Automated particle recognition for engine soot nanoparticles.

A pre-trained convolution neural network based on residual error functions (ResNet) was applied to the classification of soot and non-soot carbon nanoparticles in TEM images. Two depths of ResNet, one 18 layers deep and the other 50 layers deep, were trained using training-validation sets of increasing size (containing 100, 400 and 1400 images) and were assessed using an independent test set of 200 images. Network training was optimised in terms of mini-batch size, learning rate and training length. In all tests, ResNet18 and ResNet50 had statistically similar performances, though ResNet18 required only 25-35% of the training time of ResNet50. Training using the 100-, 400- and 1400-image training-validation sets led to classification accuracies of 84%, 88% and 95%, respectively. ResNet18 and ResNet50 were also compared for their ability to categorise soot and non-soot nanoparticles via a fivefold cross-validation experiment using the entire set of 800 images of soot and 800 images of non-soot. Cross-validation was repeated 3 times with different training durations. For all cross-validation experiments, classification accuracy exceeded 91%, with no statistical differences between any of the network trainings. The most efficient network was ResNet18 trained for 5 epochs, which reached 91.2% classification after only 84 s of training on 1600 images. Use of ResNet for classification of 1000 images, the amount suggested for reliable characterisation of soot sample, requires <4 s, compared with >30 min for a skilled operator classifying images manually. Use of convolution neural networks for classification of soot and non-soot nanoparticles in TEM images is highly promising, particularly when manually classified data sets have already been established.

Effect of non-insulin-dependent diabetes mellitus on myocardial insulin responsiveness in patients with ischemic heart disease.

BACKGROUND: Patients with non-insulin-dependent diabetes mellitus (NIDDM) exhibit poor clinical outcomes from myocardial ischemia. This may reflect an impairment in their cardiac insulin-response system. METHODS AND RESULTS: We used AV balance and intracoronary infusion techniques to compare the intrinsic cardiac responsiveness to insulin in 26 coronary disease patients with (n=13) and without (n=13) NIDDM. During fasting, NIDDM hearts demonstrated lower fractional extraction of glucose from arterial plasma than controls (1.0+/-0.5% versus 2.1+/-0.5%, P<0.05) despite higher circulating insulin levels (26+/-5 versus 13+/-4 microU. mL, P<0.05). This was compensated for by higher circulating glucose levels, so that net cardiac glucose uptake in the 2 groups was equivalent (5.2+/-1.1 versus 5.3+/-1.1 micromol. min). Intracoronary insulin infusion produced an approximately 3-fold increase in fractional extraction and net uptake of glucose across the heart in both groups (to 3.7+/-0.4% and 18.3+/-3.5 micromol. min in NIDDM and to 5.4+/-0.7% and 17.7+/-4.3 micromol. min in controls) accompanied by an approximately 30% increase in net lactate uptake, suggesting preserved insulin action on both glucose uptake and glucose oxidation in the NIDDM heart. In nondiabetics, insulin consistently increased coronary blood flow, but this effect was absent in NIDDM. CONCLUSIONS: In contrast to their peripheral tissues and coronary vasculature, the myocardium of patients with NIDDM expresses a competent insulin-response system with respect to glucose metabolism. This suggests that insulin resistance is mediated at the level of individual organs and that different mechanisms are involved in muscle and vascular tissue.

Serum complement activation in congestive heart failure.

BACKGROUND: Although activation of the complement system in myocardial infarction and cardiopulmonary bypass has been shown to contribute to myocardial injury, its role in congestive heart failure (CHF) is unknown. The purpose of this study was to determine the presence of terminal complement activation and its relation to clinical outcomes in patients with CHF. METHODS: We measured serum levels of the terminal complement complex C5b-9 in 36 patients with symptomatic heart failure and left ventricular ejection fraction <40%. We compared the serum C5b-9 levels of these patients with CHF with a group of 12 age-matched control patients. Combined clinical outcomes (death, urgent heart transplantation, or hospitalization with worsening heart failure) at 6 months were determined. RESULTS: The serum C5b-9 [median (25th to 75th percentiles)] levels in 36 patients with CHF [101.5 ng/mL (40 to 164)] were significantly (P =.003) higher than in the 12 control patients [36.5 ng/mL (22 to 50)]. Significantly more of the patients with CHF with the highest levels of C5b-9 (highest 50th percentile) had New York Heart Association class IV symptoms (67% vs 33%; P =.04) and adverse clinical outcomes by 6 months (56% vs 17%; P =.02) compared with the patients with CHF with lower levels (lowest 50th percentile). CONCLUSIONS: We have described a significant elevation in circulating C5b-9, the terminal complement complex, in patients with symptomatic heart failure and have observed an association between high levels of C5b-9 and near-term adverse events.

Myocardial blood-flow response during mental stress in patients with coronary artery disease.

Positron emission tomography was used to quantify changes in myocardial blood flow during mental stress in patients with and without coronary artery disease. Blunted augmentation of myocardial blood flow during mental stress was observed in regions without significant epicardial stenosis.

Effect of plasma insulin level on myocardial blood flow and its mechanism of action.

Considerable evidence suggests that coronary endothelium regulates myocardial blood flow and metabolism by elaborating vasoactive substances. The physiologic signals mediating this process are uncertain. To test the hypothesis that the process is influenced by physiologic variation in local insulin concentration, we examined the effect of direct intracoronary insulin infusion on myocardial blood flow and oxidative substrate metabolism in 10 patients with coronary heart disease. Ten men (aged 51 to 68 years) who were fasting received a 60-minute intracoronary infusion of insulin at a rate (10 mU/min) sufficient to raise coronary venous plasma insulin from 12+/-4 to 133+/-17 mU/ml without increasing the systemic insulin level. Local coronary hyperinsulinemia increased coronary sinus blood flow in every subject, from 50+/-4 to 61+/-6 ml/min (p<0.01). Insulin also increased myocardial uptake of glucose (from 6+/-1 to 17+/-6 mmol/min) and lactate (from 8+/-2 to 12+/-5 mmol/min), resulting in approximately 30% increase in total oxidative substrate uptake, but without increasing myocardial oxygen consumption (7.0+/-0.7 vs. 7.1+/-0.8 ml/min). Thus, physiologic elevation in the local plasma insulin concentration increases coronary blood flow in the absence of any increase in myocardial oxygen demand or consumption, suggesting a primary reduction in coronary tone, while simultaneously restraining the oxidation of imported substrates. These observations are consistent with insulin-mediated elaboration of vasoactive and/or paracrine factors within the coronary circulation.

Placement of an intracoronary stent via translumbar puncture.

Translumbar puncture for access to the central arterial tree has been used for aortography and for selective coronary angiography in selected patients where conventional peripheral access is not possible. A single case report of coronary angioplasty through this access technique is available in the literature. We report a case of coronary stent placement using the translumbar approach to access the coronary arteries.

Modulation of circulating cellular adhesion molecules in postmenopausal women with coronary artery disease.

OBJECTIVES: The present study examined the association of estrogen (E2) and the inflammatory response of endothelium in coronary artery disease (CAD) by measuring circulating cellular adhesion molecules (cCAMs) in subjects with atherosclerosis. BACKGROUND: Atherosclerotic plaque demonstrates features similar to inflammation. Endothelial cell activation by inflammatory cytokines induces expression of cellular adhesion molecules (CAMs), thereby perhaps augmenting leukocyte adhesion and recruitment and subsequent development of atherosclerosis. The incidence of CAD is lower in women; this may be due to the cardioprotective effects of E2. METHODS: Consecutive eligible subjects with CAD admitted for cardiac catheterization were studied. The groups evaluated were men, postmenopausal women receiving E2 replacement therapy (ERT), postmenopausal women not receiving ERT and premenopausal women. Control groups included men and women without CAD. Preprocedural blood samples were drawn from all groups. Measurements of cCAMs, E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 were performed by enzyme-linked immunoabsorbant assay. E2 levels were assessed by radioimmunoassay. RESULTS: We observed a statistically significant increase in all cCAMs in men with CAD and postmenopausal women with CAD not receiving ERT compared with postmenopausal women with CAD receiving ERT. Premenopausal women with CAD and postmenopausal women with CAD receiving ERT had a significant increase in VCAM-1 alone compared with the female control group. CONCLUSIONS: A possible mechanism by which E2 exerts one of its cardioprotective effects is by limiting the inflammatory response to injury by modulating the expression of CAMs from the endothelium.

Acute cocaine effects on absolute cerebral blood flow.

Cocaine use has been associated with vasoconstriction and stroke, and several studies have demonstrated that it decreases relative cerebral blood flow (rCBF) in humans. However, rCBF has not been quantitated. We compared 40 mg IV cocaine hydrochloride to placebo effects on absolute rCBF in four cocaine users using 99mTc-HMPAO SPECT with a modified microsphere model for CBF quantitation. Cocaine produced significant decreases in rCBF in all regions studied with a mean decrease of 30% in absolute whole brain blood flow (P = 0.002) which was 3-fold greater than relative blood flow changes.

IL-8 and angiogenesis: evidence that human endothelial cells lack receptors and do not respond to IL-8 in vitro.

It has been established that IL-8 triggers angiogenesis in vivo, but this effect may be mediated either by IL-8-recruited leukocytes or by direct actions of IL-8 upon endothelial cells (EC). We have approached this question by examining interactions of recombinant human IL-8 with cultured large vessel and microvascular human EC. We are unable to detect specific IL-8 binding to cultured human umbilical vein endothelial cells (HUVEC) or leukocyte-like IL-8 receptor mRNA expression by either cultured HUVEC or human dermal microvascular endothelial cells (DMEC). We find no alteration of cytoplasmic calcium concentration ([Ca2+]i) in either cell type in response to IL-8 treatment. Finally, we find no IL-8-induced change in EC proliferative rates in the presence or absence of endothelial cell growth factor. Our data favour an indirect action for IL-8 as an angiogenic factor.

Lymphocyte adhesion-dependent calcium signaling in human endothelial cells.

Vascular endothelial cells (ECs) can undergo dramatic phenotypic and functional alterations in response to humoral and cellular stimuli. These changes promote endothelial participation in the inflammatory response through active recruitment of immune effector cells, increased vascular permeability, and alteration in vascular tone. In an attempt to define early events in lymphocyte-mediated EC signaling, we investigated cytosolic-free calcium (Ca2+) changes in single, Fluo-3-labeled human umbilical vein ECs (HUVECs), using an ACAS interactive laser cytometer. Of all lymphocyte subsets tested, allogeneic CD3-, CD56+ natural killer (NK) cells uniquely elicited oscillatory EC Ca2+ signals in cytokine (interleukin [IL]-1- or tumor necrosis factor [TNF])-treated ECs. The induction of these signals required avid intercellular adhesion, consisted of both Ca2+ mobilization and extracellular influx, and was associated with EC inositol phosphate (IP) generation. Simultaneous recording of NK and EC Ca2+ signals using two-color fluorescence detection revealed that, upon adhesion, NK cells flux prior to EC. Lymphocyte Ca2+ buffering with 1,2-bis-5-methyl-amino-phenoxylethane-N,N,N'-tetra-acetoxymethyl acetate (MAPTAM) demonstrated that lymphocyte fluxes are, in fact, prerequisites for the adhesion-dependent EC signals. mAb studies indicate that the beta 2 integrin-intercellular adhesion molecule (ICAM)-1 adhesion pathway is critically involved. However, ICAM-1 antisense oligonucleotide inhibition of IL-1-mediated ICAM-1 hyperinduction had no effect on EC Ca2+ signaling in lymphocyte-EC conjugates, indicating that additional cytokine-induced EC alteration is required. These experiments combine features of lymphocyte-endothelial interactions, intercellular adhesion, EC cytokine activation and transmembrane signaling. The results implicate the IP/Ca2+ second messenger pathway in EC outside-in signaling induced by cytotoxic lymphocytes, and suggest that these signals may play a role in EC alteration by lymphocyte adhesion.

Effect of injectate temperature and thermistor position on reproducibility of thermodilution cardiac output determinations.

STUDY OBJECTIVE: The purpose of this study was to evaluate the effect of thermistor position with varying injectate temperatures on the reproducibility of thermodilution cardiac output determination. The key hypothesis to be tested was that the positioning of the proximal thermistor at the right atrial port would improve the reproducibility of thermodilution cardiac output determination, independent of injectate temperature. DESIGN: Prospective randomized trial. SETTING: The study was performed in the cardiac catheterization laboratory of the West Haven Veterans Affairs Medical Center. PARTICIPANTS: Twenty consecutive patients undergoing right and left heart catheterizations were enrolled in the study. INTERVENTIONS: Each patient underwent triplicate determination of thermodilution cardiac output measurements under four experimental conditions: (1) ambient or room temperature injectate using an external thermistor in the injectate reservoir; (2) iced injectate using an external thermistor; (3) room temperature injectate using an internal right atrium (RA) thermistor; and (4) iced injectate using an RA thermistor. Reproducibility was assessed by the coefficient of variation (CV) and standard error of the mean percent (SEM%), of the triplicate measurements. MEASUREMENTS AND RESULTS: Using an internal RA thermistor improved the reproducibility of cardiac output determinations independent of injectate temperature. Using room temperature injectate, the CV was 12.8 percent using an external thermistor and 7.9 percent using an internal RA thermistor (p < 0.05). Using iced injectate, the CV was 10.2 percent using an external thermistor and 5.5 percent using an internal RA thermistor (p < 0.05). CONCLUSIONS: Reproducibility of thermodilution cardiac output determinations is improved when injectate temperature is measured internally, at the RA, as opposed to externally in the reservoir. This has clinical implications for determining significant changes in serial cardiac output determinations.

Heterogeneous activation thresholds to cytokines in genetically distinct endothelial cells: evidence for diverse transcriptional responses.

It is well accepted that the induction of endothelial cell (EC) adhesion molecules is a critical component in acute inflammatory responses as well as allogeneic interactions in vascularized allografts and, possibly, atherogenesis. The "inflammatory triad" of interleukin 1 (IL-1), tumor necrosis factor, and lipopolysaccharide are potent stimulators of the EC activation/adhesion molecules intercellular adhesion molecule 1 (ICAM-1), endothelial-leukocyte adhesion molecule 1 (ELAM-1), and vascular cell adhesion molecule 1 (VCAM-1). To address whether there exist differing thresholds to cytokine-mediated EC activation, we utilized a panel of genetically distinct human umbilical vein EC lines, assessing their modulated EC surface expression and transcriptional responses to cytokines, with regard to the cell adhesion molecules (CAMs) ELAM-1, ICAM-1, and VCAM-1. With submaximal concentrations of cytokine, EC ELAM-1 surface expression varied from negligible to marked. This phenotypic response was maintained over numerous passages in culture and was observed in ex vivo organ culture analyses with cytokine-treated umbilical vein sections. Relative patterns of ELAM-1, ICAM-1, and VCAM-1 induction were similar in response to multiple stimuli (IL-1, tumor necrosis factor, and lipopolysaccharide, but not phorbol 12-myristate 13-acetate). Nuclear run-off experiments demonstrated that the "high responder" phenotype is a consequence of enhanced transcriptional activation of the CAM genes in response to IL-1 (1 unit/ml), whereas transcriptional responses in "low responders" are minimal. Despite the known involvement of NF-kappa B in endothelial CAM transcription, gel shift assays failed to demonstrate a correlation between the levels of IL-1-mediated nuclear NF-kappa B expression and CAM induction in high and low responding lines. We postulate that varying EC activation thresholds to cytokines observed here, in vitro, may be a critical determinant in the susceptibility to vasculopathic states.

Initiation of cardiac allograft rejection New developments in cellular and molecular mechanisms.

Although cardiac transplantation has become a viable therapeutic option for end-stage heart disease, long-term success is limited by rejection and medical complications of immunosuppression. Earliest events in rejection revolve around T-lymphocyte recognition of foreign antigen through a panel of specialized cell surface proteins. This is followed by a cascade of signaling events, transcription of multiple genes, and cytokine production. The central importance of these events, collectively known as T-cell activation, to the rejection process is underscored by the clinical utility of cyclosporine, which prevents T-cell activation. The specific site of action of cyclosporine within the T cell has recently been elucidated at the molecular level, revealing important details of T-cell biology. Once the immune response has been initiated, localization of inflammatory cells to the graft requires alteration in the functional state of the vascular endothelium, which can then express adhesive ligands for immune effector cells. Recent evidence suggests that host natural killer lymphocytes may play a role in this process. Understanding the molecular details of these and other early events in rejection will facilitate the development of more specific and effective immunosuppressive therapies.