Mitochondrial transplant after ischemia reperfusion promotes cellular salvage and improves lung function during ex-vivo lung perfusion.

BACKGROUND: In lung transplantation, ischemia-reperfusion injury associated with mitochondrial damage can lead to graft rejection. Intact, exogenous mitochondria provide a unique treatment option to salvage damaged cells within lung tissue. METHODS: We developed a novel method to freeze and store allogeneic mitochondria isolated from porcine heart tissue. Stored mitochondria were injected into a model of induced ischemia-reperfusion injury using porcine ex-vivo lung perfusion. Treatment benefits to immune modulation, antioxidant defense, and cellular salvage were evaluated. These findings were corroborated in human lungs undergoing ex-vivo lung perfusion. Lung tissue homogenate and primary lung endothelial cells were then used to address underlying mechanisms. RESULTS: Following cold ischemia, mitochondrial transplant reduced lung pulmonary vascular resistance and tissue pro-inflammatory signaling and cytokine secretion. Further, exogenous mitochondria reduced reactive oxygen species by-products and promoted glutathione synthesis, thereby salvaging cell viability. These results were confirmed in a human model of ex-vivo lung perfusion wherein transplanted mitochondria decreased tissue oxidative and inflammatory signaling, improving lung function. We demonstrate that transplanted mitochondria induce autophagy and suggest that bolstered autophagy may act upstream of the anti-inflammatory and antioxidant benefits. Importantly, chemical inhibitors of the MEK autophagy pathway blunted the favorable effects of mitochondrial transplant. CONCLUSIONS: These data provide direct evidence that mitochondrial transplant improves cellular health and lung function when administered during ex-vivo lung perfusion and suggest the mechanism of action may be through promotion of cellular autophagy. Data herein contribute new insights into the therapeutic potential of mitochondrial transplant to abate ischemia-reperfusion injury during lung transplant, and thus reduce graft rejection.

Catheter-based intramural delivery of red blood cells in an animal model of atherosclerosis.

This report demonstrates intramural red blood cell (RBC) delivery in an atherosclerotic rabbit aorta model and validates the ability of fluoroscopy and computed tomography to verify RBC deposition. A microinfusion catheter with a 35-gauge needle delivered RBCs mixed with iodinated contrast agent to the aorta wall. Six rabbits were sacrificed after injection to confirm RBC delivery. Iron deposition was examined in four additional rabbits 3-7 weeks after injection. Imaging demonstrated 86% sensitivity and 100% specificity for the detection of RBC deposition (n = 25 injection attempts). Iron deposits were found in all intraplaque injection sites 3-7 weeks after injection.

Non-invasive vascular imaging is associated with cardiovascular risk factors among adolescents with bipolar disorder.

Cardiovascular disease (CVD) is exceedingly prevalent among adults with bipolar disorder (BD), implicating BD adolescents as a high-risk group for CVD. Non-invasive ultrasound measures of vascular structure (via carotid intima media thickness [cIMT]) and function (via flow-mediated dilation [FMD]) predict future CVD, and are associated with traditional CVD risk factors among adolescents without mood disorders. This study examined, for the first time, the association of cIMT and FMD with CVD risk factors among adolescents with BD. The presence of multiple potential confounds among adolescents with BD, including various medications and mood states, informs the need to demonstrate whether cIMT and FMD are associated with CVD risk factors in this population specifically. Participants were 30 adolescents, 13-19 years old, with BD, without CVD. High-resolution ultrasonography was used to evaluate vascular structure (cIMT) and function (FMD). Analyses examined associations of cIMT and FMD with traditional CVD risk factors. cIMT was significantly positively associated with systolic blood pressure and waist circumference. FMD was significantly negatively associated with waist circumference, body mass index, triglycerides, and glucose, and positively associated with high-density lipoprotein. cIMT and FMD are associated with traditional CVD risk factors among adolescents with BD. Irrespective of numerous potential confounds, non-invasive vascular ultrasound approaches may be used as CVD risk proxies among adolescents with BD as they are for other adolescents.

Late stage complicated atheroma in low-grade stenotic carotid disease: MR imaging depiction--prevalence and risk factors.

PURPOSE: To determine if complicated plaque can be found by using magnetic resonance (MR) imaging-depicted intraplaque hemorrhage (IPH), even among symptomatic patients with low-grade (≤50%) carotid stenosis. MATERIALS AND METHODS: The institutional ethics review board approved this retrospective study and waived requirements for written informed consent. Symptomatic patients with bilateral 0%-50% carotid stenosis referred for carotid MR imaging were considered. Risk factors (age, sex, hypertension, diabetes, hyperlipidemia, myocardial infarction, atrial fibrillation, smoking, coronary artery disease, and cerebrovascular disease), medications (antihypertensive drugs, diabetes drugs, statins, and aspirin), and the brain side causing symptoms were recorded. MR-depicted IPH prevalence in the carotid arteries ipsilateral and contralateral to the symptomatic side was compared by using the Fisher exact test. Multivariable regression was used to compare the MR-depicted IPH prevalence, while adjusting for risk factors and medications. RESULTS: A total of 217 patients (434 carotid arteries) were included. MR-depicted IPH was found in 13% (31 of 233) of carotid arteries ipsilateral and 7% (14 of 201) of arteries contralateral to symptoms (P < .05). Male sex (P < .05) and increasing age (P < .05) were associated with MR-depicted IPH after controlling for risk factors and medications. CONCLUSION: Complicated carotid atheroma can be found among symptomatic patients with low-grade (≤50%) stenosis, and this is associated with male sex and increasing age. MR-depicted IPH may be useful to stratify risk for patients with low-grade carotid stenosis.

Moderate carotid artery stenosis: MR imaging-depicted intraplaque hemorrhage predicts risk of cerebrovascular ischemic events in asymptomatic men.

PURPOSE: To investigate the association between magnetic resonance (MR) imaging-depicted intraplaque hemorrhage (IPH) in the carotid artery wall and the risk of future ipsilateral cerebrovascular events in men with asymptomatic moderate carotid stenosis by using a rapid three-dimensional T1-weighted fat-suppressed spoiled gradient-echo sequence. MATERIALS AND METHODS: The institutional ethics review board approved this retrospective chart review and waived the requirement for written informed consent. All patients gave informed verbal consent at follow-up telephone interviews. Ninety-one men (mean age, 74.8 years; range, 47-88 years) who attended a vascular clinic between 2003 and 2006, who had asymptomatic carotid stenosis (50%-70% at Doppler ultrasonography), and who had undergone MR imaging for IPH detection were retrospectively identified. Seventy-five men with 98 eligible carotid arteries were included in the study. Patients were followed for a minimum of 1 year (mean follow-up, 24.92 months; range, 12-43 months). Kaplan-Meier survival and univariate Cox regression analyses were conducted to compare future ipsilateral cerebrovascular event rates between carotid arteries with and those without MR-depicted IPH. RESULTS: Of the 98 carotid arteries included, 36 (36.7%) had MR-depicted IPH. Six cerebrovascular events (two strokes and four transient ischemic attacks) occurred in the carotid arteries with IPH, as compared with no clinical events in the carotid arteries without IPH. Univariate Cox regression analysis confirmed that MR-depicted IPH was associated with an increased risk of cerebrovascular events (hazard ratio, 3.59; 95% confidence interval: 2.48, 4.71; P < .001). MR-depicted IPH negatively predicted outcomes (negative predictive value = 100%). CONCLUSION: In this cohort with asymptomatic moderate carotid stenosis, MR-depicted IPH was associated with future ipsilateral cerebrovascular events. Conversely, patients without MR-depicted IPH remained asymptomatic during follow-up. The absence of IPH at MR imaging, therefore, may be a reassuring marker of plaque stability and of a lower risk of thromboembolism.

Potent nonpeptide antagonists of the bradykinin B1 receptor: structure-activity relationship studies with novel diaminochroman carboxamides.

The bradykinin B1 receptor is induced following tissue injury and/or inflammation. Antagonists of this receptor have been studied as promising candidates for treatment of chronic pain. We have identified aryl sulfonamides containing a chiral chroman diamine moiety that are potent antagonists of the human B1 receptor. Our previously communicated lead, compound 2, served as a proof-of-concept molecule, but suffered from poor pharmacokinetic properties. With guidance from metabolic profiling, we performed structure-activity relationship studies and have identified potent analogs of 2. Variation of the sulfonamide moiety revealed a preference for 3- and 3,4-disubstituted aryl sulfonamides, while bulky secondary and tertiary amines were preferred at the benzylic amine position for potency at the B1 receptor. Modifying the beta-amino acid core of the molecule lead to the discovery of highly potent compounds with improved in vitro pharmacokinetic properties. The most potent analog at the human receptor, compound 38, was also active in a rabbit B1 receptor cellular assay. Furthermore, compound 38 displayed in vivo activity in two rabbit models, a pharmacodynamic model with a blood pressure readout and an efficacy model of inflammatory pain.

Identification of a nonpeptidic and conformationally restricted bradykinin B1 receptor antagonist with anti-inflammatory activity.

We report the discovery of chroman 28, a potent and selective antagonist of human, nonhuman primate, rat, and rabbit bradykinin B1 receptors (0.4-17 nM). At 90 mg/kg s.c., 28 decreased plasma extravasation in two rodent models of inflammation. A novel method to calculate entropy is introduced and ascribed approximately 30% of the gained affinity between "flexible" 4 (Ki = 132 nM) and "rigid" 28 (Ki = 0.77 nM) to decreased conformational entropy.

A new class of bradykinin 1 receptor antagonists containing the piperidine acetic acid tetralin core.

The bradykinin 1 (B1) receptor is upregulated during times of inflammation and is important for maintaining inflamed and chronic pain states. Blocking this receptor has been shown to reverse and/or ameliorate pain and inflammation in animal models. In this report, we describe a new class of B1 receptor antagonists that contain the piperidine acetic acid tetralin core. A structure-activity relationship for these analogs is described in this paper. The most potent compounds from this class have IC50s<20 nM in a B1 receptor functional assay. One of these compounds, 13g, shows modest oral bioavailability in rats.