Search

In recent years, the improvement of cloud computing and mobile computing techniques has led to the availability of a variety of mobile applications (‘apps’) in the app store. For instance, a garbage truck app that can provide the immediate location of a garbage truck, the location of collection points, and forecasted arrival times of garbage trucks would be useful for mobile users. Since the power consumption of apps on mobile devices if of concern to mobile users, an optimised power-saving mechanism for updating messages, which is based on location information, for a proposed garbage truck fleet management system (GTFMS) is proposed and implemented in this paper. The GTFMS is a three- component system that includes the on-board units on garbage trucks, a fleet management system, and a garbage truck app. In this study, an arrival time forecasting method is designed and implemented in the fleet management system, so that the garbage truck app can retrieve the forecasted arrival time via web services. A message updating event is then triggered that reports the location of garbage truck and the forecasted arrival time. In experiments conducted on case studies, the results showed that the mean accuracy of predicted arrival time by the proposed method is about 81.45 per cent. As for power consumption, the cost of traditional mobile apps is 2,880 times that of the mechanism proposed in this study. Consequently, the GTFMS can provide the precise forecasted arrival time of garbage trucks to mobile users, while consuming less power.

This work is a study of phase transformation in (001)-cut Pb(Fe1/2Nb1/2)1-xTixO3 (x=48%) single crystals by means of dielectric permittivity, domain structure, and in situ x-ray diffraction. A first-order T(TNT)-C(TNT) phase transition was observed at the Curie temperature TC≅518 K upon zero-field heating. T, TNT, and C are tetragonal, tetragonal nanotwin, and cubic phases, respectively. T(TNT) and C(TNT) indicate that minor TNT domains reside in the T and C matrices. Nanotwins, which can cause broad diffraction peak, remain above TC≅518 K and give an average microscopic cubic symmetry in the polarizing microscopy. Colossal dielectric permittivity (>104) was observed above room temperature with strong frequency dispersion. This study suggests that nanotwins can play an important role in relaxor ferroelectric crystals while phase transition takes place. The Fe ion is a potential candidate as a B-site dopant for enhancing dielectric permittivity. [ABSTRACT FROM AUTHOR]

This work is a study of the thermal stability of (001)-cut Pb(Mg1/3Nb2/3)1-xTixO3 (x=0.30) single crystals before and after an electric (E)-field poling by means of dielectric permittivity, hysteresis loop, domain structure, polarization current, and x-ray diffraction. An R(RNT)-R(RNT)/T(TNT)-T(TNT)-C transition sequence was observed upon heating in the unpoled sample. R, RNT, T, TNT, and C are the rhombohedral, rhombohedral nanotwin, tetragonal, tetragonal nanotwin, and cubic phases, respectively. R/T indicates coexistence of the R and T phases. R(RNT) and T(TNT) indicate that the RNT and TNT structures mimic monoclinic phases in the R and T matrices, respectively. After a prior E-field poling, an R(RNT)-T(TNT)-C phase sequence takes place upon heating. The dielectric permittivity and current density evidenced an additional polarization at 355 K, which is associated with the vanishing of the dielectric dispersion, which reappears near 410 K and remains up to the Burns temperature TB=510 K. This study suggests that nanotwins (RNT and TNT) can play an important role in high-strain piezoelectric crystals while phase transition takes place. Under E=38 kV/cm, [001] T domains randomly appeared in the matrix, suggesting that the matrix consists of a glassy matrix and ferroelectric nanoclusters. [ABSTRACT FROM AUTHOR]

The dielectric permittivity, domain structure, and in situ x-ray diffraction techniques are used to study the phase transformation in (001)-cut Pb[([Fe.sub.1/2][Nb.sub.1/2]).sub.1-x][Ti.sub.x[O.sub.3] (x=48%) single crystals. The results provide insight into the significant role of nanotwins in relaxor ferroelectric crystals during phase transition and the Fe ion as potential candidate at a B-site dopant for enhancing dielectric permittivity.

The thermal stability of (001)-cut Pb[([Mg.sub.1/3][Nb.sub.2/3]).sub.1-x][Ti.sub.x][O.sub.3] (x = 0.30) single crystals before and after an electric (E)-field poling is analyzed by using dielectric permittivity, hysteresis loop, domain structure, polarization current and x-ray diffraction. The study has shown that nanotwins have played a vital role in high-strain piezoelectric crystals while phase transition has taken place.

This study aims to develop an approach which integrates the concept of climate-sensitive design with housing community design for the design of better residential buildings fitting into local wind environment. Employing research methods involving field investigation and computational fluid dynamics (CFD) simulation, the ventilation environment of four typical community atrium spaces were examined and compared in this study. Through a combination of CFD simulation and scenario-based design studies, the research results reveal that the typical long-shape building blocks with high-density housing development lead to poor natural ventilation in atrium open spaces. The research also indicates that if the openings of building masses and atrium can match the prevailing wind direction, the ventilation in community atrium open spaces will be significantly improved, but the overall effects are also affected by several design factors such as the distance between buildings, the size and shape of urban blocks, organization of building masses, and whether or not the wind corridor effect is effectively utilized. It is suggested for further research that more detailed issued-based CFD simulations (e.g., various land use controls or user preferences) of the relationship between wind environment and various design strategies should be conducted in the early stage of the entire design process to obtain the improvement in ventilation environments. [ABSTRACT FROM AUTHOR]

To find a sustainable living environment, ancestors looked to the site selection method of traditional Feng Shui from ancient China, evaluating and selecting the place most suitable to live in. The current location, environmental conditions, and building clusters of the Hua Zhai Settlement meet the most fundamental criteria of site selection. Further analysis of the local weather data from the last thirty years highlighted the most significant weather factor impacting Hua Zhai Settlement: wind. This factor is used in Computational Fluid Dynamics (CFD) wind environmental simulation, the results of which show that settlements that adhere to the most basic site selection principles are able to provide residents with a more comfortable living environment, allowing people to live sustainably. This proves that Feng Shui is indeed the accumulation of rules governing our living experiences. [ABSTRACT FROM AUTHOR]

Cell refractive index is a key biophysical parameter, which has been extensively studied. It is correlated with other cell biophysical properties including mechanical, electrical and optical properties, and not only represents the intracellular mass and concentration of a cell, but also provides important insight for various biological models. Measurement techniques developed earlier only measure the effective refractive index of a cell or a cell suspension, providing only limited information on cell refractive index and hence hindering its in-depth analysis and correlation. Recently, the emergence of microfluidic, photonic and imaging technologies has enabled the manipulation of a single cell and the 3D refractive index of a single cell down to sub-micron resolution, providing powerful tools to study cells based on refractive index. In this review, we provide an overview of cell refractive index models and measurement techniques including microfluidic chip-based techniques for the last 50 years, present the applications and significance of cell refractive index in cell biology, hematology, and pathology, and discuss future research trends in the field, including 3D imaging methods, integration with microfluidics and potential applications in new and breakthrough research areas. [ABSTRACT FROM AUTHOR]

The gas solubility of pure oxygen and of pure carbon dioxide as well as of their gaseous mixture are measured in the ternary liquid mixture cyclohexane + cyclohexanone + cyclohexanol at 313.6 K with a high-pressure view-cell technique using the synthetic method. The new experimental data are used to assess the capability of molecular simulation and conductor-like screening model (COSMO)-SAC to predict multicomponent fluid-phase coexistence behavior. These methods are also compared systematically on the basis of experimental binary fluid-phase coexistence data. In that comparison also the Peng-Robinson (PR) equation of state is included as a reference. Molecular simulation and COSMO-SAC yield good results and are found to be far superior to the PR equation of state both in predictive and in adjusted mode. © 2013 American Institute of Chemical Engineers AIChE J, 59: 2236-2250, 2013 [ABSTRACT FROM AUTHOR]

The purpose of the present study is to develop a numerical model for the investigation of water waves propagating over a pair of impermeable submerged obstacles. The mathematic model is formulated by coupling solutions of the Navier–Stokes equations and transport equations for the surface elevation using the volume of fluid method. Based on a staggered computational mesh, an explicit numerical algorithm is employed with a predictor–corrector procedure of pressure and velocity field. The proposed model provides good agreement with other experimental results and validates its good performance. Regarding the spatial harmonic evolutions of various cases, it is noted that the present fluctuating mode of harmonic amplitudes exists upstream and at the gap between obstacles. The results show that the nonlinearity of propagating waves becomes stronger than the initial wave in such areas, and reveals much steeper wave profiles compared to the initial ones. The fluctuating harmonic amplitudes vary with the gap width and form two hydrodynamic cycles. The vortices play an important role in the wave reflection as they form a water column wall to reflect the incoming waves. The reflection ratio depends on the extent of vortex development near the upstream obstacle. The maximum wave reflection occurs in cases with dimensionless gap width S/L equal to 3/8 and 7/8 in this study. [ABSTRACT FROM AUTHOR]

This paper tests the arguments put forth by Morgan (2004) and several others that the CRACK/Project Prevention programme, which offers substance-abusing women a monetary incentive to choose long-term or permanent birth control, is racist and unethical. Findings from a sample of 521 participants in the programme show that the majority of women chose a birth control method that was not permanent, and race did not play a statistically significant role in explaining the birth control choices among the women. Rather, the participant's age and the number of pregnancies she had experienced were more influential. The findings suggest that the prior criticisms of the programme for targeting minority and inherently vulnerable, addicted women for sterilisation are unsupported. Women in this study appeared to follow the larger trends among women of all races and socioeconomic backgrounds in choosing permanent birth control: they were older and had more children. [ABSTRACT FROM AUTHOR]

The article presents a comparative study of three theoretical approaches including molecular simulation, the Peng-Robinson equation of state (PR EOS), and COSMO-SAC with the oxidation of cyclohexane.

Patient treatment in a medical linear accelerator is characterized by many angular and translational movements of the gantry and couch. The direction and orientation of each treatment beam is specified by a set of gantry, turntable, and collimator angles. It is possible that some selected treatment field configurations will result in gantry/couch or gantry/patient collisions that remain undetected during the treatment planning process. In this work, a digital camera has been used to record all the workable gantry/ patient set-up images, and a Windows programming language is used to edit and display these images on a personal computer for the treatment planner to screen the treatment plans. These graphical displays enable the planner to be aware of any potential collision hazards by an actual visualization of each selected gantry/turntable or gantry/patient angle configuration.

Aortic carboxypeptidase-like protein (ACLP) is a member of a diverse group of proteins that contain a domain with similarity to that of the Dictyostelium discoideum protein discoidin I. The discoidin domain has been identified in mammalian milk fat globule membrane proteins, blood coagulation factors, and receptor tyrosine kinases, where it may facilitate cell aggregation, adhesion, or cell-cell recognition. Here we show that ACLP is a secreted protein that associates with the extracellular matrix (ECM). During mouse embryogenesis, ACLP is abundantly expressed in the ECM of collagen-rich tissues, including the vasculature, dermis, and the developing skeleton. We deleted the ACLP gene in mice by homologous recombination. The majority of ACLP(-/-) mice die perinatally due to gastroschisis, a severe disruption of the anterior abdominal wall and herniation of the abdominal organs. ACLP(-/-) mice that survived to adulthood developed nonhealing skin wounds. Following injury by a dermal punch biopsy, ACLP(-/-) mice exhibited deficient wound healing compared with controls. In addition, dermal fibroblasts isolated from ACLP(-/-) 18.5-day-postconception embryos exhibited a reduced proliferative capacity compared with wild-type cells. These results indicate that ACLP is an ECM protein that is essential for embryonic development and dermal wound healing processes.

Chronic hypoxia causes pulmonary hypertension with smooth muscle cell proliferation and matrix deposition in the wall of the pulmonary arterioles. We demonstrate here that hypoxia also induces a pronounced inflammation in the lung before the structural changes of the vessel wall. The proinflammatory action of hypoxia is mediated by the induction of distinct cytokines and chemokines and is independent of tumor necrosis factor-alpha signaling. We have previously proposed a crucial role for heme oxygenase-1 (HO-1) in protecting cardiomyocytes from hypoxic stress, and potent anti-inflammatory properties of HO-1 have been reported in models of tissue injury. We thus established transgenic mice that constitutively express HO-1 in the lung and exposed them to chronic hypoxia. HO-1 transgenic mice were protected from the development of both pulmonary inflammation as well as hypertension and vessel wall hypertrophy induced by hypoxia. Significantly, the hypoxic induction of proinflammatory cytokines and chemokines was suppressed in HO-1 transgenic mice. Our findings suggest an important protective function of enzymatic products of HO-1 activity as inhibitors of hypoxia-induced vasoconstrictive and proinflammatory pathways.

Inactivation of glycogen synthase kinase 3beta (GSK3beta) and the resulting stabilization of free beta-catenin are critical steps in the activation of Wnt target genes. While Akt regulates GSK3alpha/beta in the phosphatidylinositide 3-OH kinase signaling pathway, its role in Wnt signaling is unknown. Here we report that expression of Wnt or Dishevelled (Dvl) increased Akt activity. Activated Akt bound to the Axin-GSK3beta complex in the presence of Dvl, phosphorylated GSK3beta and increased free beta-catenin levels. Furthermore, in Wnt-overexpressing PC12 cells, dominant-negative Akt decreased free beta-catenin and derepressed nerve growth factor-induced differentiation. Therefore, Akt acts in association with Dvl as an important regulator of the Wnt signaling pathway.

The objectives of this study are to 1) assess the effects of major correlates of global subjective well-being on financial satisfaction, and 2) use empirical data to present the consequences of violating basic regression assumptions. Analyzing data from the General Social Surveys, 1972-1996 (Davis & Smith, 1996a) this study found that among Americans age forty-five and above, most of the major correlates of global subjective well-being show similar effects on financial satisfaction. The study's findings confirm a nonlinear effect of income on financial satisfaction. Comparing results from different analytical methods, this study also alerts researchers to the importance of taking into account the level of measurements of study variables, which have tended to be overlooked by previous subjective well-being research.

Aortic preferentially expressed gene (APEG)-1 is a 1.4-kilobase pair (kb) mRNA expressed in vascular smooth muscle cells and is down-regulated by vascular injury. An APEG-1 5'-end cDNA probe identified three additional isoforms. The 9-kb striated preferentially expressed gene (SPEG)alpha and the 11-kb SPEGbeta were found in skeletal muscle and heart. The 4-kb brain preferentially expressed gene was detected in the brain and aorta. We report here cloning of the 11-kb SPEGbeta cDNA. SPEGbeta encodes a 355-kDa protein that contains two serine/threonine kinase domains and is homologous to proteins of the myosin light chain kinase family. At least one kinase domain is active and capable of autophosphorylation. In the genome, all four isoforms share the middle three of the five exons of APEG-1, and they differ from each other by using different 5'- and 3'-ends and alternative splicing. We show that the expression of SPEGalpha and SPEGbeta is developmentally regulated in the striated muscle during C2C12 myoblast to myotube differentiation in vitro and cardiomyocyte maturation in vivo. This developmental regulation suggests that both SPEGalpha and SPEGbeta can serve as sensitive markers for striated muscle differentiation and that they may be important for adult striated muscle function.

The onset of myocardial infarction occurs frequently in the early morning, and it may partly result from circadian variation of fibrinolytic activity. Plasminogen activator inhibitor-1 activity shows a circadian oscillation and may account for the morning onset of myocardial infarction. However, the molecular mechanisms regulating this circadian oscillation remain unknown. Recent evidence indicates that basic helix-loop-helix (bHLH)/PAS domain transcription factors play a crucial role in controlling the biological clock that controls circadian rhythm. We isolated a novel bHLH/PAS protein, cycle-like factor (CLIF) from human umbilical vein endothelial cells. CLIF shares high homology with Drosophila CYCLE, one of the essential transcriptional regulators of circadian rhythm. CLIF is expressed in endothelial cells and neurons in the brain, including the suprachiasmatic nucleus, the center of the circadian clock. In endothelial cells, CLIF forms a heterodimer with CLOCK and up-regulates the PAI-1 gene through E-box sites. Furthermore, Period2 and Cryptochrome1, whose expression show a circadian oscillation in peripheral tissues, inhibit the PAI-1 promoter activation by the CLOCK:CLIF heterodimer. These results suggest that CLIF regulates the circadian oscillation of PAI-1 gene expression in endothelial cells. In addition, the results potentially provide a molecular basis for the morning onset of myocardial infarction.

Heme oxygenase (HO)-1 is a stress response protein that is regulated by oxidative stress. HO-1 catalyzes the generation of biliverdin, carbon monoxide, and iron from heme. Lipopolysaccharide (LPS) and interleukin (IL)-1beta induce HO-1 through the binding of nuclear proteins to AP-1 motifs in enhancer regions upstream from the transcription start site. The DNA binding activity of AP-1 proteins depends on the reduction of cysteines in their DNA-binding domains. We found that agents that disrupt free sulfhydryl groups abolish AP-1 binding activity in nuclear proteins obtained from rat aortic smooth muscle cells and macrophages stimulated with IL-1beta or LPS. Thioredoxin (TRX) may regulate the redox status of nuclear transcription factors in response to oxidative stimuli, thus we determined the role of TRX in the physiologic regulation of HO-1. TRX underwent nuclear translocation in cells stimulated with IL-1beta and LPS. We transfected macrophages with a heterologous promoter construct containing two AP-1 sites from an upstream enhancer region in the HO-1 promoter. Recombinant TRX induced promoter activity to a level analogous to that induced by LPS, and this TRX response was abolished by mutation of the AP-1 sites. An inhibitor of TRX reductase, used to prevent TRX translocation in the reduced state, decreased HO-1 induction by IL-1beta and LPS. These data provide the first evidence that TRX contributes to the induction of HO-1 by inflammatory mediators.

We investigated the role of heme oxygenase (HO)-1 in the development of hypoxia-induced pulmonary hypertension. HO catalyzes the breakdown of heme to the antioxidant bilirubin and the vasodilator carbon monoxide. Hypoxia is a potent but transient inducer of HO-1 in vascular smooth muscle cells in vitro and in the lung in vivo. By using agonists of HO-1, we sustained a high expression of HO-1 in the lungs of rats for 1 week. We report that this in vivo enhancement of HO-1 in the lung prevented the development of hypoxic pulmonary hypertension and inhibited the structural remodeling of the pulmonary vessels. The mechanism(s) underlying this effect may involve a direct vasodilating and antiproliferative action of endogenous carbon monoxide, as well as an indirect effect of carbon monoxide on the production of vasoconstrictors. These results provide evidence that enhancement of endogenous adaptive responses may be used to prevent hypoxia-induced pulmonary hypertension.

We have cloned a cardiovascular-restricted basic helix-loop-helix factor that interacts with arylhydrocarbon receptor nuclear translocator (ARNT) in a yeast two-hybrid screen. Cardiovascular helix-loop-helix factor 1 (CHF1) is distantly related to the hairy family of transcriptional repressors. We analyzed its expression pattern during mouse embryo development. At day 8.5, the expression of CHF1 is first detected in the primitive ventricle of the primordial heart tube and persists throughout gestation. In rat hearts, this expression is down-regulated after birth, concurrent with terminal differentiation of cardiomyocytes. In the developing vasculature, CHF1 first appears in the dorsal aorta at day 9.0, which precedes the reported expression of smooth muscle cell markers, and persists into adulthood. In an in vitro system of smooth muscle cell differentiation, CHF1 mRNA was barely detectable in undifferentiated cells but was induced highly in differentiated smooth muscle cells. To determine whether CHF1 might affect the function of ARNT, we performed transfection studies. Co-transfection of CHF1 inhibited ARNT/EPAS1-dependent transcription by 85%, and this inhibition is dose-dependent. In electrophoretic mobility studies, CHF1 inhibited the binding of the ARNT/EPAS1 heterodimer to its target site. Our data suggest that CHF1 functions as a transcriptional repressor and may play an important role in cardiovascular development.

Using data from the General Social Surveys (1972-1996), this study decomposes the trends in financial satisfaction into intercohort and intracohort patterns to assess the intracohort change and cohort replacement effects on financial satisfaction. The results suggest that a positive intracohort component of financial satisfaction trends, indicating more financial satisfaction with time; and a negative intercohort component, indicating that younger cohorts are less satisfied financially. The multivariate analysis further suggests that the change in financial satisfaction trends is mostly due to a strong intercohort replacement effect. That is, the change in financial satisfaction trends can be largely accounted for by the intercohort replacement effect of younger cohorts' being less satisfied financially.

Proliferation of vascular smooth muscle cells (VSMCs) is a hallmark of arteriosclerosis. Architectural transcription factors of the high mobility group (HMG)-I family have been implicated in the control of cell proliferation and gene expression. We studied the pattern of HMG-I mRNA and protein expression in proliferating VSMCs. HMG-I(Y) and HMGI-C mRNAs were barely detectable by Northern analysis in samples prepared from uninjured rat carotid arteries. In contrast, these mRNAs were induced dramatically in carotid arteries 2 and 5-6 days after balloon injury. By in situ hybridization at 6 days after injury, the induced mRNAs localized to smooth muscle cells of the developing neointima, and immunocytochemical analysis showed that HMG-I(Y) protein was expressed in the nuclei of these cells. To confirm this association between HMG-I protein induction and cell growth, we assessed HMG-I(Y) and HMGI-C mRNA expression in rat aortic smooth muscle cells (RASMCs) in primary culture. The HMG-I mRNAs were barely detectable in quiescent RASMCs but were induced markedly by serum stimulation. This induction of mRNA by serum was time dependent and peaked at 9 h. Western blot analysis confirmed that HMG-I(Y) protein induction also occurred in vitro. To our knowledge, this is the first demonstration of induction of HMG-I protein expression in proliferating RASMCs in vivo and in vitro. This demonstration suggests that the HMG-I proteins may play an important role in smooth muscle cell proliferation., (Copyright 1999 Academic Press.)

In order to apply a set of nine STR loci and the amelogenin locus in forensic testing, we have performed a population study on individuals from the Philippines and Thailand living in Taiwan (273 Philippine and 146 Thai individuals were typed by commercially available kits and an automated sequencer). A total of 73 alleles for all systems for both populations could be observed in these two populations. No new intermediate fragments were found. Allele frequencies showed no deviation from Hardy-Weinberg equilibrium. The mean exclusion power (MEP) ranged from 0.327 (TPOX) to 0.706 (FGA), the discriminating power (DP) ranged from 0.790 (TPOX) to 0.963 (FGA) for Philippinos, MEP ranged from 0.247 (TPOX) to 0.723 (FGA), DP ranged from 0.761 (TPOX) to 0.968 (FGA) for Thais, the combined MEP is > 0.9988 and the combined DP is > 0.9999999993 for both Philippinos and Thais.

Endothelial PAS domain protein 1 (EPAS1) is a basic helix-loop-helix/PAS domain transcription factor that is preferentially expressed in vascular endothelial cells. EPAS1 shares high homology with hypoxia-inducible factor-1alpha (HIF-1alpha) and, like HIF-1alpha, has been shown to bind to the HIF-1-binding site and to activate its downstream genes such as vascular endothelial growth factor (VEGF) and erythropoietin. In this report, we show that EPAS1 increased VEGF gene expression through the HIF-1-binding site. This transactivation was enhanced further by cotransfection of an aryl hydrocarbon receptor nuclear translocator expression plasmid. Deletion analysis of EPAS1 revealed a potent activation domain (amino acids 486-639) essential for EPAS1 to transactivate the VEGF promoter. We confirmed the ability of this domain to activate transcription using a Gal4 fusion protein system. Because a truncated EPAS1 protein lacking the transactivation domain at amino acids 486-639 eliminated induction of the VEGF promoter by wild-type EPAS1, the truncated protein functions as a dominant-negative mutant. Most important, infection of the cells with an adenoviral construct expressing this mutant inhibited the induction of VEGF mRNA under conditions that mimic hypoxia. Our results suggest that EPAS1 is an important regulator of VEGF gene expression. Since VEGF plays a crucial role in angiogenesis, the ability of dominant-negative EPAS1 to inhibit VEGF promoter activity raises the possibility of a novel approach to inhibiting pathological angiogenesis.

Aortic preferentially expressed gene-1 (APEG-1) was originally identified as a 1.4-kilobase (kb) transcript preferentially expressed in differentiated vascular smooth muscle cells (VSMC). Its expression is markedly down-regulated in de-differentiated VSMC, suggesting a role for APEG-1 in VSMC differentiation. We have now determined that APEG-1 is a single-copy gene in the human, rat, and mouse genomes and have mapped human APEG-1 to chromosome 2q34. To study the molecular mechanisms regulating its expression, we characterized the genomic organization and promoter of mouse APEG-1. APEG-1 spans 4.5 kb in the mouse genome and is composed of five exons. Using reporter gene transfection analysis, we found that a 2. 7-kb APEG-1 5'-flanking sequence directed a high level of promoter activity only in VSMC. Its activity was minimal in five other cell types. A repressor region located within an upstream 685-base pair sequence suppressed the activity of this 2.7-kb promoter. Further deletion and mutation analyses identified an E box motif as a positive regulatory element, which was bound by nuclear protein prepared from VSMC. In conjunction with its flanking sequence, this E box motif confers VSMC-specific enhancer activity to a heterologous SV40 promoter. To our knowledge, this is the first demonstration of an E box motif that mediates gene expression restricted to VSMC.

Heme oxygenase (HO) catalyzes the oxidation of heme to generate carbon monoxide (CO) and bilirubin. CO increases cellular levels of cGMP, which regulates vascular tone and smooth muscle development. Bilirubin is a potent antioxidant. Hypoxia increases expression of the inducible HO isoform (HO-1) but not the constitutive isoform (HO-2). To determine whether HO-1 affects cellular adaptation to chronic hypoxia in vivo, we generated HO-1 null (HO-1(-/-)) mice and subjected them to hypoxia (10% oxygen) for five to seven weeks. Hypoxia caused similar increases in right ventricular systolic pressure in wild-type and HO-1(-/-) mice. Although ventricular weight increased in wild-type mice, the increase was greater in HO-1(-/-) mice. Similarly, the right ventricles were more dilated in HO-1(-/-) mice. After seven weeks of hypoxia, only HO-1(-/-) mice developed right ventricular infarcts with organized mural thrombi. No left ventricular infarcts were observed. Lipid peroxidation and oxidative damage occurred in right ventricular cardiomyocytes in HO-1(-/-), but not wild-type, mice. We also detected apoptotic cardiomyocytes surrounding areas of infarcted myocardium by terminal deoxynucleotide transferase-mediated dUTP nick end-labeling (TUNEL) assays. Our data suggest that in the absence of HO-1, cardiomyocytes have a maladaptive response to hypoxia and subsequent pulmonary hypertension. J.Clin. Invest. 103:R23-R29 (1999).

Nitric oxide (NO), a free radical gas whose production is catalyzed by the enzyme NO synthase, participates in the regulation of multiple organ systems. The inducible isoform of NO synthase (iNOS) is transcriptionally up-regulated by inflammatory stimuli; a critical mediator of this process is nuclear factor (NF)-kappaB. Our objective was to determine which regulatory elements other than NF-kappaB binding sites are important for activation of the iNOS promoter/enhancer. We also wanted to identify transcription factors that may be functioning in conjunction with NF-kappaB (subunits p50 and p65) to drive iNOS transcription. Deletion analysis of the iNOS promoter/enhancer revealed that an AT-rich sequence (-61 to -54) downstream of the NF-kappaB site (-85 to -76) in the 5'-flanking sequence was important for iNOS induction by interleukin-1beta and endotoxin in vascular smooth muscle cells. This AT-rich sequence, corresponding to an octamer (Oct) binding site, bound the architectural transcription factor high mobility group (HMG)-I(Y) protein. Electrophoretic mobility shift assays showed that HMG-I(Y) and NF-kappaB subunit p50 bound to the iNOS promoter/enhancer to form a ternary complex. The formation of this complex required HMG-I(Y) binding at the Oct site. The location of an HMG-I(Y) binding site typically overlaps that of a recruited transcription factor. In the iNOS promoter/enhancer, however, HMG-I(Y) formed a complex with p50 while binding downstream of the NF-kappaB site. Furthermore, overexpression of HMG-I(Y) potentiated iNOS promoter/enhancer activity by p50 and p65 in transfection experiments, suggesting that HMG-I(Y) contributes to the transactivation of iNOS by NF-kappaB.