Sickle-to-C57BL/6J chimeras as shown by the reduced maximum pressure of the RV (22.1 ± 3.6 vs. 28.1 ± 8.8 mmHg, p=0.013) and mean pulmonary artery pressure (15.3 ± 2.6 vs. 18.8 ± 5.7 mmHg, p=0.020). The afterload of the Sickle-to-CD47KO chimeras was lower compared to Sickle-to-C57BL/6J chimeras as shown by the diminished pulmonary vascular resistance (1.2 ± 0.7 vs. 2.4 ± 2.2 Wood units, p=0.024) and RV effective arterial elastance (1.5 ± 0.9 vs. 2.7 ± 2.6 mmHg/µL, p=0.052). The RV diastolic function as measured by the RV dP/dtmin also showed a trend towards improvement as compared to Sickle-to-C57BL/6J chimeras (-1297.0 ± 318.2 vs. -1604.0 ± 668.2 mmHg/s, p=0.059). Finally, the Sickle-to-CD47KO chimeras had a tendency for less RV hypertrophy as evidenced by lower RV free wall weight as compared to Sickle-to-C57BL/6J chimeras (21.45 ± 5.0 vs. 24.63 ± 6.0 mg, p=0.056). Interestingly, plasma levels of soluble TSP1 were reduced in Sickle-to-CD47KO chimeras (61.45 ± 10.11 vs. 76.89 ± 10.84 pg/mL, p=0.012). On myography, aortic segments from Sickle-to-CD47KO chimeras had improved relaxation to the endothelial activator acetylcholine. We hypothesized that in SCD TSP1-CD47 signaling promotes PH, in part, by increasing ROS generation. Treatment with exogenous TSP1 (2.2x10-9 M, a concentration found in the plasma of SCD patients) stimulated increased levels of superoxide and hydrogen peroxide in human pulmonary artery endothelial cells by cytochrome C and Amplex Red assays, respectively. Furthermore, lungs of CD47KO chimeras had decreased oxidative damage as measured by reduced 4-hydroxynonenal and 3-nitrotyrosine staining vs. Sickle-to-C57BL/6J chimeras. Finally, suppression of CD47 with a morpholino (10 mg/kg body weight i.p. weekly) for 8 weeks (n=8 animals) also reduced pulmonary pressures in BERK mice. Our results show that genetic deletion or suppression of CD47 ameliorates SCD-associated PH, which may be due, in part, to decreased ROS levels. Targeting TSP1-CD47 may provide a new molecular approach to the treatment of SCD-associated PH. Disclosures Isenberg: Tioma Therapeutics: Equity Ownership; Radiation Control Technologies, Inc.: Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

Right ventricular (RV) failure in response to pulmonary hypertension (PH) is a severe disease that remains poorly understood. PH-induced pressure overload leads to changes in the RV free wall (RVFW) that eventually results in RV failure. While the development of computational models can benefit our understanding of the onset and progression of PH-induced pressure overload, detailed knowledge of the underlying structural and biomechanical events remains limited. The goal of the present study was to elucidate the structural and biomechanical adaptations of RV myocardium subjected to sustained pressure overload in a rat model. Hemodynamically confirmed severe chronic RV pressure overload was induced in Sprague-Dawley rats via pulmonary artery banding. Extensive tissue-level biaxial mechanical and histomorphological analyses were conducted to assess the remodeling response in the RV free wall. Simultaneous myofiber hypertrophy and longitudinal re-orientation of myo- and collagen fibers were observed, with both fiber types becoming more highly aligned. Transmural myo- and collagen fiber orientations were co-aligned in both the normal and diseased state. The overall tissue stiffness increased, with larger increases in longitudinal vs. circumferential stiffness. The latter was attributed to longitudinal fiber re-orientation, which increased the degree of anisotropy. Increased mechanical coupling between the two axes was attributed to the increased fiber alignment. Interestingly, estimated myofiber stiffness increased while the collagen fiber stiffness remained unchanged.The increased myofiber stiffness was consistent with clinical results showing titin-associated increased sarcomeric stiffening observed in PH patients. These results further our understanding of the underlying adaptive and maladaptive remodeling mechanisms and may lead to improved techniques for prognosis, diagnosis, and treatment for PH.

Objective Prognosis in pulmonary hypertension (PH) is largely determined by RV function. However, uncertainty remains about what metrics of RV function might be most clinically relevant. The purpose of this study was to assess the clinical relevance of metrics of RV functional adaptation to increased afterload. Methods Patients referred for PH underwent right heart catheterisation and RV volumetric assessment within 48 h. A RV maximum pressure (Pmax) was calculated from the RV pressure curve. The adequacy of RV systolic functional adaptation to increased afterload was estimated either by a stroke volume (SV)/end-systolic volume (ESV) ratio, a Pmax/mean pulmonary artery pressure (mPAP) ratio, or by EF (RVEF). Diastolic function of the RV was estimated by a diastolic elastance coefficient β. Survival analysis was via Cox proportional HR, and Kaplan–Meier with the primary outcome of time to death or lung transplant. Results Patients (n=50; age 58±13 yrs) covered a range of mPAP (13–79 mm Hg) with an average RVEF of 39±17% and ESV of 143±89 mL. Average estimates of the ratio of end-systolic ventricular to arterial elastance were 0.79±0.67 (SV/ESV) and 2.3±0.65 (Pmax/mPAP-1). Transplantation-free survival was predicted by right atrial pressure, mPAP, pulmonary vascular resistance, β, SV, ESV, SV/ESV and RVEF, but after controlling for right atrial pressure, mPAP, and SV, SV/ESV was the only independent predictor. Conclusions The adequacy of RV functional adaptation to afterload predicts survival in patients referred for PH. Whether this can simply be evaluated using RV volumetric imaging will require additional confirmation.

Extracorporeal membrane oxygenation (ECMO) is rarely used in patients with severe pulmonary hypertension (PH) as a bridge to lung transplantation. In this study, we assess the blood biocompatibility of the integrated CentriMag-Novalung ECMO system (venoarterial) in an acute model of PH. Severe PH (≥2/3 systemic) was induced in eight sheep through progressive ligation of the main pulmonary artery. System performance, platelet activation, thromboelastography (TEG) parameters, fibrinogen, plasma-free hemoglobin, and total plasma protein were measured at initiation, 3, and 6 hr of support in the ECMO (N = 4) and sham (N = 4) groups. A stable ECMO flow (2.2 ± 0.1 L/min), low transmembrane pressure gradient, and steady blood O2 and CO2 levels were maintained. Platelet activation was low (

Background Left ventricular heart failure ( LVHF ) remains progressive and fatal and is a formidable health problem because ever‐larger numbers of people are diagnosed with this disease. Therapeutics, while relieving symptoms and extending life in some cases, cannot resolve this process and transplant remains the option of last resort for many. Our team has described a widely expressed cell surface receptor ( CD 47) that is activated by its high‐affinity secreted ligand, thrombospondin 1 ( TSP 1), in acute injury and chronic disease; however, a role for activated CD 47 in LVHF has not previously been proposed. Methods and Results In experimental LVHF TSP 1‐ CD 47 signaling is increased concurrent with up‐regulation of cardiac histone deacetylase 3 ( HDAC 3). Mice mutated to lack CD 47 displayed protection from transverse aortic constriction ( TAC )‐driven LVHF with enhanced cardiac function, decreased cellular hypertrophy and fibrosis, decreased maladaptive autophagy, and decreased expression of HDAC 3. In cell culture, treatment of cardiac myocyte CD 47 with a TSP 1‐derived peptide, which binds and activates CD 47, increased HDAC 3 expression and myocyte hypertrophy in a Ca 2+ /calmodulin protein kinase II (Ca MKII )‐dependent manner. Conversely, antibody blocking of CD 47 activation, or pharmacologic inhibition of Ca MKII , suppressed HDAC 3 expression, decreased myocyte hypertrophy, and mitigated established LVHF . Downstream gene suppression of HDAC 3 mimicked the protective effects of CD 47 blockade and decreased hypertrophy in myocytes and mitigated LVHF in animals. Conclusions These data identify a proximate role for the TSP 1‐ CD 47 axis in promoting LVHF by Ca KMII ‐mediated up‐regulation of HDAC 3 and suggest novel therapeutic opportunities.

Attentional Control Theory (ACT) predicts that trait anxiety and situation stress combine to reduce performance efficiency on tasks requiring rapid shifts in attention. Recent evidence has also suggested that working memory capacity (WMC) might moderate this relationship. We controlled for methodological difficulties in the existing literature to investigate the relationships between trait anxiety, situational stress, and WMC on attentional shifting.Seventy undergraduate students participated in the study. Trait anxiety was operationalized using questionnaire scores, situational stress was manipulated through a pressured counting task, and WMC was based on performance on the Automated Operation Span Task (AOSPAN). The shifting task involved a modified version of the Sternberg paradigm as the primary task and an oddball tone-discrimination task as the secondary task. Dependent variables were performance effectiveness (accuracy) and processing efficiency (accuracy divided by response time) on the secondary task.There was no effect of anxiety, stress, or WMC in predicting performance effectiveness; however, a significant three-way interaction on processing efficiency was observed. At higher WMC, anxiety and situational stress were not associated with processing efficiency. Conversely, at lower WMC, higher trait anxiety was associated with poorer efficiency but only for those who reported higher situational stress; for those who reported lower situational stress higher trait anxiety predicted facilitated efficiency.Results are interpreted with respect to ACT and directions for future research are discussed.

Electronic channel sounding plays a vital role in developing wireless communication systems. It is critical for transceivers' equalization and filtering operations. However, cur- rent pure electronic channel sounding techniques are not well- suited for emerging scenarios such as opportunistic spectrum access, channel impulse response (CIR) based wireless positioning, and wireless security applications, which demand rapid, high- resolution, and spectrum agile channel measurements on com- mercial off-the-shelf (COTS) devices. To address these critical issues, this paper proposes EV-Sounding, a novel methodology for visual assisted electronic channel sounding. Based on frequency domain channel sounding, EV-Sounding leverages cameras for visual estimation of sparsity locations to reduce the number of frequency samples, thus speeding up the sounding process. EV-Sounding achieves both high-resolution CIR measurements and spectrum agility. We prototype an EV-Sounding system on COTS devices. Our real-world experimental results and extensive simulations validate EV-Sounding's performance. Index Terms—Channel sounding, wireless measurement. I. INTRODUCTION A. Motivation

Background: Pulmonary hypertension (PH) has a poor outcome overwhelmingly related to right ventricular (RV) failure. Augmentation area index (AI) could capture hemodynamic effects of vessel stiffen...

Introduction: Misclassification of patients with pulmonary hypertension may occur due to reliance on automated pulmonary wedge pressure (PWP) measurements instead of end-expiration measurements. The long-term outcomes of patients who are misclassified are unclear. We performed a retrospective study to characterize misclassified patients and to assess their outcomes. Methods: Review of clinical and hemodynamic data was performed for all patients who underwent right heart catheterization at University of Pittsburgh Medical Center with a mean pulmonary arterial pressure>25mmHg and PWP

The most recent upgrades of the electromagnetic (EM) physics “standard” and “low energy” sub-libraries of the general purpose Geant4 Monte Carlo simulation toolkit are described. These upgrades are relevant to different application domains including high energy physics, medical physics and space science. Validation results are presented and discussed.

International audience; Chronic obstructive lung disease (COPD) and diffuse parenchymal lung diseases (DPLD), including idiopathic pulmonary fibrosis (IPF) and sarcoidosis, are associated with a high incidence of pulmonary hypertension (PH), which is linked with exercise limitation and a worse prognosis. Patients with combined pulmonary fibrosis and emphysema (CPFE) are particularly prone to the development of PH. Echocardiography and right heart catheterization are the principal modalities for the diagnosis of COPD and DPLD. For discrimination between group 1 PH patients with concomitant respiratory abnormalities and group 3 PH patients (PH caused by lung disease), patients should be transferred to a center with expertise in both PH and lung diseases for comprehensive evaluation. The task force encompassing the authors of this article provided criteria for this discrimination and suggested using the following definitions for group 3 patients, as exemplified for COPD, IPF, and CPFE: COPD/IPF/CPFE without PH (mean pulmonary artery pressure [mPAP] = 25 mm Hg); PH-COPD, PH-IPF, and PH-CPFE); COPD/IPF/CPFE with severe PH (mPAP >= 35 mm Hg or mPAP >= 25 mm Hg with low cardiac index [CI

Aims Obesity is a risk factor for diabetes and cardiovascular diseases, with the incidence of these disorders becoming epidemic. Pathogenic responses to obesity have been ascribed to adipose tissue (AT) dysfunction that promotes bioactive mediator secretion from visceral AT and the initiation of pro-inflammatory events that induce oxidative stress and tissue dysfunction. Current understanding supports that suppressing pro-inflammatory and oxidative events promotes improved metabolic and cardiovascular function. In this regard, electrophilic nitro-fatty acids display pleiotropic anti-inflammatory signalling actions. Methods and results It was hypothesized that high-fat diet (HFD)-induced inflammatory and metabolic responses, manifested by loss of glucose tolerance and vascular dysfunction, would be attenuated by systemic administration of nitrooctadecenoic acid (OA-NO2). Male C57BL/6j mice subjected to a HFD for 20 weeks displayed increased adiposity, fasting glucose, and insulin levels, which led to glucose intolerance and pulmonary hypertension, characterized by increased right ventricular (RV) end-systolic pressure (RVESP) and pulmonary vascular resistance (PVR). This was associated with increased lung xanthine oxidoreductase (XO) activity, macrophage infiltration, and enhanced expression of pro-inflammatory cytokines. Left ventricular (LV) end-diastolic pressure remained unaltered, indicating that the HFD produces pulmonary vascular remodelling, rather than LV dysfunction and pulmonary venous hypertension. Administration of OA-NO2 for the final 6.5 weeks of HFD improved glucose tolerance and significantly attenuated HFD-induced RVESP, PVR, RV hypertrophy, lung XO activity, oxidative stress, and pro-inflammatory pulmonary cytokine levels. Conclusions These observations support that the pleiotropic signalling actions of electrophilic fatty acids represent a therapeutic strategy for limiting the complex pathogenic responses instigated by obesity. Fil: Kelley, Eric E.. University of Pittsburgh; Estados Unidos Fil: Baust, Jeff. University of Pittsburgh; Estados Unidos Fil: Bonacci, Gustavo Roberto. University of Pittsburgh; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina Fil: Golin Bisello, Franca. University of Pittsburgh; Estados Unidos Fil: Devlin, Jason E.. University of Pittsburgh; Estados Unidos Fil: Croix, Claudette M. St.. University of Pittsburgh; Estados Unidos Fil: Watkins, Simon C.. University of Pittsburgh; Estados Unidos Fil: Gor, Sonia. University of Pittsburgh; Estados Unidos Fil: Cantu Medellin, Nadiezhda. University of Pittsburgh; Estados Unidos Fil: Weidert, Eric R.. University of Pittsburgh; Estados Unidos Fil: Frisbee,Jefferson C.. University of Virginia; Estados Unidos Fil: Gladwin, Mark T.. University of Pittsburgh; Estados Unidos Fil: Champion, Hunter C.. University of Pittsburgh; Estados Unidos Fil: Freeman, Bruce A.. University of Pittsburgh; Estados Unidos Fil: Khoo, Nicholas K.H.. University of Pittsburgh; Estados Unidos

Prolific sensing devices like mobile phones and video cameras can sense electronic and visual information, respectively, which can help detect and verify a person. Accurate detection and verification of a person's identity from this sensed data is important for enabling applications such as surveillance and e-health. This is challenging due to sensed data heterogeneity, measurement noise, and the prohibitive cost of specialized equipment. This paper proposes R-Focus, a rotating platform with electronic and visual sensors that can detect and verify a person of interest in an area. R-Focus performs electronic and visual data collection and rotates based on the collected data. R-Focus uses the electronic identity information for a person to gather visual identity information for the person, who is verified and tracked. We implement R-Focus on commercial off-the-shelf hardware and software. Our experimental evaluation shows R-Focus's promise for detecting and verifying a person of interest.

Erectile dysfunction in the aging male is caused, in part, by inadequate relaxation of the corpora cavernosal smooth musculature. Calcitonin gene-related peptide (CGRP), a peptide neurotrasmitter localized in the corpora cavernosa, is down-regulated in the aging rat penis. We examined the hypothesis that this reduction in CGRP may contribute to decreased cavernosal smooth muscle relaxation. Therefore, we sought to determine whether adenoviral-mediated gene transfer of prepro-CGRP (AdRSVCGRP) could enhance erectile responses in aged rats. We found a significant decrease in CGRP concentrations and in cAMP and cGMP levels in aged rat cavernosal tissue compared to younger rats. Aged rats also had significantly lower erectile function as determined by cavernosal nerve stimulation compared to younger rats. Five days after transfection with AdRSVCGRP, these aged rats had an approximately threefold increase in cavernosal CGRP levels compared to animals transfected with adenoviruses encoding nuclear-targeted beta-galactosidase (AdRSV beta gal). The AdRSVCGRP-transfected animals also demonstrated an increase in CGRP mRNA and immunohistochemical localization of CGRP in the smooth muscle of the corpora cavernosa. In addition, cAMP levels in the corpora cavernosa were significantly increased, whereas cGMP levels remained unchanged. Adenoviral transduction efficiency of beta-galactosidase reporter gene was measured by chemiluminescence and was observed in cavernosal tissue 5 days after transfection with AdRSV beta gal. More importantly, 5 days after administration of AdRSVCGRP, a significant increase was observed in the erectile response to cavernosal nerve stimulation in the aged rat, similar to the response observed in younger rats. These data suggest that in vivo adenoviral gene transfer of CGRP can physiologically improve erectile function in the aged rat.

Responses to human calcitonin gene-related peptide (hCGRP) and human adrenomedullin (hADM) hAmylin were investigated in isolated mesenteric resistance arteries from the rat. The results of the present investigation show that hCGRP, hAmylin, and hADM induce dose-related vasodilator responses in isolated resistance arteries from the rat mesenteric vascular bed. Vasodilator responses to hCGRP and hAmylin were not altered after denuding the vascular endothelium, after administration of the nitric oxide synthase inhibitor L-NA, or after administration of the soluble guanylate cyclase inhibitor ODQ, suggesting that vasodilator responses to hCGRP and hAmylin are not mediated by the release of nitric oxide from the vascular endothelium and the subsequent increase in cGMP. Vasodilator responses to hCGRP, hAmylin, and hADM were not altered by the vascular selective K+ATP channel antagonist U-37883A. The role of the CGRP1 receptor was investigated and responses to hCGRP and hAmylin, but not hADM, were significantly reduced following administration of hCGRP-(8–37). Moreover, vasodilator responses to hCGRP and hAmylin, but not hADM, were significantly reduced by hAmylin-(8–37), suggesting that an hAmylin-(8–37)-sensitive receptor mediates responses to hCGRP and hAmylin in the rat mesenteric artery. These data suggest that hCGRP and hAmylin have direct vasodilator effects in the isolated mesenteric resistance artery that are mediated by hAmylin-(8–37)- and hCGRP-(8–37)-sensitive receptors.

Background: This study was designed to examine the responses to and the mechanism by which purinergic agonists decrease vascular resistance in the mesenteric vascular bed of the cat. Methods and Results: Injections of ATP, UTP, and 2-MethylThioATP (2-MetSATP) into the mesenteric perfusion circuit elicited dose-dependent decreases in perfusion pressure while injections of P,y-MethylATP (f3,y-MetATP) produced a biphasic response with an initial vasopressor response followed by a vasodilator response. The order of potency of the vasodilator response was 2-MetSATP > ATP > UTP > P,y-MetATP. The vasodilator responses to ATP, UTP, 2-MetSATP, and P,y-MetATP were increased in duration by the cAMP phosphodiesterase inhibitor, rolipram. However, vasodilator responses were not altered by the adminstration of a nitric oxide synthase inhibitor, a cGMP phosphodiesterase inhibitor, or a cyclooxygenase inhibitor. Treatment with PPADS, a P2X,, P2Y, and P2Y4 receptor antagonist, did not alter vasodilator responses to the purinergic agonists; however, the vasopressor component of the response to P,y-MetATP was decreased. Conclusions: These data suggest that ATP, UTP, 2-MetSATP, and P,Y-MetATP dilate the mesentary vascular bed in the cat by a cAMP dependent mechanism, and that nitric oxide or prostaglandin release, cGMP accumulation, or activation of P2X,, P2Y, or P2Y4 receptors play little or no role in mediating vasodilator responses to the purinergic agonists in this regional vascular bed. In addition, these results suggest that the pressor component of the response to P,y-MetATP is mediated by the activation of P2X, receptors.

Separate granite plutons in southeastern Australia can commonly be grouped into suites on the basis of shared similarities in field, petrographic and compositional data. Granites in different plutons of the same suite share common properties or exhibit a sequence of such features. Rocks of the same suite are cogenetic, but the details of their genesis need not be known or agreed on, to group granite units in such a way. These rocks are cogenetic in the sense that they shared a similar petrogenesis and were derived from source materials of essentially the same composition, whereas differences between suites reflect analogous differences in their source rocks. The term suite is lithologic or lithodemic in a stratigraphic sense and is closely analogous to the lithostratigraphic term group. As such, the plutons within a suite need not be of the same age, and age is not a factor in recognising a suite. However, the fact that the petrogenesis of the components of a suite resulted in such similar products means that their ages are likely to be similar. Granite plutons that share many similar features, but which also show distinct differences and which may be assigned to more than one suite, may be grouped into supersuites. The allocation of granites to suites is fundamental to understanding their petrogenesis. Suites vary in the complexity of their compositional variation. Simple suites show variations in element abundances that are highly correlated and the dispersion of composition within such suites is considered to result from varying degrees of fractionation of entrained restite from a melt. Intricate suites vary in composition in more complex ways and their variation is considered to be a consequence of processes such as fractional crystallisation. Any mineralisation is generally associated with intricate suites, and the occurrence of mineralisation and its precise character is generally specific to particular suites.

Controlled release of nitric oxide (NO*) may be useful in the treatment of a variety of vascular disorders. NO* donors of the diazeniumdiolate family with different rates of spontaneous NO* release have been synthesized. In the current study responses to seven diazeniumdiolate NO* donors (DEA/NO*, DETA/NO*, OXI/NO*, PIPERAZI/NO*, PROLI/NO*, SPER/NO*, and SULFI/NO*) were investigated in the hindquarters vascular bed of the cat. Intravenous injections of all NO* donors caused dose-dependent decreases in systemic arterial pressure and the rank order of potency was SNP > DEA/NO* > PIPERAZI/NO* > SPER/NO* > PROLI/NO* > OXI/NO*. Injections of all NO* donors into the hindlimb perfusion circuit caused dose-related decreases in hindquarters perfusion pressure that were similar to the order of potency in decreasing systemic arterial pressure. The rank order of the time required for the response to return to 50% of the maximal decrease in pressure (T(1/2)) and total duration of action of the NO* donors was SPER/NO* > PIPERAZI/NO* > DEA/NO* > OXI/NO* > DETA/NO* > PROLI/NO* > SULFI/NO*. After treatment with the NO* synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (100 mg/kg, i.v.), hindlimb vasodilator responses to the NO* donors were not significantly different, but vasodilator responses to acetylcholine were significantly reduced. After treatment with zaprinast (2 mg/kg, i.v.), a type V cyclic 3',5'-guanosine monophosphate-specific phosphodiesterase inhibitor, the duration of vasodilator responses to the NO* donors, as measured by T(1/2), was increased significantly, whereas the duration of the response to the beta2-adrenergic receptor agonist albuterol was unchanged. These data suggest that diazeniumdiolate NO* donors are endothelium-independent, directly stimulate soluble guanylate cyclase, and decrease vascular resistance by increasing cyclic 3',5'-guanosine monophosphate levels in the hindquarters vascular bed of the cat.

Purpose: Zaprinast, dipyridamole and sildenafil were injected into the corpora cavernosa of cats to determine whether changes in the steady state level of cyclic guanosine monophosphate (cGMP) induced by inhibiting type 5 phosphodiesterase would cause an erectile response.Materials and Methods: Increases in intracavernous pressure, penile length and erectile response duration were determined after intracavernous injection of the type 5 cGMP specific phosphodiesterase inhibitors zaprinast, dipyridamole and sildenafil as well as combined zaprinast and prostaglandin E1 (PGE1), and zaprinast and sodium nitroprusside. Systemic arterial pressure was concurrently assessed in these experiments. All responses to phosphodiesterase inhibitors were compared to a control triple drug combination of 1.65 mg. papaverine, 0.5 μg. PGE1 and 25 μg. phentolamine.Results: Each selective type 5 phosphodiesterase inhibitor caused dose related increases in intracorporeal pressure and penile length. However, none of the compounds ...

appears to require a subduction environment, many Archaean TTG The 2·95 Ga Pilbara high-Mg diorite suite intrudes the central suites show no clear chemical evidence of having interacted with a part of the Archaean granite–greenstone terrain of the Pilbara mantle wedge, and on that basis are more likely to represent partial Craton, Western Australia, and shows many features typical of melts of basaltic lower crust rather than of subducted slab. Highhigh-Mg diorite (sanukitoid) suites from other late Archaean terrains. Mg diorite suites appear to concentrate in the Late Archaean, Such suites form a minor component of Archaean felsic crust. They suggesting that subduction may have become an important process are typically emplaced in lateto post-kinematic settings, sometimes only after >3·0 Ga. in association with felsic alkaline magmatism, and are either unaccompanied by, or post-date, tonalite–trondhjemite–granodiorite (TTG) magmatism, which comprises a much greater proportion of Archaean felsic crust. The TTG series comprises sodic, Sr-rich

Responses to the nonpeptide angiotensin II agonist 5, 7-Dimethyl-2-ethyl-3-[[2'-([butyloxycarbonyl) aminosulfonyl]-5'-(3-methyoxybenzyl)-[1, 1'-biphenyl]-4-yl]methyl]-3H-imidazo[4,5-b]pyridine (L-163,491) were investigated and compared with responses to angiotensin II, angiotensin IV and norepinephrine in the hindquarters vascular bed of the cat under constant-flow conditions. Injections of L-163,491 into the hindquarter perfusion circuit caused dose-related increases in hindquarters perfusion pressure. In relative terms, angiotensin II was more potent than norepinephrine, which was more potent than angiotensin IV and L-163,491 in increasing hindlimb vascular resistance. The slope of the dose-response curve for L-163,491 was flat, while the apparent affinity of the compound for angiotensin AT(1) receptors was slightly greater than angiotensin IV. Responses to L-163,491 were inhibited by the angiotensin AT(1) receptor antagonist DuP 532 (2-propyl-4-pentafluoroethyl-1-[2'-(1H-tetrazol-5-yl)bipheny l-4-yl)me thyl]imidazole-5-carboxylic acid) and were not altered by the angiotensin AT(2) receptor antagonist PD123,319 (S(+)-1-[[4-(Dimethylamino)-3-methylphenyl]methyl]-5-(diphenylacetyl+ ++) -4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid ditribluoroacetate). However, the increase in hindlimb perfusion pressure in response to angiotensin II and angiotensin IV was significantly decreased following injection of L-163,491. These data suggest that the nonpeptide angiotensin analog L-163,491 has partial agonist activity, which is dependent on the stimulation of angiotensin AT(1) receptors in the hindquarters vascular bed of the cat.