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Sample records for hif driver hilda

  1. Muuseum korraldab noorteõhtu Hilda Gleserist

    Index Scriptorium Estoniae

    2006-01-01

    16. märtsil toimub Eesti Teatri- ja Muusikamuuseumis noorteõhtu näitleja, lavastaja ja teatripedagoog Hilda Gleserist. Etendus kannab nime "Hilda Nukitsamees" ja selle korraldab Assauwe Teater eesotsas lavastajatudengi Jaanika Juhansoniga

  2. Muuseum korraldab noorteõhtu Hilda Gleserist

    Index Scriptorium Estoniae

    2006-01-01

    16. märtsil toimub Eesti Teatri- ja Muusikamuuseumis noorteõhtu näitleja, lavastaja ja teatripedagoog Hilda Gleserist. Etendus kannab nime "Hilda Nukitsamees" ja selle korraldab Assauwe Teater eesotsas lavastajatudengi Jaanika Juhansoniga

  3. Hilda Cid: physicist, crystallographer, structural biologist

    Science.gov (United States)

    Gutiérrez, Gonzalo

    2016-05-01

    A brief review of Dr. Hilda Cid, Chilean scientist who excelled in the field of crystallography, is presented. Dr. Cid was born in 1933 in Talcahuano, where she completed her primary and secondary education, graduating as a Teacher of mathematics and physics from the University of Chile in 1958. She continued graduate studies at MIT, USA, where obtained the PhD in 1964. Upon her return to Chile, she joined the Austral University in Valdivia, where she dedicated to characterize, by X-ray, biological material. During that time, she was actively involved in the social changes taking place in the university and in the country. Thus, following the bloody coup of 1973, she and her family were persecuted and forced to go into exile to Sweden. In the decade of the 1980 she returns to Chile, and settles at University of Concepción, where she teaches, conducts research and organizes a number of latino-american courses and symposiums. Knowing that Hilda Cid formed a real school, where her students -now distinguished professionals and researchers- cast their teachings through Chile and other countries, we ended the article with a reflection about why Hilda Cid, despite its obvious scientific achievements, remains ignored by most of the scientific establishment in Chile.

  4. Hilda family contribution to the Jupiter Family Comets (JFC)

    Science.gov (United States)

    di Sisto, R. P.; Brunini, A.; Dirani, L. D.; Orellana, R. B.

    The distinction between asteroids and comets, is based in their observational qualities rather than in their orbital characteristics. Comets show activity when they reach the interior Solar System. Asteroids from the outer Belt, may have compound of the same volatile material, dust and organic molecules than comets, but they didn't approach enough to the Sun, to show activity. From the compositional point of view, it is a bit arbitrary or at least difficult to distinguish between asteroids from the external main belt and comets. The firsts may be very similar to comets, or at least be objects of intermediate characteristics. The Hildas asteroids, in 3:2 mean motion with Jupiter, have great quantity of volatiles. The main source of Jupiter Family Comets (JFC) is the transneptunian region, but less than 10 % of them comes from the Troyan swarms. In this article we study the Hilda family as another probable source of JFC. We perform numerical simulations and follow the dynamical evolution of Hildas escaped from the resonance. From the 391 particles that escaped from the resonance, 386 (98.7 %) live at least for 1000 years as JFC. The mean life time in this zone is 1.4 × 106 years. The escape rate of an Hilda asteroid, with diameter D greater than 1 km. Is 1.1 × 10-4, so, there is 65 Hildas with D > 1km. (the typical size of a comet) in the JFC region. Therefore, the contribution of Hilda asteroids to the population of comets is important.

  5. Focusing magnets for HIF based on racetracks

    Energy Technology Data Exchange (ETDEWEB)

    Martovetsky, N N; Manahan, R R

    2000-09-11

    Heavy Ion Fusion (HIF) is considered a promising path to a practical fusion reactor. A driver for a HIF reactor will require a large number of quadrupole arrays to focus heavy ion beams. A conceptual design, and trade off studies of the quadrupole array based on racetracks are presented. A comparison with a conventional shell magnet is given and advantages and disadvantages are discussed. A more detailed design of a single quadrupole for the High Current experiment (HCX) is presented and discussed.

  6. Workshop on transport for a common ion driver

    Energy Technology Data Exchange (ETDEWEB)

    Olson, C.C. [Sandia National Labs., Albuquerque, NM (United States); Lee, E. [Lawrence Berkeley Lab., CA (United States); Langdon, B. [Lawrence Livermore National Lab., CA (United States)

    1994-12-31

    This report contains research in the following areas related to beam transport for a common ion driver: multi-gap acceleration; neutralization with electrons; gas neutralization; self-pinched transport; HIF and LIF transport, and relevance to common ion driver; LIF and HIF reactor concepts and relevance to common ion driver; atomic physics for common ion driver; code capabilities and needed improvement.

  7. Workshop on transport for a common ion driver

    Energy Technology Data Exchange (ETDEWEB)

    Olson, C.C. [Sandia National Labs., Albuquerque, NM (United States); Lee, E. [Lawrence Berkeley Lab., CA (United States); Langdon, B. [Lawrence Livermore National Lab., CA (United States)

    1994-12-31

    This report contains research in the following areas related to beam transport for a common ion driver: multi-gap acceleration; neutralization with electrons; gas neutralization; self-pinched transport; HIF and LIF transport, and relevance to common ion driver; LIF and HIF reactor concepts and relevance to common ion driver; atomic physics for common ion driver; code capabilities and needed improvement.

  8. Poesia no espelho: Yona Wollach e Hilda Hilst

    Directory of Open Access Journals (Sweden)

    Lucius de Mello

    2013-03-01

    Full Text Available Elas estilhaçaram a própria identidade para atingir o estado mais puro e subjetivo da Poesia. O diálogo entre os poemas subversivos da israelense Yona Wollach e da brasileira Hilda Hilst. Desvendar os mundos poéticos e narrativos dessas duas poetisas é um desafio de fôlego. Este artigo não tem a pretensão de fazer uma análise profunda. Pretendemos trazer à luz apenas algumas semelhanças e diferenças entre Wollach e Hilst, duas mulheres sempre à deriva, sempre “estrangeiras” mesmo dentro da própria casa.

  9. Tööst Venemaa arhiivides aastail 1989-1994 : Hilda Sabbo / Peep Varju

    Index Scriptorium Estoniae

    Varju, Peep

    2007-01-01

    Ida-eestlaste Seltsi represseeritute komisjoni juhina alustas Hilda Sabbo andmete kogumist Venemaal hukatud eestlaste kohta. Pr. Sabbo tööst Moskva arhiivides. 1994. aasta alguseks oli Hilda Sabbol kogutud uskumatult rikkalik materjal. Ilmus 7-köiteline sari "Võimatu vaikida" (1996-2005). Kõige märkimisväärsemana võib esile tuua juuniküüditamise täpse kajastamise algallikate järgi

  10. 0.7-2.5 μm Spectra of Hilda Asteroids

    Science.gov (United States)

    Wong, Ian; Brown, Michael E.; Emery, Joshua P.

    2017-09-01

    The Hilda asteroids are primitive bodies in resonance with Jupiter whose origin and physical properties are not well understood. Current models posit that these asteroids formed in the outer solar system and were scattered along with the Jupiter Trojans into their present-day positions during a chaotic episode of dynamical restructuring. In order to explore the surface composition of these enigmatic objects in comparison with an analogous study of Trojans, we present new near-infrared spectra (0.7-2.5 μm) of 25 Hilda asteroids. No discernible absorption features are apparent in the data. Synthesizing the bimodalities in optical color and infrared reflectivity reported in previous studies, we classify 26 of the 28 Hildas in our spectral sample into the so-called less-red and red sub-populations and find that the two sub-populations have distinct average spectral shapes. Combining our results with visible spectra, we find that Trojans and Hildas possess similar overall spectral shapes, suggesting that the two minor body populations share a common progenitor population. A more detailed examination reveals that while the red Trojans and Hildas have nearly identical spectra, less-red Hildas are systematically bluer in the visible and redder in the near-infrared than less-red Trojans, indicating a putative broad, shallow absorption feature between 0.5 and 1.0 μm. We argue that the less-red and red objects found in both Hildas and Trojans represent two distinct surface chemistries and attribute the small discrepancy between less-red Hildas and Trojans to the difference in surface temperatures between the two regions.

  11. Tööst Venemaa arhiivides aastail 1989-1994 : Hilda Sabbo / Peep Varju

    Index Scriptorium Estoniae

    Varju, Peep

    2007-01-01

    Ida-eestlaste Seltsi represseeritute komisjoni juhina alustas Hilda Sabbo andmete kogumist Venemaal hukatud eestlaste kohta. Pr. Sabbo tööst Moskva arhiivides. 1994. aasta alguseks oli Hilda Sabbol kogutud uskumatult rikkalik materjal. Ilmus 7-köiteline sari "Võimatu vaikida" (1996-2005). Kõige märkimisväärsemana võib esile tuua juuniküüditamise täpse kajastamise algallikate järgi

  12. The Color–Magnitude Distribution of Hilda Asteroids: Comparison with Jupiter Trojans

    Science.gov (United States)

    Wong, Ian; Brown, Michael E.

    2017-02-01

    Current models of solar system evolution posit that the asteroid populations in resonance with Jupiter are comprised of objects scattered inward from the outer solar system during a period of dynamical instability. In this paper, we present a new analysis of the absolute magnitude and optical color distribution of Hilda asteroids, which lie in 3:2 mean-motion resonance with Jupiter, with the goal of comparing the bulk properties with previously published results from an analogous study of Jupiter Trojans. We report an updated power-law fit of the Hilda magnitude distribution through H = 14. Using photometric data listed in the Sloan Moving Object Catalog, we confirm the previously reported strong bimodality in visible spectral slope distribution, indicative of two subpopulations with differing surface compositions. When considering collisional families separately, we find that collisional fragments follow a unimodal color distribution with spectral slope values consistent with the bluer of the two subpopulations. The color distributions of Hildas and Trojans are comparable and consistent with a scenario in which the color bimodality in both populations developed prior to emplacement into their present-day locations. We propose that the shallower magnitude distribution of the Hildas is a result of an initially much larger Hilda population, which was subsequently depleted as smaller bodies were preferentially ejected from the narrow 3:2 resonance via collisions. Altogether, these observations provide a strong case supporting a common origin for Hildas and Trojans as predicted by current dynamical instability theories of solar system evolution.

  13. A Comparison of Hildas and Jupiter Trojans Using Photometry, Spectroscopy, and Size Distributions

    Science.gov (United States)

    Wong, Ian; Brown, Michael E.

    2016-10-01

    The current paradigm of Solar System evolution describes a scenario in which Jupiter and Saturn crossed their mutual 2:1 mean-motion resonance, leading to a period of dynamical instability and significant restructuring of the orbital architecture throughout the middle and outer Solar System. Simulations have shown that the initial minor body populations in resonance with Jupiter (Hildas and Jupiter Trojans) were first emptied during this chaotic episode, and then replaced primarily with objects scattered inward from the trans-Neptunian region. The major implication of these models is that Kuiper Belt objects, Trojans, and Hildas are expected to share a common progenitor population in the outer Solar System. By comparing the properties of Hildas and Trojans, we can evaluate their similarities and/or differences and thereby empirically test current dynamical instability models.Our present understanding of Hildas and Trojans reveals many notable similarities. Beyond sharing the general characteristics of reddish colors and very low albedos, both minor body populations have been shown to display a color bimodality. Building on previously published works, we have derived spectral slopes from the Sloan Moving Object Catalog for both Hildas and Trojans, which reveal a robust bifurcation in the optical color distribution over a wide range of sizes and indicate the presence of two classes of objects within the Hildas and Trojans, referred to as the less-red and red sub-populations. We present the first direct comparison between the Hilda and Trojan magnitude distributions, as well as the individual less-red and red population magnitude distributions; we discuss these results in the context of collisional processes and surface properties. We have also obtained new near-infrared spectra of Hildas from the Infrared Telescope Facility and Keck Observatory, covering the wavelength range 0.8–4.0 microns, which supplement previously-obtained spectra for Trojans in the same

  14. The sublime and the grotesque: obscenity in Hilda Hilst

    Directory of Open Access Journals (Sweden)

    Jo A-mi

    2016-08-01

    Full Text Available The present work aims to discuss the dialogical relationship between sublime and grotesque in the work A obscena Senhora D by Hilda Hilst, based on its status of contemporary fiction permeated by reconditioning of the language in its performance in the non-linear time. In this sense, it was used studies of the sublime, as a historical and philosophical and literary concept in the works of Longinus, Edmund Burke, Immanuel Kant, Victor Hugo and François Lyotard; issues relating to discussions on the grotesque and its dialogue with the sublime ways, the works of Mikhail Bakhtin and Michel Maffesoli had great importance - the poetic and obscene relationship of the work discussed in convergent concepts such as obscenity, eroticism, sacred and profane. From these analyzes, it was concluded that in A obscena senhora D the sublime and the grotesque show in a multifaceted character, contradictory, interlocutory, and therefore not exclusionary: solidifying the hybrid linguistic-imagistic flow of the hilstiana literature.

  15. Drift en koers : de levens van Hilda Verwey-Jonker (1908-2004)

    NARCIS (Netherlands)

    Steen, Margaretha Wilhelmina Francisca van der

    2011-01-01

    Drift en koers (Passion and control) is the first scientific biography on the Dutch socialist, sociologist and feminist Hilda Verwey-Jonker (1908-2004). She is best known for the introduction of the word allochtonen (foreigner/alien) in the Dutch discourse and has very been influential in improving

  16. Zschokkella hildae Auerbach, 1910: phylogenetic position, morphology, and location in cultured Atlantic cod.

    Science.gov (United States)

    Holzer, Astrid Sibylle; Wootten, Rod; Sommerville, Christina

    2010-06-01

    The myxozoan Zschokkella hildae Auerbach, 1910, was detected with a prevalence of 100% in cultured Atlantic cod, Gadus morhua L. aged 1+ from a culture facility on the west coast of Scotland. Sporogonic stages of Z. hildae, plasmodia producing 2-5 mature spores, were located predominantly in the collecting ducts and ureters of the kidney, and spores were present in the urine collected from the bladder. Less frequently, plasmodia were detected in the interstitial tissue of the kidney. The parasite prevalence in cultured fish was considerably higher than reported in wild fish but no obvious signs of pathology were detected. SSU rDNA sequencing and phylogenetic analysis showed that Z. hildae is closely related to a Sinuolinea sp. from the urinary system of turbot, Psetta maxima (L.), and that these two species, together with other myxozoans from the urinary system of marine fish cluster together in a sub-clade of the recognised marine clade of myxozoans. This sub-clade is characterised by a specific linear expansion segment, helix E23_15 in the secondary structure of variable region V4 of the SSU rDNA. Z. hildae and Sinuolinea sp. show extraordinary large linear expansion segment in both V4 and V7 and an important number of complementary base changes in the conservative regions of the SSU rDNA, indicating considerable evolutionary changes in the SSU rDNA of these species when compared with other myxozoans from the marine environment.

  17. Did the Hilda collisional family form during the late heavy bombardment?

    CERN Document Server

    Brož, M; Morbidelli, A; Nesvorný, D; Bottke, W F; 10.1111/j.1365-2966.2011.18587.x

    2011-01-01

    We model the long-term evolution of the Hilda collisional family located in the 3/2 mean-motion resonance with Jupiter. Its eccentricity distribution evolves mostly due to the Yarkovsky/YORP effect and assuming that: (i) impact disruption was isotropic, and (ii) albedo distribution of small asteroids is the same as for large ones, we can estimate the age of the Hilda family to be $4_{-1}^{+0}\\,{\\rm Gyr}$. We also calculate collisional activity in the J3/2 region. Our results indicate that current collisional rates are very low for a 200\\,km parent body such that the number of expected events over Gyrs is much smaller than one. The large age and the low probability of the collisional disruption lead us to the conclusion that the Hilda family might have been created during the Late Heavy Bombardment when the collisions were much more frequent. The Hilda family may thus serve as a test of orbital behavior of planets during the LHB. We tested the influence of the giant-planet migration on the distribution of the ...

  18. Drift en koers : de levens van Hilda Verwey-Jonker (1908-2004)

    NARCIS (Netherlands)

    Steen, Margaretha Wilhelmina Francisca van der

    2011-01-01

    Drift en koers (Passion and control) is the first scientific biography on the Dutch socialist, sociologist and feminist Hilda Verwey-Jonker (1908-2004). She is best known for the introduction of the word allochtonen (foreigner/alien) in the Dutch discourse and has very been influential in improving

  19. Observations of Candidate Binary Asteroids in the Jovian Trojan and Hilda Populations

    Science.gov (United States)

    Sonnett, Sarah M.; Mainzer, Amy K.; Grav, Tommy; Masiero, Joseph R.; Bauer, James M.; Kramer, Emily A.

    2016-10-01

    Jovian Trojans (hereafter, Trojans) are asteroids in stable orbits at Jupiter's L4 and L5 Lagrange points, and Hilda asteroids are inwards of the Trojans in 3:2 mean-motion resonance with Jupiter. Due to their special dynamical properties, observationally constraining the formation location and dynamical histories of Trojans and HIldas offers key input for giant planet migration models. A fundamental parameter in assessing formation location is the bulk density - with low-density objects associated with an ice-rich formation environment in the outer solar system and high-density objects typically linked to the warmer inner solar system. Bulk density can only be directly measured during a close fly-by or by determining the mutual orbits of binary asteroid systems. With the aim of determining densities for a statistically significant sample of Trojans and Hildas, we are undertaking an observational campaign to confirm and characterize candidate binary asteroids published in Sonnett et al. (2015). These objects were flagged as binary candidates because their large NEOWISE brightness variations imply shapes so elongated that they are not likely explained by a singular equilibrium rubble pile and instead may be two elongated, gravitationally bound asteroids. We are obtaining densely sampled rotational light curves of these possible binaries to search for light curve features diagnostic of binarity and to determine the orbital properties of any confirmed binary systems by modeling the light curve. We present preliminary results from the follow-up campaign of these candidates, including estimates on the densities of objects that appear to be in binary systems and the binary fraction for Trojans and Hildas.

  20. Inhibition of HIF-2.alpha. heterodimerization with HIF1.beta. (ARNT)

    Energy Technology Data Exchange (ETDEWEB)

    Bruick, Richard K.; Caldwell, Charles G.; Frantz, Doug E.; Gardner, Kevin H.; MacMillan, John B.; Scheuermann, Thomas H.; Tambar, Uttam K.

    2017-09-12

    Provided is a method of inhibiting heterodimerization of HIF-2.alpha. to HIF1.beta. (ARNT) comprising binding certain small molecules to the HIF-2.alpha. PAS-B domain cavity but not to HIF1.alpha. and inhibiting HIF-2.alpha. heterodimerization to HIF1.beta. (ARNT) but not inhibiting HIF1.alpha. heterodimerization to HIF1.beta. (ARNT). Those certain small molecules are also referenced synonymously as HIF2-HDI and HIF2.alpha. heterodimerization inhibitors and also simply as certain small molecules.

  1. Non-electron transfer chain mitochondrial defects differently regulate HIF-1α degradation and transcription

    Directory of Open Access Journals (Sweden)

    Antonina N. Shvetsova

    2017-08-01

    Full Text Available Mitochondria are the main consumers of molecular O2 in a cell as well as an abundant source of reactive oxygen species (ROS. Both, molecular oxygen and ROS are powerful regulators of the hypoxia-inducible factor-1α-subunit (HIF-α. While a number of mechanisms in the oxygen-dependent HIF-α regulation are quite well known, the view with respect to mitochondria is less clear. Several approaches using pharmacological or genetic tools targeting the mitochondrial electron transport chain (ETC indicated that ROS, mainly formed at the Rieske cluster of complex III of the ETC, are drivers of HIF-1α activation. However, studies investigating non-ETC located mitochondrial defects and their effects on HIF-1α regulation are scarce, if at all existing. Thus, in the present study we examined three cell lines with non-ETC mitochondrial defects and focused on HIF-1α degradation and transcription, target gene expression, as well as ROS levels. We found that cells lacking the key enzyme 2-enoyl thioester reductase/mitochondrial enoyl-CoA reductase (MECR, and cells lacking manganese superoxide dismutase (MnSOD showed a reduced induction of HIF-1α under long-term (20 h hypoxia. By contrast, cells lacking the mitochondrial DNA depletion syndrome channel protein Mpv17 displayed enhanced levels of HIF-1α already under normoxic conditions. Further, we show that ROS do not exert a uniform pattern when mediating their effects on HIF-1α, although all mitochondrial defects in the used cell types increased ROS formation. Moreover, all defects caused a different HIF-1α regulation via promoting HIF-1α degradation as well as via changes in HIF-1α transcription. Thereby, MECR- and MnSOD-deficient cells showed a reduction in HIF-1α mRNA levels whereas the Mpv17 lacking cells displayed enhanced HIF-1α mRNA levels under normoxia and hypoxia. Altogether, our study shows for the first time that mitochondrial defects which are not related to the ETC and Krebs cycle

  2. Shapes and binary fractions of Jovian Trojans and Hildas through NEOWISE

    Science.gov (United States)

    Sonnett, S.; Mainzer, A.; Grav, T.; Bauer, J.; Masiero, J.; Stevenson, R.; Nugent, C.

    2014-07-01

    Jovian Trojans (hereafter, Trojans) and Hildas are indicative of planetary migration patterns since their capture and physical state must be explained by dynamical evolution models. Early models of minimal planetary migration necessitate that Trojans were dynamically captured from the giant planet region (e.g., Marzari & Scholl 1998). The Nice model instead suggests that Trojans were injected from the outer solar system during a period of significant giant planet migration (e.g., Morbidelli et al. 2005). A more recent version of the Nice model suggests that asymmetric scatterings and collisions would have taken place, producing dissimilar L4 and L5 clouds (Nesvorny et al. 2013). Each of these formation scenarios predicts a different origin and/or collisional evolution for Trojans, which can be inferred from rotation properties. Namely, the physical shape as a function of size helps determine the degree of collisional processing (Farinella et al. 1992). Also, the binary fraction as a function of separation between the two components can be used to determine the dominant binary formation mechanism and thus helps characterize the dynamical environment (e.g., Kern & Elliot 2006). Rotational variation usually corresponds to elongated shapes, but high amplitudes (> 0.9 magnitudes; Sheppard & Jewitt 2004) can only be explained by close or contact binaries. Therefore, rotational lightcurves can be used to infer both shape and the presence of a close companion. Motivated by the need for more observational constraints on solar system formation models and a poor understanding of the rotation properties and binary fraction of Trojans and Hildas, we are studying their rotational lightcurve amplitudes using infrared photometry from NEOWISE (Mainzer et al. 2011; Grav et al. 2011) in order to determine debiased rotational lightcurve amplitude distributions for various Trojan subpopulations and for Trojans compared to Hildas. Preliminary amplitude distributions show a large

  3. A advertência poética de Hilda Hilst em As aves da noite

    OpenAIRE

    Cunha,Rubens da

    2017-01-01

    resumo Entre 1967 e 1969, Hilda Hilst escreveu oito peças de teatro, entre elas, As aves da noite, que relata os últimos momentos de seis prisioneiros na cela da fome, num campo nazista. Neste artigo, analisamos As aves da noite como uma peça de advertência sobre o terror imposto por qualquer estado totalitário, não apenas o nazista. A análise foi fundamentada, entre outros, por Alain Badiou, para quem o século XX não cumpriu a promessa da modernidade e a vida só cumpriu seu destino e seu des...

  4. La advertencia poética de Hilda Hilst en As aves da noite

    OpenAIRE

    Cunha,Rubens da

    2017-01-01

    Entre 1967 y 1969, Hilda Hilst escribió ocho piezas de teatro, entre ellas, As aves da noite, en la cual relata los últimos momentos de seis prisioneros en la celda del hambre, en un campo de concentración nazi. En este artículo, analizamos As aves da noite como una pieza de advertencia sobre el terror impuesto por cualquier estado totalitario, no solamente el instaurado por el nacionalsocialismo alemán. El análisis fue fundamentado, entre otros pensadores, en Alain Badiou, para quien el sigl...

  5. VizieR Online Data Catalog: Reflectance spectra of 12 Trojans and Hildas (Marsset+, 2014)

    Science.gov (United States)

    Marsset, M.; Vernazza, P.; Gourgeot, F.; Dumas, C.; Birlan, M.; Lamy, P.; Binzel, R. P.

    2014-07-01

    We present 17 reflectance spectra of 12 high albedo (pv>0.14) Trojans (8 objects) and Hildas (4 objects) obtained with the ESO/VLT Echelle spectrograph X-SHOOTER in the 0.3-2.2um spectral range (14 spectra) and with the NASA/IRTF spectrograph SpeX in the 0.8-2.5um spectral range (3 spectra). X-SHOOTER spectra were normalized to unity at 0.55um and SpeX spectra were normalized to unity at 2.2um . The spectra presented in this work were collected between April and December 2013. (18 data files).

  6. Professional Development: What Works? Q&A with Dr. Hilda Borko. REL Mid-Atlantic Teacher Effectiveness Webinar Series

    Science.gov (United States)

    Regional Educational Laboratory Mid-Atlantic, 2013

    2013-01-01

    In this webinar, Dr. Hilda Borko, professor, Stanford University Graduate School of Education, presented examples of promising models of professional development, including the Problem-Solving Cycle, Learning and Teaching Geometry, and the use of videos as tools for teacher learning. This Q&A addressed the questions participants had for Dr.…

  7. Differential roles of Sirt1 in HIF-1α and HIF-2α mediated hypoxic responses

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Haejin; Shin, Seung-Hyun; Shin, Dong Hoon; Chun, Yang-Sook; Park, Jong-Wan, E-mail: parkjw@snu.ac.kr

    2014-01-31

    Highlights: • Roles of SIRT1 in HIF-1α and HIF-2α regulations are reevaluated using specific antibodies and Gal4 reporters. • SIRT1 represses the HIF-1α-driven transcription constantly in ten cancer cell-lines. • SIRT1 regulates the HIF-2α-driven transcription cell context-dependently. • SIRT1 determines cell growth under hypoxia by regulating HIF-1α and HIF-2α activities. - Abstract: Hypoxia-inducible factors 1α and 2α (HIF-1α and HIF-2α) determine cancer cell fate under hypoxia. Despite the similarities of their structures, HIF-1α and HIF-2α have distinct roles in cancer growth under hypoxia, that is, HIF-1α induces growth arrest whereas HIF-2α promotes cell growth. Recently, sirtuin 1 (Sirt1) was reported to fine-tune cellular responses to hypoxia by deacetylating HIF-1α and HIF-2α. Yet, the roles of Sirt1 in HIF-1α and HIF-2α functions have been controversial. We here investigated the precise roles of Sirt1 in HIF-1α and HIF-2α regulations. Immunological analyses revealed that HIF-1α K674 and HIF-2α K741 are acetylated by PCAF and CBP, respectively, but are deacetylated commonly by Sirt1. In the Gal4 reporter systems, Sirt1 was found to repress HIF-1α activity constantly in ten cancer cell-lines but to regulate HIF-2α activity cell type-dependently. Moreover, Sirt1 determined cell growth under hypoxia depending on HIF-1α and HIF-2α. Under hypoxia, Sirt1 promoted cell proliferation of HepG2, in which Sirt1 differentially regulates HIF-1α and HIF-2α. In contrast, such an effect of Sirt1 was not shown in HCT116, in which Sirt1 inactivates both HIF-1α and HIF-2α because conflicting actions of HIF-1α and HIF-2α on cell growth may be offset. Our results provide a better understanding of the roles of Sirt1 in HIF-mediated hypoxic responses and also a basic concept for developing anticancer strategy targeting Sirt1.

  8. Cobalt stimulates HIF-1-dependent but inhibits HIF-2-dependent gene expression in liver cancer cells

    Science.gov (United States)

    Befani, Christina; Mylonis, Ilias; Gkotinakou, Ioanna-Maria; Georgoulias, Panagiotis; Hu, Cheng-Jun; Simos, George; Liakos, Panagiotis

    2013-01-01

    Hypoxia-inducible factors (HIFs) are transcriptional regulators that mediate the cellular response to low oxygen. Although HIF-1 is usually considered as the principal mediator of hypoxic adaptation, several tissues and different cell types express both HIF-1 and HIF-2 isoforms under hypoxia or when treated with hypoxia mimetic chemicals such as cobalt. However, the similarities or differences between HIF-1 and HIF-2, in terms of their tissue- and inducer-specific activation and function, are not adequately characterized. To address this issue, we investigated the effects of true hypoxia and hypoxia mimetics on HIF-1 and HIF-2 induction and specific gene transcriptional activity in two hepatic cancer cell lines, Huh7 and HepG2. Both hypoxia and cobalt caused rapid induction of both HIF-1α and HIF-2α proteins. Hypoxia induced erythropoietin (EPO) expression and secretion in a HIF-2-dependent way. Surprisingly, however, EPO expression was not induced when cells were treated with cobalt. In agreement, both HIF-1- and HIF-2-dependent promoters (of PGK and SOD2 genes, respectively) were activated by hypoxia while cobalt only activated the HIF-1-dependent PGK promoter. Unlike cobalt, other hypoxia mimetics such as DFO and DMOG activated both types of promoters. Furthermore, cobalt impaired the hypoxic stimulation of HIF-2, but not HIF-1, activity and cobalt-induced HIF-2α interacted poorly with USF-2, a HIF-2-specific co-activator. These data show that, despite similar induction of HIF-1α and HIF-2α protein expression, HIF-1 and HIF-2 specific gene activating functions respond differently to different stimuli and suggest the operation of oxygen-independent and gene- or tissue-specific regulatory mechanisms involving additional transcription factors or co-activators. PMID:23958427

  9. A advertência poética de Hilda Hilst em As aves da noite

    OpenAIRE

    Cunha, Rubens da

    2017-01-01

    Entre 1967 e 1969, Hilda Hilst escreveu oito peças de teatro, entre elas, As aves da noite, que relata os últimos momentos de seis prisioneiros na cela da fome, num campo nazista. Neste artigo, analisamos As aves da noite como uma peça de advertência sobre o terror imposto por qualquer estado totalitário, não apenas o nazista. A análise foi fundamentada, entre outros, por Alain Badiou, para quem o século XX não cumpriu a promessa da modernidade e a vida só cumpriu seu destino e seu desígnio p...

  10. Hilda Anna Krisch: pioneira na enfermagem catarinense - formação e contribuição

    Directory of Open Access Journals (Sweden)

    Miriam Süsskind Borenstein

    2004-06-01

    Full Text Available Trata-se de pesquisa sócio-histórica cujo objetivo é desvelar a historicidade da primeira enfermeira catarinense formada pela Escola de Enfermagem Anna Nery, com atuação de destaque no cenário nacional. Na coleta de dados foram utilizadas entrevistas arquivadas realizadas com Hilda Anna Krisch e com outras personagens que fizeram parte do cenário da época. Além destas, utilizou-se outras fontes documentais como: periódicos, livros, cartas, relatórios, histórico escolar e fotografias. Os dados permitiram identificar a origem e a trajetória da enfermeira, bem como os trabalhos desenvolvidos por esta, durante sua vida profissional. Os achados possibilitam inferir que a enfermeira Hilda Anna Krisch, foi um baluarte na enfermagem catarinense e brasileira por ter desempenhado relevante papel dentro e fora da profissão.

  11. Inertially confined fusion using heavy ion drivers

    Energy Technology Data Exchange (ETDEWEB)

    Herrmannsfeldt, W.B. (Stanford Linear Accelerator Center, Menlo Park, CA (United States)); Bangerter, R.O. (Lawrence Berkeley Lab., CA (United States)); Bock, R. (Gesellschaft fuer Schwerionenforschung mbH, Darmstadt (Germany)); Hogan, W.J.; Lindl, J.D. (Lawrence Livermore National Lab., CA (United States))

    1991-10-01

    The various technical issues of HIF will be briefly reviewed in this paper. It will be seen that there are numerous areas in common in all the approaches to HIF. In the recent International Symposium on Heavy Ion Inertial Fusion, the attendees met in specialized workshop sessions to consider the needs for research in each area. Each of the workshop groups considered the key questions of this report: (1) Is this an appropriate time for international collaboration in HIF (2) Which problems are most appropriate for such collaboration (3) Can the sharing of target design information be set aside until other driver and systems issues are better resolved, by which time it might be supposed that there could be a relaxation of classification of target issues (4) What form(s) of collaboration are most appropriate, e.g., bilateral or multilateral (5) Can international collaboration be sensibly attempted without significant increases in funding for HIF The authors of this report share the conviction that collaboration on a broad scale is mandatory for HIF to have the resources, both financial and personnel, to progress to a demonstration experiment. Ultimately it may be possible for a single driver with the energy, power, focusibility, and pulse shape to satisfy the needs of the international community for target physics research. Such a facility could service multiple experimental chambers with a variety of beam geometries and target concepts.

  12. Inertially confined fusion using heavy ion drivers

    Energy Technology Data Exchange (ETDEWEB)

    Herrmannsfeldt, W.B. [Stanford Linear Accelerator Center, Menlo Park, CA (United States); Bangerter, R.O. [Lawrence Berkeley Lab., CA (United States); Bock, R. [Gesellschaft fuer Schwerionenforschung mbH, Darmstadt (Germany); Hogan, W.J.; Lindl, J.D. [Lawrence Livermore National Lab., CA (United States)

    1991-10-01

    The various technical issues of HIF will be briefly reviewed in this paper. It will be seen that there are numerous areas in common in all the approaches to HIF. In the recent International Symposium on Heavy Ion Inertial Fusion, the attendees met in specialized workshop sessions to consider the needs for research in each area. Each of the workshop groups considered the key questions of this report: (1) Is this an appropriate time for international collaboration in HIF? (2) Which problems are most appropriate for such collaboration? (3) Can the sharing of target design information be set aside until other driver and systems issues are better resolved, by which time it might be supposed that there could be a relaxation of classification of target issues? (4) What form(s) of collaboration are most appropriate, e.g., bilateral or multilateral? (5) Can international collaboration be sensibly attempted without significant increases in funding for HIF? The authors of this report share the conviction that collaboration on a broad scale is mandatory for HIF to have the resources, both financial and personnel, to progress to a demonstration experiment. Ultimately it may be possible for a single driver with the energy, power, focusibility, and pulse shape to satisfy the needs of the international community for target physics research. Such a facility could service multiple experimental chambers with a variety of beam geometries and target concepts.

  13. A angústia diante da morte em “Lázaro”, de Hilda Hilst

    Directory of Open Access Journals (Sweden)

    Willian André

    2016-01-01

    Full Text Available Este artículo tiene como objetivo construir una reflexión sobre el tema de “la angustia ante la mue rte” a partir del análisis de la narrativa “Lázaro” (1970, de la escritora brasileña Hilda Hilst. Al proponer una nueva lectura del episodio bíblico que narra la muerte y resurrección de Lázaro, el texto en cuestión nos permite establecer un diálogo con e l concepto de ser para la muerte , de Martin Heidegger, así como con la teoría de lo sublime , de Edmund Burke, en cuyo núcleo se encuentra la manifestación de la muerte. Al delinear dichos diálogos, tenemos la intención de observar cómo el narrador - protagon ista del texto de Hilst puede ser pensado como una encarnación del ser para la muerte heideggeriano.

  14. Narciso e seu reino de sombra em Cantares, de Hilda Hilst

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    Souza, Enivalda Nunes Freitas e

    2009-01-01

    Full Text Available Os poemas de Cantares, de Hilda Hilst, convidam ao desvelamento do mito do amor, empreitada que procuro enriquecer, neste artigo, recorrendo à matriz ovidiana do mito de Narciso e seus tantos mitemas, como a busca do conhecimento, a questão da sombra e do duplo e a intrigante relação entre amor e ódio, amor e morte. Como afirma Durand, os mitos são variações de arquétipos, o que provoca a aproximação e o entrelaçamento dos conteúdos míticos. De todos os mitos sobre os desencontros amorosos e a consequente solidão humana, poucos têm a beleza e a força psíquica desencadeada pelo mito de Narciso elaborado por Ovídio. Além do mito de Narciso, este artigo resgata a genealogia de Eros e os andróginos desafortunados, relatados por Platão, e a história de Salomão e Sulamita elaborada nos Cânticos bíblicos. O aporte teórico envolve Gilbert Durand, Jung, Freud e Kristeva

  15. Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance.

    Science.gov (United States)

    Vukovic, Milica; Guitart, Amelie V; Sepulveda, Catarina; Villacreces, Arnaud; O'Duibhir, Eoghan; Panagopoulou, Theano I; Ivens, Alasdair; Menendez-Gonzalez, Juan; Iglesias, Juan Manuel; Allen, Lewis; Glykofrydis, Fokion; Subramani, Chithra; Armesilla-Diaz, Alejandro; Post, Annemarie E M; Schaak, Katrin; Gezer, Deniz; So, Chi Wai Eric; Holyoake, Tessa L; Wood, Andrew; O'Carroll, Dónal; Ratcliffe, Peter J; Kranc, Kamil R

    2015-12-14

    Leukemogenesis occurs under hypoxic conditions within the bone marrow (BM). Knockdown of key mediators of cellular responses to hypoxia with shRNA, namely hypoxia-inducible factor-1α (HIF-1α) or HIF-2α, in human acute myeloid leukemia (AML) samples results in their apoptosis and inability to engraft, implicating HIF-1α or HIF-2α as therapeutic targets. However, genetic deletion of Hif-1α has no effect on mouse AML maintenance and may accelerate disease development. Here, we report the impact of conditional genetic deletion of Hif-2α or both Hif-1α and Hif-2α at different stages of leukemogenesis in mice. Deletion of Hif-2α accelerates development of leukemic stem cells (LSCs) and shortens AML latency initiated by Mll-AF9 and its downstream effectors Meis1 and Hoxa9. Notably, the accelerated initiation of AML caused by Hif-2α deletion is further potentiated by Hif-1α codeletion. However, established LSCs lacking Hif-2α or both Hif-1α and Hif-2α propagate AML with the same latency as wild-type LSCs. Furthermore, pharmacological inhibition of the HIF pathway or HIF-2α knockout using the lentiviral CRISPR-Cas9 system in human established leukemic cells with MLL-AF9 translocation have no impact on their functions. We therefore conclude that although Hif-1α and Hif-2α synergize to suppress the development of AML, they are not required for LSC maintenance.

  16. Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance

    Science.gov (United States)

    Vukovic, Milica; Guitart, Amelie V.; Sepulveda, Catarina; Villacreces, Arnaud; O'Duibhir, Eoghan; Panagopoulou, Theano I.; Ivens, Alasdair; Menendez-Gonzalez, Juan; Iglesias, Juan Manuel; Allen, Lewis; Glykofrydis, Fokion; Subramani, Chithra; Armesilla-Diaz, Alejandro; Post, Annemarie E.M.; Schaak, Katrin; Gezer, Deniz; So, Chi Wai Eric; Holyoake, Tessa L.; Wood, Andrew; O'Carroll, Dónal; Ratcliffe, Peter J.

    2015-01-01

    Leukemogenesis occurs under hypoxic conditions within the bone marrow (BM). Knockdown of key mediators of cellular responses to hypoxia with shRNA, namely hypoxia-inducible factor-1α (HIF-1α) or HIF-2α, in human acute myeloid leukemia (AML) samples results in their apoptosis and inability to engraft, implicating HIF-1α or HIF-2α as therapeutic targets. However, genetic deletion of Hif-1α has no effect on mouse AML maintenance and may accelerate disease development. Here, we report the impact of conditional genetic deletion of Hif-2α or both Hif-1α and Hif-2α at different stages of leukemogenesis in mice. Deletion of Hif-2α accelerates development of leukemic stem cells (LSCs) and shortens AML latency initiated by Mll-AF9 and its downstream effectors Meis1 and Hoxa9. Notably, the accelerated initiation of AML caused by Hif-2α deletion is further potentiated by Hif-1α codeletion. However, established LSCs lacking Hif-2α or both Hif-1α and Hif-2α propagate AML with the same latency as wild-type LSCs. Furthermore, pharmacological inhibition of the HIF pathway or HIF-2α knockout using the lentiviral CRISPR-Cas9 system in human established leukemic cells with MLL-AF9 translocation have no impact on their functions. We therefore conclude that although Hif-1α and Hif-2α synergize to suppress the development of AML, they are not required for LSC maintenance. PMID:26642852

  17. HIF transcription factors, inflammation, and immunity.

    Science.gov (United States)

    Palazon, Asis; Goldrath, Ananda W; Nizet, Victor; Johnson, Randall S

    2014-10-16

    The hypoxic response in cells and tissues is mediated by the family of hypoxia-inducible factor (HIF) transcription factors; these play an integral role in the metabolic changes that drive cellular adaptation to low oxygen availability. HIF expression and stabilization in immune cells can be triggered by hypoxia, but also by other factors associated with pathological stress: e.g., inflammation, infectious microorganisms, and cancer. HIF induces a number of aspects of host immune function, from boosting phagocyte microbicidal capacity to driving T cell differentiation and cytotoxic activity. Cellular metabolism is emerging as a key regulator of immunity, and it constitutes another layer of fine-tuned immune control by HIF that can dictate myeloid cell and lymphocyte development, fate, and function. Here we discuss how oxygen sensing in the immune microenvironment shapes immunological response and examine how HIF and the hypoxia pathway control innate and adaptive immunity.

  18. HIF-1α Promotes A Hypoxia-Independent Cell Migration.

    Science.gov (United States)

    Li, Liyuan; Madu, Chikezie O; Lu, Andrew; Lu, Yi

    2010-01-01

    Hypoxia-inducible factor-1α (HIF-1α) is known as a transactivator for VEGF gene promoter. It can be induced by hypoxia. However, no study has been done so far to dissect HIF-1α-mediated effects from hypoxia or VEGF-mediated effects. By using a HIF-1α knockout (HIF-1α KO) cell system in mouse embryonic fibroblast (MEF) cells, this study analyzes cell migration and HIF-1α, hypoxia and VEGF activation. A hypoxia-mediated HIF-1α induction and VEGF transactivation were observed: both HIF-1α WT lines had significantly increased VEGF transactivation, as an indicator for HIF-1α induction, in hypoxia compared to normoxia; in contrast, HIF-1α KO line had no increased VEGF transactivation under hypoxia. HIF-1α promotes cell migration: HIF-1α-KO cells had a significantly reduced migration compared to that of the HIF-1α WT cells under both normoxia and hypoxia. The significantly reduced cell migration in HIF-1α KO cells can be partially rescued by the restoration of WT HIF-1α expression mediated by adenoviral-mediated gene transfer. Interestingly, hypoxia has no effect on cell migration: the cells had a similar cell migration rate under hypoxic and normoxic conditions for both HIF-1α WT and HIF-1α KO lines, respectively. Collectively, these data suggest that HIF-1α plays a role in MEF cell migration that is independent from hypoxia-mediated effects.

  19. Differential regulation of HIF-1α and HIF-2α in neuroblastoma: Estrogen-related receptor alpha (ERRα) regulates HIF2A transcription and correlates to poor outcome

    Energy Technology Data Exchange (ETDEWEB)

    Hamidian, Arash; Stedingk, Kristoffer von; Munksgaard Thorén, Matilda; Mohlin, Sofie; Påhlman, Sven, E-mail: sven.pahlman@med.lu.se

    2015-06-05

    Hypoxia-inducible factors (HIFs) are differentially regulated in tumor cells. While the current paradigm supports post-translational regulation of the HIF-α subunits, we recently showed that hypoxic HIF-2α is also transcriptionally regulated via insulin-like growth factor (IGF)-II in the childhood tumor neuroblastoma. Here, we demonstrate that transcriptional regulation of HIF-2α seems to be restricted to neural cell-derived tumors, while HIF-1α is canonically regulated at the post-translational level uniformly across different tumor forms. Enhanced expression of HIF2A mRNA at hypoxia is due to de novo transcription rather than increased mRNA stability, and chemical stabilization of the HIF-α proteins at oxygen-rich conditions unexpectedly leads to increased HIF2A transcription. The enhanced HIF2A levels do not seem to be dependent on active HIF-1. Using a transcriptome array approach, we identified members of the Peroxisome proliferator-activated receptor gamma coactivator (PGC)/Estrogen-related receptor (ERR) complex families as potential regulators of HIF2A. Knockdown or inhibition of one of the members, ERRα, leads to decreased expression of HIF2A, and high expression of the ERRα gene ESRRA correlates with poor overall and progression-free survival in a clinical neuroblastoma material consisting of 88 tumors. Thus, targeting of ERRα and pathways regulating transcriptional HIF-2α are promising therapeutic avenues in neuroblastoma. - Highlights: • Transcriptional control of HIF-2α is restricted to neural cell-derived tumors. • Enhanced transcription of HIF2A is not due to increased mRNA stability. • Chemical stabilization of the HIF-α subunits leads to increased HIF2A transcription. • ERRα regulates HIF2A mRNA expression in neuroblastoma. • High expression of ESRRA correlates to poor outcome in neuroblastoma.

  20. Spiral wobbling beam illumination uniformity in HIF fuel target implosion

    Directory of Open Access Journals (Sweden)

    Kawata S.

    2013-11-01

    Full Text Available A few % wobbling-beam illumination nonuniformity is realized in heavy ion inertial confinement fusion (HIF throughout the heavy ion beam (HIB driver pulse by a newly introduced spiraling beam axis motion in the first two rotations. The wobbling HIB illumination was proposed to realize a uniform implosion in HIF. However, the initial imprint of the wobbling HIBs was a serious problem and introduces a large unacceptable energy deposition nonuniformity. In the wobbling HIBs illumination, the illumination nonuniformity oscillates in time and space. The oscillating-HIB energy deposition may produce a time-dependent implosion acceleration, which reduces the Rayleigh-Taylor (R-T growth [Laser Part. Beams 11, 757 (1993, Nuclear Inst. Methods in Phys. Res. A 606, 152 (2009, Phys. Plasmas 19, 024503 (2012] and the implosion nonuniformity. The wobbling HIBs can be generated in HIB accelerators and the oscillating frequency may be several 100 MHz ∼ 1 GHz [Phys. Rev. Lett. 104, 254801 (2010]. Three-dimensional HIBs illumination computations present that the few % wobbling HIBs illumination nonuniformity oscillates with the same wobbling HIBs frequency.

  1. Differential regulation of HIF-1α and HIF-2α in neuroblastoma: Estrogen-related receptor alpha (ERRα) regulates HIF2A transcription and correlates to poor outcome.

    Science.gov (United States)

    Hamidian, Arash; von Stedingk, Kristoffer; Munksgaard Thorén, Matilda; Mohlin, Sofie; Påhlman, Sven

    2015-06-01

    Hypoxia-inducible factors (HIFs) are differentially regulated in tumor cells. While the current paradigm supports post-translational regulation of the HIF-α subunits, we recently showed that hypoxic HIF-2α is also transcriptionally regulated via insulin-like growth factor (IGF)-II in the childhood tumor neuroblastoma. Here, we demonstrate that transcriptional regulation of HIF-2α seems to be restricted to neural cell-derived tumors, while HIF-1α is canonically regulated at the post-translational level uniformly across different tumor forms. Enhanced expression of HIF2A mRNA at hypoxia is due to de novo transcription rather than increased mRNA stability, and chemical stabilization of the HIF-α proteins at oxygen-rich conditions unexpectedly leads to increased HIF2A transcription. The enhanced HIF2A levels do not seem to be dependent on active HIF-1. Using a transcriptome array approach, we identified members of the Peroxisome proliferator-activated receptor gamma coactivator (PGC)/Estrogen-related receptor (ERR) complex families as potential regulators of HIF2A. Knockdown or inhibition of one of the members, ERRα, leads to decreased expression of HIF2A, and high expression of the ERRα gene ESRRA correlates with poor overall and progression-free survival in a clinical neuroblastoma material consisting of 88 tumors. Thus, targeting of ERRα and pathways regulating transcriptional HIF-2α are promising therapeutic avenues in neuroblastoma.

  2. DRIVER INATTENTION

    Directory of Open Access Journals (Sweden)

    Richard TAY

    2004-01-01

    Full Text Available Driver inattention, especially driver distraction, is an extremely influential but generally neglected contributing factor of road crashes. This paper explores some of the common behaviours associated with several common forms of driver inattention, with respect to their perceived crash risks, rates of self-reported behaviours and whether drivers regulate such behaviours depending on the road and traffic environment, and provides some policy recommendations to address issues raised.

  3. HIF-2α- a mediator of osteoarthritis?

    Institute of Scientific and Technical Information of China (English)

    Christopher L Murphy

    2010-01-01

    @@ Two recent publications in Nature Medicine (June 2010) have focused at-tention on the role ofhypoxia inducible factor HIF-2a in cartilage [ 1,2]. Using similar mouse models, both groups provide striking evidence implicating HIF-2a in experimentally induced os-teoarthritis (OA). While undoubtedly representing significant advances in the field, care must be taken in interpreting these results, in particular in extrapolat-ing the findings to humans.

  4. INTERTEXTUALIDAD EN "VOCALES PARA HILDA" DE GONZALO ROJAS: UN DIÁLOGO ABIERTO CON RIMBAUD Y CÉSAR VALLEJO

    Directory of Open Access Journals (Sweden)

    Héctor Veliz Gatica

    2009-11-01

    Full Text Available El presente trabajo se propone demostrar la existencia de una intertextualidad entre el poema "Vocales para Hilda" de Gonzalo Rojas y los sonetos "Vocales" ("Voyelles" y "El dolor de las cinco vocales" de Arthur Rimbaud y César Vallejo, respectivamente. Para ello, se procedió con la recolección bibliográfica y revisión crítica de textos, además de un registro de los datos intratextuales (nivel fónico, gramatical y semántico presentes en el poema. Los resultados obtenidos demuestran la existencia de dicha relación intertextual, por lo que concluimos que el texto de Rojas no se reduce a un poema de corte privativamente amoroso, ya que al plantearse en diálogo con los autores aquí considerados, se amplía y enriquece en su potencial de sentidos.The present work proposes to demonstrate the existence of an intertextuality between the poem " Vocales para Hilda " of Gonzalo Rojas and the sonnets " Voyelles " and "El dolor de las cinco vocales " of Arthur Rimbaud and César Vallejo respectively. To achieve this purpose a bibliographic recollection and a critical review of the texts has been done, and a register of the intertextual data present in the poem (phonic, grammatical, and semantic level was also establihed. The results demostrated the presence ofthe above mentioned intertextual relationship, that is why it may be assumed that Gonzalo Rojas' text is not only a love poem; it also establishes a dialogue with the already mentioned Rimbaud and Vallejo, which means thispoem has a much wider potential meaning.

  5. A compendium of proteins that interact with HIF-1α.

    Science.gov (United States)

    Semenza, Gregg L

    2017-07-15

    Hypoxia-inducible factor 1 (HIF-1) is the founding member of a family of transcription factors that function as master regulators of oxygen homeostasis. HIF-1 is composed of an O2-regulated HIF-1α subunit and a constitutively expressed HIF-1β subunit. This review provides a compendium of proteins that interact with the HIF-1α subunit, many of which regulate HIF-1 activity in either an O2-dependent or O2-independent manner. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Endothelial HIF-1α Enables Hypothalamic Glucose Uptake to Drive POMC Neurons.

    Science.gov (United States)

    Varela, Luis; Suyama, Shigetomo; Huang, Yan; Shanabrough, Marya; Tschöp, Matthias H; Gao, Xiao-Bing; Giordano, Frank J; Horvath, Tamas L

    2017-06-01

    Glucose is the primary driver of hypothalamic proopiomelanocortin (POMC) neurons. We show that endothelial hypoxia-inducible factor 1α (HIF-1α) controls glucose uptake in the hypothalamus and that it is upregulated in conditions of undernourishment, during which POMC neuronal activity is decreased. Endothelium-specific knockdown of HIF-1α impairs the ability of POMC neurons to adapt to the changing metabolic environment in vivo, resulting in overeating after food deprivation in mice. The impaired functioning of POMC neurons was reversed ex vivo or by parenchymal glucose administration. These observations indicate an active role for endothelial cells in the central control of metabolism and suggest that central vascular impairments may cause metabolic disorders. © 2017 by the American Diabetes Association.

  7. Nuclear HIF1A expression is strongly prognostic in sporadic but not familial male breast cancer.

    Science.gov (United States)

    Deb, Siddhartha; Johansson, Ida; Byrne, David; Nilsson, Cecilia; Investigators, kConFab; Constable, Leonie; Fjällskog, Marie-Louise; Dobrovic, Alexander; Hedenfalk, Ingrid; Fox, Stephen B

    2014-09-01

    Male breast cancer is poorly understood with a large proportion arising in the familial context particularly with the BRCA2 germline mutation. As phenotypic and genotypic differences between sporadic and familial male breast cancers have been noted, we investigated the importance of a hypoxic drive in these cancers as this pathway has been shown to be of importance in familial female breast cancer. Expression of two major hypoxia-induced proteins, the hypoxia-inducible factor-1α (HIF1A) and the carbonic anhydrase IX (CA9), examined within a large cohort including 61 familial (3 BRCA1, 28 BRCA2, 30 BRCAX) and 225 sporadic male breast cancers showed that 31% of all male breast cancers expressed either HIF1A (25%) and/or CA9 (8%) in the combined cohort. Expression of HIF1A correlated with an increased incidence of a second-major malignancy (P=0.04), histological tumor type (P=0.005) and basal phenotype (P=0.02). Expression of CA9 correlated with age (P=0.004) in sporadic cases and an increased tumor size (P=0.003). Expression of HIF1A was prognostic for disease-specific survival in sporadic male breast cancers (HR: 3.8, 95% CI: 1.5-9.8, P=0.006) but not within familial male breast cancer, whereas CA9 was only prognostic in familial male breast cancers (HR: 358.0, 95% CI: 9.3-13781.7, P=0.002) and not in sporadic male breast cancer. This study found that hypoxic drive is less prevalent in male breast cancer compared with female breast cancer, possibly due to a different breast microenvironment. The prognostic impact of HIF1A is greatest in sporadic male breast cancers with an alternate dominant mechanism for the oncogenic drivers suggested in high risk familial male breast cancers.

  8. HIF1α and HIF2α exert distinct nutrient preferences in renal cells.

    Directory of Open Access Journals (Sweden)

    Alexandra Arreola

    Full Text Available BACKGROUND: Hypoxia Inducible Factors (HIF1α and HIF2α are commonly stabilized and play key roles related to cell growth and metabolic programming in clear cell renal cell carcinoma. The relationship of these factors to discretely alter cell metabolic activities has largely been described in cancer cells, or in hypoxic conditions, where other confounding factors undoubtedly compete. These transcription factors and their specific roles in promoting cancer metabolic phenotypes from the earliest stages are poorly understood in pre-malignant cells. METHODS: We undertook an analysis of SV40-transformed primary kidney epithelial cells derived from newborn mice genetically engineered to express a stabilized HIF1α or HIF2α transgene. We examined the metabolic profile in relation to each gene. RESULTS: Although the cells proliferated similarly, the metabolic profile of each genotype of cell was markedly different and correlated with altered gene expression of factors influencing components of metabolic signaling. HIF1α promoted high levels of glycolysis as well as increased oxidative phosphorylation in complete media, but oxidative phosphorylation was suppressed when supplied with single carbon source media. HIF2α, in contrast, supported oxidative phosphorylation in complete media or single glucose carbon source, but these cells were not responsive to glutamine nutrient sources. This finding correlates to HIF2α-specific induction of Glul, effectively reducing glutamine utilization by limiting the glutamate pool, and knockdown of Glul allows these cells to perform oxidative phosphorylation in glutamine media. CONCLUSION: HIF1α and HIF2α support highly divergent patterns of kidney epithelial cell metabolic phenotype. Expression of these factors ultimately alters the nutrient resource utilization and energy generation strategy in the setting of complete or limiting nutrients.

  9. A compreensão é uma grande porca acinzentada – Uma leitura sobre a busca da linguagem em A obscena senhora D, de Hilda Hilst.

    Directory of Open Access Journals (Sweden)

    Tatiana Franca Rodrigues

    2012-09-01

    Full Text Available The aim of this work is, by reading A obscena senhora D, of Hilda Hilst, to understand  how the theme of eroticism is connected to questions related to metalanguage and to the issue of the process of writing itself. Under these circumstances, the study intends to apprehend pornography as a speech through which critical thoughts regarding contemporaneity are taken into consideration.

  10. A compreensão é uma grande porca acinzentada – Uma leitura sobre a busca da linguagem em A obscena senhora D, de Hilda Hilst.

    Directory of Open Access Journals (Sweden)

    Tatiana Franca Rodrigues

    2012-09-01

    Full Text Available The aim of this work is, by reading A obscena senhora D, of Hilda Hilst, to understand  how the theme of eroticism is connected to questions related to metalanguage and to the issue of the process of writing itself. Under these circumstances, the study intends to apprehend pornography as a speech through which critical thoughts regarding contemporaneity are taken into consideration.

  11. Effects of transforming growth factor beta, tumor necrosis factor alpha, interferon gamma and LIF-HILDA on the proliferation of acute myeloid leukemia cells.

    Science.gov (United States)

    Kerangueven, F; Sempere, C; Tabilio, A; Mannoni, P

    1990-01-01

    A group of polypeptide factors that regulate cell growth and differentiation has been tested for their biological activities on the growth and differentiation of leukemic cells isolated from patients with Acute Myeloid Leukemias (AML). The effects of Transforming Growth Factor beta 1 (TGF beta), Tumor Necrosis Factor alpha (TNF alpha), Interferon gamma (IFN gamma) and LIF-HILDA were compared on leukemic cells cultured in vitro for seven days. Spontaneously growing leukemic cells were selected in order to study either inhibition or enhancement of proliferation induced by these factors. Only TGF beta 1 was found to induce a clear inhibition of leukemic proliferation in all cases tested. Recombinant TNF alpha and IFN gamma were found to induce either inhibition or enhancement of the proliferation on separate specimens. Under the conditions of culture, it was not possible to document any effect of LIF-HILDA. Cell differentiation and cell maturation were documented studying the modulation of cell surface antigens. TGF beta did not modify antigen expression on the cells surviving after 3 days in culture. Both TNF alpha and IFN gamma were found to enhance the expression of adhesion molecules and to a lesser extent, the expression of some lineage associated antigens. No effect of LIF-HILDA on antigen modulation was documented in the cases tested. These data confirm that TGF beta is by itself a potent inhibitor of the myeloid leukemia cells proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Opposite prognostic roles of HIF1β and HIF2β expressions in bone metastatic clear cell renal cell cancer

    DEFF Research Database (Denmark)

    Szendroi, Attila; Szász, A. Marcell; Kardos, Magdolna

    2016-01-01

    BACKGROUND: Prognostic markers of bone metastatic clear cell renal cell cancer (ccRCC) are poorly established. We tested prognostic value of HIF1β/HIF2β and their selected target genes in primary tumors and corresponding bone metastases. RESULTS: Expression of HIF2β was lower in mRCC both at mRNA...

  13. Loss of Hif-2α Rescues the Hif-1α Deletion Phenotype of Neonatal Respiratory Distress In Mice.

    Directory of Open Access Journals (Sweden)

    Yogesh Saini

    Full Text Available Hypoxia is a state of decreased oxygen reaching the tissues of the body. During prenatal development, the fetus experiences localized occurrences of hypoxia that are essential for proper organogenesis and survival. The response to decreased oxygen availability is primarily regulated by hypoxia-inducible factors (HIFs, a family of transcription factors that modulate the expression of key genes involved in glycolysis, angiogenesis, and erythropoiesis. HIF-1α and HIF-2α, two key isoforms, are important in embryonic development, and likely are involved in lung morphogenesis. We have recently shown that the inducible loss of Hif-1α in lung epithelium starting at E4.5 leads to death within an hour of parturition, with symptoms similar to neonatal respiratory distress syndrome (RDS. In addition to Hif-1α, Hif-2α is also expressed in the developing lung, although the overlapping roles of Hif-1α and Hif-2α in this context are not fully understood. To further investigate the independent role of Hif-2α in lung epithelium and its ability to alter Hif-1α-mediated lung maturation, we generated two additional lung-specific inducible Hif-α knockout models (Hif-2α and Hif-1α+Hif-2α. The intrauterine loss of Hif-2α in the lungs does not lead to decreased viability or observable phenotypic changes in the lung. More interestingly, survivability observed after the loss of both Hif-1α and Hif-2α suggests that the loss of Hif-2α is capable of rescuing the neonatal RDS phenotype seen in Hif-1α-deficient pups. Microarray analyses of lung tissue from these three genotypes identified several factors, such as Scd1, Retlnγ, and Il-1r2, which are differentially regulated by the two HIF-α isoforms. Moreover, network analysis suggests that modulation of hormone-mediated, NF-κB, C/EBPα, and c-MYC signaling are central to HIF-mediated changes in lung development.

  14. Older Drivers

    Science.gov (United States)

    ... in this topic was provided by the National Highway Traffic Safety Administration Topic last reviewed: March 2015 For ... see Traffic Safety Facts 2012: Older Population. (National Highway Traffic Safety Administration). Crashes Down Among Older Drivers Fortunately, ...

  15. A hypoxia-inducible factor (HIF)-3α splicing variant, HIF-3α4 impairs angiogenesis in hypervascular malignant meningiomas with epigenetically silenced HIF-3α4

    Energy Technology Data Exchange (ETDEWEB)

    Ando, Hitoshi [Department of Neurosurgery, Nagoya University School of Medicine, Nagoya (Japan); Department of Neurosurgery, Fukushima Medical University School of Medicine, Fukushima (Japan); Natsume, Atsushi, E-mail: anatsume@med.nagoya-u.ac.jp [Department of Neurosurgery, Nagoya University School of Medicine, Nagoya (Japan); Iwami, Kenichiro; Ohka, Fumiharu [Department of Neurosurgery, Nagoya University School of Medicine, Nagoya (Japan); Kuchimaru, Takahiro; Kizaka-Kondoh, Shinae [Department of Biomolecular Engineering, Tokyo Institute of Technology Graduate School of Bioscience and Biotechnology, Yokohama (Japan); Ito, Kengo [National Center for Geriatrics and Gerontology, Aichi (Japan); Saito, Kiyoshi [Department of Neurosurgery, Fukushima Medical University School of Medicine, Fukushima (Japan); Sugita, Sachi; Hoshino, Tsuneyoshi [MICRON Inc.Medical Facilities Support Department, Aichi (Japan); Wakabayashi, Toshihiko [Department of Neurosurgery, Nagoya University School of Medicine, Nagoya (Japan)

    2013-03-29

    Highlights: ► HIF-3α4 is silenced by DNA methylation in meningiomas. ► Induction of HIF-3α4 impaired angiogenesis in meningiomas. ► Induction of HIF-3α4 impaired proliferation and oxygen-dependent metabolism. -- Abstract: Hypoxia inducible factor is a dominant regulator of adaptive cellular responses to hypoxia and controls the expression of a large number of genes regulating angiogenesis as well as metabolism, cell survival, apoptosis, and other cellular functions in an oxygen level-dependent manner. When a neoplasm is able to induce angiogenesis, tumor progression occurs more rapidly because of the nutrients provided by the neovasculature. Meningioma is one of the most hypervascular brain tumors, making anti-angiogenic therapy an attractive novel therapy for these tumors. HIF-3α has been conventionally regarded as a dominant-negative regulator of HIF-1α, and although alternative HIF-3α splicing variants are extensively reported, their specific functions have not yet been determined. In this study, we found that the transcription of HIF-3α4 was silenced by the promoter DNA methylation in meningiomas, and inducible HIF-3α4 impaired angiogenesis, proliferation, and metabolism/oxidation in hypervascular meningiomas. Thus, HIF-3α4 could be a potential molecular target in meningiomas.

  16. [Effect of dominant negative HIF-1alpha (dn HIF-1alpha) on biological characteristics of uterine cervix cancer cells].

    Science.gov (United States)

    Tang, Bin-Zhi; Zhao, Feng-Yan; Wei, Ting; Mu, De-Zhi; Mao, Meng; Fu, Qiang; Zhang, Lin; Qu, Yia

    2008-05-01

    To explore the effect of dominant negative HIF-1alpha (dn HIF-1alpha) on biological characteristics of uterine cervix cancer cell SiHa and elucidate the related mechanism. pcDNA3. 1-dn HIF-1alpha was transfected into SiHa cells. The expression of HIF-1alpha and VEGF protein were detected by immunocytochemical method and Western Blotting. The growth proliferation of cells was surveyed by the MTT assay and cell apoptosis was detected through TUNEL after treated with CoCl2, meanwhile the results were compared with the group transfected with mock plasmid and untransfected group. After successfully transfected with relevant plasmid, there's no obvious difference of expression of HIF-1alpha among dn HIF-1alpha group, pcDNA3. 1 group, and untransfected group, however the expression of VEGF of dn HIF-1alpha group was significantly lower than that of the others (P dn HIF-1alpha group was obviously lower than that of the other two (P dn HIF-1alpha group among these three (P < 0.05). Domain negative HIF-1alpha can inhibit the proliferation of uterine cervix cancer cell and accelerate its apoptosis under hypoxia induced by CoCl2, as well as decrease the expression of VEGF protein. The implications of all this were that the domain negative HIF-1alpha may play an important role in the therapy of uterine cervix cancer.

  17. Endothelial HIF signaling regulates pulmonary fibrosis-associated pulmonary hypertension.

    Science.gov (United States)

    Bryant, Andrew J; Carrick, Ryan P; McConaha, Melinda E; Jones, Brittany R; Shay, Sheila D; Moore, Christy S; Blackwell, Thomas R; Gladson, Santhi; Penner, Niki L; Burman, Ankita; Tanjore, Harikrishna; Hemnes, Anna R; Karwandyar, Ayub K; Polosukhin, Vasiliy V; Talati, Megha A; Dong, Hui-Jia; Gleaves, Linda A; Carrier, Erica J; Gaskill, Christa; Scott, Edward W; Majka, Susan M; Fessel, Joshua P; Haase, Volker H; West, James D; Blackwell, Timothy S; Lawson, William E

    2016-02-01

    Pulmonary hypertension (PH) complicating chronic parenchymal lung disease, such as idiopathic pulmonary fibrosis, results in significant morbidity and mortality. Since the hypoxia-inducible factor (HIF) signaling pathway is important for development of pulmonary hypertension in chronic hypoxia, we investigated whether HIF signaling in vascular endothelium regulates development of PH related to pulmonary fibrosis. We generated a transgenic model in which HIF is deleted within vascular endothelial cells and then exposed these mice to chronic intraperitoneal bleomycin to induce PH associated with lung fibrosis. Although no differences in the degree of fibrotic remodeling were observed, we found that endothelial HIF-deficient mice were protected against development of PH, including right ventricle and pulmonary vessel remodeling. Similarly, endothelial HIF-deficient mice were protected from PH after a 4-wk exposure to normobaric hypoxia. In vitro studies of pulmonary vascular endothelial cells isolated from the HIF-targeted mice and controls revealed that endothelial HIF signaling increases endothelial cell expression of connective tissue growth factor, enhances vascular permeability, and promotes pulmonary artery smooth muscle cell proliferation and wound healing ability, all of which have the potential to impact the development of PH in vivo. Taken together, these studies demonstrate that vascular endothelial cell HIF signaling is necessary for development of hypoxia and pulmonary fibrosis associated PH. As such, HIF and HIF-regulated targets represent a therapeutic target in these conditions.

  18. The relationship between factor inhibiting HIF-1-alpha (HIF1AN( expression and vascular invasion in colon cancer

    Directory of Open Access Journals (Sweden)

    Reza Najafipour

    2016-10-01

    Full Text Available Background: Hypoxia is a common phenomenon in human solid tumors which by increasing in angiogenesis induction cause tumor growth survival and metastasis. Inhibitory factor hypoxia regulatory factor (HIF1AN by binding to transcription co activators CBP/P300(, inhibits hypoxia inducible factor (HIF1α. Objective: The relationship between HIF1AN expression and vascular invasion in colon tumors. Methods: The study included 101 patients with colon adenocarcinoma which were divided to vascular invasion and non-vascular invasion groups. Tumor paraffin blocks were immunohistochemistry stained for HIF1AN and were assessed for intensity and extent of positivity. Statistical relation of marker expression and clinic pathologic findings were assessed. Data were analyzed by SPSS 21 software and logistic regression and chi-square test. Findings: Nuclear immunoreactivity of HIF1AN was different between two groups. Statistical relation between low HIF1AN expression and tumor vascular invasion were seen (P=0.01. No relation was found between tumor differentiation, depth and HIF1AN. Conclusion: Evidence showed that the low expression or incorrect position of HIF1AN in nucleus of tumor cells was effective on HIF1α inhibition failure and factors associated angiogenesis increased. The HIF1AN played an tumor suppressor gene (TSG( role in colon tumors and decreased protein in the nucleus of colon cancer cells increased the expression of angiogenesis factors and vascular invasion.

  19. HIF-1alpha and HIF-2alpha are differentially activated in distinct cell populations in retinal ischaemia.

    Directory of Open Access Journals (Sweden)

    Freya M Mowat

    Full Text Available BACKGROUND: Hypoxia plays a key role in ischaemic and neovascular disorders of the retina. Cellular responses to oxygen are mediated by hypoxia-inducible transcription factors (HIFs that are stabilised in hypoxia and induce the expression of a diverse range of genes. The purpose of this study was to define the cellular specificities of HIF-1alpha and HIF-2alpha in retinal ischaemia, and to determine their correlation with the pattern of retinal hypoxia and the expression profiles of induced molecular mediators. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the tissue distribution of retinal hypoxia during oxygen-induced retinopathy (OIR in mice using the bio-reductive drug pimonidazole. We measured the levels of HIF-1alpha and HIF-2alpha proteins by Western blotting and determined their cellular distribution by immunohistochemistry during the development of OIR. We measured the temporal expression profiles of two downstream mediators, vascular endothelial growth factor (VEGF and erythropoietin (Epo by ELISA. Pimonidazole labelling was evident specifically in the inner retina. Labelling peaked at 2 hours after the onset of hypoxia and gradually declined thereafter. Marked binding to Müller glia was evident during the early hypoxic stages of OIR. Both HIF-1alpha and HIF-2alpha protein levels were significantly increased during retinal hypoxia but were evident in distinct cellular distributions; HIF-1alpha stabilisation was evident in neuronal cells throughout the inner retinal layers whereas HIF-2alpha was restricted to Müller glia and astrocytes. Hypoxia and HIF-alpha stabilisation in the retina were closely followed by upregulated expression of the downstream mediators VEGF and EPO. CONCLUSIONS/SIGNIFICANCE: Both HIF-1alpha and HIF-2alpha are activated in close correlation with retinal hypoxia but have contrasting cell specificities, consistent with differential roles in retinal ischaemia. Our findings suggest that HIF-2alpha activation

  20. Analysis list: Hif1a [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Hif1a Blood,Embryo + mm9 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/Hif1...a.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/target/Hif1a.5.tsv http://dbarchive.biosciencedbc.jp/...kyushu-u/mm9/target/Hif1a.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/Hif1a.Blood.tsv,http://dbarchive.bioscience...dbc.jp/kyushu-u/mm9/colo/Hif1a.Embryo.tsv http://dbarchive.bioscience...dbc.jp/kyushu-u/mm9/colo/Blood.gml,http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/colo/Embryo.gml ...

  1. DNA methylation and gene expression of HIF3A

    DEFF Research Database (Denmark)

    Main, Ailsa Maria; Gillberg, Linn; Jacobsen, Anna Louisa;

    2016-01-01

    from 48 families, from whom we had SAT and muscle biopsies. DNA methylation of four CpG sites in the HIF3A promoter was analyzed in the blood and SAT by pyrosequencing, and HIF3A gene expression was analyzed in SAT and muscle by qPCR. An index of whole-body insulin sensitivity was estimated from oral....... Interestingly, HIF3A expression in SAT, but not in muscle, associated negatively with BMI and whole-body insulin resistance. We found a significant effect of familiality on HIF3A methylation levels in the blood and HIF3A expression levels in skeletal muscle. CONCLUSIONS: Our findings are in line...... individuals, and whether HIF3A gene expression in SAT and skeletal muscle biopsies showed associations with BMI and insulin resistance. Furthermore, we aimed to investigate gender specificity and heritability of these traits. METHODS: We studied 137 first-degree relatives of type 2 diabetes (T2D) patients...

  2. STAT3 or USF2 Contributes to HIF Target Gene Specificity

    Science.gov (United States)

    Pawlus, Matthew R.; Wang, Liyi; Murakami, Aya; Dai, Guanhai; Hu, Cheng-Jun

    2013-01-01

    The HIF1- and HIF2-mediated transcriptional responses play critical roles in solid tumor progression. Despite significant similarities, including their binding to promoters of both HIF1 and HIF2 target genes, HIF1 and HIF2 proteins activate unique subsets of target genes under hypoxia. The mechanism for HIF target gene specificity has remained unclear. Using siRNA or inhibitor, we previously reported that STAT3 or USF2 is specifically required for activation of endogenous HIF1 or HIF2 target genes. In this study, using reporter gene assays and chromatin immuno-precipitation, we find that STAT3 or USF2 exhibits specific binding to the promoters of HIF1 or HIF2 target genes respectively even when over-expressed. Functionally, HIF1α interacts with STAT3 to activate HIF1 target gene promoters in a HIF1α HLH/PAS and N-TAD dependent manner while HIF2α interacts with USF2 to activate HIF2 target gene promoters in a HIF2α N-TAD dependent manner. Physically, HIF1α HLH and PAS domains are required for its interaction with STAT3 while both N- and C-TADs of HIF2α are involved in physical interaction with USF2. Importantly, addition of functional USF2 binding sites into a HIF1 target gene promoter increases the basal activity of the promoter as well as its response to HIF2+USF2 activation while replacing HIF binding site with HBS from a HIF2 target gene does not change the specificity of the reporter gene. Importantly, RNA Pol II on HIF1 or HIF2 target genes is primarily associated with HIF1α or HIF2α in a STAT3 or USF2 dependent manner. Thus, we demonstrate here for the first time that HIF target gene specificity is achieved by HIF transcription partners that are required for HIF target gene activation, exhibit specific binding to the promoters of HIF1 or HIF2 target genes and selectively interact with HIF1α or HIF2α protein. PMID:23991099

  3. Inhibition of hypoxia-inducible factor (HIF) hydroxylases by citric acid cycle intermediates: possible links between cell metabolism and stabilization of HIF

    National Research Council Canada - National Science Library

    Koivunen, Peppi; Hirsilä, Maija; Remes, Anne M; Hassinen, Ilmo E; Kivirikko, Kari I; Myllyharju, Johanna

    ... (HIF-P4Hs) and one HIF asparaginyl hydroxylase (FIH). We have studied possible links between metabolic pathways and HIF hydroxylases by analyzing the abilities of citric acid cycle intermediates to inhibit purified human HIF-P4Hs and FIH...

  4. Intracellular ascorbate enhances hypoxia-inducible factor (HIF)-hydroxylase activity and preferentially suppresses the HIF-1 transcriptional response.

    Science.gov (United States)

    Kuiper, Caroline; Dachs, Gabi U; Currie, Margaret J; Vissers, Margreet C M

    2014-04-01

    Hypoxia-inducible factor (HIF)-1 drives the transcription of hundreds of genes to support cell survival under conditions of microenvironmental and metabolic stress. HIF-1 is downregulated by iron-containing 2-oxoglutarate-dependent enzymes that require ascorbate as a cofactor. The HIF hydroxylases control both protein stability and the formation of an active transcription complex and, consequently, ascorbate could affect HIF-1α stabilization and/or gene expression, but the relative effect of ascorbate on these separate processes has not been well characterized. In this study we examined the effects of known intracellular ascorbate concentrations on both processes in response to various means of hydroxylase inhibition, including CoCl2, NiCl2, desferrioxamine, dimethyloxalylglycine, and hypoxia. Ascorbate inhibited HIF-1 activity most dramatically with all mechanisms of iron competition. In addition, HIF-1-dependent gene expression was effectively prevented by ascorbate and was inhibited even under conditions that allowed HIF-1α protein stabilization. This suggests that (1) ascorbate acts primarily to stabilize and reduce the iron atom in the hydroxylase active site and (2) the asparagine hydroxylase controlling HIF-1 transcriptional activity is particularly susceptible to fluctuations in intracellular ascorbate. These findings suggest that ascorbate plays a significant role in supporting HIF-hydroxylase function and that it could thereby modulate the cell survival response.

  5. Differential HIF-1α and HIF-2α Expression in Mammary Epithelial Cells during Fat Pad Invasion, Lactation, and Involution.

    Directory of Open Access Journals (Sweden)

    Sven Påhlman

    Full Text Available The development and functional cycle of the mammary gland involves a number of processes that are caricatured by breast cancer cells during invasion and metastasis. Expression of the hypoxia-inducible transcription factors HIF-1 and HIF-2 has been associated with metastatic, poor prognosis, and high-grade breast cancers. Since hypoxia affects normal epithelial differentiation, we hypothesise that HIFs are important for normal breast epithelial development and regeneration as well as cancer initiation and progression. Here, we investigated the expression of the oxygen-sensitive HIF-alpha subunits during mouse mammary gland development, lactation, and involution. In breast epithelial cells, HIF-1α was expressed during early development, prior to cell polarisation. In contrast, expression of HIF-2α occurred later and was restricted to a subpopulation of luminal epithelial cells in the lactating gland. Mammary gland involution is a developmental stage that involves extensive tissue remodelling with cell death but survival of tissue stem/progenitor cells. At this stage, HIF-2α, but little HIF-1α, was expressed in CK14-positive epithelial cells. The temporal but differential expression of the HIF-alpha subunits during the mammary gland life cycle indicates that their expression is controlled by additional factors to hypoxia. Further functional studies of the roles of these proteins in the mammary gland and breast cancer are warranted.

  6. Discovery of Indenopyrazoles as a New Class of Hypoxia Inducible Factor (HIF)-1 Inhibitors

    Science.gov (United States)

    2013-01-01

    The indenopyrazole framework was investigated as a new class of HIF-1α inhibitors. Indenopyrazole 2l was found to most strongly inhibit the hypoxia-induced HIF-1α transcriptional activity (IC50 = 0.014 μM) among all of the known compounds having relatively simple structures, unlike manassantins. Indenopyrazole 2l suppressed HIF-1α transcriptional activity without affecting both HIF-1α protein accumulation and HIF-1α/HIF-1β heterodimerization in nuclei under the hypoxic conditions, suggesting that 2l probably affected the transcriptional pathway induced by the HIF-1α/HIF-1β heterodimer. PMID:24900662

  7. The validity of the SF-36 in an Australian National Household Survey: demonstrating the applicability of the Household Income and Labour Dynamics in Australia (HILDA Survey to examination of health inequalities

    Directory of Open Access Journals (Sweden)

    Crosier Timothy

    2004-10-01

    Full Text Available Abstract Background The SF-36 is one of the most widely used self-completion measures of health status. The inclusion of the SF-36 in the first Australian national household panel survey, the Household, Income and Labour Dynamics in Australia (HILDA Survey, provides an opportunity to investigate health inequalities. In this analysis we establish the psychometric properties and criterion validity of the SF-36 HILDA Survey data and examine scale profiles across a range of measures of socio-economic circumstance. Methods Data from 13,055 respondents who completed the first wave of the HILDA Survey were analysed to determine the psychometric properties of the SF-36 and the relationship of the SF-36 scales to other measures of health, disability, social functioning and demographic characteristics. Results Results of principle components analysis were similar to previous Australian and international reports. Survey scales demonstrated convergent and divergent validity, and different markers of social status demonstrated unique patterns of outcomes across the scales. Conclusion Results demonstrated the validity of the SF-36 data collected during the first wave of the HILDA Survey and support its use in research examining health inequalities and population health characteristics in Australia.

  8. On Archetypical Images in Hilda Doolittle' s Oread%杜丽特尔·希尔达《山林女神》中意象的原型视角阐释

    Institute of Scientific and Technical Information of China (English)

    黄艳芳

    2012-01-01

    This thesis tries to study the images and their characteristics in Hilda Doolittle's poem Oread from the perspective of archetype.%本论文尝试从原型的角度对杜立特尔·希尔达的《山林女神》中的意象进行研究。

  9. Analysis list: HIF1A [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available HIF1A Blood,Breast + hg19 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/target/HI...F1A.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/target/HIF1A.5.tsv http://dbarchive.biosciencedbc....jp/kyushu-u/hg19/target/HIF1A.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/HIF1A.Blood.tsv,http://dbarchive.bioscience...dbc.jp/kyushu-u/hg19/colo/HIF1A.Breast.tsv http://dbarchive.bioscience...dbc.jp/kyushu-u/hg19/colo/Blood.gml,http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/Breast.gml ...

  10. NAS TRAMAS DO “SEU DEUS”: CONSIDERAÇÕES ACERCA DA POÉTICA VITIMOLÓGICA DE HILDA HILST

    OpenAIRE

    Sampaio, Higor Alberto; UNESP

    2015-01-01

    Esta reflexão tem por objetivo perscrutar um dos temas caros à poesia de Hilda Hilst: a representação do Sagrado, no intuito de estabelecer um conhecimento acerca da construção da subjetividade  presente nessa poesia. O que me interessa aqui é a importância do papel da vítima para a construção de um espaço sagrado, e a leitura de quatro poemas da autora evidenciou que o conhecimento de uma certa “ideia de Deus”, tramada no discurso poético, acaba por delinear um universo vitimológico, erótico...

  11. A impossibilidade de se dizer o indizível: reflexões sobre o duplo na novela “O unicórnio”, de Hilda Hilst

    Directory of Open Access Journals (Sweden)

    Willian André

    2014-01-01

    Full Text Available This study aims at reflecting on the raise of the double in the novella “O unicórnio”, by Hilda Hilst, as an evidence of one’s impossibility to express what cannot be expressed. Echoing Gregor Samsa’s uncanny experience, the novella’s narrator suddenly becomes an awkward, absurd creature. A double of the narrator, this new “I” she becomes makes explicit the devious relation, built throughout the text, between a deep necessity of comprehending the world and a world that is closed in its silence, denying to be comprehended. Interpreting the raise of this double-unicorn as the main evidence of this confront, we hope to draw, in the following lines, some reflections on the limits of the language in face of the unspeakable.

  12. Homology modeling and molecular dynamics simulation of the HIF2α degradation-related HIF2α-VHL complex.

    Science.gov (United States)

    Dong, Xiaotian; Su, Xiaoru; Yu, Jiong; Liu, Jingqi; Shi, Xiaowei; Pan, Qiaoling; Yang, Jinfeng; Chen, Jiajia; Li, Lanjuan; Cao, Hongcui

    2017-01-01

    Hypoxia-inducible factor 2 alpha (HIF2α), prolyl hydroxylase domain protein 2 (PHD2), and the von Hippel Lindau tumor suppressor protein (pVHL) are three principal proteins in the oxygen-sensing pathway. Under normoxic conditions, a conserved proline in HIF2α is hydroxylated by PHD2 in an oxygen-dependent manner, and then pVHL binds and promotes the degradation of HIF2α. However, the crystal structure of the HIF2α-pVHL complex has not yet been established, and this has limited research on the interaction between HIF and pVHL. Here, we constructed a structural model of a 23-residue HIF2α peptide (528-550)-pVHL-ElonginB-ElonginC complex by using homology modeling and molecular dynamics simulations. We also applied these methods to HIF2α mutants (HYP531PRO, F540L, A530 V, A530T, and G537R) to reveal structural defects that explain how these mutations weaken the interaction with pVHL. Homology modeling and molecular dynamics simulations were used to construct a three-dimensional (3D) structural model of the HIF2α-VHL complex. Subsequently, MolProbity, an active validation tool, was used to analyze the reliability of the model. Molecular mechanics energies combined with the generalized Born and surface area continuum solvation (MM-GBSA) and solvated interaction energy (SIE) methods were used to calculate the binding free energy between HIF2a and pVHL, and the stability of the simulation system was evaluated by using root mean square deviation (RMSD) analysis. We also determined the secondary structure of the system by using the definition of secondary structure of proteins (DSSP) algorithm. Finally, we investigated the structural significance of specific point mutations known to have clinical implications. We established a reliable structural model of the HIF2α-pVHL complex, which is similar to the crystal structure of HIF1α in 1LQB. Furthermore, we compared the structural model of the HIF2α-pVHL complex and the HIF2α (HYP531P, F540L, A530V, A530T, and G537

  13. HIF hydroxylase pathways in cardiovascular physiology and medicine.

    Science.gov (United States)

    Bishop, Tammie; Ratcliffe, Peter J

    2015-06-19

    Hypoxia inducible factors (HIFs) are α/β heterodimeric transcription factors that direct multiple cellular and systemic responses in response to changes in oxygen availability. The oxygen sensitive signal is generated by a series of iron and 2-oxoglutarate-dependent dioxygenases that catalyze post-translational hydroxylation of specific prolyl and asparaginyl residues in HIFα subunits and thereby promote their destruction and inactivation in the presence of oxygen. In hypoxia, these processes are suppressed allowing HIF to activate a massive transcriptional cascade. Elucidation of these pathways has opened several new fields of cardiovascular research. Here, we review the role of HIF hydroxylase pathways in cardiac development and in cardiovascular control. We also consider the current status, opportunities, and challenges of therapeutic modulation of HIF hydroxylases in the therapy of cardiovascular disease.

  14. HIF Signaling Pathway in Pheochromocytoma and Other Neuroendocrine Tumors

    National Research Council Canada - National Science Library

    I Jochmanová; T Zelinka; J Widimský Jr; K Pacak

    2014-01-01

    .... Dysregulation of these proteins contributes to tumorigenesis and cancer progression. Recent findings revealed the important role of HIFs in the pathogenesis of neuroendocrine tumors, especially pheochromocytoma (PHEO) and paraganglioma (PGL...

  15. Evolutionary conserved regulation of HIF-1β by NF-κB.

    Science.gov (United States)

    van Uden, Patrick; Kenneth, Niall S; Webster, Ryan; Müller, H Arno; Mudie, Sharon; Rocha, Sonia

    2011-01-27

    Hypoxia Inducible Factor-1 (HIF-1) is essential for mammalian development and is the principal transcription factor activated by low oxygen tensions. HIF-α subunit quantities and their associated activity are regulated in a post-translational manner, through the concerted action of a class of enzymes called Prolyl Hydroxylases (PHDs) and Factor Inhibiting HIF (FIH) respectively. However, alternative modes of HIF-α regulation such as translation or transcription are under-investigated, and their importance has not been firmly established. Here, we demonstrate that NF-κB regulates the HIF pathway in a significant and evolutionary conserved manner. We demonstrate that NF-κB directly regulates HIF-1β mRNA and protein. In addition, we found that NF-κB-mediated changes in HIF-1β result in modulation of HIF-2α protein. HIF-1β overexpression can rescue HIF-2α protein levels following NF-κB depletion. Significantly, NF-κB regulates HIF-1β (tango) and HIF-α (sima) levels and activity (Hph/fatiga, ImpL3/ldha) in Drosophila, both in normoxia and hypoxia, indicating an evolutionary conserved mode of regulation. These results reveal a novel mechanism of HIF regulation, with impact in the development of novel therapeutic strategies for HIF-related pathologies including ageing, ischemia, and cancer.

  16. HIF1α protein stability is increased by acetylation at lysine 709.

    Science.gov (United States)

    Geng, Hao; Liu, Qiong; Xue, Changhui; David, Larry L; Beer, Tomasz M; Thomas, George V; Dai, Mu-Shui; Qian, David Z

    2012-10-12

    Lysine acetylation regulates protein stability and function. p300 is a component of the HIF-1 transcriptional complex and positively regulates the transactivation of HIF-1. Here, we show a novel molecular mechanism by which p300 facilitates HIF-1 activity. p300 increases HIF-1α (HIF1α) protein acetylation and stability. The regulation can be opposed by HDAC1, but not by HDAC3, and is abrogated by disrupting HIF1α-p300 interaction. Mechanistically, p300 specifically acetylates HIF1α at Lys-709, which increases the protein stability and decreases polyubiquitination in both normoxia and hypoxia. Compared with the wild-type protein, a HIF1α K709A mutant protein is more stable, less polyubiquitinated, and less dependent on p300. Overexpression of the HIF1α wild-type or K709A mutant in cancer cells lacking the endogenous HIF1α shows that the K709A mutant is transcriptionally more active toward the HIF-1 reporter and some endogenous target genes. Cancer cells containing the K709A mutant are less sensitive to hypoxia-induced growth arrest than the cells containing the HIF1α wild-type. Taken together, these data demonstrate a novel biological consequence upon HIF1α-p300 interaction, in which HIF1α can be stabilized by p300 via Lys-709 acetylation.

  17. Adult hematopoietic stem cells lacking Hif-1α self-renew normally.

    Science.gov (United States)

    Vukovic, Milica; Sepulveda, Catarina; Subramani, Chithra; Guitart, Amélie V; Mohr, Jasmine; Allen, Lewis; Panagopoulou, Theano I; Paris, Jasmin; Lawson, Hannah; Villacreces, Arnaud; Armesilla-Diaz, Alejandro; Gezer, Deniz; Holyoake, Tessa L; Ratcliffe, Peter J; Kranc, Kamil R

    2016-06-09

    The hematopoietic stem cell (HSC) pool is maintained under hypoxic conditions within the bone marrow microenvironment. Cellular responses to hypoxia are largely mediated by the hypoxia-inducible factors, Hif-1 and Hif-2. The oxygen-regulated α subunits of Hif-1 and Hif-2 (namely, Hif-1α and Hif-2α) form dimers with their stably expressed β subunits and control the transcription of downstream hypoxia-responsive genes to facilitate adaptation to low oxygen tension. An initial study concluded that Hif-1α is essential for HSC maintenance, whereby Hif-1α-deficient HSCs lost their ability to self-renew in serial transplantation assays. In another study, we demonstrated that Hif-2α is dispensable for cell-autonomous HSC maintenance, both under steady-state conditions and following transplantation. Given these unexpected findings, we set out to revisit the role of Hif-1α in cell-autonomous HSC functions. Here we demonstrate that inducible acute deletion of Hif-1α has no impact on HSC survival. Notably, unstressed HSCs lacking Hif-1α efficiently self-renew and sustain long-term multilineage hematopoiesis upon serial transplantation. Finally, Hif-1α-deficient HSCs recover normally after hematopoietic injury induced by serial administration of 5-fluorouracil. We therefore conclude that despite the hypoxic nature of the bone marrow microenvironment, Hif-1α is dispensable for cell-autonomous HSC maintenance.

  18. Evolutionary conserved regulation of HIF-1β by NF-κB.

    Directory of Open Access Journals (Sweden)

    Patrick van Uden

    Full Text Available Hypoxia Inducible Factor-1 (HIF-1 is essential for mammalian development and is the principal transcription factor activated by low oxygen tensions. HIF-α subunit quantities and their associated activity are regulated in a post-translational manner, through the concerted action of a class of enzymes called Prolyl Hydroxylases (PHDs and Factor Inhibiting HIF (FIH respectively. However, alternative modes of HIF-α regulation such as translation or transcription are under-investigated, and their importance has not been firmly established. Here, we demonstrate that NF-κB regulates the HIF pathway in a significant and evolutionary conserved manner. We demonstrate that NF-κB directly regulates HIF-1β mRNA and protein. In addition, we found that NF-κB-mediated changes in HIF-1β result in modulation of HIF-2α protein. HIF-1β overexpression can rescue HIF-2α protein levels following NF-κB depletion. Significantly, NF-κB regulates HIF-1β (tango and HIF-α (sima levels and activity (Hph/fatiga, ImpL3/ldha in Drosophila, both in normoxia and hypoxia, indicating an evolutionary conserved mode of regulation. These results reveal a novel mechanism of HIF regulation, with impact in the development of novel therapeutic strategies for HIF-related pathologies including ageing, ischemia, and cancer.

  19. Basal HIF-1a expression levels are not predictive for radiosensitivity of human cancer cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Schilling, D.; Multhoff, G. [Klinikum rechts der Isar der Technischen Univ. Muenchen (Germany). Dept. of Radiation Oncology; Helmholtz Center Munich, CCG - Innate Immunity in Tumor Biology, Munich (Germany). German Research Center for Environmental Health - Inst. of Pathology; Bayer, C.; Emmerich, K.; Molls, M.; Vaupel, P. [Klinikum rechts der Isar der Technischen Univ. Muenchen (Germany). Dept. of Radiation Oncology; Huber, R.M. [Klinikum der Univ. Muenchen (Germany). Dept. of Pneumology

    2012-04-15

    High levels of hypoxia inducible factor (HIF)-1a in tumors are reported to be associated with tumor progression and resistance to therapy. To examine the impact of HIF-1a on radioresistance under normoxia, the sensitivity towards irradiation was measured in human tumor cell lines that differ significantly in their basal HIF-1a levels. HIF-1a levels were quantified in lysates of H1339, EPLC-272H, A549, SAS, XF354, FaDu, BHY, and CX- tumor cell lines by ELISA. Protein levels of HIF-1a, HIF-2a, carbonic anhydrase IX (CA IX), and GAPDH were assessed by Western blot analysis. Knock-down experiments were performed using HIF-1a siRNA. Clonogenic survival after irradiation was determined by the colony forming assay. According to their basal HIF-1a status, the tumor cell lines were divided into low (SAS, XF354, FaDu, A549, CX-), intermediate (EPLC-272H, BHY), and high (H1339) HIF-1a expressors. The functionality of the high basal HIF-1a expression in H1339 cells was proven by reduced CA IX expression after knocking-down HIF-1a. Linear regression analysis revealed no correlation between basal HIF-1a levels and the survival fraction at either 2 or 4 Gy in all tumor cell lines investigated. Our data suggest that basal HIF-1a levels in human tumor cell lines do not predict their radiosensitivity under normoxia. (orig.)

  20. Development of Superconducting Focusing Quadrupoles for Heavy Ion Drivers

    Energy Technology Data Exchange (ETDEWEB)

    Martovetsky, N; Manahan, R; Lietzke, A F

    2001-09-10

    Heavy Ion Fusion (HIF) is exploring a promising path to a practical inertial-confinement fusion reactor. The associated heavy ion driver will require a large number of focusing quadrupole magnets. A concept for a superconducting quadrupole array, using many simple racetrack coils, was developed at LLNL. Two, single-bore quadrupole prototypes of the same design, with distinctly different conductor, were designed, built, and tested. Both prototypes reached their short sample currents with little or no training. Magnet design, and test results, are presented and discussed.

  1. The role of factor inhibiting HIF (FIH-1 in inhibiting HIF-1 transcriptional activity in glioblastoma multiforme.

    Directory of Open Access Journals (Sweden)

    Enfeng Wang

    Full Text Available Glioblastoma multiforme (GBM accounts for about 38% of primary brain tumors in the United States. GBM is characterized by extensive angiogenesis induced by vascular growth factors and cytokines. The transcription of these growth factors and cytokines is regulated by the Hypoxia-Inducible-Factor-1(HIF-1, which is a key regulator mediating the cellular response to hypoxia. It is known that Factor Inhibiting HIF-1, or FIH-1, is also involved in the cellular response to hypoxia and has the capability to physically interact with HIF-1 and block its transcriptional activity under normoxic conditions. Delineation of the regulatory role of FIH-1 will help us to better understand the molecular mechanism responsible for tumor growth and progression and may lead to the design of new therapies targeting cellular pathways in response to hypoxia. Previous studies have shown that the chromosomal region of 10q24 containing the FIH-1 gene is often deleted in GBM, suggesting a role for the FIH-1 in GBM tumorigenesis and progression. In the current study, we found that FIH-1 is able to inhibit HIF-mediated transcription of GLUT1 and VEGF-A, even under hypoxic conditions in human glioblastoma cells. FIH-1 has been found to be more potent in inhibiting HIF function than PTEN. This observation points to the possibility that deletion of 10q23-24 and loss or decreased expression of FIH-1 gene may lead to a constitutive activation of HIF-1 activity, an alteration of HIF-1 targets such as GLUT-1 and VEGF-A, and may contribute to the survival of cancer cells in hypoxia and the development of hypervascularization observed in GBM. Therefore FIH-1 can be potential therapeutic target for the treatment of GBM patients with poor prognosis.

  2. Upregulation of the Chemokine Receptor CCR7 expression by HIF-1αand HIF-2α in non-small cell lung cancer

    Directory of Open Access Journals (Sweden)

    Yang LI

    2008-10-01

    Full Text Available Background and objective CCR7 is closely related with the lymph node metastasis of non-small cell lung cancer. The objective of this work is to investigate the expressions of chemokine receptor CCR7, hypoxiainducible factor 1α (HIF-1α and hypoxia inducible factor 2α (HIF-2α protein in non small cell lung cancer and the relationships of their expression, and to study the mechanism of CCR7 upregulation in NSCLC. Methods T he levels of expressions of CCR7, HIF-1α and HIF-2α protein were detected in 94 specimens of human primary non small cell lung cancer by immunohistochemical S-P method. Human lung adenocarcinoma cell line A549 cells were transfected by lipofection with HIF-1α siRNA、HIF-2α siRNA, the change of CCR7 was observed by RT-PCR and immunofluorescence staining. Correlations between the expression of CCR7 and HIF-1α, HIF-2α were respectively analyzed. Results Immunohistochemistry showed that CCR7 was distributed in cytoplasm and/or membrane of tumor cells, HIF-1α, HIF-2α was distributed in nucleus and/or cytoplasm of tumor cells. The levels of expressions of CCR7, HIF-1α and HIF-2α protein were found to be 75.53% (71/94, 54.25% (51/ 94 and 70.21% (66/94 in non small celllung cancer, respectively. the levels of expression of CCR7 protein were closely related to the clinical stages (P 0.05. Furthermore, A significant correlation were found among CCR7, Hif-1α and HIF-2α (r =0.272, P <0.01 (r=0.225, P <0.05. In addition, the expression of CCR7 mRNA and protein levels were decreased in the transfected specificHIF-1α, HIF-2αsiRNA group (P <0.05. Conclusion CCR7 expression is significantly associated with non small cell lung cancer invasion and metastasis. The upregulation of CCR7 is regulated by HIF-1α and HIF-2α in non small cell lung cancer.

  3. Design of 3x3 focusing array for heavy ion driver. Final report on CRADA

    Energy Technology Data Exchange (ETDEWEB)

    Martovetsky, N N

    2005-03-30

    This memo presents a design of a 3 x 3 quadrupole array for HIF. It contains 3 D magnetic field computations of the array build with racetrack coils with and without different shields. It is shown that it is possible to have a low error magnetic field in the cells and shield the stray fields to acceptable levels. The array design seems to be a practical solution to any size array for future multi-beam heavy ion fusion drivers.

  4. EPIDERMAL DELETION OF HIF-2α STIMULATES WOUND CLOSURE

    Science.gov (United States)

    Cowburn, Andrew S.; Crotty Alexander, Laura E.; Southwood, Mark; Nizet, Victor; Chilvers, Edwin R.; Johnson, Randall S.

    2013-01-01

    Wound closure requires a complex series of micro-environmentally influenced events. A key aspect of wound closure is the migration of keratinocytes across the open wound. It has been found previously that the response to hypoxia via the HIF-1α transcription factor is a key feature of wound closure. The need for hypoxic response is likely due to interrupted wound vasculature as well as infection, and in this work, we investigated the need for a highly related hypoxic response transcription factor, HIF-2α. This factor was deleted tissue-specifically in mice, and the resulting mice were found to have an accelerated rate of wound closure. This is correlated with a reduced bacterial load and inflammatory response in these mice. This indicates that manipulating or reducing the HIF-2α response in keratinocytes could be a useful means to accelerate wound healing and tissue repair. PMID:24037341

  5. Cytoplasmic polyadenylation-element-binding protein (CPEB)1 and 2 bind to the HIF-1alpha mRNA 3'-UTR and modulate HIF-1alpha protein expression.

    Science.gov (United States)

    Hägele, Sonja; Kühn, Uwe; Böning, Melanie; Katschinski, Dörthe M

    2009-01-01

    The heterodimeric HIF (hypoxia-inducible factor)-1 is a transcriptional master regulator of several genes involved in mammalian oxygen homoeostasis. Besides the well described regulation of the HIF-1alpha subunit via hydroxylation-mediated protein stability in hypoxia, there are several indications of an additional translational control of the HIF-1alpha mRNA, especially after growth factor stimulation. We identified an interaction of CPEB (cytoplasmic polyadenylation-element-binding protein) 1 and CPEB2 with the 3'-UTR (untranslated region) of HIF-1alpha mRNA. Overexpression of CPEB1 and CPEB2 affected HIF-1alpha protein levels mediated by the 3'-UTR of HIF-1alpha mRNA. Stimulation of neuroblastoma SK-N-MC cells with insulin and thus activation of endogenous CPEBs increased the expression of a luciferase reporter gene fused to the 3'-UTR of HIF-1alpha as well as endogenous HIF-1alpha protein levels. This could be abrogated by treating the cells with CPEB1 or CPEB2 siRNAs (short interfering RNAs). Injection of HIF-1alpha cRNA into Xenopus oocytes verified the elongation of the poly(A)+ (polyadenylated) tail by cytoplasmic polyadenylation. Thus CPEB1 and CPEB2 are involved in the regulation of HIF-1alpha following insulin stimulation.

  6. Inhibition of hypoxia-inducible factor (HIF) hydroxylases by citric acid cycle intermediates: possible links between cell metabolism and stabilization of HIF.

    Science.gov (United States)

    Koivunen, Peppi; Hirsilä, Maija; Remes, Anne M; Hassinen, Ilmo E; Kivirikko, Kari I; Myllyharju, Johanna

    2007-02-16

    The stability and transcriptional activity of the hypoxia-inducible factors (HIFs) are regulated by two oxygen-dependent events that are catalyzed by three HIF prolyl 4-hydroxylases (HIF-P4Hs) and one HIF asparaginyl hydroxylase (FIH). We have studied possible links between metabolic pathways and HIF hydroxylases by analyzing the abilities of citric acid cycle intermediates to inhibit purified human HIF-P4Hs and FIH. Fumarate and succinate were identified as in vitro inhibitors of all three HIF-P4Hs, fumarate having K(i) values of 50-80 microM and succinate 350-460 microM, whereas neither inhibited FIH. Oxaloacetate was an additional inhibitor of all three HIF-P4Hs with K(i) values of 400-1000 microM and citrate of HIF-P4H-3, citrate being the most effective inhibitor of FIH with a K(i) of 110 microM. Culturing of cells with fumarate diethyl or dimethyl ester, or a high concentration of monoethyl ester, stabilized HIF-1alpha and increased production of vascular endothelial growth factor and erythropoietin. Similar, although much smaller, changes were found in cultured fibroblasts from a patient with fumarate hydratase (FH) deficiency and upon silencing FH using small interfering RNA. No such effects were seen upon culturing of cells with succinate diethyl or dimethyl ester. As FIH was not inhibited by fumarate, our data indicate that the transcriptional activity of HIF is quite high even when binding of the coactivator p300 is prevented. Our data also support recent suggestions that the increased fumarate and succinate levels present in the FH and succinate dehydrogenase-deficient tumors, respectively, can inhibit the HIF-P4Hs with consequent stabilization of HIF-alphas and effects on tumor pathology.

  7. Oncogenic kinase NPM/ALK induces expression of HIF1a mRNA

    DEFF Research Database (Denmark)

    Marzec, M; Liu, X; Wong, W;

    2011-01-01

    to the HIF1a gene promoter as shown by the chromatin immunoprecipitation assay and is required for HIF1a gene expression as demonstrated by its small interfering RNA-mediated depletion. In turn, depletion of HIF1a increases mammalian target of rapamycin complex 1 activation, cell growth and proliferation...

  8. HIF2α–arginase axis is essential for the development of pulmonary hypertension

    Science.gov (United States)

    Cowburn, Andrew S.; Crosby, Alexi; Macias, David; Branco, Cristina; Colaço, Renato D. D. R.; Southwood, Mark; Toshner, Mark; Crotty Alexander, Laura E.; Morrell, Nicholas W.; Chilvers, Edwin R.; Johnson, Randall S.

    2016-01-01

    Hypoxic pulmonary vasoconstriction is correlated with pulmonary vascular remodeling. The hypoxia-inducible transcription factors (HIFs) HIF-1α and HIF-2α are known to contribute to the process of hypoxic pulmonary vascular remodeling; however, the specific role of pulmonary endothelial HIF expression in this process, and in the physiological process of vasoconstriction in response to hypoxia, remains unclear. Here we show that pulmonary endothelial HIF-2α is a critical regulator of hypoxia-induced pulmonary arterial hypertension. The rise in right ventricular systolic pressure (RVSP) normally observed following chronic hypoxic exposure was absent in mice with pulmonary endothelial HIF-2α deletion. The RVSP of mice lacking HIF-2α in pulmonary endothelium after exposure to hypoxia was not significantly different from normoxic WT mice and much lower than the RVSP values seen in WT littermate controls and mice with pulmonary endothelial deletion of HIF-1α exposed to hypoxia. Endothelial HIF-2α deletion also protected mice from hypoxia remodeling. Pulmonary endothelial deletion of arginase-1, a downstream target of HIF-2α, likewise attenuated many of the pathophysiological symptoms associated with hypoxic pulmonary hypertension. We propose a mechanism whereby chronic hypoxia enhances HIF-2α stability, which causes increased arginase expression and dysregulates normal vascular NO homeostasis. These data offer new insight into the role of pulmonary endothelial HIF-2α in regulating the pulmonary vascular response to hypoxia. PMID:27432976

  9. Melatonin and the von Hippel-Lindau/HIF-1 oxygen sensing mechanism: A review.

    Science.gov (United States)

    Vriend, Jerry; Reiter, Russel J

    2016-04-01

    There are numerous reports that melatonin inhibits the hypoxia-inducible factor, HIF-1α, and the HIF-1α-inducible gene, VEGF, both in vivo and in vitro. Through the inhibition of the HIF-1-VEGF pathway, melatonin reduces hypoxia-induced angiogenesis. Herein we discuss the interaction of melatonin with HIF-1α and HIF-1α-inducible genes in terms of what is currently known concerning the HIF-1α hypoxia response element (HIF-1α-HRE) pathway. The von Hippel-Lindau protein (VHL), also known as the VHL tumor suppressor, functions as part of a ubiquitin ligase complex which recognizes HIF-1α as a substrate. As such, VHL is part of the oxygen sensing mechanism of the cell. Under conditions of hypoxia, HIF-1α stimulates the transcription of numerous HIF-1α-induced genes, including EPO, VEGF, and PFKFB3; the latter is an enzyme which regulates glycolysis. Data from several studies show that ROS generated in mitochondria under conditions of hypoxia stimulate HIF-1α. Since melatonin acts as an antioxidant and reduces ROS, these data suggest that the antioxidant action of melatonin could account for reduced HIF-1, less VEGF, and reduced glycolysis in cancer cells (Warburg effect). A direct or indirect inhibitory action (via the reduction in ROS) of melatonin on proteasome activity would account for much of the published data.

  10. File list: Oth.Bld.05.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.05.HIF1A.AllCell hg19 TFs and others HIF1A Blood SRX212354,SRX212361,SRX212...353,SRX212360,SRX212352,SRX212351,SRX212359,SRX212362 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.05.HIF1A.AllCell.bed ...

  11. File list: Oth.ALL.05.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.05.HIF1A.AllCell hg19 TFs and others HIF1A All cell types SRX666556,SRX1576...28430 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.ALL.05.HIF1A.AllCell.bed ...

  12. File list: Oth.Bld.50.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.50.HIF1A.AllCell hg19 TFs and others HIF1A Blood SRX212353,SRX212351,SRX212...359,SRX212354,SRX212361,SRX212360,SRX212362,SRX212352 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.50.HIF1A.AllCell.bed ...

  13. File list: Oth.Bld.10.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.10.HIF1A.AllCell hg19 TFs and others HIF1A Blood SRX212354,SRX212361,SRX212...353,SRX212352,SRX212351,SRX212360,SRX212359,SRX212362 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.10.HIF1A.AllCell.bed ...

  14. File list: Oth.ALL.20.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.20.HIF1A.AllCell hg19 TFs and others HIF1A All cell types SRX157608,SRX1576...12351 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.ALL.20.HIF1A.AllCell.bed ...

  15. File list: Oth.Bld.20.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: Oth.ALL.10.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.10.HIF1A.AllCell hg19 TFs and others HIF1A All cell types SRX666556,SRX1576...28430 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.ALL.10.HIF1A.AllCell.bed ...

  17. File list: Oth.ALL.50.HIF1A.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.50.HIF1A.AllCell hg19 TFs and others HIF1A All cell types SRX157608,SRX6665...12352 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.ALL.50.HIF1A.AllCell.bed ...

  18. Driver Behavior and Motivation.

    Science.gov (United States)

    Thomas, Patricia

    School bus driver behavior and motivation are continuing concerns for leaders/administrators in the field of transportation. Motivation begins with selection of a potential new driver. Drivers must like children and be patient, loyal, and punctual. The applicant's background must be verified, in view of the national concern for child safety.…

  19. Differential contribution of key metabolic substrates and cellular oxygen in HIF signalling

    Energy Technology Data Exchange (ETDEWEB)

    Zhdanov, Alexander V., E-mail: a.zhdanov@ucc.ie [School of Biochemistry and Cell Biology, University College Cork, Cavanagh Pharmacy Building, College Road, Cork (Ireland); Waters, Alicia H.C. [School of Biochemistry and Cell Biology, University College Cork, Cavanagh Pharmacy Building, College Road, Cork (Ireland); Golubeva, Anna V. [Alimentary Pharmabiotic Centre, University College Cork, Bioscience Institute, Western Road, Cork (Ireland); Papkovsky, Dmitri B. [School of Biochemistry and Cell Biology, University College Cork, Cavanagh Pharmacy Building, College Road, Cork (Ireland)

    2015-01-01

    Changes in availability and utilisation of O{sub 2} and metabolic substrates are common in ischemia and cancer. We examined effects of substrate deprivation on HIF signalling in PC12 cells exposed to different atmospheric O{sub 2}. Upon 2–4 h moderate hypoxia, HIF-α protein levels were dictated by the availability of glutamine and glucose, essential for deep cell deoxygenation and glycolytic ATP flux. Nuclear accumulation of HIF-1α dramatically decreased upon inhibition of glutaminolysis or glutamine deprivation. Elevation of HIF-2α levels was transcription-independent and associated with the activation of Akt and Erk1/2. Upon 2 h anoxia, HIF-2α levels strongly correlated with cellular ATP, produced exclusively via glycolysis. Without glucose, HIF signalling was suppressed, giving way to other regulators of cell adaptation to energy crisis, e.g. AMPK. Consequently, viability of cells deprived of O{sub 2} and glucose decreased upon inhibition of AMPK with dorsomorphin. The capacity of cells to accumulate HIF-2α decreased after 24 h glucose deprivation. This effect, associated with increased AMPKα phosphorylation, was sensitive to dorsomorphin. In chronically hypoxic cells, glutamine played no major role in HIF-2α accumulation, which became mainly glucose-dependent. Overall, the availability of O{sub 2} and metabolic substrates intricately regulates HIF signalling by affecting cell oxygenation, ATP levels and pathways involved in production of HIF-α. - Highlights: • Gln and Glc regulate HIF levels in hypoxic cells by maintaining low O{sub 2} and high ATP. • HIF-α levels under anoxia correlate with cellular ATP and critically depend on Glc. • Gln and Glc modulate activity of Akt, Erk and AMPK, regulating HIF production. • HIF signalling is differentially inhibited by prolonged Glc and Gln deprivation. • Unlike Glc, Gln plays no major role in HIF signalling in chronically hypoxic cells.

  20. Arylsulfonamide KCN1 inhibits in vivo glioma growth and interferes with HIF signaling by disrupting HIF-1α interaction with co-factors p300/CBP

    Science.gov (United States)

    Yin, Shaoman; Kaluz, Stefan; Devi, Narra S.; Jabbar, Adnan A.; de Noronha, Rita G.; Mun, Jiyoung; Zhang, Zhaobin; Boreddy, Purushotham R.; Wang, Wei; Wang, Zhibo; Abbruscato, Thomas; Chen, Zhengjia; Olson, Jeffrey J.; Zhang, Ruiwen; Goodman, Mark M.; Nicolaou, K.C.; Van Meir, Erwin G.

    2012-01-01

    Purpose The hypoxia inducible factor-1 (HIF-1) plays a critical role in tumor adaptation to hypoxia, and its elevated expression correlates with poor prognosis and treatment failure in cancer patients. In this study, we determined whether 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide, KCN1, the lead inhibitor in a novel class of arylsulfonamide inhibitors of the HIF-1 pathway, had anti-tumorigenic properties in vivo and further defined its mechanism of action. Experimental Design We studied the inhibitory effect of systemic KCN1 delivery on the growth of human brain tumors in mice. To define mechanisms of KCN1 anti-HIF activities, we examined its influence on the assembly of a functional HIF1α/HIF1β/p300 transcription complex. Results KCN1 specifically inhibited HIF reporter gene activity in several glioma cell lines at the nanomolar level. KCN1 also downregulated transcription of endogenous HIF-1 target genes, such as VEGF, Glut-1 and carbonic anhydrase 9, in an HRE-dependent manner. KCN1 potently inhibited the growth of subcutaneous malignant glioma tumor xenografts with minimal adverse effects on the host. It also induced a temporary survival benefit in an intracranial model of glioma but had no effect in a model of melanoma metastasis to the brain. Mechanistically, KCN1 did not down-regulate levels of HIF-1α or other components of the HIF transcriptional complex; rather, it antagonized hypoxia-inducible transcription by disrupting the interaction of HIF-1α with transcriptional co-activators p300/CBP. Conclusions Our results suggest that the new HIF pathway inhibitor KCN1 has antitumor activity in mouse models, supporting its further translation for the treatment of human tumors displaying hypoxia or HIF overexpression. PMID:22923450

  1. HIF induces human embryonic stem cell markers in cancer cells.

    Science.gov (United States)

    Mathieu, Julie; Zhang, Zhan; Zhou, Wenyu; Wang, Amy J; Heddleston, John M; Pinna, Claudia M A; Hubaud, Alexis; Stadler, Bradford; Choi, Michael; Bar, Merav; Tewari, Muneesh; Liu, Alvin; Vessella, Robert; Rostomily, Robert; Born, Donald; Horwitz, Marshall; Ware, Carol; Blau, C Anthony; Cleary, Michele A; Rich, Jeremy N; Ruohola-Baker, Hannele

    2011-07-01

    Low oxygen levels have been shown to promote self-renewal in many stem cells. In tumors, hypoxia is associated with aggressive disease course and poor clinical outcomes. Furthermore, many aggressive tumors have been shown to display gene expression signatures characteristic of human embryonic stem cells (hESC). We now tested whether hypoxia might be responsible for the hESC signature observed in aggressive tumors. We show that hypoxia, through hypoxia-inducible factor (HIF), can induce an hESC-like transcriptional program, including the induced pluripotent stem cell (iPSC) inducers, OCT4, NANOG, SOX2, KLF4, cMYC, and microRNA-302 in 11 cancer cell lines (from prostate, brain, kidney, cervix, lung, colon, liver, and breast tumors). Furthermore, nondegradable forms of HIFα, combined with the traditional iPSC inducers, are highly efficient in generating A549 iPSC-like colonies that have high tumorigenic capacity. To test potential correlation between iPSC inducers and HIF expression in primary tumors, we analyzed primary prostate tumors and found a significant correlation between NANOG-, OCT4-, and HIF1α-positive regions. Furthermore, NANOG and OCT4 expressions positively correlated with increased prostate tumor Gleason score. In primary glioma-derived CD133 negative cells, hypoxia was able to induce neurospheres and hESC markers. Together, these findings suggest that HIF targets may act as key inducers of a dynamic state of stemness in pathologic conditions.

  2. Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF-2α expression in lung cancer cells under normoxia and hypoxia

    Science.gov (United States)

    AQUINO-GÁLVEZ, ARNOLDO; GONZÁLEZ-ÁVILA, GEORGINA; DELGADO-TELLO, JAVIER; CASTILLEJOS-LÓPEZ, MANUEL; MENDOZA-MILLA, CRISELDA; ZÚÑIGA, JOAQUÍN; CHECA, MARCO; MALDONADO-MARTÍNEZ, HÉCTOR AQUILES; TRINIDAD-LÓPEZ, AXEL; CISNEROS, JOSÉ; TORRES-ESPÍNDOLA, LUZ MARÍA; HERNÁNDEZ-JIMÉNEZ, CLAUDIA; SOMMER, BETTINA; CABELLO-GUTIÉRREZ, CARLOS; GUTIÉRREZ-GONZÁLEZ, LUIS H.

    2016-01-01

    Hypoxic tumor cells are known to be more resistant to conventional chemotherapy and radiation than normoxic cells. However, the effects of 2-methoxyestradiol (2-ME), an anti-angiogenic, antiproliferative and pro-apoptotic drug, on hypoxic lung cancer cells are unknown. The aim of the present study was to compare the effects of 2-ME on cell growth, apoptosis, hypoxia-inducible factor 1α (HIF-1α) and HIF-2α gene and protein expression in A549 cells under normoxic and hypoxic conditions. To establish the optimal 2-ME concentration with which to carry out the apoptosis assay and to examine mRNA and protein expression of HIFs, cell growth analysis was carried out through N-hexa-methylpararosaniline staining assays in A549 cell cultures treated with one of five different 2-ME concentrations at different times under normoxic or hypoxic growth conditions. The 2-ME concentration of 10 mM at 72 h was selected to perform all further experiments. Apoptotic cells were analyzed by flow cytometry. Western blotting was used to determine HIF-1α and HIF-2α protein expression in total cell extracts. Cellular localization of HIF-1α and HIF-2α was assessed by immunocytochemistry. HIF-1α and HIF-2α gene expression was determined by real-time PCR. A significant increase in the percentage of apoptosis was observed when cells were treated with 2-ME under a normoxic but not under hypoxic conditions (p=0.006). HIF-1α and HIF-2α protein expression levels were significantly decreased in cells cultured under hypoxic conditions and treated with 2-ME (p<0.001). Furthermore, 2-ME decreased the HIF-1α and HIF-2α nuclear staining in cells cultured under hypoxia. The HIF-1α and HIF-2α mRNA levels were significantly lower when cells were exposed to 2-ME under normoxia and hypoxia. Our results suggest that 2-ME could have beneficial results when used with conventional chemotherapy in an attempt to lower the invasive and metastatic processes during cancer development due to its effects on

  3. Effects of HIF-1 and HIF2 on Growth and Metabolism of Clear-Cell Renal Cell Carcinoma 786-0 Xenografts

    Directory of Open Access Journals (Sweden)

    Swethajit Biswas

    2010-01-01

    Full Text Available In cultured clear-cell renal carcinoma (CCRCC 786-0 cells transfected with HIF1 (HIF-1+, HIF-2 (HIF-2+, or empty vector (EV, no significant differences were observed in the growth rates in vitro, but when grown in vivo as xenografts HIF-2 significantly increased, and HIF-1 significantly decreased growth rates, compared to EV tumors. Factors associated with proliferation were increased and factors associated with cell death were decreased in HIF-2+ tumors. Metabolite profiles showed higher glucose and lower lactate and alanine levels in the HIF-2+ tumors whilst immunostaining demonstrated higher pyruvate dehydrogenase and lower pyruvate dehydrogenase kinase 1, compared to control tumors. Taken together, these results suggest that overexpression of HIF-2 in CCRCC 786-0 tumors regulated growth both by maintaining a low level of glycolysis and by allowing more mitochondrial metabolism and tolerance to ROS induced DNA damage. The growth profiles observed may be mediated by adaptive changes to a more oxidative phenotype.

  4. A therapeutic role for targeting c-Myc/Hif-1-dependent signaling pathways.

    Science.gov (United States)

    Podar, Klaus; Anderson, Kenneth C

    2010-05-01

    Deregulated c-Myc occurs in approximately 30% of human cancers. Similarly, hypoxia-inducible factor (HIF) is commonly overexpressed in a variety of human malignancies. Under physiologic conditions, HIF inhibits c-Myc activity; however, when deregulated oncogenic c-Myc collaborates with HIF in inducing the expression of VEGF, PDK1 and hexokinase 2. Most of the knowledge of HIF derives from studies investigating a role of HIF under hypoxic conditions, however, HIF-1alpha stabilization is also found in normoxic conditions. Specifically, under hypoxic conditions HIF-1-mediated regulation of oncogenic c-Myc plays a pivotal role in conferring metabolic advantages to tumor cells as well as adaptation to the tumorigenic micromilieu. In addition, our own results show that under normoxic conditions oncogenic c-Myc is required for constitutive high HIF-1 protein levels and activity in Multiple Myeloma (MM) cells, thereby influencing VEGF secretion and angiogenic activity within the bone marrow microenvironment. Further studies are needed to delineate the functional relevance of HIF, MYC, and the HIF-MYC collaboration in MM and other malignancies, also integrating the tumor microenvironment and the cellular context. Importantly, early studies already demonstrate promising preclinical of novel agents, predominantly small molecules, which target c-Myc, HIF or both.

  5. Technology and teen drivers.

    Science.gov (United States)

    Lee, John D

    2007-01-01

    The rapid evolution of computing, communication, and sensor technology is likely to affect young drivers more than others. The distraction potential of infotainment technology stresses the same vulnerabilities that already lead young drivers to crash more frequently than other drivers. Cell phones, text messaging, MP3 players, and other nomadic devices all present a threat because young drivers may lack the spare attentional capacity for vehicle control and the ability to anticipate and manage hazards. Moreover, young drivers are likely to be the first and most aggressive users of new technology. Fortunately, emerging technology can also support safe driving. Electronic stability control, collision avoidance systems, intelligent speed adaptation, and vehicle tracking systems can all help mitigate the threats to young drivers. However, technology alone is unlikely to make young drivers safer. One promising approach to tailoring technology to teen drivers is to extend proven methods for enhancing young driver safety. The success of graduated drivers license programs (GDL) and the impressive safety benefit of supervised driving suggest ways of tailoring technology to the needs of young drivers. To anticipate the effects of technology on teen driving it may be useful to draw an analogy between the effects of passengers and the effects of technology. Technology can act as a teen passenger and undermine safety or it can act as an adult passenger and enhance safety. Rapidly developing technology may have particularly large effects on teen drivers. To maximize the positive effects and minimize the negative effects will require a broad range of industries to work together. Ideally, vehicle manufacturers would work with infotainment providers, insurance companies, and policy makers to craft new technologies so that they accommodate the needs of young drivers. Without such collaboration young drivers will face even greater challenges to their safety as new technologies emerge.

  6. Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α upregulation in human tumours.

    Directory of Open Access Journals (Sweden)

    Cameron E Snell

    Full Text Available The stabilisation of HIF-α is central to the transcriptional response of animals to hypoxia, regulating the expression of hundreds of genes including those involved in angiogenesis, metabolism and metastasis. HIF-α is degraded under normoxic conditions by proline hydroxylation, which allows for recognition and ubiquitination by the von-Hippel-Lindau (VHL E3 ligase complex. The aim of our study was to investigate the posttranslational modification of HIF-1α in tumours, to assess whether there are additional mechanisms besides reduced hydroxylation leading to stability. To this end we optimised antibodies against the proline-hydroxylated forms of HIF-1α for use in formalin fixed paraffin embedded (FFPE immunohistochemistry to assess effects in tumour cells in vivo. We found that HIF-1α proline-hydroxylated at both VHL binding sites (Pro402 and Pro564, was present in hypoxic regions of a wide range of tumours, tumour xenografts and in moderately hypoxic cells in vitro. Staining for hydroxylated HIF-1α can identify a subset of breast cancer patients with poorer prognosis and may be a better marker than total HIF-1α levels. The expression of unhydroxylated HIF-1α positively correlates with VHL in breast cancer suggesting that VHL may be rate-limiting for HIF degradation. Our conclusions are that the degradation of proline-hydroxylated HIF-1α may be rate-limited in tumours and therefore provides new insights into mechanisms of HIF upregulation. Persistence of proline-hydroxylated HIF-1α in perinecrotic areas suggests there is adequate oxygen to support prolyl hydroxylase domain (PHD activity and proline-hydroxylated HIF-1α may be the predominant form associated with the poorer prognosis that higher levels of HIF-1α confer.

  7. HIF1α deficiency reduces inflammation in a mouse model of proximal colon cancer

    Directory of Open Access Journals (Sweden)

    Dessislava N. Mladenova

    2015-09-01

    Full Text Available Hypoxia-inducible factor 1α (HIF1α is a transcription factor that regulates the adaptation of cells to hypoxic microenvironments, for example inside solid tumours. Stabilisation of HIF1α can also occur in normoxic conditions in inflamed tissue or as a result of inactivating mutations in negative regulators of HIF1α. Aberrant overexpression of HIF1α in many different cancers has led to intensive efforts to develop HIF1α-targeted therapies. However, the role of HIF1α is still poorly understood in chronic inflammation that predisposes the colon to carcinogenesis. We have previously reported that the transcription of HIF1α is upregulated and that the protein is stabilised in inflammatory lesions that are caused by the non-steroidal anti-inflammatory drug (NSAID sulindac in the mouse proximal colon. Here, we exploited this side effect of long-term sulindac administration to analyse the role of HIF1α in colon inflammation using mice with a Villin-Cre-induced deletion of Hif1α exon 2 in the intestinal epithelium (Hif1αΔIEC. We also analysed the effect of sulindac sulfide on the aryl hydrocarbon receptor (AHR pathway in vitro in colon cancer cells. Most sulindac-treated mice developed visible lesions, resembling the appearance of flat adenomas in the human colon, surrounded by macroscopically normal mucosa. Hif1αΔIEC mice still developed lesions but they were smaller than in the Hif1α-floxed siblings (Hif1αF/F. Microscopically, Hif1αΔIEC mice had significantly less severe colon inflammation than Hif1αF/F mice. Molecular analysis showed reduced MIF expression and increased E-cadherin mRNA expression in the colon of sulindac-treated Hif1αΔIEC mice. However, immunohistochemistry analysis revealed a defect of E-cadherin protein expression in sulindac-treated Hif1αΔIEC mice. Sulindac sulfide treatment in vitro upregulated Hif1α, c-JUN and IL8 expression through the AHR pathway. Taken together, HIF1α expression augments inflammation

  8. A human tRNA methyltransferase 9-like protein prevents tumour growth by regulating LIN9 and HIF1-α

    Science.gov (United States)

    Begley, Ulrike; Sosa, Maria Soledad; Avivar-Valderas, Alvaro; Patil, Ashish; Endres, Lauren; Estrada, Yeriel; Chan, Clement TY; Su, Dan; Dedon, Peter C; Aguirre-Ghiso, Julio A; Begley, Thomas

    2013-01-01

    Emerging evidence points to aberrant regulation of translation as a driver of cell transformation in cancer. Given the direct control of translation by tRNA modifications, tRNA modifying enzymes may function as regulators of cancer progression. Here, we show that a tRNA methyltransferase 9-like (hTRM9L/KIAA1456) mRNA is down-regulated in breast, bladder, colorectal, cervix and testicular carcinomas. In the aggressive SW620 and HCT116 colon carcinoma cell lines, hTRM9L is silenced and its re-expression and methyltransferase activity dramatically suppressed tumour growth in vivo. This growth inhibition was linked to decreased proliferation, senescence-like G0/G1-arrest and up-regulation of the RB interacting protein LIN9. Additionally, SW620 cells re-expressing hTRM9L did not respond to hypoxia via HIF1-α-dependent induction of GLUT1. Importantly, hTRM9L-negative tumours were highly sensitive to aminoglycoside antibiotics and this was associated with altered tRNA modification levels compared to antibiotic resistant hTRM9L-expressing SW620 cells. Our study links hTRM9L and tRNA modifications to inhibition of tumour growth via LIN9 and HIF1-α-dependent mechanisms. It also suggests that aminoglycoside antibiotics may be useful to treat hTRM9L-deficient tumours. PMID:23381944

  9. Erythrocytosis-associated HIF-2α Mutations Demonstrate a Critical Role for Residues C-terminal to the Hydroxylacceptor Proline*

    OpenAIRE

    2009-01-01

    A classic physiologic response to hypoxia in humans is the up-regulation of the ERYTHROPOIETIN (EPO) gene, which is the central regulator of red blood cell mass. The EPO gene, in turn, is activated by hypoxia inducible factor (HIF). HIF is a transcription factor consisting of an α subunit (HIF-α) and a β subunit (HIF-β). Under normoxic conditions, prolyl hydroxylase domain protein (PHD, also known as HIF prolyl hydroxylase and egg laying-defective nine protein) site sp...

  10. Increased size of solid organs in patients with Chuvash polycythemia and in mice with altered expression of HIF-1α and HIF-2α

    Science.gov (United States)

    Yoon, Donghoon; Okhotin, David V.; Kim, Bumjun; Okhotina, Yulia; Okhotin, Daniel J.; Miasnikova, Galina Y.; Sergueeva, Adelina I.; Polyakova, Lydia A.; Maslow, Alexei; Lee, Yonggu; Semenza, Gregg L.; Prchal, Josef T.

    2010-01-01

    Chuvash polycythemia, the first hereditary disease associated with dysregulated oxygen-sensing to be recognized, is characterized by a homozygous germ-line loss-of-function mutation of the VHL gene (VHLR200W) resulting in elevated hypoxia inducible factor (HIF)-1α and HIF-2α levels, increased red cell mass and propensity to thrombosis. Organ volume is determined by the size and number of cells, and the underlying molecular control mechanisms are not fully elucidated. Work from several groups has demonstrated that the proliferation of cells is regulated in opposite directions by HIF-1α and HIF-2α. HIF-1α inhibits cell proliferation by displacing MYC from the promoter of the gene encoding the cyclin-dependent kinase inhibitor, p21Cip1, thereby inducing its expression. In contrast, HIF-2α promotes MYC activity and cell proliferation. Here we report that the volumes of liver, spleen, and kidneys relative to body mass were larger in 30 individuals with Chuvash polycythemia than in 30 matched Chuvash controls. In Hif1a+/− mice, which are heterozygous for a null (knockout) allele at the locus encoding HIF-1α, hepatic HIF-2α mRNA was increased (2-fold) and the mass of the liver was increased, compared with wild-type littermates, without significant difference in cell volume. Hepatic p21Cip1 mRNA levels were 9.5-fold lower in Hif1a+/− mice compared with wild-type littermates. These data suggest that, in addition to increased red cell mass, the sizes of liver, spleen, and kidneys are increased in Chuvash polycythemia. At least in the liver, this phenotype may result from increased HIF-2α and decreased p21Cip1 levels leading to increased hepatocyte proliferation. PMID:20140661

  11. Comparative analysis of Haemophilus influenzae hifA (pilin) genes.

    Science.gov (United States)

    Clemans, D L; Marrs, C F; Patel, M; Duncan, M; Gilsdorf, J R

    1998-02-01

    Adherence of Haemophilus influenzae to epithelial cells plays a central role in colonization and is the first step in infection with this organism. Pili, which are large polymorphic surface proteins, have been shown to mediate the binding of H. influenzae to cells of the human respiratory tract. Earlier experiments have demonstrated that the major epitopes of H. influenzae pili are highly conformational and immunologically heterogenous; their subunit pilins are, however, immunologically homogenous. To define the extent of structural variation in pilins, which polymerize to form pili, the pilin genes (hifA) of 26 type a to f and 16 nontypeable strains of H. influenzae were amplified by PCR and subjected to restriction fragment length polymorphism (RFLP) analysis with AluI and RsaI. Six different RFLP patterns were identified. Four further RFLP patterns were identified from published hifA sequences from five nontypeable H. influenzae strains. Two patterns contained only nontypeable isolates; one of these contained H. influenzae biotype aegyptius strains F3031 and F3037. Another pattern contained predominantly H. influenzae type f strains. All other patterns were displayed by a variety of capsular and noncapsular types. Sequence analysis of selected hifA genes confirmed the 10 RFLP patterns and showed strong identity among representatives displaying the same RFLP patterns. In addition, the immunologic reactivity of pili with antipilus antisera correlated with the groupings of strains based on hifA RFLP patterns. Those strains that show greater reactivity with antiserum directed against H. influenzae type b strain M43 pili tend to fall into one RFLP pattern (pattern 3); while those strains that show equal or greater reactivity with antiserum directed against H. influenzae type b strain Eagan pili tend to fall in a different RFLP pattern (pattern 1). Sequence analysis of representative HifA pilins from typeable and nontypeable H. influenzae identified several highly

  12. National Driver Register (NDR) -

    Data.gov (United States)

    Department of Transportation — Information regarding individuals who have had their driver licenses revoked, suspended or otherwise denied for cause, or who have been convicted of certain traffic...

  13. Disulfiram deregulates HIF-α subunits and blunts tumor adaptation to hypoxia in hepatoma cells

    Science.gov (United States)

    Park, Hye-joon; Kim, Min-sung; Cho, Kumsun; Yun, Jang-hyuk; Choi, Yong-joon; Cho, Chung-hyun

    2013-01-01

    Aim: Disulfiram is an aldehyde dehydrogenase inhibitor that was used to treat alcoholism and showed anticancer activity, but its anticancer mechanism remains unclear. The aim of this study was to investigate the effects of disulfiram on the hypoxia-inducible factor (HIF)-driven tumor adaptation to hypoxia in vitro. Methods: Hep3B, Huh7 and HepG2 hepatoma cells were incubated under normoxic (20% O2) or hypoxic (1% O2) conditions for 16 h. The expression and activity of HIF-1α and HIF-2α proteins were evaluated using immunoblotting and luciferase reporter assay, respectively. Semi-quantitative RT-PCR was used to analyze HIF-mediated gene expression. Endothelial tubule formation assay was used to evaluate the anti-angiogenic effect. Results: Hypoxia caused marked expression of HIF-1α and HIF-1α in the 3 hepatoma cell lines, dramatically increased HIF activity and induced the expression of HIF downstream genes (EPO, CA9, VEGF-A and PDK1) in Hep3B cells. HIF-2α expression was positively correlated with the induction of hypoxic genes (CA9, VEGF-A and PDK1). Moreover, hypoxia markedly increased VEGF production and angiogenic potential of Hep3B cells. Disulfiram (0.3 to 2 μmol/L) inhibited hypoxia-induced gene expression and HIF activity in a dose-dependent manner. Disulfiram more effectively suppressed the viability of Hep3B cells under hypoxia, but it did not affect the cell cycle. Overexpression of HIF-2α in Hep3B cells reversed the inhibitory effects of disulfiram on hypoxia-induced gene expression and cell survival under hypoxia. Conclusion: Disulfiram deregulates the HIF-mediated hypoxic signaling pathway in hepatoma cells, which may contribute to its anticancer effect. Thus, disulfiram could be used to treat solid tumors that grow in a HIF-dependent manner. PMID:23852087

  14. HIF-2alpha-dependent PAI-1 induction contributes to angiogenesis in hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Geis, Theresa, E-mail: geis@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Döring, Claudia, E-mail: C.Doering@em.uni-frankfurt.de [Dr. Senckenberg Institute of Pathology, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Popp, Rüdiger, E-mail: popp@vrc.uni-frankfurt.de [Institute for Vascular Signalling, Centre for Molecular Medicine, Faculty of Medicine Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60596 Frankfurt am Main (Germany); Grossmann, Nina, E-mail: grossmann@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Fleming, Ingrid, E-mail: fleming@vrc.uni-frankfurt.de [Institute for Vascular Signalling, Centre for Molecular Medicine, Faculty of Medicine Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60596 Frankfurt am Main (Germany); Hansmann, Martin-Leo, E-mail: m.l.hansmann@em.uni-frankfurt.de [Dr. Senckenberg Institute of Pathology, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Dehne, Nathalie, E-mail: dehne@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany); Brüne, Bernhard, E-mail: b.bruene@biochem.uni-frankfurt.de [Institute of Biochemistry I—Pathobiochemistry, Faculty of Medicine, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt am Main (Germany)

    2015-02-01

    Hypoxia promotes progression of hepatocellular carcinoma (HCC), not only affecting tumor cell proliferation and invasion, but also angiogenesis and thus, increasing the risk of metastasis. Hypoxia inducible factors (HIF)-1α and -2α cause adaptation of tumors to hypoxia, still with uncertainties towards the angiogenic switch. We created a stable knockdown of HIF-1α and HIF-2α in HepG2 cells and generated cocultures of HepG2 spheroids with embryonic bodies as an in vitro tumor model mimicking the cancer microenvironment. The naturally occuring oxygen and nutrient gradients within the cocultures allow us to question the role of distinct HIF isoforms in regulating HCC angiogenesis. In cocultures with a HIF-2α knockdown, angiogenesis was attenuated, while the knockdown of HIF-1α was without effect. Microarray analysis identified plasminogen activator inhibitor 1 (PAI-1) as a HIF-2α target gene in HepG2 cells. The knockdown of PAI-1 in HepG2 cells also lowered angiogenesis. Blocking plasmin, the downstream target of PAI-1, with aprotinin in HIF-2α knockdown (k/d) cells proved a cause–effect relation and restored angiogenesis, with no effect on control cocultures. Suggestively, HIF-2α increases PAI-1 to lower concentrations of active plasmin, thereby supporting angiogenesis. We conclude that the HIF-2α target gene PAI-1 favors the angiogenic switch in HCC. - Highlights: • HepG2 were cocultured with stem cells to mimic a cancer microenvironment in vitro. • A knockdown of HIF-2α reduces angiogenesis. • PAI-1 was identified as a HIF-2α target gene in HCC by microarray analysis. • HIF-2α induces the angiogenic switch via inhibition of plasmin.

  15. Myeloid HIF-1 is protective in Helicobacter pylori-mediated gastritis.

    Science.gov (United States)

    Matak, Pavle; Heinis, Mylène; Mathieu, Jacques R R; Corriden, Ross; Cuvellier, Sylvain; Delga, Stéphanie; Mounier, Rémi; Rouquette, Alexandre; Raymond, Josette; Lamarque, Dominique; Emile, Jean-François; Nizet, Victor; Touati, Eliette; Peyssonnaux, Carole

    2015-04-01

    Helicobacter pylori infection triggers chronic inflammation of the gastric mucosa that may progress to gastric cancer. The hypoxia-inducible factors (HIFs) are the central mediators of cellular adaptation to low oxygen levels (hypoxia), but they have emerged recently as major transcriptional regulators of immunity and inflammation. No studies have investigated whether H. pylori affects HIF signaling in immune cells and a potential role for HIF in H. pylori-mediated gastritis. HIF-1 and HIF-2 expression was examined in human H. pylori-positive gastritis biopsies. Subsequent experiments were performed in naive and polarized bone marrow-derived macrophages from wild-type (WT) and myeloid HIF-1α-null mice (HIF-1(Δmyel)). WT and HIF-1(Δmyel) mice were inoculated with H. pylori by oral gavage and sacrificed 6 mo postinfection. HIF-1 was specifically expressed in macrophages of human H. pylori-positive gastritis biopsies. Macrophage HIF-1 strongly contributed to the induction of proinflammatory genes (IL-6, IL-1β) and inducible NO synthase in response to H. pylori. HIF-2 expression and markers of M2 macrophage differentiation were decreased in response to H. pylori. HIF-1(Δmyel) mice inoculated with H. pylori for 6 mo presented with a similar bacterial colonization than WT mice but, surprisingly, a global increase of inflammation, leading to a worsening of the gastritis, measured by an increased epithelial cell proliferation. In conclusion, myeloid HIF-1 is protective in H. pylori-mediated gastritis, pointing to the complex counterbalancing roles of innate immune and inflammatory phenotypes in driving this pathology. Copyright © 2015 by The American Association of Immunologists, Inc.

  16. Identification of small molecule compounds that inhibit the HIF-1 signaling pathway

    Directory of Open Access Journals (Sweden)

    Sun Yi

    2009-12-01

    Full Text Available Abstract Background Hypoxia-inducible factor-1 (HIF-1 is the major hypoxia-regulated transcription factor that regulates cellular responses to low oxygen environments. HIF-1 is composed of two subunits: hypoxia-inducible HIF-1α and constitutively-expressed HIF-1β. During hypoxic conditions, HIF-1α heterodimerizes with HIF-1β and translocates to the nucleus where the HIF-1 complex binds to the hypoxia-response element (HRE and activates expression of target genes implicated in cell growth and survival. HIF-1α protein expression is elevated in many solid tumors, including those of the cervix and brain, where cells that are the greatest distance from blood vessels, and therefore the most hypoxic, express the highest levels of HIF-1α. Therapeutic blockade of the HIF-1 signaling pathway in cancer cells therefore provides an attractive strategy for development of anticancer drugs. To identify small molecule inhibitors of the HIF-1 pathway, we have developed a cell-based reporter gene assay and screened a large compound library by using a quantitative high-throughput screening (qHTS approach. Results The assay is based upon a β-lactamase reporter under the control of a HRE. We have screened approximate 73,000 compounds by qHTS, with each compound tested over a range of seven to fifteen concentrations. After qHTS we have rapidly identified three novel structural series of HIF-1 pathway Inhibitors. Selected compounds in these series were also confirmed as inhibitors in a HRE β-lactamase reporter gene assay induced by low oxygen and in a VEGF secretion assay. Three of the four selected compounds tested showed significant inhibition of hypoxia-induced HIF-1α accumulation by western blot analysis. Conclusion The use of β-lactamase reporter gene assays, in combination with qHTS, enabled the rapid identification and prioritization of inhibitors specific to the hypoxia induced signaling pathway.

  17. Young novice drivers.

    NARCIS (Netherlands)

    2013-01-01

    In The Netherlands, young novice drivers (18-24 years of age) show a crash rate that is five times higher than that of experienced drivers (30-59 years of age). The rate of young males is even seven times as high. The main reasons are lack of driving experience and hazardous behaviour typical of ado

  18. A Simple Wave Driver

    Science.gov (United States)

    Temiz, Burak Kagan; Yavuz, Ahmet

    2015-01-01

    This study was done to develop a simple and inexpensive wave driver that can be used in experiments on string waves. The wave driver was made using a battery-operated toy car, and the apparatus can be used to produce string waves at a fixed frequency. The working principle of the apparatus is as follows: shortly after the car is turned on, the…

  19. Zinc downregulates HIF-1α and inhibits its activity in tumor cells in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Lavinia Nardinocchi

    Full Text Available BACKGROUND: Hypoxia inducible factor-1α (HIF-1α is responsible for the majority of HIF-1-induced gene expression changes under hypoxia and for the "angiogenic switch" during tumor progression. HIF-1α is often upregulated in tumors leading to more aggressive tumor growth and chemoresistance, therefore representing an important target for antitumor intervention. We previously reported that zinc downregulated HIF-1α levels. Here, we evaluated the molecular mechanisms of zinc-induced HIF-1α downregulation and whether zinc affected HIF-1α also in vivo. METHODOLOGY/PRINCIPAL FINDINGS: Here we report that zinc downregulated HIF-1α protein levels in human prostate cancer and glioblastoma cells under hypoxia, whether induced or constitutive. Investigations into the molecular mechanisms showed that zinc induced HIF-1α proteasomal degradation that was prevented by treatment with proteasomal inhibitor MG132. HIF-1α downregulation induced by zinc was ineffective in human RCC4 VHL-null renal carcinoma cell line; likewise, the HIF-1αP402/P564A mutant was resistant to zinc treatment. Similarly to HIF-1α, zinc downregulated also hypoxia-induced HIF-2α whereas the HIF-1β subunit remained unchanged. Zinc inhibited HIF-1α recruitment onto VEGF promoter and the zinc-induced suppression of HIF-1-dependent activation of VEGF correlated with reduction of glioblastoma and prostate cancer cell invasiveness in vitro. Finally, zinc administration downregulated HIF-1α levels in vivo, by bioluminescence imaging, and suppressed intratumoral VEGF expression. CONCLUSIONS/SIGNIFICANCE: These findings, by demonstrating that zinc induces HIF-1α proteasomal degradation, indicate that zinc could be useful as an inhibitor of HIF-1α in human tumors to repress important pathways involved in tumor progression, such as those induced by VEGF, MDR1, and Bcl2 target genes, and hopefully potentiate the anticancer therapies.

  20. Silencing HIF-1α reduces the adhesion and secretion functions of acute leukemia hBMSCs

    Directory of Open Access Journals (Sweden)

    Zeng Dong-Feng

    2012-10-01

    Full Text Available Hypoxia inducible factor-1α (HIF-1α is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenviroment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF and stromal cell-derived factor-1α (SDF-1α were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system was constructed by putting the two cells into the same suitable cultured media and conditions. Cell adhesion and secretion functions of stromal cells were evaluated after transfection with the lentiviral RNAi vector of HIF-1α. Increased HIF-1α mRNA and protein was detected in the nucleus of the acute myeloblastic and acute lymphoblastic leukemia compared with normal BMSCs. The lentiviral RANi vector for HIF-1α was successfully constructed and was applied to block the expression of HIF-1α. When HIF-1α of BMSCs was blocked, the expression of VEGF and SDF-1 secreted by stromal cells were decreased. When HIF-1α was blocked, the co-cultured Jurkat cell’s adhesion and migration functions were also decreased. Taken together, these results suggest that HIF-1α acts as an important transcription factor and can significantly affect the secretion and adhesion functions of leukemia BMSCs.

  1. Silencing HIF-1α reduces the adhesion and secretion functions of acute leukemia hBMSCs

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Dong-Feng [Department of Hematology, XinQiao Hospital, Third Military Medical University, ChongQing (China); Liu, Ting [Department of Ophthalmology, DaPing Hospital, Third Military Medical University, ChongQing (China); Chang, Cheng; Zhang, Xi; Liang, Xue; Chen, Xing-Hua; Kong, Pei-Yan [Department of Hematology, XinQiao Hospital, Third Military Medical University, ChongQing (China)

    2012-06-29

    Hypoxia inducible factor-1α (HIF-1α) is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenvironment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs) and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system was constructed by putting the two cells into the same suitable cultured media and conditions. Cell adhesion and secretion functions of stromal cells were evaluated after transfection with the lentiviral RNAi vector of HIF-1α. Increased HIF-1α mRNA and protein was detected in the nucleus of the acute myeloblastic and acute lymphoblastic leukemia compared with normal BMSCs. The lentiviral RANi vector for HIF-1α was successfully constructed and was applied to block the expression of HIF-1α. When HIF-1α of BMSCs was blocked, the expression of VEGF and SDF-1α secreted by stromal cells was decreased. When HIF-1α was blocked, the co-cultured Jurkat cell's adhesion and migration functions were also decreased. Taken together, these results suggest that HIF-1α acts as an important transcription factor and can significantly affect the secretion and adhesion functions of leukemia BMSCs.

  2. Hostile takeover: Manipulation of HIF-1 signaling in pathogen-associated cancers (Review).

    Science.gov (United States)

    Zhu, Caixia; Zhu, Qing; Wang, Chong; Zhang, Liming; Wei, Fang; Cai, Qiliang

    2016-10-01

    Hypoxia-inducible factor (HIF)-1 is a central regulator in the adaptation process of cell response to hypoxia (low oxygen). Emerging evidence has demonstrated that HIF-1 plays an important role in the development and progression of many types of human diseases, including pathogen-associated cancers. In the present review, we summarize the recent understandings of how human pathogenic agents including viruses, bacteria and parasites deregulate cellular HIF-1 signaling pathway in their associated cancer cells, and highlight the common molecular mechanisms of HIF-1 signaling activated by these pathogenic infection, which could act as potential diagnostic markers and new therapeutic strategies against human infectious cancers.

  3. Tumor Cells Upregulate Normoxic HIF-1α in Response to Doxorubicin

    Science.gov (United States)

    Cao, Yiting; Eble, Joseph M.; Moon, Ejung; Yuan, Hong; Weitzel, Douglas H.; Landon, Chelsea D.; Nien, Charleen Yu-Chih; Hanna, Gabi; Rich, Jeremy N.; Provenzale, James M.; Dewhirst, Mark W.

    2013-01-01

    Hypoxia-inducible factor 1 (HIF-1) is a master transcription factor that controls cellular homeostasis. While its activation benefits normal tissue, HIF-1 activation in tumors is a major risk factor for angiogenesis, therapeutic resistance and poor prognosis. HIF-1 activity is usually suppressed under normoxic conditions because of rapid oxygen-dependent degradation of HIF-1α. Here we show that under normoxic conditions HIF-1α is upregulated in tumor cells in response to doxorubicin, a chemotherapy used to treat many cancers. Doxorubicin also enhanced VEGF secretion by normoxic tumor cells and stimulated tumor angiogenesis. Doxorubicin-induced accumulation of HIF-1α in normoxic cells was caused by increased expression and activation of STAT1, the activation of which stimulated expression of iNOS and its synthesis of NO in tumor cells. Mechanistic investigations established that blocking NO synthesis or STAT1 activation was sufficient to attenuate the HIF-1α accumulation induced by doxorubicin in normoxic cancer cells. To our knowledge, this is the first report that a chemotherapeutic drug can induce HIF-1α accumulation in normoxic cells, an efficacy-limiting activity. Our results argue that HIF-1α targeting strategies may enhance doxorubicin efficacy. More generally, they suggest a broader perspective on the design of combination chemotherapy approaches with immediate clinical impact. PMID:23959856

  4. Gefitinib circumvents hypoxia-induced drug resistance by the modulation of HIF-1alpha.

    Science.gov (United States)

    Rho, Jin Kyung; Choi, Yun Jung; Lee, Jin Kyung; Ryoo, Baek-Yeol; Na, Im Ii; Yang, Sung Hyun; Kim, Cheol Hyeon; Yoo, Young Do; Lee, Jae Cheol

    2009-03-01

    Hypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor which is activated by hypoxia and associated with cell survival, proliferation and drug resistance. Recent studies have shown that the down-stream molecules of the epidermal growth factor receptor (EGFR) signal are involved in the hypoxia-dependent or -independent HIF-1alpha protein accumulation. Thus, we hypothesized that an EGFR-TK inhibitor, gefitinib, might circumvent the hypoxia-induced drug resistance via the regulation of HIF-1alpha expression. In our data, treatment of gefitinib suppressed induced HIF-1alpha by hypoxia. This action of gefitinib was caused by reduced protein stability without any change in the level of HIF-1alpha mRNA. The effect of gefitinib on down-regulation of HIF-1alpha was reversed by transfection of constitutively active form of Akt. The cellular response to gefitinib was similar in both normoxia and hypoxia condition. However, the response to conventional chemotherapeutic drugs decreased >50% under hypoxia condition and they did not change HIF-1alpha expression. In addition, the suppression of HIF-1alpha using siRNA overcame partially hypoxia-induced drug resistance. In conclusion, gefitinib was able to circumvent hypoxia-induced drug resistance suggesting that the effective suppression of HIF-1alpha by the inhibition of EGFR-Akt pathway may overcome the hypoxia-induced drug resistance.

  5. 神经干细胞移植对脑缺血/再灌注损伤大鼠HIF-1α和HIF-3α表达的影响%Effects of Neural Stem Cells Transplantation on the Expression of HIF-1α and HIF-3α after Cerebral Ischemia-Reperfusion in Rats

    Institute of Scientific and Technical Information of China (English)

    王莹; 李文媛; 刘艳翠; 丁利; 赵斯达

    2012-01-01

    Objective To investigate the effects of transplantation of neural stem cells( NSCs )on the expression of HIF-1 α and HIF-3 α in hippocampus region after cerebral ischemia-reperfusion in rats. Methods The rat model of middle cerebral artery occlusion was established by the suture method. Rats were randomly divided into sham operation group, model group and NSCs group. 72h after reperfusion, the neurological function was investigated by NSS method, the expression of HIF-1 α and HIF-3 α in hippocampus region was detected by immunohistochemical method. Results PKH-26 labeled NSCs were observed in hippocampus region in NSCs group. Compared with model group,the NSS of NSCs group was decreased( P <0. 05 ),and the expression of HIF-1 α and HIF-3 α in hippocampus region was increased( P <0.05 ). Conclusion NSCs transplantation could promote the nerve functional recovery of cerebral ischemia injury by elevating HIF-1 α and HIF-3 α in hippocampus region of rats.%目的 探讨神经干细胞(NSCs)移植对大鼠脑缺血/再灌注损伤海马区低氧诱导因子1α(HIF-1α)和低氧诱导因子3α(HIF-3α)表达的影响.方法 线栓法制作大鼠大脑中动脉缺血/再灌注模型,大鼠随机分为假手术组、模型组、NSCs组.再灌注72 h后神经功能损害评分表(NSS)法行神经功能评分,免疫组化法检测海马区HIF-1α和HIF-3α蛋白表达.结果 NSCs移植后可见PKH26标记的NSCs在海马区有表达.与模型组比较,NSCs组NSS评分显著降低(P<0.05),海马区HIF-1α和HIF-3α蛋白表达显著上调(P<0.05).结论 神经干细胞移植可上调脑缺血/再灌注损伤大鼠海马区HIF-1α和HIF-3α表达,促进脑缺血损伤神经功能恢复.

  6. Compression-induced HIF-1 enhances thrombosis and PAI-1 expression in mouse skin.

    Science.gov (United States)

    Kaneko, Maki; Minematsu, Takeo; Yoshida, Mikako; Nishijima, Yoshimi; Noguchi, Hiroshi; Ohta, Yasunori; Nakagami, Gojiro; Mori, Taketoshi; Sanada, Hiromi

    2015-09-01

    Pressure ulcers result from tissue hypoxia caused by external forces. Thrombosis due to external forces is considered important, and hypoxia inducible factor-1 (HIF-1) is a master regulator of pressure ulcer development. To date, however, their causal relationship has not been determined. This study therefore investigated the mutual relationship between thrombosis and HIF-1 activation in compressed mouse skin, based on a hypothesis that HIF-1 regulation by plasminogen activator inhibitor-1 (PAI-1) enhances thrombosis. Compression of mouse skin significantly increased the numbers of thrombi and HIF-1α-positive cells compared with control skin. A thrombosis inhibitor significantly reduced the numbers of HIF-1α-positive cells and an HIF-1 inhibitor significantly inhibited thrombosis in compressed skin tissue, suggesting a mutual relationship between thrombosis and HIF-1 activation. Compression of mouse skin also enhanced the level of Pai-1 messenger RNA expression, but this increase was significantly reduced by treatment with an HIF-1 inhibitor, whereas a thrombosis inhibitor had no effect. These results suggested the involvement of PAI-1 in HIF-1-enhanced thrombosis and that an additional factor participates in regulating Pai-1 expression in compressed skin. These findings may suggest new strategies in pressure ulcer management.

  7. Overcoming cisplatin resistance of ovarian cancer cells by targeting HIF-1-regulated cancer metabolism.

    Science.gov (United States)

    Ai, Zhihong; Lu, Yang; Qiu, Songbo; Fan, Zhen

    2016-04-01

    Cisplatin is currently one of the most effective chemotherapeutic drugs used for treating ovarian cancer; however, resistance to cisplatin is common. In this study, we explored an experimental strategy for overcoming cisplatin resistance of human ovarian cancer from the new perspective of cancer cell metabolism. By using two pairs of genetically matched cisplatin-sensitive and cisplatin-resistant ovarian cancer cell lines, we tested the hypothesis that downregulating hypoxia-inducible factor-1 (HIF-1), which regulates metabolic enzymes involved in glycolysis, is a promising strategy for overcoming cisplatin resistance of human ovarian cancer cells. We found that cisplatin downregulated the level of the regulatable α subunit of HIF-1, HIF-1α, in cisplatin-sensitive ovarian cancer cells through enhancing HIF-1α degradation but did not downregulate HIF-1α in their cisplatin-resistant counterparts. Overexpression of a degradation-resistant HIF-1α (HIF-1α ΔODD) reduced cisplatin-induced apoptosis in cisplatin-sensitive cells, whereas genetic knockdown of HIF-1α or pharmacological promotion of HIF-1α degradation enhanced response to cisplatin in both cisplatin-sensitive and cisplatin-resistant ovarian cancer cells. We further demonstrated that knockdown of HIF-1α improved the response of cisplatin-resistant ovarian cancer cells to cisplatin by redirecting the aerobic glycolysis in the resistant cancer cells toward mitochondrial oxidative phosphorylation, leading to cell death through overproduction of reactive oxygen species. Our findings suggest that the HIF-1α-regulated cancer metabolism pathway could be a novel target for overcoming cisplatin resistance in ovarian cancer.

  8. Drivers 65 Plus

    Science.gov (United States)

    ... see, hear, and feel in relation to other cars and drivers, traffic signs and signals, conditions of the highway, and the performance of your car. These decisions are usually made close to other ...

  9. Proactive driver training program

    Energy Technology Data Exchange (ETDEWEB)

    Vossler, W. [Kinetic Safety Consulting Inc., Grande Prairie, AB (Canada)

    2005-07-01

    Skid avoidance training is a recent approach to driver training and has been employed in various countries with a high degree of success. Among top ranked countries, motor vehicle incidents trends indicate higher incident rates among drivers are often due to lack of knowledge, experience and risk awareness. If lowered age limit experience is attained under direct supervision and in safe training conditions, it was suggested, incident frequency is reduced. A Norway study confirmed an increase in vehicle incident rates after drivers had received skid control training. The drivers were unable to maintain skill levels needed to react to critical driving tasks and had unrealistic expectations of skill after training. However, a skid avoidance training program launched in Sweden in 1999 has resulted in a 50 per cent reduction of vehicle incidents in the last 2 years. Details of the Skidcar System were presented, including details of the driving simulator, where simulation of actual driving situations is achieved by simply adjusting the amount of grip the vehicle has with the driving surface. Instructors modify driving behaviors based upon the driver's ability to maintain grip. There are over 200 units in North America. In addition, a Proactive Light Vehicle Driver Training/ Heavy Vehicle Assessment Program was initiated in 2003, with a motor vehicle incident rate reduction of 50 per cent at the end of 2004. Various examples of situations in which drivers have used their skid avoidance skills to avoid incidents were included. It was noted that the trend among driver training professionals has been towards decision-based rather than skills-based training, as skills-based training will diminish over time, and requires frequent re-training periods. Cognitive and perceptual skills were examined, as well as cognitive, associative and autonomous learning phases. It was concluded that skid avoidance is largely a decision-based skill. tabs, figs.

  10. Older drivers, crashes and injuries.

    Science.gov (United States)

    Koppel, Sjaanie; Bohensky, Megan; Langford, Jim; Taranto, David

    2011-10-01

    This article aimed to identify the main features of older driver casualty crashes, including detailed descriptions of injury outcomes. Data were obtained from the Transport Accident Commission insurance claims database for 2 groups of drivers: aged 41 to 55 years (middle-aged drivers) and aged 65 years and older (older drivers). In terms of crash circumstances, the majority of crashes involved a collision with another vehicle (70.0% of middle-aged drivers and 68.7% of older drivers). The 2 main maneuvers at the time of crash were driving straight ahead (44.6% of middle-aged drivers and 42.8% of older drivers) and turning right (equivalent of left turn in North America; 15.2% of middle-aged drivers and 17.6% of older drivers). In terms of injury outcomes, older drivers sustained a significantly higher proportion of injuries to the thorax (30.9% compared to 18.5% of middle-aged drivers). Conversely, a significantly higher proportion of middle-aged drivers sustained some form of injury to the neck (30.6% compared to 12.1% of older drivers). These findings highlight the situations that are particularly risky for older drivers and provide important insights for developing solutions to reduce older driver crash and injury risk.

  11. Characteristics of Chinese Driver Behavior

    NARCIS (Netherlands)

    Li, J.

    2014-01-01

    The high growth rate of vehicle ownership and many novel drivers in China determine the special features of Chinese driver behavior. This thesis introduces a comparative study on driver behavior by the analysis of saturation flow at urban intersections, Driver Behavior Questionnaire surveys, focus g

  12. Copy number of pilus gene clusters in Haemophilus influenzae and variation in the hifE pilin gene.

    Science.gov (United States)

    Read, T D; Satola, S W; Opdyke, J A; Farley, M M

    1998-04-01

    Brazilian purpuric fever (BPF)-associated Haemophilus influenzae biogroup aegyptius strain F3031 contains two identical copies of a five gene cluster (hifA to hifE) encoding pili similar to well-characterized Hif fimbriae of H. influenzae type b. HifE, the putative pilus tip adhesin of F3031, shares only 40% amino acid sequence similarity with the same molecule from type b strains, whereas the other four proteins have 75 to 95% identity. To determine whether pilus cluster duplication and the hifE(F3031) allele were special features of BPF-associated bacteria, we analyzed a collection of H. influenzae strains by PCR with hifA- and hifE-specific oligonucleotides, by Southern hybridization with a hifC gene probe, and by nucleotide sequencing. The presence of two pilus clusters was limited to some H. influenzae biogroup aegyptius strains. The hifE(F3031) allele was limited to H. influenzae biogroup aegyptius. Two strains contained one copy of hifE(F3031) and one copy of a variant hifE allele. We determined the nucleotide sequences of four hifE genes from H. influenzae biogroup aegyptius and H. influenzae capsule serotypes a and c. The predicted proteins produced by these genes demonstrated only 35 to 70% identity to the three published HifE proteins from nontypeable H. influenzae, serotype b, and BPF strains. The C-terminal third of the molecules implicated in chaperone binding was the most highly conserved region. Three conserved domains in the otherwise highly variable N-terminal putative receptor-binding region of HifE were similar to conserved portions in the N terminus of Neisseria pilus adhesin PilC. We concluded that two pilus clusters and hifE(F3031) were not specific for BPF-causing H. influenzae, and we also identified portions of HifE possibly involved in binding mammalian cell receptors.

  13. Drosophila genome-wide RNAi screen identifies multiple regulators of HIF-dependent transcription in hypoxia.

    Directory of Open Access Journals (Sweden)

    Andrés Dekanty

    2010-06-01

    Full Text Available Hypoxia-inducible factors (HIFs are a family of evolutionary conserved alpha-beta heterodimeric transcription factors that induce a wide range of genes in response to low oxygen tension. Molecular mechanisms that mediate oxygen-dependent HIF regulation operate at the level of the alpha subunit, controlling protein stability, subcellular localization, and transcriptional coactivator recruitment. We have conducted an unbiased genome-wide RNA interference (RNAi screen in Drosophila cells aimed to the identification of genes required for HIF activity. After 3 rounds of selection, 30 genes emerged as critical HIF regulators in hypoxia, most of which had not been previously associated with HIF biology. The list of genes includes components of chromatin remodeling complexes, transcription elongation factors, and translational regulators. One remarkable hit was the argonaute 1 (ago1 gene, a central element of the microRNA (miRNA translational silencing machinery. Further studies confirmed the physiological role of the miRNA machinery in HIF-dependent transcription. This study reveals the occurrence of novel mechanisms of HIF regulation, which might contribute to developing novel strategies for therapeutic intervention of HIF-related pathologies, including heart attack, cancer, and stroke.

  14. Correlation between BOLD-MRI and HIF expression level in renal carcinoma.

    Science.gov (United States)

    Li, Dong; Wang, Xingming; Wang, Shuai; Cheng, Jie

    2015-01-01

    Occupying about 2%~3% of all malignant tumors, renal carcinoma is the most common primary cancer in kidney. The oxidative level of tumor cells is of vital role for optimizing treatment plan, evaluating efficacy and predicting prognosis. This study thus investigated the R2(*) value in mouse renal carcinoma model and the correlation between tumor hypoxia and expression level of hypoxia inducible factor-1 (HIF-1). A total of 20 BALB/C nude mice (4~6 weeks old) were inoculated with human ACHN renal carcinoma cells to generate renal cancer model. After the tumor diameter reached 0.5 cm, all animals were examined by BOLD-MRI, both under normal inhalation (R2a(*)) and carbogen treatment (R2b(*)). The alternation of R2(*) values (ΔR2(*)=R2a(*) - R2b(*)) was calculated. Mice were then sacrificed for Immunohistochemical (IHC) staining targeting HIF-1α and HIF-2α. The positive score of HIF was then analyzed for its correlation with R2(*) value. In 18 mice finished both experiments, Pearson correlation analysis revealed significant negative correlation between R2a(*) and ΔR2(*) (r=-0.48, Pcorrelated with tumor R(*) values. The positive correlation between ΔR2(*) and HIF-2α, but not HIF-1α, suggested potential role of combined BOLD-MRI technique and HIF-1α staining in clinical diagnosis of renal carcinoma. HIF-2α may work as biological marker for renal cancer.

  15. Drosophila genome-wide RNAi screen identifies multiple regulators of HIF-dependent transcription in hypoxia.

    Directory of Open Access Journals (Sweden)

    Andrés Dekanty

    2010-06-01

    Full Text Available Hypoxia-inducible factors (HIFs are a family of evolutionary conserved alpha-beta heterodimeric transcription factors that induce a wide range of genes in response to low oxygen tension. Molecular mechanisms that mediate oxygen-dependent HIF regulation operate at the level of the alpha subunit, controlling protein stability, subcellular localization, and transcriptional coactivator recruitment. We have conducted an unbiased genome-wide RNA interference (RNAi screen in Drosophila cells aimed to the identification of genes required for HIF activity. After 3 rounds of selection, 30 genes emerged as critical HIF regulators in hypoxia, most of which had not been previously associated with HIF biology. The list of genes includes components of chromatin remodeling complexes, transcription elongation factors, and translational regulators. One remarkable hit was the argonaute 1 (ago1 gene, a central element of the microRNA (miRNA translational silencing machinery. Further studies confirmed the physiological role of the miRNA machinery in HIF-dependent transcription. This study reveals the occurrence of novel mechanisms of HIF regulation, which might contribute to developing novel strategies for therapeutic intervention of HIF-related pathologies, including heart attack, cancer, and stroke.

  16. Hypoxia and HIFs in regulating the development of the hematopoietic system.

    Science.gov (United States)

    Imanirad, Parisa; Dzierzak, Elaine

    2013-12-01

    Many physiologic processes during the early stages of mammalian ontogeny, particularly placental and vascular development, take place in the low oxygen environment of the uterus. Organogenesis is affected by hypoxia inducible factor (HIF) transcription factors that are sensors of hypoxia. In response to hypoxia, HIFs activate downstream target genes - growth and metabolism factors. During hematopoietic system ontogeny, blood cells and hematopoietic progenitor/stem cells are respectively generated from mesodermal precursors, hemangioblasts, and from a specialized subset of endothelial cells that are hemogenic. Since HIFs are known to play a central role in vascular development, and hematopoietic system development occurs in parallel to that of the vascular system, several studies have examined the role of HIFs in hematopoietic development. The response to hypoxia has been examined in early and mid-gestation mouse embryos through genetic deletion of HIF subunits. We review here the data showing that hematopoietic tissues of the embryo are hypoxic and express HIFs and HIF downstream targets, and that HIFs regulate the development and function of hematopoietic progenitor/stem cells.

  17. Analysis of HIF-1 inhibition by manassantin A and analogues with modified tetrahydrofuran configurations

    OpenAIRE

    Kasper, Amanda C.; Moon, Eui Jung; Hu, Xiangqian; Park, Yongho; Wooten, Ceshea M.; Kim, Hyoungsu; Yang, Weitao; Dewhirst, Mark W.; Hong, Jiyong

    2009-01-01

    We have shown that manassantin A downregulated the HIF-1α expression and inhibited the secretion of VEGF. We have also demonstrated that the 2,3-cis-3,4-trans-4,5-cis-configuration of the tetrahydrofuran is critical to the HIF-1 inhibition of manassantin A.

  18. Analysis of HIF-1 inhibition by manassantin A and analogues with modified tetrahydrofuran configurations.

    Science.gov (United States)

    Kasper, Amanda C; Moon, Eui Jung; Hu, Xiangqian; Park, Yongho; Wooten, Ceshea M; Kim, Hyoungsu; Yang, Weitao; Dewhirst, Mark W; Hong, Jiyong

    2009-07-15

    We have shown that manassantin A downregulated the HIF-1alpha expression and inhibited the secretion of VEGF. We have also demonstrated that the 2,3-cis-3,4-trans-4,5-cis-configuration of the tetrahydrofuran is critical to the HIF-1 inhibition of manassantin A.

  19. Association of HIF- expression and cell apoptosis after traumatic brain injury in the rat

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective: To explore the expression of hypoxia inducible factor-1α (HIF-1~) and the correlation between HIF-1α and apoptosis after traumatic brain injury.Methods: Using experimental traumatic brain injury in the rats, the expression of HIF-1α was studied by immunohisto-chemistry in cerebral tissue, apoptotic cell death was evaluated with TUNEL (transferase-mediated XdUTP nick end labeling ), and double-labeled immunohistochemistry and TUNEL methods were used to investigate the relationship between HIF-1α and apoptosis.Results: There was remarkable difference in the expression of HIF-1α between the experimental groups and the control groups (P < 0.01), in the experimental groups,the expression of HIF-1α at 48 hours was highest; the evidence of apoptotic cell death after experimental traumatic brain injury was found by TUNEL; the apoptotic percentage increased or decreased according to the changes of the positive expression of HIF-1α (r = 0.99).Conclusions: The results suggest that secondary brain ischemia plays a crucial role in apoptotic cell death after traumatic brain injury; HIF-1α can prompt apoptotic cell death after experimental traumatic brain injury.e expres

  20. Single nucleotide polymorphisms in the HIF-1α gene and chemoradiotherapy of locally advanced rectal cancer

    DEFF Research Database (Denmark)

    Havelund, Birgitte Mayland; Spindler, Karen-Lise Garm; Ploen, John

    2012-01-01

    The aim of this study was to investigate the predictive impact of polymorphisms in the HIF-1α gene on the response to chemoradiotherapy (CRT) in rectal cancer. This study included two cohorts of patients with locally advanced rectal cancer receiving long-course CRT. The HIF-1α C1772T (rs11549465...

  1. CypA, a gene downstream of HIF-1α, promotes the development of PDAC.

    Directory of Open Access Journals (Sweden)

    Huan Zhang

    Full Text Available Hypoxia-inducible factor-1α (HIF-1α is a highly important transcription factor involved in cell metabolism. HIF-1α promotes glycolysis and inhibits of mitochondrial respiration in pancreatic ductal adenocarcinoma (PDAC. In response to tumor hypoxia, cyclophilin A (CypA is over-expressed in various cancer types, and is associated with cell apoptosis, tumor invasion, metastasis, and chemoresistance in PDAC. In this study, we showed that both HIF-1α and CypA expression were significantly associated with lymph node metastasis and tumor stage. The expression of CypA was correlated with HIF-1α. Moreover, the mRNA and protein expression of CypA markedly decreased or increased following the suppression or over-expression of HIF-1α in vitro. Chromatin immunoprecipitation analysis showed that HIF-1α could directly bind to the hypoxia response element (HRE in the CypA promoter regions and regulated CypA expression. Consistent with other studies, HIF-1α and CypA promoted PDAC cell proliferation and invasion, and suppressed apoptosis in vitro. Furthermore, we proved the combination effect of 2-methoxyestradiol and cyclosporin A both in vitro and in vivo. These results suggested that,CypA, a gene downstream of HIF-1α, could promote the development of PDAC. Thus, CypA might serve as a potential therapeutic target for PDAC.

  2. Characteristics of Chinese Driver Behavior

    OpenAIRE

    LI J

    2014-01-01

    The high growth rate of vehicle ownership and many novel drivers in China determine the special features of Chinese driver behavior. This thesis introduces a comparative study on driver behavior by the analysis of saturation flow at urban intersections, Driver Behavior Questionnaire surveys, focus group discussion, and in-car tests. The main characteristics of Chinese driver behavior have been identified. A new method is developed for a simulation model calibration based on the study results.

  3. Towards "bionic" proteins: replacement of continuous sequences from HIF-1α with proteomimetics to create functional p300 binding HIF-1α mimics.

    Science.gov (United States)

    Burslem, George M; Kyle, Hannah F; Breeze, Alexander L; Edwards, Thomas A; Nelson, Adam; Warriner, Stuart L; Wilson, Andrew J

    2016-04-01

    Using the HIF-1α transcription factor as a model, this manuscript illustrates how an extended sequence of α-amino acids in a polypeptide can be replaced with a non-natural topographical mimic of an α-helix comprised from an aromatic oligoamide. The resultant hybrid is capable of reproducing the molecular recognition profile of the p300 binding sequence of HIF-1α from which it is derived.

  4. Hypoxia inducible factor-1-alpha (HIF-1α) is related to both angiogenesis and inflammation in rheumatoid arthritis

    NARCIS (Netherlands)

    Brouwer, Elisabeth; Gouw, Annette S.H.; Posthumus, Marcel D.; Van Leeuwen, Miek A.; Boerboom, Alexander L.; Bijzet, Johan; Bos, Reinhard; Limburg, Piet C.; Kallenberg, Cees G.M.; Westra, Johanna

    2009-01-01

    Objectives: Despite the important role of the transcription factor HIF-1α in angiogenesis and inflammation, only a few studies on HIF-1α expression have been performed in RA patients. The aim of the present study was to identify the layer in synovial tissue of RA patients where HIF1α is expressed an

  5. Hypoxia inducible factor-1-alpha (HIF-1 alpha) is related to both angiogenesis and inflammation in rheumatoid arthritis

    NARCIS (Netherlands)

    Brouwer, E.; Gouw, A. S. H.; Posthumus, M. D.; van Leeuwen, M. A.; Boerboom, A. L.; Bijzet, J.; Limburg, P. C.; Kallenberg, C. G. M.; Westra, J.; Bos, R

    2009-01-01

    Objectives Despite the important role of the transcription factor HIF-1 alpha in angiogenesis and inflammation, only a few studies on HIF-1 alpha expression have been performed in RA patients. The aim of the present study was to identify the layer in synovial tissue of RA patients where HIF1 alpha i

  6. Hypnosis and music interventions (HMIs) inactivate HIF-1: A potential curative efficacy for cancers and hypertension.

    Science.gov (United States)

    Wang, Jing-Zhang; Li, Ling; Pan, Li-Lan; Chen, Jian-Hua

    2015-11-01

    Hypnosis and music interventions (HMIs) have shown positive influence on cancers for nearly 200years, but the underlying mechanisms were rarely explored systematically. The hypothesis suggests a potential curative efficacy of HMIs on cancers by inhibiting hypoxia inducible factor-1 (HIF-1), which is a key mediator of cancer development, especially under hypoxic conditions. HMIs are sufficient to attenuate the pain and anxiety degree of individuals, improve multiple psychological and physiological parameters, and consequently, lead to increased oxygen saturation in vivo. Furthermore, abundant oxygen in vivo inhibits the activation of HIF-1 and potentially blockades kinds of HIF-1-induced oncogenic signaling pathways. The hypothesized efficacy of HMIs is very similar to anti-cancer medicines targeting HIF-1. The implication of the hypothesis in preventing hypertension is also discussed. In summary, the hypothesis clearly suggests the potential involvement of the convenient, safe, non-pharmaceutical, and low-cost HMIs in preventing HIF-1-mediated diseases, including cancers and hypertension.

  7. Noscapine sensitizes chemoresistant ovarian cancer cells to cisplatin through inhibition of HIF-1α.

    Science.gov (United States)

    Su, Wenjing; Huang, Lei; Ao, Qilin; Zhang, Qinghua; Tian, Xun; Fang, Yong; Lu, Yunping

    2011-06-01

    Hypoxia-inducible factor 1 alpha (HIF-1α) is closely related with chemoresistance of solid tumors. The purpose of this study was to investigate the ability of noscapine to inhibit HIF-1α and sensitize ovarian cancer cells to cisplatin (DDP) under hypoxic conditions. Herein, we report that noscapine sensitized cobalt-induced chemoresistant ovarian cancer cells to DDP-induced apoptosis and inhibition of cell proliferation. Noscapine also promoted proteasome-mediated degradation of cobalt-stabilized HIF-1α protein, with subsequent inhibition of HIF-1 transcriptional activity. These data establish noscapine as a small molecule inhibitor of HIF-1α and provide an evidence for its combination with DDP in combating ovarian cancer chemoresistance.

  8. HIF-1α inhibition blocks the cross talk between multiple myeloma plasma cells and tumor microenvironment

    Energy Technology Data Exchange (ETDEWEB)

    Borsi, Enrica, E-mail: enrica.borsi2@unibo.it [Department of Experimental Diagnostic and Specialty Medicine (DIMES), “L. and A. Seràgnoli”, Bologna University School of Medicine, S. Orsola' s University Hospital (Italy); Perrone, Giulia [Fondazione IRCCS Istituto Nazionale dei Tumori, Hematology Department, Via Venezian 1, 20133 Milano (Italy); Terragna, Carolina; Martello, Marina; Zamagni, Elena; Tacchetti, Paola; Pantani, Lucia; Brioli, Annamaria; Dico, Angela Flores; Zannetti, Beatrice Anna; Rocchi, Serena; Cavo, Michele [Department of Experimental Diagnostic and Specialty Medicine (DIMES), “L. and A. Seràgnoli”, Bologna University School of Medicine, S. Orsola' s University Hospital (Italy)

    2014-11-01

    Multiple myeloma (MM) is a malignant disorder of post-germinal center B cells, characterized by the clonal proliferation of malignant plasma cells (PCs) within the bone marrow (BM). The reciprocal and complex interactions that take place between the different compartments of BM and the MM cells result in tumor growth, angiogenesis, bone disease, and drug resistance. Given the importance of the BM microenvironment in MM pathogenesis, we investigated the possible involvement of Hypoxia-Inducible transcription Factor-1 alpha (HIF-1α) in the PCs-bone marrow stromal cells interplay. To test this hypothesis, we used EZN-2968, a 3rd generation antisense oligonucleotide against HIF-1α, to inhibit HIF-1α functions. Herein, we provide evidence that the interaction between MM cells and BM stromal cells is drastically reduced upon HIF-1α down-modulation. Notably, we showed that upon exposure to HIF-1α inhibitor, neither the incubation with IL-6 nor the co-culture with BM stromal cells were able to revert the anti-proliferative effect induced by EZN-2968. Moreover, we observed a down-modulation of cytokine-induced signaling cascades and a reduction of MM cells adhesion capability to the extracellular matrix proteins in EZN-2968-treated samples. Taken together, these results strongly support the concept that HIF-1α plays a critical role in the interactions between bone BM cells and PCs in Multiple Myeloma. - Highlights: • HIF-1α inhibition induces a mild apoptotic cell death. • Down-modulation of cytokine-induced signaling cascades upon HIF-1α inhibition. • Reduced interaction between MM cells and BMSCs upon HIF-1α down-modulation. • Reduced PCs adhesion to the extracellular matrix protein induced by EZN-2968. • HIF-1α inhibition may be an attractive therapeutic strategy for Multiple Myeloma.

  9. HIF-1 in cancer therapy: two decade long story of a transcription factor.

    Science.gov (United States)

    Soni, Sourabh; Padwad, Yogendra S

    2017-04-01

    Oxygen (O2) homeostasis is an indispensable requirement of eukaryotes. O2 concentration in cellular milieu is defined as normoxia (∼21% O2), physoxia (∼1-13% O2) or hypoxia (∼0.1-1% O2). Hypoxia, a striking micro-environmental feature in tumorigenesis, is countered by tumor cells via induction of O2 governed transcription factor, hypoxia inducible factor-1 (HIF-1). Post discovery, HIF-1 has emerged as a promising anticancer therapeutic target during the last two decades. Recent reports have highlighted that enhanced levels of HIF-1 correlate with tumor metastasis leading to poor patient prognosis. A systematic search in PubMed and SciFinder for the literature on HIF-1 biology and therapeutic importance in cancer was carried out. This review highlights the initial description as well as the recent insights into HIF-1 biology and regulation. We have focused on emerging data regarding varied classes of HIF-1 target genes affecting various levels of crosstalk among tumorigenic pathways. We have emphasized on the fact that HIF-1 acts as a networking hub coordinating activities of multiple signaling molecules influencing tumorigenesis. Emerging evidences indicate role of many HIF-induced proteomic and genomic alterations in malignant progression by mediating a myriad of genes stimulating angiogenesis, anaerobic metabolism and survival of cancer cells in O2-deficient microenvironment. Better understanding of the crucial role of HIF-1 in carcinogenesis could offer promising new avenues to researchers and aid in elucidating various open issues regarding the use of HIF-1 as an anticancer therapeutic target. In spite of large efforts in this field, many questions still remain unanswered. Hence, future investigations are necessary to devise, assess and refine methods for translating previous research efforts into novel clinical practices in cancer treatment.

  10. Importance of the HIF pathway in cobalt nanoparticle-induced cytotoxicity and inflammation in human macrophages.

    Science.gov (United States)

    Nyga, Agata; Hart, Alister; Tetley, Teresa D

    2015-01-01

    Recent, unexpected high failure rates of metal-on-metal hip implants have reintroduced the issue of cobalt toxicity. An adverse reaction to cobalt ions and cobalt-induced lung injury occurs during environmental exposure and is now strictly controlled. Currently adverse reaction occurs to cobalt nanoparticles during wear and tear of metal-on-metal hip implants of which the underlying mechanism is not fully understood. The putative role of the hypoxia-inducible factor (HIF) pathway in the mechanism of cobalt nanoparticle (Co-NPs) toxicity was examined using the U937 cell line, human alveolar macrophages and monocyte-derived macrophages. Co-NPs (5-20 μg/ml)-induced cytotoxicity (viability ranged from 75% to cobalt ions (Co(II); up to 350 μM) did not. Co-NPs induced HIF-1α stabilization. Addition of ascorbic acid (100 µM) and glutathione (1 mM) both prevented the increased ROS. However, only treatment with ascorbic acid reduced HIF-1α levels and prevented cell death, indicating that a ROS-independent pathway is involved in Co-NPs-induced cytotoxicity. Replenishing intracellular ascorbate, which is crucial in preventing HIF pathway activation, modified Co-induced HIF target gene expression and the inflammatory response, by decreasing interleukin-1 beta (IL-1β) mRNA and protein expression. Addition of glutathione had no effect on Co-NPs-induced HIF target gene expression or inflammatory response. Thus, Co-NPs induce the HIF pathway by depleting intracellular ascorbate, leading to HIF stabilization and pathway activation. This suggests a strong, ROS-independent role for HIF activation in Co-NPs-induced cytotoxicity and a possible role for HIF in metal-on-metal hip implant pathology.

  11. Desafiando o campo literário: Bufólicas e as estratégias de produção da crença em Hilda Hilst

    Directory of Open Access Journals (Sweden)

    Clovis Carvalho Britto

    2016-01-01

    Full Text Available El artículo analiza las estrategias de producción de creencia en Hilda Hilst (1930-2004 a través de las tensiones generadas por Bufólicas (1992 en el campo literario brasilero. Basado en referencias a Pierre Bourdieu, el articulo pone de relieve cómo la escritora promueve medios en favor de su distinción en ese espacio social al establecer un proyecto basado en la obscenidad, la risa y una fuerte crítica del mercado editorial. Con este fin, investiga cómo el libro consistió en un proyecto político, dirigiendo la producción de repertorios específicos acerca de Hilst en el mercado de bienes simbólicos, y al re-actualizar y ritualizar versiones construidas por la autora y por otros agentes.

  12. HIF-1α和HIF-2α上调非小细胞肺癌中CCR7的表达%Upregulation of the Chemokine Receptor CCR7 expression by HIF-1α and HIF-2α in non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    李洋; 张清富; 江黎黎; 邱雪杉; 王恩华

    2008-01-01

    背景与目的 CCR7与非小细胞肺癌的淋巴结转移密切相关,但CCR7的上调机制却不是很清楚.本研究通过观察趋化因子受体CCR7和缺氧诱导因子1α(HIF-1α)、缺氧诱导因子2α(HIF-2α)在非小细胞肺癌组织中的表达及相互关系,探讨非小细胞肺癌CCR7上调机制.方法 应用免疫组织化学染色(SP法)检测94例非小细胞肺癌组织中CCR7、HIF-1α和HIF-2α的表达,并联合运用RNAi技术沉默人肺腺癌A549细胞中HIF-1α、HIF-2α表达,采用RT-PCR和免疫荧光方法观察CCR7的变化情况,分析CCR7表达与HIF-1α、HIF-2α之间的关系:结果免疫组织化学结果显示:CCR7主要表达于癌细胞质和(或)胞膜,HIF-1α、HIF-2α主要表达于癌细胞核和(或)细胞质,非小细胞肺癌中CCR7、HIF-1α和HIF-2α的表达率分别为75.53%(71/94)、54.25%(51/94)和70.21%(66194),χ2榆验显示CCR7表达与非小细胞肺癌的临床病理分期(P0.05).此外.CCR7和HIF-1α、HIF-2α表达正相关(r=0.272,P<0.01)(r=0.225,P<0.05).向A549细胞中转染HIF-1α、HIF-2α特异性siRNA.分别抑制HIF-1α、HIF-2α表达后发现CCR7的mRNA和蛋白水平均下调(P<0.05).结论 CCR7表达与非小细胞肺癌侵袭转移密切相关,CCR7在NSCLC中过表达受HIF-1α和H1F-2α调控.

  13. Real-time imaging of HIF-1alpha stabilization and degradation.

    Directory of Open Access Journals (Sweden)

    Ekaterina Moroz

    Full Text Available HIF-1alpha is overexpressed in many human cancers compared to normal tissues due to the interaction of a multiplicity of factors and pathways that reflect specific genetic alterations and extracellular stimuli. We developed two HIF-1alpha chimeric reporter systems, HIF-1alpha/FLuc and HIF-1alpha(DeltaODDD/FLuc, to investigate the tightly controlled level of HIF-1alpha protein in normal (NIH3T3 and HEK293 and glioma (U87 cells. These reporter systems provided an opportunity to investigate the degradation of HIF-1alpha in different cell lines, both in culture and in xenografts. Using immunofluorescence microscopy, we observed different patterns of subcellular localization of HIF-1alpha/FLuc fusion protein between normal cells and cancer cells; similar differences were observed for HIF-1alpha in non-transduced, wild-type cells. A dynamic cytoplasmic-nuclear exchange of the fusion protein and HIF-1alpha was observed in NIH3T3 and HEK293 cells under different conditions (normoxia, CoCl2 treatment and hypoxia. In contrast, U87 cells showed a more persistent nuclear localization pattern that was less affected by different growing conditions. Employing a kinetic model for protein degradation, we were able to distinguish two components of HIF-1alpha/FLuc protein degradation and quantify the half-life of HIF-1alpha fusion proteins. The rapid clearance component (t(1/2 approximately 4-6 min was abolished by the hypoxia-mimetic CoCl2 MG132 treatment and deletion of ODD domain, and reflects the oxygen/VHL-dependent degradation pathway. The slow clearance component (t(1/2 approximately 200 min is consistent with other unidentified non-oxygen/VHL-dependent degradation pathways. Overall, the continuous bioluminescence readout of HIF-1alpha/FLuc stabilization in vitro and in vivo will facilitate the development and validation of therapeutics that affect the stability and accumulation of HIF-1alpha.

  14. NF-{kappa}B suppresses HIF-1{alpha} response by competing for P300 binding

    Energy Technology Data Exchange (ETDEWEB)

    Mendonca, Daniela B.S., E-mail: daniela_mendonca@dentistry.unc.edu [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Av. W5 Norte, 70790-160 Brasilia, DF (Brazil); Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 330 Brauer Hall, CB 7450, Chapel Hill, NC 27599 (United States); Mendonca, Gustavo [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Av. W5 Norte, 70790-160 Brasilia, DF (Brazil); Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 330 Brauer Hall, CB 7450, Chapel Hill, NC 27599 (United States); Aragao, Francisco J.L. [Universidade Catolica de Brasilia, Pos-Graduacao em Ciencias Genomicas e Biotecnologia, SGAN Quadra 916, Av. W5 Norte, 70790-160 Brasilia, DF (Brazil); Embrapa Recursos Geneticos e Biotecnologia, Laboratorio de Introducao e Expressao de Genes, PqEB W5 Norte, 70770-900 Brasilia, DF (Brazil); Cooper, Lyndon F., E-mail: lyndon_cooper@dentistry.unc.edu [Bone Biology and Implant Therapy Laboratory, Department of Prosthodontics, University of North Carolina at Chapel Hill, 330 Brauer Hall, CB 7450, Chapel Hill, NC 27599 (United States)

    2011-01-28

    Research highlights: {yields} p65 completely blocked HIF-1{alpha} activity at the HRE on different cell lines. {yields} p65 caused minor changes in HIF-1{alpha} and HIF-1{alpha} target genes mRNA expression. {yields} p65 reduced transcription of VEGF promoter. {yields} p65 competes with HIF-1{alpha} for p300. -- Abstract: Hypoxia has emerged as a key determinant of osteogenesis. HIF-1{alpha} is the transcription factor mediating hypoxia responses that include induction of VEGF and related bone induction. Inflammatory signals antagonize bone repair via the NF-{kappa}B pathway. The present investigation explored the functional relationship of hypoxia (HIF-1{alpha} function) and inflammatory signaling (NF-{kappa}B) in stem like and osteoprogenitor cell lines. The potential interaction between HIF-1{alpha} and NF-{kappa}B signaling was explored by co-transfection studies in hFOB with p65, HIF-1{alpha} and 9x-HRE-luc or HIF-1{alpha} target genes reporter plasmids. Nuclear cross-talk was directly tested using the mammalian Gal4/VP16 two-hybrid, and confirmed by co-immunoprecipitation/western blotting assays. The results show that inflammatory stimulation (TNF-{alpha} treatment) causes a marked inhibition of HIF-1{alpha} function at the HRE in all cell lines studied. Also, co-transfection with p65 expression vector leads to reduced hVEGFp transcription after DFO-induced hypoxia. However, TNF-{alpha} treatment had little effect on HIF-1{alpha} mRNA levels. The functional interaction of Gal4-HIF-1{alpha} and VP16-p300 fusion proteins is effectively blocked by expression of p65 in a dose dependent manner. It was concluded that NF-{kappa}B-mediated inflammatory signaling is able to block HIF-1{alpha} transactivation at HRE-encoding genes by direct competition for p300 binding at the promoter. Inflammation may influence the stem cell niche and tissue regeneration by influencing cellular responses to hypoxia.

  15. Alternate laser fusion drivers

    Energy Technology Data Exchange (ETDEWEB)

    Pleasance, L.D.

    1979-11-01

    Over the past few years, several laser systems have been considered as possible laser fusion drivers. Recently, there has been an increasing effort to evaluate these systems in terms of a reactor driver application. The specifications for such a system have become firmer and generally more restrictive. Several of the promising candidates such as the group VI laser, the metal vapor excimers and some solid state lasers can be eliminated on the basis of inefficiency. New solid state systems may impact the long range development of a fusion driver. Of the short wavelength gas lasers, the KrF laser used in conjunction with Raman compression and pulse stacking techniques is the most promising approach. Efficiencies approaching 10% may be possible with this system. While technically feasible, these approaches are complex and costly and are unsatisfying in an aethetic sense. A search for new lasers with more compelling features is still needed.

  16. Really Scary Drivers

    Institute of Scientific and Technical Information of China (English)

    马莲花

    2005-01-01

    A new wave of "road killers", or new drivers, on Beijing's streets has prompted traffic authorities to do something to make driving tests more difficult. This year, the move has targeted new drivers to keep them from posing a threat, the Beijing Traffic Management Bureau says. The new test has been adopted citywide and the average pass rate is down to 50 per cent from a previous 80 per cent, at the city's 22 test centers, said Jiang Jing, a bureau press officer. The test now has six mandatory items chosen r...

  17. Myc post-transcriptionally induces HIF1 protein and target gene expression in normal and cancer cells

    Science.gov (United States)

    Doe, Megan R.; Ascano, Janice; Kaur, Mandeep; Cole, Michael D.

    2012-01-01

    c-Myc is frequently overexpressed in tumors and plays an important role in the regulation of cancer metabolism. Hypoxia-inducible factor-1 (HIF1), the master regulator of the hypoxic response, enhances tumorigenesis and influences metabolism via upregulation of the glycolytic pathway and suppression of mitochondrial respiration. Together, deregulated Myc and HIF1 cooperate to lend metabolic advantages to proliferating cancer cells and contribute to the Warburg Effect. Here we show that overexpression of Myc significantly stabilizes the alpha subunit of HIF1 (HIF1alpha) under normoxic conditions and enhances HIF1alpha accumulation under hypoxic conditions in cells. Post-transcriptional regulation of HIF1α by Myc led to the induction of HIF1α gene targets. Normoxic HIF1α protein expression was also dependent on Myc. Functionally; HIF1α expression was required for Myc-induced anchorage-independent growth and cell proliferation. Myc-dependent stabilization of HIF1α involved either disruption of binding to the VHL complex or post-translational protein modifications. Taken together, our findings uncover a previously uncharacterized regulatory relationship between Myc and HIF1 that has important implications for cancer metabolism and development. PMID:22186139

  18. Castration Therapy of Prostate Cancer Results in Downregulation of HIF-1{alpha} Levels

    Energy Technology Data Exchange (ETDEWEB)

    Al-Ubaidi, Firas L.T. [Department of Genetics, Microbiology and Toxicology, Stockholm University, Stockholm (Sweden); Department of Urology, Central Hospital, Vaesteras (Sweden); Schultz, Niklas [Department of Genetics, Microbiology and Toxicology, Stockholm University, Stockholm (Sweden); Egevad, Lars [Department of Oncology-Pathology, Karolinska Institutet, Stockholm (Sweden); Granfors, Torvald [Department of Urology, Central Hospital, Vaesteras (Sweden); Helleday, Thomas, E-mail: helleday@gmt.su.se [Department of Genetics, Microbiology and Toxicology, Stockholm University, Stockholm (Sweden); Gray Institute for Radiation Oncology and Biology, University of Oxford, Oxford (United Kingdom); Science for Life Laboratory, Stockholm University, Solna (Sweden)

    2012-03-01

    Background and Purpose: Neoadjuvant androgen deprivation in combination with radiotherapy of prostate cancer is used to improve radioresponsiveness and local tumor control. Currently, the underlying mechanism is not well understood. Because hypoxia causes resistance to radiotherapy, we wanted to test whether castration affects the degree of hypoxia in prostate cancer. Methods and Materials: In 14 patients with locally advanced prostate cancer, six to 12 prostatic needle core biopsy specimens were taken prior to castration therapy. Bilateral orchidectomy was performed in 7 patients, and 7 were treated with a GnRH-agonist (leuprorelin). After castrationm two to four prostatic core biopsy specimens were taken, and the level of hypoxia-inducible factor-1{alpha} (HIF-1{alpha}) in cancer was determined by immunofluorescence. Results: Among biopsy specimens taken before castration, strong HIF-1{alpha} expression (mean intensity above 30) was shown in 5 patients, weak expression (mean intensity 10-30) in 3 patients, and background levels of HIF-1{alpha} (mean intensity 0-10) in 6 patients. Downregulation of HIF-1{alpha} expression after castration was observed in all 5 patients with strong HIF-1{alpha} precastration expression. HIF-1{alpha} expression was also reduced in 2 of 3 patients with weak HIF-1{alpha} precastration expression. Conclusions: Our data suggest that neoadjuvant castration decreases tumor cell hypoxia in prostate cancer, which may explain increased radiosensitivity after castration.

  19. HIF1-Alpha Expression Predicts Survival of Patients with Squamous Cell Carcinoma of the Oral Cavity

    Science.gov (United States)

    dos Santos, Marcelo; Mercante, Ana Maria da Cunha; Louro, Iúri Drumond; Gonçalves, Antônio José; de Carvalho, Marcos Brasilino; da Silva, Eloiza Helena Tajara; da Silva, Adriana Madeira Álvares

    2012-01-01

    Background Oral squamous cell carcinoma is an important cause of death and morbidity wordwide and effective prognostic markers are still to be discovered. HIF1α protein is associated with hypoxia response and neovascularization, essential conditions for solid tumors survival. The relationship between HIF1α expression, tumor progression and treatment response in head and neck cancer is still poorly understood. Patients and Methods In this study, we investigated HIF1α expression by immunohistochemistry in tissue microarrays and its relationship with clinical findings, histopathological results and survival of 66 patients with squamous cell carcinoma of the lower mouth. Results Our results demonstrated that high HIF1α expression is associated with local disease-free survival, independently from the choice of treatment. Furthermore, high expression of HIF1α in patients treated with postoperative radiotherapy was associated with survival, therefore being a novel prognostic marker in squamous cell carcinoma of the mouth. Additionally, our results showed that MVD was associated with HIF1α expression and local disease relapse. Conclusion These findings suggest that HIF1α expression can be used as a prognostic marker and predictor of postoperative radiotherapy response, helping the oncologist choose the best treatment for each patient. PMID:23028863

  20. Expression and significance of HIF-1α and VEGF in rats with diabetic retinopathy

    Institute of Scientific and Technical Information of China (English)

    Hong-Tao Yan; Guan-Fang Su

    2014-01-01

    Objective:To investigate the expression of hypoxia inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VEGF) in diabetic retinopathy(DR) rats and its effect on theDR occurrence and development.Methods:A total of120SD rats were randomly divided into trial group and control group with60 in each.STZi.p. was used in the trial group to establish theDM model, citrate buffer salt of same amount was usedi.p. to the control group.1,3 and6 months after injection, respective20 rats were sacrificed in each group to observe expression ofHIF-1α andVEGF in the rat retina tissue at different time points.Results:Expression ofHIF-1α andVEGF were negative in the control group; expression ofHIF-1α andVEGF protein in retinal tissue were weak after1 month ofDR mold formation.It showed progressive enhancement along with the progression in different organizations, differences between groups were significant (P<0.05).Conclusions:Expressions ofHIF-1α andVEGF were correlated with disease progression in early diabetic retinopathy.Retinal oxygen can induce over-expression ofHIF-1α andVEGF.It shows thatHIF-1α andVEGF play an important role in the pathogenesis ofDR.

  1. Myeloid cell-derived HIF attenuates inflammation in UUO-induced kidney injury

    Science.gov (United States)

    Kobayashi, Hanako; Gilbert, Victoria; Liu, Qingdu; Kapitsinou, Pinelopi P.; Unger, Travis L.; Rha, Jennifer; Rivella, Stefano; Schlöndorff, Detlef; Haase, Volker H.

    2012-01-01

    Renal fibrosis and inflammation are associated with hypoxia, and tissue pO2 plays a central role in modulating the progression of chronic kidney disease. Key mediators of cellular adaptation to hypoxia are hypoxia-inducible factor (HIF)-1 and -2. In the kidney they are expressed in a cell type-specific manner; to what degree activation of each homolog modulates renal fibrogenesis and inflammation has not been established. To address this issue, we used Cre-loxP recombination to activate or to delete both Hif-1 and Hif-2 either globally or cell type-specifically in myeloid cells. Global activation of Hif suppressed inflammation and fibrogenesis in mice subjected to unilateral ureteral obstruction, while activation of Hif in myeloid cells suppressed inflammation only. Suppression of inflammatory cell infiltration was associated with down-regulation of CC chemokine receptors in renal macrophages. Conversely, global deletion or myeloid-specific inactivation of Hif promoted inflammation. Furthermore, prolonged hypoxia suppressed the expression of multiple inflammatory molecules in non-injured kidneys. Collectively, we provide experimental evidence that hypoxia and/or myeloid cell-specific HIF activation attenuates renal inflammation associated with chronic kidney injury. PMID:22490864

  2. The HIF/VHL pathway: from oxygen sensing to innate immunity.

    Science.gov (United States)

    Walmsley, Sarah R; McGovern, Naomi N; Whyte, Moira K B; Chilvers, Edwin R

    2008-03-01

    In aerobic organisms, all cells have the capacity to respond to changes in oxygenation through the stabilization and transcriptional activation of hypoxia-inducible factor (HIF). At sites of tissue injury, oxygen delivery to individual cells may be compromised or insufficient due to increased metabolic demands, and it is to these areas that immune cells, including neutrophils, must migrate and operate effectively. In addition to the role of HIF to regulate the adaptive metabolic and survival responses of these cells at sites of reduced oxygenation, more complex interactions between HIF and pro-inflammatory pathways are now emerging. The mechanisms by which HIF modulates pro-inflammatory myeloid cell lifespan and function remain to be fully characterized, but roles for the oxygen-sensing hydroxylase enzymes through direct hydroxylation of NF-kappaB and its repressor protein IkappaBalpha have been suggested. The ability of HIF to modulate cellular glucose utilization is also thought to be important, with the maintenance of intracellular ATP pools linked to enhanced myeloid cell aggregation, motility, invasiveness, and bacterial killing. Additional non-hypoxia-mediated routes to up-regulate HIF are also now recognized. In this review we describe the role of HIF in the oxygen-sensing response, and the oxygen-dependent and -independent regulation of myeloid cell function and longevity. Understanding these processes and the role they play in regulating innate immune responses within inflamed sites, both hypoxic and normoxic, may offer new opportunities for therapeutic intervention.

  3. Cloning and characterization of the rat HIF-1 alpha prolyl-4-hydroxylase-1 gene.

    Science.gov (United States)

    Cobb, Ronald R; McClary, John; Manzana, Warren; Finster, Silke; Larsen, Brent; Blasko, Eric; Pearson, Jennifer; Biancalana, Sara; Kauser, Katalin; Bringmann, Peter; Light, David R; Schirm, Sabine

    2005-08-01

    Prolyl-4-hydroxylase domain-containing enzymes (PHDs) mediate the oxygen-dependent regulation of the heterodimeric transcription factor hypoxia-inducible factor-1 (HIF-1). Under normoxic conditions, one of the subunits of HIF-1, HIF-1alpha, is hydroxylated on specific proline residues to target HIF-1alpha for degradation by the ubiquitin-proteasome pathway. Under hypoxic conditions, the hydroxylation by the PHDs is attenuated by lack of the oxygen substrate, allowing HIF-1 to accumulate, translocate to the nucleus, and mediate HIF-mediated gene transcription. In several mammalian species including humans, three PHDs have been identified. We report here the cloning of a full-length rat cDNA that is highly homologous to the human and murine PHD-1 enzymes and encodes a protein that is 416 amino acids long. Both cDNA and protein are widely expressed in rat tissues and cell types. We demonstrate that purified and crude baculovirus-expressed rat PHD-1 exhibits HIF-1alpha specific prolyl hydroxylase activity with similar substrate affinities and is comparable to human PHD-1 protein.

  4. Rapamycin Inhibits Proliferation of Hemangioma Endothelial Cells by Reducing HIF-1-Dependent Expression of VEGF

    Science.gov (United States)

    Medici, Damian; Olsen, Bjorn R.

    2012-01-01

    Hemangiomas are tumors formed by hyper-proliferation of vascular endothelial cells. This is caused by elevated vascular endothelial growth factor (VEGF) signaling through VEGF receptor 2 (VEGFR2). Here we show that elevated VEGF levels produced by hemangioma endothelial cells are reduced by the mTOR inhibitor rapamycin. mTOR activates p70S6K, which controls translation of mRNA to generate proteins such as hypoxia inducible factor-1 (HIF-1). VEGF is a known HIF-1 target gene, and our data show that VEGF levels in hemangioma endothelial cells are reduced by HIF-1α siRNA. Over-expression of HIF-1α increases VEGF levels and endothelial cell proliferation. Furthermore, both rapamycin and HIF-1α siRNA reduce proliferation of hemangioma endothelial cells. These data suggest that mTOR and HIF-1 contribute to hemangioma endothelial cell proliferation by stimulating an autocrine loop of VEGF signaling. Furthermore, mTOR and HIF-1 may be therapeutic targets for the treatment of hemangiomas. PMID:22900063

  5. Radiation promotes malignant progression of glioma cells through HIF-1alpha stabilization.

    Science.gov (United States)

    Kim, Young-Heon; Yoo, Ki-Chun; Cui, Yan-Hong; Uddin, Nizam; Lim, Eun-Jung; Kim, Min-Jung; Nam, Seon-Young; Kim, In-Gyu; Suh, Yongjoon; Lee, Su-Jae

    2014-11-01

    Given its contribution to malignant phenotypes of cancer, tumor hypoxia has been considered as a potential therapeutic problem. In the stressful microenvironment condition, hypoxia inducible factor 1 (HIF1) is well known to mediate the transcriptional adaptation of cells to hypoxia and acts as a central player for the process of hypoxia-driven malignant cancer progression. Here, we found that irradiation causes the HIF1α protein to stabilize, even in normoxia condition through activation of p38 MAPK, thereby promoting angiogenesis in tumor microenvironment and infiltrative property of glioma cells. Notably, irradiation reduced hydroxylation of HIF1α through destabilization of prolyl hydroxylases (PHD)-2. Moreover, radiation also decreased the half-life of protein von Hippel-Lindau (pVHL), which is a specific E3 ligase for HIF1α. Of note, inhibition of p38 MAPK attenuated radiation-induced stabilization of HIF1α through destabilization of PHD-2 and pVHL. In agreement with these results, targeting of either p38 MAPK, HIF1α, pVHL or PHD-2 effectively mitigated the radiation-induced tube formation of human brain-derived micro-vessel endothelial cells (HB-MEC) and infiltration of glioma cells. Taken together, our findings suggest that targeting HIF1α in combination with ionizing radiation might increase the efficacy of radiotherapy for glioma treatment. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  6. Effect of inducible expression of HIF-1α on prostate cancer cell in MPPa-PDT

    Science.gov (United States)

    Tian, Y. Y.; Kong, F.; Tian, X.; Guo, Q.; Cui, F. A.

    2008-10-01

    Photodynamic therapy (PDT) is a promising treatment on neoplastic pathologic tissues, which involves the administration of a photosensitizing agent followed by exposure of the tissue to visible non-thermal light. In our previous findings MPPa was showed to be a good photosensitizer candidature, because it can lead PC-3M cell line to death mainly via apoptotic way. In this study, we studied the effect of overexpression of hypoxia inducible factor-1α (HIF-1α) on MPPa-PDT. Over expression of HIF-1α was induced by cobalt chloride (CoCl2) and then the cell viability was tested by MTT assay. After that, HIF-1α siRNA was transfected into PC-3M cells to knock down the expression of HIF-1α and the cell viability was observed. Western blot was used to determine the expression of HIF-1α. Finally, we found that the over expression of HIF-1α would reduce the photodynamic effect of MPPa on PC-3M cells, while the photodynamic effect would be enhanced by HIF-1α siRNA.

  7. Mosaicism in HIF2A-related polycythemia-paraganglioma syndrome.

    Science.gov (United States)

    Buffet, Alexandre; Smati, Sarra; Mansuy, Ludovic; Ménara, Mélanie; Lebras, Maëlle; Heymann, Marie-Françoise; Simian, Christophe; Favier, Judith; Murat, Arnaud; Cariou, Bertrand; Gimenez-Roqueplo, Anne-Paule

    2014-02-01

    HIF2A germline mutations were known to cause congenital polycythemia. Recently, HIF2A somatic mutations were found in several patients with polycythemia and paraganglioma, pheochromocytoma, or somatostatinoma, suggesting the occurrence of a de novo postzygotic HIF2A mutation that has not been demonstrated clearly. Patient 1 is a woman suffering from polycythemia diagnosed at the age of 16 years. She was operated on for a pheochromocytoma at 45 years and for two abdominal paragangliomas at 59 years. She was also diagnosed with somatostatinoma. Patient 2 is a young boy who suffered from polycythemia since infancy. He underwent surgery for a nonfunctional adrenal paraganglioma at the age of 9 years. We sequenced by Sanger and next-generation sequencing the HIF2A gene in DNA extracted from tumors, leukocytes, and buccal cells. In patient 1, we identified a somatic HIF2A mutation (c.1586T>C; p.Leu529Pro) in DNA extracted from both paragangliomas. The mutation was detected as a somatic mosaic in DNA extracted from somatostatinoma and was absent from germline DNA. In patient 2, we found an HIF2A heterozygous mutation (c.1625T>C; p.Leu542Pro) in the paraganglioma, but the mutation was also present as a mosaic in leukocyte DNA and in DNA extracted from buccal cells (3.3 and 8.96% of sequencing reads, respectively). Both mutations disrupt the hydroxylation domain of the HIF2α protein. Our study shows that HIF2A-related tumors are caused by postzygotic mutations occurring in early developmental stages. Potential germline mosaicism should be considered during the familial genetic counseling when an individual has been diagnosed with HIF2A-related polycythemia-paraganglioma syndrome.

  8. P70S6K 1 regulation of angiogenesis through VEGF and HIF-1{alpha} expression

    Energy Technology Data Exchange (ETDEWEB)

    Bian, Chuan-Xiu; Shi, Zhumei [Department of Pathology, Cancer Center, Nanjing Medical University, Nanjing 210029 (China); Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi [Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107 (United States); Jiang, Bing-Hua, E-mail: binghjiang@yahoo.com [Department of Pathology, Cancer Center, Nanjing Medical University, Nanjing 210029 (China); Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107 (United States)

    2010-07-30

    Research highlights: {yields} P70S6K1 regulates VEGF expression; {yields} P70S6K1 induces transcriptional activation through HIF-1{alpha} binding site; {yields} P70S6K1 regulates HIF-1{alpha}, but not HIF-1{beta} protein expression; {yields} P70S6K1 mediates tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression. -- Abstract: The 70 kDa ribosomal S6 kinase 1 (p70S6K1), a downstream target of phosphoinositide 3-kinase (PI3K) and ERK mitogen-activated protein kinase (MAPK), is an important regulator of cell cycle progression, and cell proliferation. Recent studies indicated an important role of p70S6K1 in PTEN-negative and AKT-overexpressing tumors. However, the mechanism of p70S6K1 in tumor angiogenesis remains to be elucidated. In this study, we specifically inhibited p70S6K1 activity in ovarian cancer cells using vector-based small interfering RNA (siRNA) against p70S6K1. We found that knockdown of p70S6K1 significantly decreased VEGF protein expression and VEGF transcriptional activation through the HIF-1{alpha} binding site at its enhancer region. The expression of p70S6K1 siRNA specifically inhibited HIF-1{alpha}, but not HIF-1{beta} protein expression. We also found that p70S6K1 down-regulation inhibited ovarian tumor growth and angiogenesis, and decreased cell proliferation and levels of VEGF and HIF-1{alpha} expression in tumor tissues. Our results suggest that p70S6K1 is required for tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression, providing a molecular mechanism of human ovarian cancer mediated by p70S6K1 signaling.

  9. ECM-dependent HIF induction directs trophoblast stem cell fate via LIMK1-mediated cytoskeletal rearrangement.

    Directory of Open Access Journals (Sweden)

    Hwa J Choi

    Full Text Available The Hypoxia-inducible Factor (HIF family of transcriptional regulators coordinates the expression of dozens of genes in response to oxygen deprivation. Mammalian development occurs in a hypoxic environment and HIF-null mice therefore die in utero due to multiple embryonic and placental defects. Mouse embryonic stem cells do not differentiate into placental cells; therefore, trophoblast stem cells (TSCs are used to study mouse placental development. Consistent with a requirement for HIF activity during placental development in utero, TSCs derived from HIF-null mice exhibit severe differentiation defects and fail to form trophoblast giant cells (TGCs in vitro. Interestingly, differentiating TSCs induce HIF activity independent of oxygen tension via unclear mechanisms. Here, we show that altering the extracellular matrix (ECM composition upon which TSCs are cultured changes their differentiation potential from TGCs to multinucleated syncytiotropholasts (SynTs and blocks oxygen-independent HIF induction. We further find that modulation of Mitogen Activated Protein Kinase Kinase-1/2 (MAP2K1/2, MEK-1/2 signaling by ECM composition is responsible for this effect. In the absence of ECM-dependent cues, hypoxia-signaling pathways activate this MAPK cascade to drive HIF induction and redirect TSC fate along the TGC lineage. In addition, we show that integrity of the microtubule and actin cytoskeleton is critical for TGC fate determination. HIF-2α ensures TSC cytoskeletal integrity and promotes invasive TGC formation by interacting with c-MYC to induce non-canonical expression of Lim domain kinase 1-an enzyme that regulates microtubule and actin stability, as well as cell invasion. Thus, we find that HIF can integrate positional and metabolic cues from within the TSC niche to regulate placental development by modulating the cellular cytoskeleton via non-canonical gene expression.

  10. HIF-1 modulates dietary restriction-mediated lifespan extension via IRE-1 in Caenorhabditis elegans.

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    Di Chen

    2009-05-01

    Full Text Available Dietary restriction (DR extends lifespan in various species and also slows the onset of age-related diseases. Previous studies from flies and yeast have demonstrated that the target of rapamycin (TOR pathway is essential for longevity phenotypes resulting from DR. TOR is a conserved protein kinase that regulates growth and metabolism in response to nutrients and growth factors. While some of the downstream targets of TOR have been implicated in regulating lifespan, it is still unclear whether additional targets of this pathway also modulate lifespan. It has been shown that the hypoxia inducible factor-1 (HIF-1 is one of the targets of the TOR pathway in mammalian cells. HIF-1 is a transcription factor complex that plays key roles in oxygen homeostasis, tumor formation, glucose metabolism, cell survival, and inflammatory response. Here, we describe a novel role for HIF-1 in modulating lifespan extension by DR in Caenorhabditis elegans. We find that HIF-1 deficiency results in extended lifespan, which overlaps with that by inhibition of the RSKS-1/S6 kinase, a key component of the TOR pathway. Using a modified DR method based on variation of bacterial food concentrations on solid agar plates, we find that HIF-1 modulates longevity in a nutrient-dependent manner. The hif-1 loss-of-function mutant extends lifespan under rich nutrient conditions but fails to show lifespan extension under DR. Conversely, a mutation in egl-9, which increases HIF-1 activity, diminishes the lifespan extension under DR. This deficiency is rescued by tissue-specific expression of egl-9 in specific neurons and muscles. Increased lifespan by hif-1 or DR is dependent on the endoplasmic reticulum (ER stress regulator inositol-requiring protein-1 (IRE-1 and is associated with lower levels of ER stress. Therefore, our results demonstrate a tissue-specific role for HIF-1 in the lifespan extension by DR involving the IRE-1 ER stress pathway.

  11. Loss of skeletal muscle HIF-1alpha results in altered exercise endurance.

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    Steven D Mason

    2004-10-01

    Full Text Available The physiological flux of oxygen is extreme in exercising skeletal muscle. Hypoxia is thus a critical parameter in muscle function, influencing production of ATP, utilization of energy-producing substrates, and manufacture of exhaustion-inducing metabolites. Glycolysis is the central source of anaerobic energy in animals, and this metabolic pathway is regulated under low-oxygen conditions by the transcription factor hypoxia-inducible factor 1alpha (HIF-1alpha. To determine the role of HIF-1alpha in regulating skeletal muscle function, we tissue-specifically deleted the gene encoding the factor in skeletal muscle. Significant exercise-induced changes in expression of genes are decreased or absent in the skeletal-muscle HIF-1alpha knockout mice (HIF-1alpha KOs; changes in activities of glycolytic enzymes are seen as well. There is an increase in activity of rate-limiting enzymes of the mitochondria in the muscles of HIF-1alpha KOs, indicating that the citric acid cycle and increased fatty acid oxidation may be compensating for decreased flow through the glycolytic pathway. This is corroborated by a finding of no significant decreases in muscle ATP, but significantly decreased amounts of lactate in the serum of exercising HIF-1alpha KOs. This metabolic shift away from glycolysis and toward oxidation has the consequence of increasing exercise times in the HIF-1alpha KOs. However, repeated exercise trials give rise to extensive muscle damage in HIF-1alpha KOs, ultimately resulting in greatly reduced exercise times relative to wild-type animals. The muscle damage seen is similar to that detected in humans in diseases caused by deficiencies in skeletal muscle glycogenolysis and glycolysis. Thus, these results demonstrate an important role for the HIF-1 pathway in the metabolic control of muscle function.

  12. Improving Driver Performance. A Curriculum for Licensed Drivers.

    Science.gov (United States)

    Highway Users Federation for Safety and Mobility, Washington, DC.

    Curriculum material presented in this manual is for use in the development of an instructional program for drivers who either want or need to improve their driving performance. Three principal units are included: man and highway transportation, driver performance, and factors influencing driver behavior. Each unit is further divided into episodes…

  13. Simulators in driver training.

    NARCIS (Netherlands)

    2009-01-01

    In 2010, about 150 driving simulators were being used for the basic driver training in the Netherlands. According to theories about how people learn, simulator training has both advantages and disadvantages. In order to be able to learn something from a simulator, its technical quality must be

  14. Simulators in driver training.

    NARCIS (Netherlands)

    2009-01-01

    In 2010, about 150 driving simulators were being used for the basic driver training in the Netherlands. According to theories about how people learn, simulator training has both advantages and disadvantages. In order to be able to learn something from a simulator, its technical quality must be adequ

  15. Seven Performance Drivers.

    Science.gov (United States)

    Ross, Linda

    2003-01-01

    Recent work with automotive e-commerce clients led to the development of a performance analysis methodology called the Seven Performance Drivers, including: standards, incentives, capacity, knowledge and skill, measurement, feedback, and analysis. This methodology has been highly effective in introducing and implementing performance improvement.…

  16. Molecular characterization of hypoxia and hypoxia-inducible factor 1 alpha (HIF-1α) from Taiwan voles (Microtus kikuchii).

    Science.gov (United States)

    Jiang, Yi-Fan; Chou, Chung-Hsi; Lin, En-Chung; Chiu, Chih-Hsien

    2011-02-01

    Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that senses and adapts cells to hypoxic environmental conditions. HIF-1 is composed of an oxygen-regulated α subunit (HIF-1α) and a constitutively expressed β subunit (HIF-1β). Taiwan voles (Microtus kikuchii) are an endemic species in Taiwan, found only in mountainous areas greater than 2000m above sea level. In this study, the full-length HIF-1α cDNA was cloned and sequenced from liver tissues of Taiwan voles. We found that HIF-1α of Taiwan voles had high sequence similarity to HIF-1α of other species. Sequence alignment of HIF-1α functional domains indicated basic helix-loop-helix (bHLH), PER-ARNT-SIM (PAS) and C-terminal transactivation (TAD-C) domains were conserved among species, but sequence variations were found between the oxygen-dependent degradation domains (ODDD). To measure Taiwan vole HIF-1α responses to hypoxia, animals were challenged with cobalt chloride, and HIF-1α mRNA and protein expression in brain, lung, heart, liver, kidney, and muscle was assessed by quantitative RT-PCR and Western blot analysis. Upon induction of hypoxic stress with cobalt chloride, an increase in HIF-1α mRNA levels was detected in lung, heart, kidney, and muscle tissue. In contrast, protein expression levels showed greater variation between individual animals. These results suggest that the regulation of HIF-1α may be important to the Taiwan vole under cobalt chloride treatments. But more details regarding the evolutionary effect of environmental pressure on HIF-1α primary sequence, HIF-1α function and regulation in Taiwan voles remain to be identified.

  17. Intermittent induction of HIF-1α produces lasting effects on malignant progression independent of its continued expression.

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    Hyunsung Choi

    Full Text Available Dysregulation of hypoxia-inducible transcription factors HIF-1α and HIF-2α correlates with poor prognosis in human cancers; yet, divergent and sometimes opposing activities of these factors in cancer biology have been observed. Adding to this complexity is that HIF-1α apparently possesses tumor-suppressing activities, as indicated by the loss-of-function mutations or even homozygous deletion of HIF1A in certain human cancers. As a step towards understanding this complexity, we employed 8-week intermittent induction of a stable HIF-1α variant, HIF1α(PP, in various cancer cell lines and examined the effects on malignant progression in xenografts of immunocompromised mice in comparison to those of HIF2α(PP. Although 8-week treatment led to eventual loss of HIF1α(PP expression, treated osteosarcoma U-2 OS cells acquired tumorigenicity in the subcutaneous tissue. Furthermore, the prior treatment resulted in widespread invasion of malignant glioma U-87 MG cells in the mouse brain and sustained growth of U-118 MG glioma cells. The lasting effects of HIF-1α on malignant progression are specific because neither HIF2α(PP nor β-galactosidase yielded similar effects. By contrast, transient expression of HIF1α(PP in U-87 MG cells or constitutive expression of HIF1α(PP but not HIF2α(PP in a patient-derived glioma sphere culture inhibited tumor growth and spread. Our results indicate that intermittent induction of HIF-1α produces lasting effects on malignant progression even at its own expense.

  18. Wogonin inhibits tumor angiogenesis via degradation of HIF-1α protein

    Energy Technology Data Exchange (ETDEWEB)

    Song, Xiuming; Yao, Jing; Wang, Fei; Zhou, Mi; Zhou, Yuxin; Wang, Hu; Wei, Libin; Zhao, Li; Li, Zhiyu; Lu, Na, E-mail: luna555@163.com; Guo, Qinglong, E-mail: anticancer_drug@yahoo.com.cn

    2013-09-01

    Wogonin, a plant-derived flavone, has been shown recently to have antitumor effects. However, the mechanisms that wogonin inhibits tumor angiogenesis are not well known. In this study, we investigated the effects of wogonin on expression of hypoxia-inducible factor-1α (HIF-1α) and secretion of vascular endothelial growth factor (VEGF) in tumor cells. We found that wogonin decreased the expression of HIF-1α by affecting its stability and reduced the secretion of VEGF, which suppressed angiogenesis in cancer. Wogonin promoted the degradation of HIF-1α by increasing its prolyl hydroxylation, which depended on prolyl hydroxylase (PHD) and the von Hippel–Lindau tumor suppressor (VHL). Intriguingly, wogonin impeded the binding between heat-shock protein 90 (Hsp90) and HIF-1α. In addition, wogonin down-regulated the Hsp90 client proteins EGFR, Cdk4 and survivin, but did not affect the level of Hsp90. Wogonin also increased ubiquitination of HIF-1α and promoted its degradation in proteasome. We also found that wogonin could inhibit nuclear translocation of HIF-1α. Electrophoresis mobility shift assay (EMSA) showed that wogonin decreased the binding activity of exogenous consensus DNA oligonucleotide with HIF-1α in nuclear extracts from MCF-7 cells. Chromatin immunoprecipitation (ChIP) assay also revealed that HIF-1α directly binded to endogenous hypoxia-responsive element (HRE) and this binding was significantly decreased in MCF-7 cells treated with wogonin. Preliminary results indicated in vivo activity of wogonin against xenograft-induced angiogenesis in nude mice. Taken together, the results suggested that wogonin was a potent inhibitor of HIF-1α and provided a new insight into the mechanisms of wogonin against cancers. - Highlights: • Wogonin is an all around inhibitor of VEGF signaling. • We firstly demonstrate that wogonin inhibits secretion of VEGF by decreasing HIF-1α. • Wogonin enhances PDH and VHL expression and inhibits Hsp90 function.

  19. Measuring and Ranking Value Drivers

    NARCIS (Netherlands)

    M.M. Akalu

    2002-01-01

    textabstractAnalysis of the strength of value drivers is crucial to understand their influence in the process of free cash flow generation. The paper addresses the issue of value driver measurement and ranking. The research reveals that, value drivers have similar pattern across industries.

  20. Measuring and Ranking Value Drivers

    NARCIS (Netherlands)

    M.M. Akalu

    2002-01-01

    textabstractAnalysis of the strength of value drivers is crucial to understand their influence in the process of free cash flow generation. The paper addresses the issue of value driver measurement and ranking. The research reveals that, value drivers have similar pattern across industries. Furtherm

  1. HIF-2α downregulation in the absence of functional VHL is not sufficient for renal cell differentiation

    Directory of Open Access Journals (Sweden)

    Burk Robert D

    2007-06-01

    Full Text Available Abstract Background Mutational inactivation of the von Hippel-Lindau (VHL tumor suppressor gene has been linked to hereditary as well as sporadic clear cell renal carcinomas. The product of the VHL gene, pVHL, acts to target hypoxia-inducible factor alpha (HIF-α subunits for ubiquitination and subsequent degradation. Using an RNA interference approach to lower levels of HIF-2α in two different renal cell lines that lack functional pVHL, we have tested the contribution of HIF-2α toward cellular pVHL activities. Results Knockdown of HIF-2α resulted in cell cycle arrest of renal cells that were grown on collagen I, indicating that this pVHL function is dependent on HIF-2α regulation. However, cellular morphological changes and downregulation of integrins α5 and β1, which were seen upon pVHL replacement, were not faithfully phenocopied by HIF-2α reduction. Moreover, fibronectin deposition and expression of renal cell differentiation markers were observed in cells containing replaced pVHL, but not in HIF-2α knockdown cells, indicating that these pVHL functions may occur independently of HIF-2α downregulation. Conclusion These results indicate that HIF-2α regulation is not sufficient for pVHL-induced renal cell differentiation. We hypothesize that in addition to HIF-2α dysregulation, abrogation of additional pVHL functions is required for the initiation of renal carcinogenesis.

  2. Expression of MDR1, HIF-1α and MRP1 in sacral chordoma and chordoma cell line CM-319

    Directory of Open Access Journals (Sweden)

    Ma Baoan

    2010-12-01

    Full Text Available Abstract Background Chordoma was a typically slow-growing tumor. The therapeutic approach to chordoma had traditionally relied mainly on surgical therapy. And the main reason for therapeutic failure was resistance to chemotherapy and radiotherapy. However the refractory mechanism was not clear. The aim of this study was to investigate the expression of three genes (MDR1, HIF-1α and MRP1 associated with resistance to chemotherapy and radiotherapy in chordoma and chordoma cell line CM-319. Materials and methods Using immunohistochemical techniques, the expression of MDR1, HIF-1α and MRP1 was investigated in 50 chordoma specimen. Using RT-PCR and Western blot, the expression of MDR1, HIF-1α and MRP1 was investigated in chordoma and chordoma cell line CM-319. Results Expression of MDR1, HIF-1α and MRP1 was observed in 10%, 80% and 74% of all cases, respectively. Expression of MRP1 was correlated with HIF-1α. On the other hand, expression of MDR1 was not correlated with the expression of HIF-1α or MRP1. The expression of HIF-1α and MRP1 was observed, but MDR1 was not observed in chordoma and CM-319. Conclusion Expression of HIF-1α and MRP1 was observed in most chordoma specimen and CM-319 cell line; expression of HIF-1α correlated with MRP1. HIF-1α and MRP1 may play a role in the multidrug resistance of chordoma to chemotherapy.

  3. Erythrocytosis-associated HIF-2α Mutations Demonstrate a Critical Role for Residues C-terminal to the Hydroxylacceptor Proline*

    Science.gov (United States)

    Furlow, Paul W.; Percy, Melanie J.; Sutherland, Scott; Bierl, Charlene; McMullin, Mary Frances; Master, Stephen R.; Lappin, Terence R. J.; Lee, Frank S.

    2009-01-01

    A classic physiologic response to hypoxia in humans is the up-regulation of the ERYTHROPOIETIN (EPO) gene, which is the central regulator of red blood cell mass. The EPO gene, in turn, is activated by hypoxia inducible factor (HIF). HIF is a transcription factor consisting of an α subunit (HIF-α) and a β subunit (HIF-β). Under normoxic conditions, prolyl hydroxylase domain protein (PHD, also known as HIF prolyl hydroxylase and egg laying-defective nine protein) site specifically hydroxylates HIF-α in a conserved LXXLAP motif (where underlining indicates the hydroxylacceptor proline). This provides a recognition motif for the von Hippel Lindau protein, a component of an E3 ubiquitin ligase complex that targets hydroxylated HIF-α for degradation. Under hypoxic conditions, this inherently oxygen-dependent modification is arrested, thereby stabilizing HIF-α and allowing it to activate the EPO gene. We previously identified and characterized an erythrocytosis-associated HIF2A mutation, G537W. More recently, we reported two additional erythrocytosis-associated HIF2A mutations, G537R and M535V. Here, we describe the functional characterization of these two mutants as well as a third novel erythrocytosis-associated mutation, P534L. These mutations affect residues C-terminal to the LXXLAP motif. We find that all result in impaired degradation and thus aberrant stabilization of HIF-2α. However, each exhibits a distinct profile with respect to their effects on PHD2 binding and von Hippel Lindau interaction. These findings reinforce the importance of HIF-2α in human EPO regulation, demonstrate heterogeneity of functional defects arising from these mutations, and point to a critical role for residues C-terminal to the LXXLAP motif in HIF-α. PMID:19208626

  4. VHL and HIF-1α: gene variations and prognosis in early-stage clear cell renal cell carcinoma.

    Science.gov (United States)

    Lessi, Francesca; Mazzanti, Chiara Maria; Tomei, Sara; Di Cristofano, Claudio; Minervini, Andrea; Menicagli, Michele; Apollo, Alessandro; Masieri, Lorenzo; Collecchi, Paola; Minervini, Riccardo; Carini, Marco; Bevilacqua, Generoso

    2014-03-01

    Von Hipple-Lindau gene (VHL) inactivation represents the most frequent abnormality in clear cell renal cell carcinoma (ccRCC). Hypoxia-inducible factor-1α (HIF-1α) expression is regulated by O2 level. In normal O2 conditions, VHL binds HIF-1α and allows HIF-1α proteasomal degradation. A single-nucleotide polymorphism (SNP) has been found located in the oxygen-dependent degradation domain at codon 582 (C1772T, rs11549465, Pro582Ser). In hypoxia, VHL/HIF-1α interaction is abolished and HIF-1α activates target genes in the nucleus. This study analyzes the impact of genetic alterations and protein expression of VHL and the C1772T SNP of HIF-1α gene (HIF-1α) on prognosis in early-stage ccRCC (pT1a, pT1b, and pT2). Mutational analysis of the entire VHL sequence and the genotyping of HIF-1α C1772T SNP were performed together with VHL promoter methylation analysis and loss of heterozygosis (LOH) analysis at (3p25) locus. Data obtained were correlated with VHL and HIF-1α protein expression and with tumor-specific survival (TSS). VHL mutations, methylation status, and LOH were detected in 51, 11, and 12% of cases, respectively. Our results support the association between biallelic alterations and/or VHL silencing with a worse TSS. Moreover, we found a significant association between the HIF-1α C1772C genotype and a worse TSS. The same association was found when testing the presence of HIF-1α protein in the nucleus. Our results highlight the role of VHL/HIF-1α pathway in RCC and support the molecular heterogeneity of early-stage ccRCC. More important, we show the involvement of HIF-1α C1772T SNP in ccRCC progression.

  5. Automobile Driver Fingerprinting

    Directory of Open Access Journals (Sweden)

    Enev Miro

    2016-01-01

    Full Text Available Today’s automobiles leverage powerful sensors and embedded computers to optimize efficiency, safety, and driver engagement. However the complexity of possible inferences using in-car sensor data is not well understood. While we do not know of attempts by automotive manufacturers or makers of after-market components (like insurance dongles to violate privacy, a key question we ask is: could they (or their collection and later accidental leaks of data violate a driver’s privacy? In the present study, we experimentally investigate the potential to identify individuals using sensor data snippets of their natural driving behavior. More specifically we record the in-vehicle sensor data on the controllerarea- network (CAN of a typical modern vehicle (popular 2009 sedan as each of 15 participants (a performed a series of maneuvers in an isolated parking lot, and (b drove the vehicle in traffic along a defined ~ 50 mile loop through the Seattle metropolitan area. We then split the data into training and testing sets, train an ensemble of classifiers, and evaluate identification accuracy of test data queries by looking at the highest voted candidate when considering all possible one-vs-one comparisons. Our results indicate that, at least among small sets, drivers are indeed distinguishable using only incar sensors. In particular, we find that it is possible to differentiate our 15 drivers with 100% accuracy when training with all of the available sensors using 90% of driving data from each person. Furthermore, it is possible to reach high identification rates using less than 8 minutes of training data. When more training data is available it is possible to reach very high identification using only a single sensor (e.g., the brake pedal. As an extension, we also demonstrate the feasibility of performing driver identification across multiple days of data collection

  6. HIF3A DNA Methylation Is Associated with Childhood Obesity and ALT

    OpenAIRE

    Shuo Wang; Jieyun Song; Yide Yang; Yining Zhang; Haijun Wang; Jun Ma

    2015-01-01

    Gene polymorphisms associated so far with body mass index (BMI) can explain only 1.18-1.45% of observed variation in BMI. Recent studies suggest that epigenetic modifications, especially DNA methylation, could contribute to explain part of the missing heritability, and two epigenetic genome-wide analysis studies (EWAS) have reported that Hypoxia Inducible Factor 3 Alpha Subunit (HIF3A) methylation was associated with BMI or BMI change. We therefore assessed whether the HIF3A methylation is as...

  7. HIF-1α contributes to proliferation and invasiveness of neuroblastoma cells via SHH signaling.

    Science.gov (United States)

    Chen, Sheng; Zhang, Min; Xing, Lili; Wang, Yue; Xiao, Yongtao; Wu, Yeming

    2015-01-01

    The aim of this study was to investigate the effects of hypoxia-inducible factor-1α (HIF-1α) on the proliferation, migration and invasion of neuroblastoma (NB) cells and the mechanisms involved. We here initially used the real-time polymerase chain reaction (real-time PCR), Western blotting and immunohistochemistry (IHC) to detect the expression of HIF-1α and components of the sonic hedgehog (SHH) signaling pathway in NB cells and human specimens. Subsequently, cell proliferation, migration and invasion were analyzed using the cell counting assay, wound healing assay and Transwell system in two types of human NB cell lines, SH-SY5Y and IMR32. In addition, the role of HIF-1α in NB cells growth was determined in a xenograft nude mouse model. We found that the level of HIF-1α was significantly upregulated during NB progression and was associated with the expression of two components of SHH signaling, SHH and GLI1. We next indicated that the proliferation, migration and invasiveness of SH-SY5Y and IMR32 cells were significantly inhibited by HIF-1α knockdown, which was mediated by small interfering RNAs (siRNAs) targeting against its mRNA. Furthermore, the growth of NB cells in vivo was also suppressed by HIF-1α inhibition. Finally, the pro-migration and proliferative effects of HIF-1α could be reversed by disrupting SHH signaling. In conclusion, our results demonstrated that upregulation of HIF-1α in NB promotes proliferation, migration and invasiveness via SHH signaling.

  8. HIF-1α contributes to proliferation and invasiveness of neuroblastoma cells via SHH signaling.

    Directory of Open Access Journals (Sweden)

    Sheng Chen

    Full Text Available The aim of this study was to investigate the effects of hypoxia-inducible factor-1α (HIF-1α on the proliferation, migration and invasion of neuroblastoma (NB cells and the mechanisms involved. We here initially used the real-time polymerase chain reaction (real-time PCR, Western blotting and immunohistochemistry (IHC to detect the expression of HIF-1α and components of the sonic hedgehog (SHH signaling pathway in NB cells and human specimens. Subsequently, cell proliferation, migration and invasion were analyzed using the cell counting assay, wound healing assay and Transwell system in two types of human NB cell lines, SH-SY5Y and IMR32. In addition, the role of HIF-1α in NB cells growth was determined in a xenograft nude mouse model. We found that the level of HIF-1α was significantly upregulated during NB progression and was associated with the expression of two components of SHH signaling, SHH and GLI1. We next indicated that the proliferation, migration and invasiveness of SH-SY5Y and IMR32 cells were significantly inhibited by HIF-1α knockdown, which was mediated by small interfering RNAs (siRNAs targeting against its mRNA. Furthermore, the growth of NB cells in vivo was also suppressed by HIF-1α inhibition. Finally, the pro-migration and proliferative effects of HIF-1α could be reversed by disrupting SHH signaling. In conclusion, our results demonstrated that upregulation of HIF-1α in NB promotes proliferation, migration and invasiveness via SHH signaling.

  9. Molecular selection and functional divergence of HIF-α proteins in vertebrates.

    Science.gov (United States)

    Zhang, Xiangzhe; Wang, Minghui; Tan, Guifang; Wang, Qishan; Zhao, Hongbo; Pan, Yuchun

    2010-12-01

    HIF-α transcription factors, as key master regulators of oxygen homeostasis, constitute a subgroup of the large bHLH-PAS transcription factor family and have been identified in many vertebrates. Although the amino acid sequences of bHLH-PAS domain are conserved, the physiological and pathological roles of this family are variable. They also have different patterns of expression. It is possible that the HIF-α copies have been retained as a consequence of adaptive amino acid replacements or relaxed selective constraint which have conferred subtle changes in function after duplications. Phylogenetic analysis indicated that at least two major duplications had occurred early in the vertebrate lineages. Analyses of the ratios of nonsynonymous/synonymous substitution rates revealed that relaxation of selective constraints might play important roles over evolutionary time and shape variation in some members of the family. The coefficients of functional divergence (θ) estimated between pairwise comparisons of gene groups from HIF-1α, HIF-2α, and HIF-3α indicated statistically significant site-specific shift of evolutionary rates between them, suggesting that altered functional constraints may have taken place at some amino acid residues after their duplications. Moreover, we also mapped sites identified to have been relaxed from purifying selection onto the three-dimensional structure of human HIF-2α. Overall, our study demonstrated that the functional diversity of HIF-αs members may be caused by relaxed negative selection on the N-terminal transactivation domains after HIF-αs duplications, which recruited new partners leading to functional specificity.

  10. Oncogenic kinase NPM/ALK induces expression of HIF1α mRNA.

    Science.gov (United States)

    Marzec, M; Liu, X; Wong, W; Yang, Y; Pasha, T; Kantekure, K; Zhang, P; Woetmann, A; Cheng, M; Odum, N; Wasik, M A

    2011-03-17

    The mechanisms of malignant cell transformation mediated by the oncogenic anaplastic lymphoma kinase (ALK) tyrosine kinase remain only partially understood. In this study, we report that T-cell lymphoma (TCL) cells carrying the nucleophosmin (NPM)/ALK fusion protein (ALK+ TCL) strongly express hypoxia-induced factor 1α (HIF1α) mRNA, even under normoxic conditions, and markedly upregulate HIF1α protein expression under hypoxia. HIF1α expression is strictly dependent on the expression and enzymatic activity of NPM/ALK, as shown in BaF3 cells transfected with wild-type NPM/ALK and kinase-inactive NPM/ALK K210R mutant and by the inhibition of the NPM/ALK function in ALK+ TCL cells by a small-molecule ALK inhibitor. NPM/ALK induces HIF1α expression by upregulating its gene transcription through its key signal transmitter signal transducer and activator of transcription 3 (STAT3), which binds to the HIF1α gene promoter as shown by the chromatin immunoprecipitation assay and is required for HIF1α gene expression as demonstrated by its small interfering RNA-mediated depletion. In turn, depletion of HIF1α increases mammalian target of rapamycin complex 1 activation, cell growth and proliferation and decreases vascular endothelial growth factor synthesis. These results identify a novel cell-transforming property of NPM/ALK, namely its ability to induce the expression of HIF1α, a protein with an important role in carcinogenesis. These results also provide another rationale to therapeutically target NPM/ALK and STAT3 in ALK+ TCL.

  11. HIF2α–arginase axis is essential for the development of pulmonary hypertension

    OpenAIRE

    Cowburn, Andrew S.; Crosby, Alexi; Macias, David; Branco, Cristina; Colaço, Renato D. D. R.; Southwood, Mark; Toshner, Mark; Crotty Alexander, Laura E.; Morrell, Nicholas W.; Chilvers, Edwin R.; Johnson, Randall S.

    2016-01-01

    The expression of hypoxia-inducible factor (HIF)-2α in pulmonary endothelium of mice influences pulmonary vascular resistance and development of hypoxic pulmonary hypertension (PH) via an arginase-1–dependent mechanism. The HIF-2α:arginase-1 axis influences the homeostatic regulation of nitric oxide synthesis in the lung. Impaired generation of this vasoactive agent contributes to the initial development and vascular remodeling process of PH.

  12. The prognosis outcome of oral squamous cell carcinoma using HIF-2α.

    Science.gov (United States)

    Lim, Elva; Kuo, Chia-Chun; Tu, Hsi-Feng; Yang, Cheng-Chieh

    2017-07-06

    Hypoxia-induced factors (HIF) has a role in angiogenesis and regulate tumorigenesis of cancer cell. The HIF is the best-identified mechanism that shows imbalance between consumption and oxygen supply in progressing tumor. This study of HIF-2α expression in oral squamous cell carcinoma (OSCC) aimed to investigate the relationship of HIF-2α and pathology characteristics related to its clinical correlation. Fifty-eight samples of OSCC and adjacent tissues were fixed in paraffin for microarray preparation. The tissue array then was stained using primary antibody HIF-2α (NB100-122) and autoprobe II ABC universal staining kit. Each tissue sample was captured using camera microscope, and images were analyzed with Photoshop 6.0 using the CMYK method. A statistical analysis was performed with the two-tailed t-test, Kaplan-Meier and log-rank test using Prism for Windows version 5.0. The samples of the non-cancerous matched tissues (NCMTs) paired with their OSCC samples showed HIF-2α overexpression with significance difference p serve as a good biomarker for cancer status for all tumor stages and may predict an early recurrence within two years. Copyright © 2017. Published by Elsevier Taiwan LLC.

  13. Low Molecular Weight Fucoidan Inhibits Tumor Angiogenesis through Downregulation of HIF-1/VEGF Signaling under Hypoxia

    Directory of Open Access Journals (Sweden)

    Meng-Chuan Chen

    2015-07-01

    Full Text Available Activation of hypoxia-induced hypoxia-inducible factors-1 (HIF-1 plays a critical role in promoting tumor angiogenesis, growth and metastasis. Low molecular weight fucoidan (LMWF is prepared from brown algae, and exhibits anticancer activity. However, whether LMWF attenuates hypoxia-induced angiogenesis in bladder cancer cells and the molecular mechanisms involved remain unclear. This is the first study to demonstrate that LMWF can inhibit hypoxia-stimulated H2O2 formation, HIF-1 accumulation and transcriptional activity vascular endothelial growth factor (VEGF secretion, and the migration and invasion in hypoxic human bladder cancer cells (T24 cells. LMWF also downregulated hypoxia-activated phosphorylation of PI3K/AKT/mTOR/p70S6K/4EBP-1 signaling in T24 cells. Blocking PI3K/AKT or mTOR activity strongly diminished hypoxia-induced HIF-1α expression and VEGF secretion in T24 cells, supporting the involvement of PI3K/AKT/mTOR in the induction of HIF-1α and VEGF. Additionally, LMWF significantly attenuated angiogenesis in vitro and in vivo evidenced by reduction of tube formation of hypoxic human umbilical vascular endothelial cells and blood capillary generation in the tumor. Similarly, administration of LMWF also inhibited the HIF-1α and VEGF expression in vivo, accompanied by a reduction of tumor growth. In summary, under hypoxia conditions, the antiangiogenic activity of LMWF in bladder cancer may be associated with suppressing HIF-1/VEGF-regulated signaling pathway.

  14. HIF1α is required for osteoclast activation by estrogen deficiency in postmenopausal osteoporosis.

    Science.gov (United States)

    Miyauchi, Yoshiteru; Sato, Yuiko; Kobayashi, Tami; Yoshida, Shigeyuki; Mori, Tomoaki; Kanagawa, Hiroya; Katsuyama, Eri; Fujie, Atsuhiro; Hao, Wu; Miyamoto, Kana; Tando, Toshimi; Morioka, Hideo; Matsumoto, Morio; Chambon, Pierre; Johnson, Randall S; Kato, Shigeaki; Toyama, Yoshiaki; Miyamoto, Takeshi

    2013-10-08

    In women, estrogen deficiency after menopause frequently accelerates osteoclastic bone resorption, leading to osteoporosis, the most common skeletal disorder. However, mechanisms underlying osteoporosis resulting from estrogen deficiency remain largely unknown. Here we show that in bone-resorbing osteoclasts, estrogen-dependent destabilization of hypoxia-inducible factor 1 alpha (HIF1α), which is unstable in the presence of oxygen, plays a pivotal role in promoting bone loss in estrogen-deficient conditions. In vitro, HIF1α was destabilized by estrogen treatment even in hypoxic conditions, and estrogen loss in ovariectomized (Ovx) mice stabilized HIF1α in osteoclasts and promoted their activation and subsequent bone loss in vivo. Osteoclast-specific HIF1α inactivation antagonized bone loss in Ovx mice and osteoclast-specific estrogen receptor alpha deficient mice, both models of estrogen-deficient osteoporosis. Oral administration of a HIF1α inhibitor protected Ovx mice from osteoclast activation and bone loss. Thus, HIF1α represents a promising therapeutic target in osteoporosis.

  15. Prognostic value of plasma levels of HIF-1a and PGC-1a in breast cancer.

    Science.gov (United States)

    Cai, Feng-Feng; Xu, Cheng; Pan, Xin; Cai, Lu; Lin, Xiao-Yan; Chen, Su; Biskup, Ewelina

    2016-11-22

    Cellular adaptive mechanisms are crucial for tumorigenesis and a common feature in solid tumor progression. Hypoxia-inducible factor-1α (HIF-1α) facilitates the biological response to hypoxia, advancing angiogenesis and metastatic potential of the tumor. The peroxisome proliferator-activated receptor γ coactivators 1α (PGC-1α) enhances mitochondrial biogenesis, favored by migratory/invasive cancer cells. We conducted a prospective, long-term follow up study to determine whether HIF-1α and PGC-1α can be implemented as predictive biomarker in breast cancer. HIF-1α and PGC-1α plasma concentrations were measured in patients and in healthy controls by enzyme linked immune sorbent assay. Breast cancer patients had significantly higher HIF-1α and PGC-1α levels, which correlated with clinicopathological features, overall with more aggressive cancer characteristics. Disease free and overall survival of breast cancer patients with high HIF-1α and PGC-1α were significantly poorer than in patients with low plasma levels. In multivariate analysis, high amount of PGC-1α showed independent prognostic value. Our data suggests that HIF-1α and PGC-1α may be promising, noninvasive, biomarkers with a high potential for future clinical implication to identify subgroups of patients with poorer prognosis and to indicate early, subclinical metastasis.

  16. Loss of hif-1 promotes resistance to the exogenous mitochondrial stressor ethidium bromide in Caenorhabditis elegans.

    Science.gov (United States)

    Kamal, Muntasir; D'Amora, Dayana R; Kubiseski, Terrance J

    2016-09-13

    Mitochondrial dysfunction is one of the leading causes of neurological disorders in humans. Mitochondrial perturbations lead to adaptive mechanisms that include HIF-1 stabilization, though the consequences of increased levels of HIF-1 following mitochondrial stress remain poorly understood. Using Caenorhabditis elegans, we show that a hif-1 loss-of-function mutation confers resistance towards the mitochondrial toxin ethidium bromide (EtBr) and suppresses EtBr-induced production of ROS. In mammals, the PD-related gene DJ-1 is known to act as a redox sensor to confer protection against antioxidants and mitochondrial inhibitors. A deletion mutant of the C. elegans homolog djr-1.1 also showed increased resistance to EtBr. Furthermore, our data implicates p38 MAP kinase as an indispensable factor for survival against mitochondrial stress in both hif-1 and djr-1.1 mutants. We propose that EtBr-induced HIF-1 activates pathways that are antagonistic in conferring protection against EtBr toxicity and that blocking HIF-1 activity may promote survival in cells with compromised mitochondrial function.

  17. Isorhamnetin Inhibits Reactive Oxygen Species-Dependent Hypoxia Inducible Factor (HIF)-1α Accumulation.

    Science.gov (United States)

    Seo, Suho; Seo, Kyuhwa; Ki, Sung Hwan; Shin, Sang Mi

    2016-01-01

    Isorhamnetin is a flavonoid metabolite of quercetin and isolated from water dropwort (Oenanthe javanica, Umbelliferae). It has been reported that isorhamnetin exerts beneficial effects including antioxidant, anti-inflammatory, and anti-proliferative activities. The present study investigated whether the antioxidant activity of isorhamnetin is correlated with its anti-cancer effects on colorectal cancer cells. Isorhamnetin significantly repressed cobalt chloride (CoCl2)- or hypoxia-induced hypoxia inducible factor-1α (HIF-1α) accumulation in HCT116 and HT29 cells. When compared with quercetin, isorhamnetin showed potent inhibition of HIF-1α. Moreover, it inhibited CoCl2-induced activity of hypoxia response element reporter gene and HIF-1α-dependent transcription of genes such as glucose transporter 1, lactate dehydrogenase A, carbonic anhydrase-IX, and pyruvate dehydrogenase kinase 1. Isorhamnetin also blocked hydrogen peroxide (H2O2)-induced HIF-1α accumulation. The antioxidant effects of isorhamnetin were confirmed by observation of CoCl2- or H2O2-induced reactive oxygen species (ROS) production. Consistently, overexpressed HIF-1α was decreased by isorhamnetin or N-acetyl-L-cysteine in HEK293 cells. In vitro migration and invasion assay further confirmed the inhibitory effects of isorhamnetin on cancer cells. Collectively, these results demonstrate that isorhamnetin inhibits ROS-mediated HIF-1α accumulation, which contributes to its anti-metastatic efficacy.

  18. Estrogen Receptor β2 Induces Hypoxia Signature of Gene Expression by Stabilizing HIF-1α in Prostate Cancer.

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    Prasenjit Dey

    Full Text Available The estrogen receptor (ER β variant ERβ2 is expressed in aggressive castration-resistant prostate cancer and has been shown to correlate with decreased overall survival. Genome-wide expression analysis after ERβ2 expression in prostate cancer cells revealed that hypoxia was an overrepresented theme. Here we show that ERβ2 interacts with and stabilizes HIF-1α protein in normoxia, thereby inducing a hypoxic gene expression signature. HIF-1α is known to stimulate metastasis by increasing expression of Twist1 and increasing vascularization by directly activating VEGF expression. We found that ERβ2 interacts with HIF-1α and piggybacks to the HIF-1α response element present on the proximal Twist1 and VEGF promoters. These findings suggest that at least part of the oncogenic effects of ERβ2 is mediated by HIF-1α and that targeting of this ERβ2 - HIF-1α interaction may be a strategy to treat prostate cancer.

  19. Estrogen Receptor β2 Induces Hypoxia Signature of Gene Expression by Stabilizing HIF-1α in Prostate Cancer

    Science.gov (United States)

    Dey, Prasenjit; Velazquez-Villegas, Laura A.; Faria, Michelle; Turner, Anthony; Jonsson, Philp; Webb, Paul; Williams, Cecilia; Gustafsson, Jan-Åke; Ström, Anders M.

    2015-01-01

    The estrogen receptor (ER) β variant ERβ2 is expressed in aggressive castration-resistant prostate cancer and has been shown to correlate with decreased overall survival. Genome-wide expression analysis after ERβ2 expression in prostate cancer cells revealed that hypoxia was an overrepresented theme. Here we show that ERβ2 interacts with and stabilizes HIF-1α protein in normoxia, thereby inducing a hypoxic gene expression signature. HIF-1α is known to stimulate metastasis by increasing expression of Twist1 and increasing vascularization by directly activating VEGF expression. We found that ERβ2 interacts with HIF-1α and piggybacks to the HIF-1α response element present on the proximal Twist1 and VEGF promoters. These findings suggest that at least part of the oncogenic effects of ERβ2 is mediated by HIF-1α and that targeting of this ERβ2 – HIF-1α interaction may be a strategy to treat prostate cancer. PMID:26010887

  20. Structural insights into yeast histone chaperone Hif1: a scaffold protein recruiting protein complexes to core histones.

    Science.gov (United States)

    Liu, Hejun; Zhang, Mengying; He, Wei; Zhu, Zhongliang; Teng, Maikun; Gao, Yongxiang; Niu, Liwen

    2014-09-15

    Yeast Hif1 [Hat1 (histone acetyltransferase 1)-interacting factor], a homologue of human NASP (nuclear autoantigenic sperm protein), is a histone chaperone that is involved in various protein complexes which modify histones during telomeric silencing and chromatin reassembly. For elucidating the structural basis of Hif1, in the present paper we demonstrate the crystal structure of Hif1 consisting of a superhelixed TPR (tetratricopeptide repeat) domain and an extended acid loop covering the rear of TPR domain, which represent typical characteristics of SHNi-TPR [Sim3 (start independent of mitosis 3)-Hif1-NASP interrupted TPR] proteins. Our binding assay indicates that Hif1 could bind to the histone octamer via histones H3 and H4. The acid loop is shown to be crucial for the binding of histones and may also change the conformation of the TPR groove. By binding to the core histone complex Hif1 may recruit functional protein complexes to modify histones during chromatin reassembly.

  1. Synthesis and biological evaluation of diaryl-substituted carboranes as inhibitors of hypoxia inducible factor (HIF)-1 transcriptional activity.

    Science.gov (United States)

    Minegishi, Hidemitsu; Matsukawa, Takuya; Nakamura, Hiroyuki

    2013-02-01

    Diaromatic-substituted ortho- and meta-carboranes were synthesized as mimics of manassantin A. Among the carboranes synthesized, compounds 1 and 2 showed significant inhibition of hypoxia-induced HIF-1 transcriptional activity, with IC(50) values of 3.2 and 2.2 μM, respectively. Compounds 1 and 2 similarly suppressed hypoxia-induced HIF-1α accumulation in a concentration-dependent manner without affecting the expression level of HIF-1α mRNA. The hypoxia-induced accumulation and translocation of HIF-1α into nuclei were not observed in HeLa cells treated with compounds 1 and 2 by immunofluorescence analysis, revealing that the inhibition of hypoxia-induced HIF-1 transcriptional activity is induced by compounds 1 and 2 through a degradation pathway of the HIF-1α protein under hypoxic conditions.

  2. Investigating the Multifaceted Impact of HIF-1 during Prostate Cancer and Its Potential Value as a Therapeutic Target

    Science.gov (United States)

    2016-10-01

    CD8+ T cell gene expression using an RNAseq approach. Figure 1. HIF1 inhibition could slow down prostate tumor growth. ProHAxTRAMP... RNAseq and network analysis reveals CD8 T cell gene expression in the setting of HIF-1 deficiency   Metabolic factors and processes play a...contribution of HIF-1 to these important agents of cell- mediated immunity. 4 To this end we utilized a whole transcriptome analysis approach ( RNASeq ) to

  3. Decreased serum glucose and glycosylated hemoglobin levels in patients with Chuvash polycythemia: a role for HIF in glucose metabolism

    OpenAIRE

    McClain, Donald A.; Abuelgasim, Khadega A; Nouraie, Mehdi; Salomon-Andonie, Juan; Niu, Xiaomei; Miasnikova, Galina; Polyakova, Lydia A.; Sergueeva, Adelina; Okhotin, Daniel J.; Cherqaoui, Rabia; Okhotin, David; Cox, James E.; Swierczek, Sabina; Song, Jihyun; Simon, M. Celeste

    2012-01-01

    In Chuvash polycythemia, a homozygous 598C>T mutation in the von Hippel-Lindau gene (VHL) leads to an R200W substitution in VHL protein, impaired degradation of α-subunits of hypoxia inducible factor (HIF)-1 and HIF-2, and augmented hypoxic responses during normoxia. Chronic hypoxia of high altitude is associated with decreased serum glucose and insulin concentrations. Other investigators reported that HIF-1 promotes cellular glucose uptake by increased expression of GLUT1 and increased glyco...

  4. The LINK-A lncRNA Activates Normoxic HIF1α Signaling in Triple-negative Breast Cancer

    OpenAIRE

    Lin, Aifu; Li, Chunlai; Xing, Zhen; Hu, Qingsong; Liang, Ke; Han, Leng; Wang, Cheng; Hawke, David H.; Wang, Shouyu; ZHANG, Yanyan; Wei, Yongkun; Ma, Guolin; Park, Peter K.; Zhou, Jianwei; Zhou, Yan

    2016-01-01

    Although long noncoding RNAs (lncRNAs) predominately reside in nuclear and exert their functions in many biological processes, their potential involvement in cytoplasmic signal transduction remains unexplored. Here, we identified a cytoplasmic lncRNA, Long-Intergenic Noncoding RNA for Kinase Activation (LINK-A), which mediates HB-EGF triggered, EGFR:GPNMB heterodimer-dependent HIF1α phosphorylation at Tyr565 and Ser797 by BRK and LRRK2 respectively. These events cause HIF1α stabilization, HIF...

  5. Extended ischemia prevents HIF1alpha degradation at reoxygenation by impairing prolyl-hydroxylation: role of Krebs cycle metabolites.

    Science.gov (United States)

    Serra-Pérez, Anna; Planas, Anna M; Núñez-O'Mara, Analía; Berra, Edurne; García-Villoria, Judit; Ribes, Antònia; Santalucía, Tomàs

    2010-06-11

    Hypoxia-inducible factor (HIF) is a heterodimeric transcription factor that activates the cellular response to hypoxia. The HIF1alpha subunit is constantly synthesized and degraded under normoxia, but degradation is rapidly inhibited when oxygen levels drop. Oxygen-dependent hydroxylation by prolyl-4-hydroxylases (PHD) mediates HIF1alpha proteasome degradation. Brain ischemia limits the availability not only of oxygen but also of glucose. We hypothesized that this circumstance could have a modulating effect on HIF. We assessed the separate involvement of oxygen and glucose in HIF1alpha regulation in differentiated neuroblastoma cells subjected to ischemia. We report higher transcriptional activity and HIF1alpha expression under oxygen deprivation in the presence of glucose (OD), than in its absence (oxygen and glucose deprivation, OGD). Unexpectedly, HIF1alpha was not degraded at reoxygenation after an episode of OGD. This was not due to impairment of proteasome function, but was associated with lower HIF1alpha hydroxylation. Krebs cycle metabolites fumarate and succinate are known inhibitors of PHD, while alpha-ketoglutarate is a co-substrate of the reaction. Lack of HIF1alpha degradation in the presence of oxygen was accompanied by a very low alpha-ketoglutarate/fumarate ratio. Furthermore, treatment with a fumarate analogue prevented HIF1alpha degradation under normoxia. In all, our data suggest that postischemic metabolic alterations in Krebs cycle metabolites impair HIF1alpha degradation in the presence of oxygen by decreasing its hydroxylation, and highlight the involvement of metabolic pathways in HIF1alpha regulation besides the well known effects of oxygen.

  6. HIF1-alpha overexpression indicates a good prognosis in early stage squamous cell carcinomas of the oral floor

    Directory of Open Access Journals (Sweden)

    Joos Ulrich

    2005-07-01

    Full Text Available Abstract Background Hypoxia-inducible factor 1 (HIF-1 is a transcription factor, which plays a central role in biologic processes under hypoxic conditions, especially concerning tumour angiogenesis. HIF-1α is the relevant, oxygen-dependent subunit and its overexpression has been associated with a poor prognosis in a variety of malignant tumours. Therefore, HIF-1α expression in early stage oral carcinomas was evaluated in relation to established clinico-pathological features in order to determine its value as a prognostic marker. Methods 85 patients with histologically proven surgically treated T1/2 squamous cell carcinoma (SCC of the oral floor were eligible for the study. Tumor specimens were investigated by means of tissue micro arrays (TMAs and immunohistochemistry for the expression of HIF-1. Correlations between clinical features and the expression of HIF-1 were evaluated by Kaplan-Meier curves, log-rank tests and multivariate Cox regression analysis. Results HIF-1α was frequently overexpressed in a probably non-hypoxia related fashion. The expression of HIF-1α was related with a significantly improved 5-year survival rate (p Conclusion HIF-1α overexpression is an indicator of favourable prognosis in T1 and T2 SCC of the oral floor. Node negative patients lacking HIF-1α expression may therefore be considered for adjuvant radiotherapy.

  7. Persistent HIF-1alpha activation in gut ischemia/reperfusion injury: potential role of bacteria and lipopolysaccharide.

    Science.gov (United States)

    Koury, Jadd; Deitch, Edwin A; Homma, Hiroshi; Abungu, Billy; Gangurde, Pranoti; Condon, Michael R; Lu, Qi; Xu, Da-Zhong; Feinman, Rena

    2004-09-01

    In both animal models of hemorrhagic shock and clinical settings, shock-induced gut ischemia has been implicated in the development of the systemic inflammatory response syndrome and distant organ injury, yet the factors transducing these events remain to be fully determined. Because hypoxia-inducible factor (HIF-1), a transcription factor composed of oxygen-labile HIF-1alpha and constitutive HIF-1beta subunits, regulates the physiologic/pathophysiologic response to hypoxia and ischemia, we examined the HIF-1 response in two rat models of gut ischemia-reperfusion. We found that ileal nuclear HIF-1alpha protein levels were induced in rats subjected to trauma (laparotomy) plus hemorrhagic shock for 90 min relative to their trauma sham-shock and naïve counterparts and that this trauma hemorrhagic shock-induced mucosal HIF-1alpha protein response persisted after 1 h and 3 h of reperfusion. Likewise, in a model of isolated gut ischemia-reperfusion injury, where the superior mesenteric artery was occluded for 45 min, nuclear HIF-1alpha were induced in the gut mucosa relative to their sham counterparts and persisted after 1 h and 3 h or reperfusion. Similar to the in vivo response, in vitro hypoxia induced HIF-alpha expression in three different enterocyte cell lines (rat IEC-6 and human Caco-2 and HT-29 cell lines). However, in contrast to the in vivo response, HIF-1 expression rapidly disappeared on subsequent reoxygenation. Because in vivo enterocytes are exposed to bacteria, we tested whether the in vitro HIF-1alpha response would persist on reoxygenation if the enterocytes were cocultured with bacteria. P. aeruginosa, an enteric bacterium, markedly induced enterocyte HIF-1alpha protein levels under normoxic conditions. Furthermore, the addition of P. aeruginosa during either the hypoxic or reoxygenation phase prevented the degradation of HIF-1alpha protein levels. Moreover, the observation that lipopolysaccharide induced HIF-1alpha expression in a time

  8. Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia

    OpenAIRE

    Syarifah Dewi; Reni Paramita; Septelia Inawati Wanandi

    2009-01-01

    Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell) deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1) is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β), the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of...

  9. Ptpmt1 induced by HIF-2α regulates the proliferation and glucose metabolism in erythroleukemia cells

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Qin-Qin [High Altitude Medicine of Ministry of Chinese Education and Research Center for High Altitude Medicine, Qinghai University, Xining, 810001 (China); Qinghai Provincial People' s Hospital, Xining (China); Xiao, Feng-Jun; Sun, Hui-Yan [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, 100850 (China); Shi, Xue-Feng [High Altitude Medicine of Ministry of Chinese Education and Research Center for High Altitude Medicine, Qinghai University, Xining, 810001 (China); Qinghai Provincial People' s Hospital, Xining (China); Wang, Hua; Yang, Yue-Feng; Li, Yu-Xiang [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, 100850 (China); Wang, Li-Sheng, E-mail: wangls@bmi.ac.cn [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, 100850 (China); Ge, Ri-Li, E-mail: geriligao@hotmail.com [High Altitude Medicine of Ministry of Chinese Education and Research Center for High Altitude Medicine, Qinghai University, Xining, 810001 (China)

    2016-03-18

    Hypoxia provokes metabolism misbalance, mitochondrial dysfunction and oxidative stress in both human and animal cells. However, the mechanisms which hypoxia causes mitochondrial dysfunction and energy metabolism misbalance still remain unclear. In this study, we presented evidence that mitochondrial phosphatase Ptpmt1 is a hypoxia response molecule that regulates cell proliferation, survival and glucose metabolism in human erythroleukemia TF-1 cells. Exposure to hypoxia or DFO treatment results in upregulation of HIF1-α, HIF-2α and Ptpmt1. Only inhibition of HIF-2α by shRNA transduction reduces Ptpmt1 expression in TF-1 cells under hypoxia. Ptpmt1 inhibitor suppresses the growth and induces apoptosis of TF-1 cells. Furthermore, we demonstrated that Ptpmt1 inhibition reduces the Glut1 and Glut3 expression and decreases the glucose consumption in TF-1 cells. In additional, Ptpmt1 knockdown also results in the mitochondrial dysfunction determined by JC1 staining. These results delineate a key role for HIF-2α-induced Ptpmt1 upregulation in proliferation, survival and glucose metabolism of erythroleukemia cells. It is indicated that Ptpmt1 plays important roles in hypoxia-induced cell metabolism and mitochondrial dysfunction. - Highlights: • Hypoxia induces upregulation of HIF-1α, HIF-2α and Ptpmt1; HIF-2a induces Ptpmt1 upregulation in TF-1 cells. • PTPMT-1 inhibition reduces growth and induces apoptosis of TF-1 cells. • PTPMT1 inhibition downregulates Glut-1, Glut-3 expression and reduces glucose consumption.

  10. Grape seed extract inhibits VEGF expression via reducing HIF-1alpha protein expression.

    Science.gov (United States)

    Lu, Jianming; Zhang, Keqiang; Chen, Shiuan; Wen, Wei

    2009-04-01

    Grape seed extract (GSE) is a widely consumed dietary supplement that has antitumor activity. Here, we have investigated the inhibitory effect of GSE on the expression of vascular endothelial growth factor (VEGF) and the mechanism underlying this action. We found that GSE inhibited VEGF messenger RNA (mRNA) and protein expression in U251 human glioma cells and MDA-MB-231 human breast cancer cells. GSE inhibited transcriptional activation of the VEGF gene through reducing protein but not mRNA expression of hypoxia-inducible factor (HIF) 1alpha. The inhibitory effect of GSE on HIF-1alpha expression was mainly through inhibiting HIF-1alpha protein synthesis rather than promoting protein degradation. Consistent with this result, GSE-suppressed phosphorylation of several important components involved in HIF-1alpha protein synthesis, such as Akt, S6 kinase and S6 protein. Furthermore, in the MDA-MB-231 tumor, we found that GSE treatment inhibited the expression of VEGF and HIF-1alpha and the phosphorylation of S6 kinase without altering the subcellular localization of HIF-1alpha, correlating with reduced vessel density and tumor size. Depletion of polyphenol with polyvinylpyrrolidone abolished the inhibitory activity of GSE, suggesting a water-soluble fraction of polyphenol in GSE is responsible for the inhibitory activity. Taken together, our results indicate that GSE inhibits VEGF expression by reducing HIF-1alpha protein synthesis through blocking Akt activation. This finding provides new insight into the mechanisms of anticancer activity of GSE and reveals a novel molecular mechanism underlying the antiangiogenic action of GSE.

  11. HIF-1 mediates pathogenic inflammatory responses to intestinal ischemia-reperfusion injury.

    Science.gov (United States)

    Feinman, Rena; Deitch, Edwin A; Watkins, Anthony C; Abungu, Billy; Colorado, Iriana; Kannan, Kolenkode B; Sheth, Sharvil U; Caputo, Francis J; Lu, Qi; Ramanathan, Madhuri; Attan, Shirhan; Badami, Chirag D; Doucet, Danielle; Barlos, Dimitrios; Bosch-Marce, Marta; Semenza, Gregg L; Xu, Da-Zhong

    2010-10-01

    Acute lung injury (ALI) and the development of the multiple organ dysfunction syndrome (MODS) are major causes of death in trauma patients. Gut inflammation and loss of gut barrier function as a consequence of splanchnic ischemia-reperfusion (I/R) have been implicated as the initial triggering events that contribute to the development of the systemic inflammatory response, ALI, and MODS. Since hypoxia-inducible factor (HIF-1) is a key regulator of the physiological and pathophysiological response to hypoxia, we asked whether HIF-1 plays a proximal role in the induction of gut injury and subsequent lung injury. Utilizing partially HIF-1α-deficient mice in a global trauma hemorrhagic shock (T/HS) model, we found that HIF-1 activation was necessary for the development of gut injury and that the prevention of gut injury was associated with an abrogation of lung injury. Specifically, in vivo studies demonstrated that partial HIF-1α deficiency ameliorated T/HS-induced increases in intestinal permeability, bacterial translocation, and caspase-3 activation. Lastly, partial HIF-1α deficiency reduced TNF-α, IL-1β, cyclooxygenase-2, and inducible nitric oxide synthase levels in the ileal mucosa after T/HS whereas IL-1β mRNA levels were reduced in the lung after T/HS. This study indicates that prolonged intestinal HIF-1 activation is a proximal regulator of I/R-induced gut mucosal injury and gut-induced lung injury. Consequently, these results provide unique information on the initiating events in trauma-hemorrhagic shock-induced ALI and MODS as well as potential therapeutic insights.

  12. Expression of HIF-1A/VEGF/ING-4 Axis in Pulmonary Sarcoidosis.

    Science.gov (United States)

    Piotrowski, W J; Kiszałkiewicz, J; Pastuszak-Lewandoska, D; Górski, P; Antczak, A; Migdalska-Sęk, M; Górski, W; Czarnecka, K H; Domańska, D; Nawrot, E; Brzeziańska-Lasota, E

    2015-01-01

    Angiogenesis/angiostasis regulated by hypoxia inducible factor-1A (HIF-1A)/vascular endothelial growth factor (VEGF)/inhibitor of growth protein 4 (ING-4) axis may be crucial for the course and outcome of sarcoidosis. Overexpression of angiogenic factors (activation of VEGF through HIF-1A) may predispose to chronic course and lung fibrosis, whereas immunoangiostasis (related to an overexpression of inhibitory ING-4) may be involved in granuloma formation in early sarcoid inflammation, or sustained or recurrent formation of granulomas. In this work we investigated gene expression of HIF-1A, VEGF and ING-4 in bronchoalveolar fluid (BALF) cells and in peripheral blood (PB) lymphocytes of sarcoidosis patients (n=94), to better understand mechanisms of the disease and to search for its biomarkers. The relative gene expression level (RQ value) was analyzed by qPCR. The results were evaluated according to the presence of lung parenchymal involvement (radiological stage I vs. II-IV), acute vs. insidious onset, lung function tests, calcium metabolism parameters, percentage of lymphocytes (BALL%) and BAL CD4+/CD8+ in BALF, age, and gender. In BALF cells, the ING-4 and VEGF RQ values were increased, while HIF-1A expression was decreased. In PB lymphocytes all studied genes were overexpressed. Higher expression of HIF-1A in PB lymphocytes of patients with abnormal spirometry, and in BALF cells of patients with lung volume restriction was found. VEGF gene expression in BALF cells was also higher in patients with abnormal spirometry. These findings were in line with previous data on the role of HIF-1A/VEGF/ING-4 axis in the pathogenesis of sarcoidosis. Up-regulated HIF-1A and VEGF genes are linked to acknowledged negative prognostics.

  13. Endothelin-1 mediates intermittent hypoxia-induced inflammatory vascular remodeling through HIF-1 activation.

    Science.gov (United States)

    Gras, Emmanuelle; Belaidi, Elise; Briançon-Marjollet, Anne; Pépin, Jean-Louis; Arnaud, Claire; Godin-Ribuot, Diane

    2016-02-15

    Obstructive sleep apnea (OSA) is a major risk factor for cardiovascular mortality, and apnea-induced intermittent hypoxia (IH) is known to promote various cardiovascular alterations such as vascular remodeling. However, the mechanisms that underlie IH remain incompletely investigated. We previously demonstrated that the hypoxia-inducible factor-1 (HIF-1) and endothelin-1 (ET-1) are involved in arterial hypertension and myocardial susceptibility to infarction induced by IH. Thus the objective of the present study was to investigate whether both ET-1 and HIF-1 were also involved in the vascular inflammatory remodeling induced by IH. Mice partially deficient for the Hif1α gene (HIF-1α(+/-)) and their wild-type equivalents, as well as C57BL/6J mice, treated or not with bosentan, a dual endothelin receptor antagonist, were exposed to IH or normoxia for 2 wk, 8 h/day. Splenocyte proliferative and secretory capacities, aortic nuclear factor-κB (NF-κB) and HIF-1 activities, and expression of cytokines and intima-media thickness (IMT) were measured. IH induced a systemic and aortic inflammation characterized by an increase in splenocyte proliferative and secretory capacities, aortic NF-κB activity, and cytokine expression in the aortic wall. This was accompanied by an increase in IMT. These modifications were prevented in HIF-1α(+/-) and bosentan-treated mice. The results of this study suggest that ET-1 is a major contributor to the vascular inflammatory remodeling induced by OSA-related IH, probably through HIF-1-dependent activation of NF-κB.

  14. Counter-driver shock tube

    Science.gov (United States)

    Tamba, T.; Nguyen, T. M.; Takeya, K.; Harasaki, T.; Iwakawa, A.; Sasoh, A.

    2015-11-01

    A "counter-driver" shock tube was developed. In this device, two counter drivers are actuated with an appropriate delay time to generate the interaction between a shock wave and a flow in the opposite direction which is induced by another shock wave. The conditions for the counter drivers can be set independently. Each driver is activated by a separate electrically controlled diaphragm rupture device, in which a pneumatic piston drives a rupture needle with a temporal jitter of better than 1.1 ms. Operation demonstrations were conducted to evaluate the practical performance.

  15. Driver Fatigue Features Extraction

    Directory of Open Access Journals (Sweden)

    Gengtian Niu

    2014-01-01

    Full Text Available Driver fatigue is the main cause of traffic accidents. How to extract the effective features of fatigue is important for recognition accuracy and traffic safety. To solve the problem, this paper proposes a new method of driver fatigue features extraction based on the facial image sequence. In this method, first, each facial image in the sequence is divided into nonoverlapping blocks of the same size, and Gabor wavelets are employed to extract multiscale and multiorientation features. Then the mean value and standard deviation of each block’s features are calculated, respectively. Considering the facial performance of human fatigue is a dynamic process that developed over time, each block’s features are analyzed in the sequence. Finally, Adaboost algorithm is applied to select the most discriminating fatigue features. The proposed method was tested on a self-built database which includes a wide range of human subjects of different genders, poses, and illuminations in real-life fatigue conditions. Experimental results show the effectiveness of the proposed method.

  16. Fast SCR Thyratron Driver

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, M.N.; /SLAC

    2007-06-18

    As part of an improvement project on the linear accelerator at SLAC, it was necessary to replace the original thyratron trigger generator, which consisted of two chassis, two vacuum tubes, and a small thyratron. All solid-state, fast rise, and high voltage thyratron drivers, therefore, have been developed and built for the 244 klystron modulators. The rack mounted, single chassis driver employs a unique way to control and generate pulses through the use of an asymmetric SCR, a PFN, a fast pulse transformer, and a saturable reactor. The resulting output pulse is 2 kV peak into 50 {Omega} load with pulse duration of 1.5 {mu}s FWHM at 180 Hz. The pulse risetime is less than 40 ns with less than 1 ns jitter. Various techniques are used to protect the SCR from being damaged by high voltage and current transients due to thyratron breakdowns. The end-of-line clipper (EOLC) detection circuit is also integrated into this chassis to interrupt the modulator triggering in the event a high percentage of line reflections occurred.

  17. Dextromethorphan in Wisconsin drivers.

    Science.gov (United States)

    Cochems, Amy; Harding, Patrick; Liddicoat, Laura

    2007-05-01

    Dextromethorphan is a synthetic analogue of codeine used in hundreds of over-the-counter medications for its antitussive effects. There have been numerous reports of dextromethorphan abuse by young adults. Dextromethorphan can produce psychoactive effects similar to that of marijuana, and higher doses will produce dissociative effects, including sensory enhancement and hallucinations. The Wisconsin State Laboratory of Hygiene examined data from blood samples submitted from January 1999 through December 2004 to determine the incidence of dextromethorphan in suspected impaired drivers. A total of 108 samples were found to be positive for dextromethorphan during this time. Dextromethorphan concentrations in these cases ranged from less than 5 to 1800 ng/mL (mean 207 ng/mL), compared to an expected therapeutic concentration range of 0.5-5.9 ng/mL. Overall, the highest dextromethorphan concentrations observed were in males aged 16-20 years. Ninety-six percent of the specimens included in this study were also found to be positive for drugs other than dextromethorphan. A review of police and drug recognition expert reports from several of these cases showed that dextromethorphan-impaired drivers exhibited poor psychomotor performance on standardized field sobriety tests, horizontal gaze nystagmus, vertical gaze nystagmus, and overall signs of central nervous system depression.

  18. Driver behaviour at roadworks.

    Science.gov (United States)

    Walker, Guy; Calvert, Malcolm

    2015-11-01

    There is an incompatibility between how transport engineers think drivers behave in roadworks and how they actually behave. As a result of this incompatibility we are losing approximately a lane's worth of capacity in addition to those closed by the roadworks themselves. The problem would have little significance were it not for the fact a lane of motorway costs approx. £30 m per mile to construct and £43 k a year to maintain, and that many more roadworks are planned as infrastructure constructed 40 or 50 years previously reaches a critical stage in its lifecycle. Given current traffic volumes, and the sensitivity of road networks to congestion, the effects of roadworks need to be accurately assessed. To do this requires a new ergonomic approach. A large-scale observational study of real traffic conditions was used to identify the issues and impacts, which were then mapped to the ergonomic knowledge-base on driver behaviour, and combined to developed practical guidelines to help in modelling future roadworks scenarios with greater behavioural accuracy. Also stemming from the work are novel directions for the future ergonomic design of roadworks themselves.

  19. Driver style and driver skills – clustering drivers differing in their potential danger in traffic

    DEFF Research Database (Denmark)

    Martinussen, Laila Marianne; Møller, Mette; Prato, Carlo Giacomo

    The Driver Behavior Questionnaire (DBQ) and the Driver Skill Inventory (DSI) are two of the most frequently used measures of driving style and driving skill. The motivation behind the present study was to test drivers’ insight into their own driving ability based on a combined use of the DBQ......, annual mileage and accident involvement. 3908 drivers aged 18–84 participated in the survey. The results suggested that the drivers have good insight into their own driving ability, as the driving skill level mirrored the frequency of aberrant driving behaviors. K-means cluster analysis revealed four...... and the DSI. Moreover, the joint use of the two instruments was applied to identify sub-groups of drivers that differ in their potential danger in traffic, as well as to test for heterogeneity across the population, namely whether the sub-groups of drivers differed in characteristics such as age, gender...

  20. Hypoxia: adapting to high altitude by mutating EPAS-1, the gene encoding HIF-2α.

    Science.gov (United States)

    van Patot, Martha C Tissot; Gassmann, Max

    2011-01-01

    Living at high altitude is demanding and thus drives adaptational mechanisms. The Tibetan population has had a longer evolutionary period to adapt to high altitude than other mountain populations such as Andeans. As a result, some Tibetans living at high altitudes do not show markedly elevated red blood cell production as compared to South American high altitude natives such as Quechuas or Aymaras, thereby avoiding high blood viscosity creating cardiovascular risk. Unexpectedly, the responsible mutation(s) reducing red blood cell production do not involve either the gene encoding the blood hormone erythropoietin (Epo), or the corresponding regulatory sequences flanking the Epo gene. Similarly, functional mutations in the hypoxia-inducible transcription factor 1α (HIF-1α) gene that represents the oxygen-dependent subunit of the HIF-1 heterodimer, the latter being the main regulator of over 100 hypoxia-inducible genes, have not been described so far. It was not until very recently that three independent groups showed that the gene encoding HIF-2α, EPAS-1 (Wenger et al. 1997), represents a key gene mutated in Tibetan populations adapted to living at high altitudes (Beall et al. 2010 , Yi et al. 2010 , Simonson et al. 2010). Hypoxia-inducible transcription factors were first identified by the description of HIF-1 (Semenza et al. 1991 , 1992), which was subsequently found to enhance transcription of multiple genes that encode proteins necessary for rescuing from hypoxic exposure, including erythropoietic, angiogenic and glycolytic proteins. Then HIF-2 was identified (Ema et al. 1997 ; Flamme et al. 1997 ; Hogenesch et al. 1997 ; and Tian et al. 1997) and although it is highly similar to HIF-1 and has the potential to bind (Camenisch et al. 2001) and mediate (Mole et al. 2009) many of the same genes as HIF-1, its biological actions in response to hypoxia are distinct from those of HIF-1 (reviewed by Loboda et al. 2010). By now, several of these HIF-2 mediated

  1. HIF3A DNA Methylation Is Associated with Childhood Obesity and ALT.

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    Full Text Available Gene polymorphisms associated so far with body mass index (BMI can explain only 1.18-1.45% of observed variation in BMI. Recent studies suggest that epigenetic modifications, especially DNA methylation, could contribute to explain part of the missing heritability, and two epigenetic genome-wide analysis studies (EWAS have reported that Hypoxia Inducible Factor 3 Alpha Subunit (HIF3A methylation was associated with BMI or BMI change. We therefore assessed whether the HIF3A methylation is associated with obesity and other obesity-related phenotypes in Chinese children. The subjects included 110 severe obese cases aged 7-17y and 110 normal-weight controls matched by age and gender for measurement of blood DNA methylation levels at the HIF3A gene locus using the Sequenom's MassARRAY system. We observed significantly higher methylation levels in obese children than in controls at positions 46801642 and 46801699 in HIF3A gene (P<0.05, and found positive associations between methylation and alanine aminotransferase (ALT levels adjusted by gender, age and BMI at the position 46801699 (r = 0.226, P = 0.007. These results suggest that HIF3A DNA methylation is associated with childhood obesity, and has a BMI-independent association with ALT. The results provide evidence for identifying epigenetic factors of elivated ALT and may be useful for risk assessment and personalized medicine of liver diseases such as non-alcoholic fatty liver disease (NAFLD.

  2. HIF-1α pathway: role, regulation and intervention for cancer therapy

    Directory of Open Access Journals (Sweden)

    Georgina N. Masoud

    2015-09-01

    Full Text Available Hypoxia-inducible factor-1 (HIF-1 has been recognized as an important cancer drug target. Many recent studies have provided convincing evidences of strong correlation between elevated levels of HIF-1 and tumor metastasis, angiogenesis, poor patient prognosis as well as tumor resistance therapy. It was found that hypoxia (low O2 levels is a common character in many types of solid tumors. As an adaptive response to hypoxic stress, hypoxic tumor cells activate several survival pathways to carry out their essential biological processes in different ways compared with normal cells. Recent advances in cancer biology at the cellular and molecular levels highlighted the HIF-1α pathway as a crucial survival pathway for which novel strategies of cancer therapy could be developed. However, targeting the HIF-1α pathway has been a challenging but promising progresses have been made in the past twenty years. This review summarizes the role and regulation of the HIF-1α in cancer, and recent therapeutic approaches targeting this important pathway.

  3. Expression of cyclooxyenase-2 protain and its relationship with HIF-1α in HCC

    Institute of Scientific and Technical Information of China (English)

    Chao Jiang; Feng Zhang; Shaojun Wang; Qifei Zou

    2005-01-01

    Objective: To investigate the clinical significance of COX-2 (Cyclooxygenase-2) expression in HCC ( Primary hepatocellular carcinoma) and clarify whether COX-2 is correlated with hypoxia-inducible factor-1α (HIF-1α) in the development of HCC.Methods: Tumor tissues were obtained from 53 patients with HCC. COX-2 and HIF-1α were determined by immunohistochemistry. All 53patients were regularly followed up and the data were collected prospectively. Results: Immunostaining showed the expression of COX-2( n = 33, 62.3 % ) and HIF-1α ( n = 36, 67.9 % ) in most tumor cells. The level of COX-2 was correlated with HIF-1α levels( r = 0.440,P <0.01 ). There were significant correlation between clinicopathological features and higher tumor cytosolic COX-2 level was in the presence of multiple tumors ( P = 0.01), venous invasion ( P = 0.03 ), advanced tumor stage ( P = 0.01), and well-different tumor grade (P = 0.03). High tumor cytosolic COX-2 level was correlated with patient's worse prognosis ( P = 0. 0085). Conclusion: Elevated tumor COX-2 level is correlated with elevated HIF-1α levels and invasiveness in HCC, suggesting COX-2 plays an important role in the progression of HCC, and may be an important therapeutic target in HCC.

  4. [Effect of HIF-1alpha on sex hormone levels and germ cell apoptosis of mice].

    Science.gov (United States)

    Chen, Xiang-Mei; Xiong, Yan-Lei; Gong, Hui; Xu, Cheng-Li

    2013-07-01

    To study the effect of hypoxia on hypothalamus-adenohypophysis-testis axis hormone levels, germ cell apoptosis and hypoxia-inducible factor-1alpha (HIF-1alpha) expression in testis of adolescent mice, and explore HIF-1alpha regulation on the reproductive function of male mice. Eighty SPF grade adolescent C57BL/6 mice were randomly divided into normoxia group, hypoxia 3, 7, 14 and 28 d groups. The level of serum testosterone (T), free testosterone (FT), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) was analyzed by ELISA. Detected the sperm count, motility rate and abnormal sperm rate of epididymal sperm suspension. The apoptosis cells in testis were determined using TUNEL method. The expression of HIF-1alpha was analyzed using Western blot. Compared with corresponding normoxia group, serum T, FT, FSH and LH concentrations in hypoxia 3 d group were significantly higher (P apoptosis index (AI) of germ cells in hypoxia 7, 14 and 28 d groups significantly increased (P levels of HIF-1alpha protein expression were significantly higher (P HIF-1alpha protein highly expressed in mice testis could induce germ cell apoptosis increased in chronic hypoxia environment.

  5. [A novel HIF-1 inhibitor--manassantin A derivative LXY6099 inhibits tumor growth].

    Science.gov (United States)

    Lai, Fang-Fang; Liu, Xiao-Yu; Niu, Fei; Lang, Li-Wei; Xie, Ping; Chen, Xiao-Guang

    2014-05-01

    Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.

  6. Immunohistochemical Expression of GLUT-1 and HIF-1α in Tooth Germ, Ameloblastoma, and Ameloblastic Carcinoma.

    Science.gov (United States)

    Sánchez-Romero, Celeste; Bologna-Molina, Ronell; Mosqueda-Taylor, Adalberto; Paes de Almeida, Oslei

    2016-08-01

    Hypoxia-inducible factor-1α (HIF-1α) promotes proteins that enable cell survival during hypoxia, such as glucose transporter 1 (GLUT-1). Their coexpression has been associated with aggressiveness in malignancies and has not been studied in odontogenic tumors. Immunohistochemical expression of HIF-1α and GLUT-1 was analyzed in 13 tooth germs (TGs), 55 ameloblastomas (AMs), and 3 ameloblastic carcinomas (ACs). HIF-1α was negative in all TGs, and just 1 case of AM and 1 of AC had nuclear positivity. GLUT-1 expressed in ameloblastic cells of all TGs, AMs, and ACs, with an increasing intensity, respectively, and was significantly higher in solid AM than in unicystic AM (P = .041). Absence of nuclear HIF-1α in TGs and most AMs suggest that GLUT-1 may be induced by alternative pathways to hypoxia. However, in ACs, HIF-1α may be activated; however, to confirm this, additional cases are needed. GLUT-1 overexpression could be related to aggressiveness in AMs and ACs and must represent a normal metabolite in TGs.

  7. The yeast histone chaperone hif1p functions with RNA in nucleosome assembly.

    Directory of Open Access Journals (Sweden)

    Amy R Knapp

    Full Text Available Hif1p is an H3/H4-specific histone chaperone that associates with the nuclear form of the Hat1p/Hat2p complex (NuB4 complex in the yeast Saccharomyces cerevisiae. While not capable of depositing histones onto DNA on its own, Hif1p can act in conjunction with a yeast cytosolic extract to assemble nucleosomes onto a relaxed circular plasmid.To identify the factor(s that function with Hif1p to carry out chromatin assembly, multiple steps of column chromatography were carried out to fractionate the yeast cytosolic extract. Analysis of partially purified fractions indicated that Hif1p-dependent chromatin assembly activity resided in RNA rather than protein. Fractionation of isolated RNA indicated that the chromatin assembly activity did not simply purify with bulk RNA. In addition, the RNA-mediated chromatin assembly activity was blocked by mutations in the human homolog of Hif1p, sNASP, that prevent the association of this histone chaperone with histone H3 and H4 without altering its electrostatic properties.These results suggest that specific RNA species may function in concert with histone chaperones to assemble chromatin.

  8. HIF-1 antagonizes p53-mediated apoptosis through a secreted neuronal tyrosinase.

    Science.gov (United States)

    Sendoel, Ataman; Kohler, Ines; Fellmann, Christof; Lowe, Scott W; Hengartner, Michael O

    2010-06-03

    Hypoxia-inducible factor (HIF) is a transcription factor that regulates fundamental cellular processes in response to changes in oxygen concentration. HIFalpha protein levels are increased in most solid tumours and correlate with patient prognosis. The link between HIF and apoptosis, a major determinant of cancer progression and treatment outcome, is poorly understood. Here we show that Caenorhabditis elegans HIF-1 protects against DNA-damage-induced germ cell apoptosis by antagonizing the function of CEP-1, the homologue of the tumour suppressor p53. The antiapoptotic property of HIF-1 is mediated by means of transcriptional upregulation of the tyrosinase family member TYR-2 in the ASJ sensory neurons. TYR-2 is secreted by ASJ sensory neurons to antagonize CEP-1-dependent germline apoptosis. Knock down of the TYR-2 homologue TRP2 (also called DCT) in human melanoma cells similarly increases apoptosis, indicating an evolutionarily conserved function. Our findings identify a novel link between hypoxia and programmed cell death, and provide a paradigm for HIF-1 dictating apoptotic cell fate at a distance.

  9. A long-term "memory" of HIF induction in response to chronic mild decreased oxygen after oxygen normalization

    Directory of Open Access Journals (Sweden)

    Green Dixy E

    2007-01-01

    Full Text Available Abstract Background Endothelial dysfunction (ED is functionally characterized by decreased vasorelaxation, increased thrombosis, increased inflammation, and altered angiogenic potential, has been intimately associated with the progression and severity of cardiovascular disease. Patients with compromised cardiac function oftentimes have a state of chronic mild decreased oxygen at the level of the vasculature and organs, which has been shown to exacerbate ED. Hypoxia inducible factor (HIF is a transcription factor complex shown to be the master regulator of the cellular response to decreased oxygen levels and many HIF target genes have been shown to be associated with ED. Methods Human endothelial and aortic smooth muscle cells were exposed either to A normoxia (21% O2 for three weeks, or to B mild decreased oxygen (15% O2 for three weeks to mimic blood oxygen levels in patients with heart failure, or to C mild decreased oxygen for two weeks followed by one week of normoxia ("memory" treatment. Levels of HIF signaling genes (HIF-1α, HIF-2α, VEGF, BNIP3, GLUT-1, PAI-1 and iNOS were measured both at the protein and mRNA levels. Results It was found that chronic exposure to mild decreased oxygen resulted in significantly increased HIF signaling. There was also a "memory" of HIF-1α and HIF target gene induction when oxygen levels were normalized for one week, and this "memory" could be interrupted by adding a small molecule HIF inhibitor to the last week of normalized oxygen. Finally, levels of ubiquitylated HIF-1α were reduced in response to chronic mild decreased oxygen and were not full restored after oxygen normalization. Conclusion These data suggest that HIF signaling may be contributing to the pathogenesis of endothelial dysfunction and that normalization of oxygen levels may not be enough to reduce vascular stress.

  10. Energy status and HIF signalling in chorionic villi show no evidence of hypoxic stress during human early placental development.

    Science.gov (United States)

    Cindrova-Davies, T; van Patot, M Tissot; Gardner, L; Jauniaux, E; Burton, G J; Charnock-Jones, D S

    2015-03-01

    Early human placental and embryonic development occurs in a physiologically low oxygen environment supported by histiotrophic secretions from endometrial glands. In this study, we compare the placental metabolomic profile in the first, second and third trimesters to determine whether the energy demands are adequately met in the first trimester. We investigated whether hypoxia-inducible factors, HIF-1α and/or HIF-2α, might regulate transcription during the first trimester. First and second trimester tissue was collected using a chorionic villus sampling-like (CVS) technique. Part of each villus sample was frozen immediately and the remainder cultured under 2 or 21% O2 ± 1 mM H2O2, and ±the p38 MAPK pathway inhibitor, PD169316. Levels of HIF-1α were assessed by western blotting and VEGFA, PlGF and GLUT3 transcripts were quantified by RT-PCR. Term samples were collected from normal elective Caesarean deliveries. There were no significant differences in concentrations of ADP, NAD(+), lactate, and glucose, and in the ATP/ADP ratio, across gestational age. Neither HIF-1α nor HIF-2α could be detected in time-zero CVS samples. However, culture under any condition (2 or 21% O2 ± 1 mM H2O2) increased HIF-1α and HIF-2α. HIF-1α and HIF-2α were additionally detected in specimens retrieved after curettage. HIF-1α stabilization was accompanied by significant increases in VEGFA and GLUT3 and a decrease in PlGF mRNAs. These effects were suppressed by PD169316. In conclusion, our data suggest that first trimester placental tissues are not energetically compromised, and that HIF-1α is unlikely to play an appreciable role in regulating transcriptional activity under steady-state conditions in vivo. However, the pathway may be activated by stress conditions.

  11. Drivers of Collaborative Advantage

    DEFF Research Database (Denmark)

    Weihe, Gudrid

    Drawing upon extant alliance literature, this article substantiates the argument that we need to look beyond mere structural and formative aspects of cooperation in order to fully understand the performance antecedents of public-private partnerships. Currently, scholarly work on operational...... processes and behavioural dimensions is practically non-existent. This article tries to remedy the current gap in the literature by reviewing research findings on interfirm collaboration (alliances). On that basis a conceptual framework for analyzing partnership processes is developed. Finally......, the antecedents of collaborative advantage are theoretically examined, and the organizational competences contributing to collaborative success are identified. The conclusion is that operational processes and social dynamics are vital drivers of collaborative advantage. Another significant conclusion...

  12. Drivers for Welfare Innovation

    DEFF Research Database (Denmark)

    Wegener, Charlotte

    2015-01-01

    Innovation has become a key goal towards which teaching and workplace learning needs to be directed. Now perceived as germane and even necessary in almost all kinds of welfare work, the innovation potential in everyday practices and ways of allowing for employer creativity have become a highly...... on the empirical material, the paper proposes a ‘driver’ model for context sensitive research of innovation in welfare workplaces. The model involves three elements which can be regarded as drivers for innovation: i) craft (i.e. professional skills and knowledge), ii) levers (i.e. experiments and adjustment...... that are not necessarily perceived, performed, and changed in phases. A pragmatic and situated perspective on welfare innovation suggests a conception of welfare innovation which is not translated from firm innovation, but derived directly from welfare contexts....

  13. A holistic perspective on corporate sustainability drivers

    NARCIS (Netherlands)

    Lozano, Rodrigo|info:eu-repo/dai/nl/36412380X

    2015-01-01

    Since company boards are increasingly discussing 'sustainability', it becomes necessary to examine the nature of sustainability drivers. Most approaches to corporate sustainability drivers have focused either on internal or external drivers. This paper is aimed at providing a more holistic

  14. Hypoxia Inducible Factor-1α (HIF-1 α and its Role in Tumour Progression to Malignancy

    Directory of Open Access Journals (Sweden)

    Gaurav Mrinal Sharma

    2008-07-01

    Full Text Available Hypoxia is a condition in which an area of the body or a tissue is deprived of sufficient supply of oxygen. The lack of nutrients in a hypoxic tissue generally causes apoptosis but some cells are able to adapt to this hypoxic environment and resist apoptosis. This adaptation occurs as a result of gene activation. Hypoxia is a characteristic feature of many cancers and is the stimulus for overexpression of HIF-1α - a basic loop-helix PAS protein family subunit of HIF, which allows the cell to adapt and survive in hostile environment. The presence of hypoxia and HIF-1α is correlated with an increased risk of metastasis and techniques that can inhibit hypoxia inducible factor may be instrumental in finding a cure for cancer.

  15. Serdemetan antagonizes the Mdm2-HIF1α axis leading to decreased levels of glycolytic enzymes.

    Directory of Open Access Journals (Sweden)

    Jason A Lehman

    Full Text Available Serdemetan (JNJ-26854165, an antagonist to Mdm2, was anticipated to promote the activation of p53. While regulation of p53 by Mdm2 is important, Mdm2 also regulates numerous proteins involved in diverse cellular functions. We investigated if Serdemetan would alter the Mdm2-HIF1α axis and affect cell survival in human glioblastoma cells independently of p53. Treatment of cells with Serdemetan under hypoxia resulted in a decrease in HIF1α levels. HIF1α downstream targets, VEGF and the glycolytic enzymes (enolase, phosphoglycerate kinase1/2, and glucose transporter 1, were all decreased in response to Serdemetan. The involvement of Mdm2 in regulating gene expression of glycolytic enzymes raises the possibility of side effects associated with therapeutically targeting Mdm2.

  16. A protective role for HIF-1 in response to redox manipulation and glucose deprivation: implications for tumorigenesis.

    NARCIS (Netherlands)

    Williams, K.J.; Telfer, B.A.; Airley, R.E.; Peters, J.P.W.; Sheridan, M.R.; Kogel, A.J. van der; Harris, A.L.; Stratford, I.J.

    2002-01-01

    We have investigated the role of HIF-1 in the cellular response to redox modulation via the inhibition of oxidative phosphorylation. We demonstrate that manipulation of redox in air, achieved by inhibiting cytochrome oxidase with cyanide, induces HIF-1 mediated transcription in wild-type CHO and HT1

  17. Transient ureteral obstruction prevents against kidney ischemia/reperfusion injury via hypoxia-inducible factor (HIF-2α activation.

    Directory of Open Access Journals (Sweden)

    Shun Zhang

    Full Text Available Although the protective effect of transient ureteral obstruction (UO prior to ischemia on subsequent renal ischemia/reperfusion (I/R injury has been documented, the underlying molecular mechanism remains to be understood. We showed in the current study that 24 h of UO led to renal tubular hypoxia in the ipsilateral kidney in mice, with the accumulation of hypoxia-inducible factor (HIF-2α, which lasted for a week after the release of UO. To address the functions of HIF-2α in UO-mediated protection of renal IRI, we utilized the Mx-Cre/loxP recombination system to knock out target genes. Inactivation of HIF-2α, but not HIF-1α blunted the renal protective effects of UO, as demonstrated by much higher serum creatinine level and severer histological damage. UO failed to prevent postischemic neutrophil infiltration and apoptosis induction in HIF-2α knockout mice, which also diminished the postobstructive up-regulation of the protective molecule, heat shock protein (HSP-27. The renal protective effects of UO were associated with the improvement of the postischemic recovery of intra-renal microvascular blood flow, which was also dependent on the activation of HIF-2α. Our results demonstrated that UO protected the kidney via activation of HIF-2α, which reduced tubular damages via preservation of adequate renal microvascular perfusion after ischemia. Thus, preconditional HIF-2α activation might serve as a novel therapeutic strategy for the treatment of ischemic acute renal failure.

  18. Co-expression of CXCL8 and HIF-1α is associated with metastasis and poor prognosis in hepatocellular carcinoma.

    Science.gov (United States)

    Li, Xian-Peng; Yang, Xiao-Yu; Biskup, Ewelina; Zhou, Jiang; Li, Hong-Liang; Wu, Yi-Feng; Chen, Ming-Liang; Xu, Feng

    2015-09-08

    Hypoxia inducible factor-1α (HIF-1α), induces cytokines such as CXCL8 and tumor dissemination, chemo- and radio-resistance. We analyzed correlation between HIF-1α and CXCL8 levels, tumor characteristics and overall survival in 102 hepatocellular carcinoma (HCC) patients. Levels of HIF-1α and CXCL8 were increased in HCC tissues and cell lines. Patients with high levels of HIF-1α and CXCL8 had worse outcome and poorer prognosis than those with lower levels. Co-overexpression of HIF-1α and CXCL8 was an independent negative prognostic factor for overall and disease-free survival. HIF-1α silencing and CXCL8 siRNA decreased migration under hypoxic conditions in vitro. Hypoxia-induced activation of AKT/mTOR/STAT3 pathways was reversed by depletion of CXCL8. We conclude that HIF-1α and CXCL8 induce HCC progression and metastasis, associated with activation of AKT/mTOR/STAT3. Co-expression of HIF-1α and CXCL8 is a prognostic marker and a potential therapeutic target in HCC.

  19. Effect of HIF-1α on VEGF-C Induced Lymphangiogenesis and Lymph Nodes Metastases of Pancreatic Cancer

    Institute of Scientific and Technical Information of China (English)

    TAO Jing; LI Tao; LI Kai; XIONG Jiongxin; YANG Zhiyong; WU Heshui; WANG Chunyou

    2006-01-01

    The effect of hypoxia inducible factor-1 α (HIF-1α) on vascular endothelial growth factor C (VEGF-C) and the correlation between HIF-1α and lymphangiogenesis and lymph nodes metastases (LNM) in pancreatic cancer were investigated. Immunohistochemical SP method was used to detect the protein expression of HIF-1α and VEGF-C, and Lymphatic vessel density (LVD) was determined by stain of VEGFR-3, collagen type Ⅳ in 75 pancreatic head cancers from regional pancreatectomy (RP) during Dec. 2001 to Dec. 2003. The relationship between HIF-1α and VEGF-C, lymphangiogenesis, LNM was analyzed statistically. The results showed that the positive expression rate of HIF-1α and VEGF-C in pancreatic cancer tissues was 48.00 % (36/75) and 65.33 % (49/75) respectively. In positive group of HIF-1α, the positive rate of VEGF-C and LVD, and LVD rate was 80.56 % (29/36), 13.22±3.76 and 88.89 % (32/36) respectively, and in negative group of HIF-1α,positive rate of VEGF-C and LVD was 51.28 % (20/39), 5.98±2.17 and 66.67 % (26/39) respectively (P<0.01 or P<0.05). It was suggested that HIF-1α could promote the expression of VEGF-C, lymphangiogenesis and LNM in pancreatic cancer.

  20. p300 and p53 levels determine activation of HIF-1 downstream targets in invasive breast cancer

    NARCIS (Netherlands)

    Vleugel, M.M.; Shvarts, D.; Wall, E. van der; Diest, P.J. van

    2006-01-01

    In previous studies, we noted that overexpression of hypoxia-inducible factor (HIF)–1a in breast cancer, especially the diffuse form, does not always lead to functional activation of its downstream genes. Transcriptional activity of HIF-1 may be repressed by p53 through competition for transcription

  1. HIF-1调控小鼠bace1基因的转录表达%Transcriptional Regulation of Mouse bace1 Gene by HIF-1

    Institute of Scientific and Technical Information of China (English)

    王瑞山; 张云武; 张弦; 洪水根; 许华曦

    2007-01-01

    阿尔茨海默症的一个关键致病原因是大脑中淀粉样蛋白(Aβ)的过度产生或堆积.Aβ是由其前体蛋白APP经β-和γ-分泌酶依次水解而生成的,但对于这些分泌酶基因表达的转录调控的了解还很少.由于大脑发生缺氧/缺血会造成Aβ的产量增加,而缺氧时所激活的转录因子HIF-1(Hypoxia-Inducible Factor 1) 会调控下游多种基因的表达,我们对HIF-1是否参与调控β-分泌酶的表达进行了研究.对小鼠β-分泌酶基因bace1的调控序列进行分析,发现其中含有一个缺氧应答元件(Hypoxia-Responsive Element,HRE).我们的数据显示,HRE突变片段启动报告基因荧光素酶表达的活性比正常序列片段的启动活性明显降低.电泳迁移率的变动分析(EMSA)实验进一步证实,HIF-1可以与小鼠bace1中HRE元件相互作用.当在哺乳动物细胞中过度表达HIF-1时,BACE1的mRNA水平和蛋白质水平均有明显的增高.这些结果表明小鼠bace1基因的表达受转录因子HIF-1的调控.鉴于目前阿尔茨海默症研究领域都把抑制BACE1作为首选治疗靶位,HIF-1因而有可能成为治疗AD的一种药物靶点.

  2. HIF-VEGF pathways are critical for chronic otitis media in Junbo and Jeff mouse mutants.

    Directory of Open Access Journals (Sweden)

    Michael T Cheeseman

    2011-10-01

    Full Text Available Otitis media with effusion (OME is the commonest cause of hearing loss in children, yet the underlying genetic pathways and mechanisms involved are incompletely understood. Ventilation of the middle ear with tympanostomy tubes is the commonest surgical procedure in children and the best treatment for chronic OME, but the mechanism by which they work remains uncertain. As hypoxia is a common feature of inflamed microenvironments, moderation of hypoxia may be a significant contributory mechanism. We have investigated the occurrence of hypoxia and hypoxia-inducible factor (HIF mediated responses in Junbo and Jeff mouse mutant models, which develop spontaneous chronic otitis media. We found that Jeff and Junbo mice labeled in vivo with pimonidazole showed cellular hypoxia in inflammatory cells in the bulla lumen, and in Junbo the middle ear mucosa was also hypoxic. The bulla fluid inflammatory cell numbers were greater and the upregulation of inflammatory gene networks were more pronounced in Junbo than Jeff. Hif-1α gene expression was elevated in bulla fluid inflammatory cells, and there was upregulation of its target genes including Vegfa in Junbo and Jeff. We therefore investigated the effects in Junbo of small-molecule inhibitors of VEGFR signaling (PTK787, SU-11248, and BAY 43-9006 and destabilizing HIF by inhibiting its chaperone HSP90 with 17-DMAG. We found that both classes of inhibitor significantly reduced hearing loss and the occurrence of bulla fluid and that VEGFR inhibitors moderated angiogenesis and lymphangiogenesis in the inflamed middle ear mucosa. The effectiveness of HSP90 and VEGFR signaling inhibitors in suppressing OM in the Junbo model implicates HIF-mediated VEGF as playing a pivotal role in OM pathogenesis. Our analysis of the Junbo and Jeff mutants highlights the role of hypoxia and HIF-mediated pathways, and we conclude that targeting molecules in HIF-VEGF signaling pathways has therapeutic potential in the treatment of

  3. HIF-VEGF pathways are critical for chronic otitis media in Junbo and Jeff mouse mutants.

    Directory of Open Access Journals (Sweden)

    Michael T Cheeseman

    2011-10-01

    Full Text Available Otitis media with effusion (OME is the commonest cause of hearing loss in children, yet the underlying genetic pathways and mechanisms involved are incompletely understood. Ventilation of the middle ear with tympanostomy tubes is the commonest surgical procedure in children and the best treatment for chronic OME, but the mechanism by which they work remains uncertain. As hypoxia is a common feature of inflamed microenvironments, moderation of hypoxia may be a significant contributory mechanism. We have investigated the occurrence of hypoxia and hypoxia-inducible factor (HIF mediated responses in Junbo and Jeff mouse mutant models, which develop spontaneous chronic otitis media. We found that Jeff and Junbo mice labeled in vivo with pimonidazole showed cellular hypoxia in inflammatory cells in the bulla lumen, and in Junbo the middle ear mucosa was also hypoxic. The bulla fluid inflammatory cell numbers were greater and the upregulation of inflammatory gene networks were more pronounced in Junbo than Jeff. Hif-1α gene expression was elevated in bulla fluid inflammatory cells, and there was upregulation of its target genes including Vegfa in Junbo and Jeff. We therefore investigated the effects in Junbo of small-molecule inhibitors of VEGFR signaling (PTK787, SU-11248, and BAY 43-9006 and destabilizing HIF by inhibiting its chaperone HSP90 with 17-DMAG. We found that both classes of inhibitor significantly reduced hearing loss and the occurrence of bulla fluid and that VEGFR inhibitors moderated angiogenesis and lymphangiogenesis in the inflamed middle ear mucosa. The effectiveness of HSP90 and VEGFR signaling inhibitors in suppressing OM in the Junbo model implicates HIF-mediated VEGF as playing a pivotal role in OM pathogenesis. Our analysis of the Junbo and Jeff mutants highlights the role of hypoxia and HIF-mediated pathways, and we conclude that targeting molecules in HIF-VEGF signaling pathways has therapeutic potential in the treatment of

  4. Acetaminophen hepatotoxicity and HIF-1α induction in acetaminophen toxicity in mice occurs without hypoxia.

    Science.gov (United States)

    Chaudhuri, Shubhra; McCullough, Sandra S; Hennings, Leah; Letzig, Lynda; Simpson, Pippa M; Hinson, Jack A; James, Laura P

    2011-05-01

    HIF-1α is a nuclear factor important in the transcription of genes controlling angiogenesis including vascular endothelial growth factor (VEGF). Both hypoxia and oxidative stress are known mechanisms for the induction of HIF-1α. Oxidative stress and mitochondrial permeability transition (MPT) are mechanistically important in acetaminophen (APAP) toxicity in the mouse. MPT may occur as a result of oxidative stress and leads to a large increase in oxidative stress. We previously reported the induction of HIF-1α in mice with APAP toxicity and have shown that VEGF is important in hepatocyte regeneration following APAP toxicity. The following study was performed to examine the relative contribution of hypoxia versus oxidative stress to the induction of HIF-1α in APAP toxicity in the mouse. Time course studies using the hypoxia marker pimonidazole showed no staining for pimonidazole at 1 or 2h in B6C3F1 mice treated with APAP. Staining for pimonidazole was present in the midzonal to periportal regions at 4, 8, 24 and 48h and no staining was observed in centrilobular hepatocytes, the sites of the toxicity. Subsequent studies with the MPT inhibitor cyclosporine A showed that cyclosporine A (CYC; 10mg/kg) reduced HIF-1α induction in APAP treated mice at 1 and 4h and did not inhibit the metabolism of APAP (depletion of hepatic non-protein sulfhydryls and hepatic protein adduct levels). The data suggest that HIF-1α induction in the early stages of APAP toxicity is secondary to oxidative stress via a mechanism involving MPT. In addition, APAP toxicity is not mediated by a hypoxia mechanism.

  5. Exosomal HIF1α supports invasive potential of nasopharyngeal carcinoma-associated LMP1-positive exosomes.

    Science.gov (United States)

    Aga, M; Bentz, G L; Raffa, S; Torrisi, M R; Kondo, S; Wakisaka, N; Yoshizaki, T; Pagano, J S; Shackelford, J

    2014-09-11

    It has emerged recently that exosomes are potential carriers of pro-tumorigenic factors that participate in oncogenesis. However, whether oncogenic transcription factors are transduced by exosomes is unknown. Hypoxia-inducible factor-1α (HIF1α) transcriptionally regulates numerous key aspects of tumor development and progression by promoting a more aggressive tumor phenotype, characterized by increased proliferation and invasiveness coupled with neoangiogenesis. It has been shown that the principal oncoprotein of Epstein-Barr virus (EBV), latent membrane protein 1 (LMP1), drives oncogenic processes and tumor progression of the highly invasive EBV malignancy, nasopharyngeal carcinoma (NPC). We now demonstrate that endogenous HIF1α is detectable in exosomes and that LMP1 significantly increases levels of HIF1α in exosomes. HIF1 recovered from exosomes retains DNA-binding activity and is transcriptionally active in recipient cells after exosome uptake. We also show that treatment of EBV-negative cells with LMP1-exosomes increases migration and invasiveness of NP cell lines in functional assays, which correlates with the phenotype associated with epithelial-mesenchymal transition (EMT). In addition, we provide evidence that HIF1α itself participates in exosome-mediated pro-metastatic effects in recipient cells, as exosome-mediated delivery of active and inactive forms of HIF1α results in reciprocal changes in the expression of E- and N-cadherins associated with EMT. Further, immunohistochemical analysis of NPC tumor tissues revealed direct correlation between protein levels of LMP1 and of the endosome/exosome marker tetraspanin, CD63, which suggests an increase in exosome formation in this EBV-positive malignancy. We hypothesize that exosome-mediated transfer of functional pro-metastatic factors by LMP1-positive NPC cells to surrounding tumor cells promotes cancer progression.

  6. Suppression of Akt-HIF-1α signaling axis by diacetyl atractylodiol inhibits hypoxia-induced angiogenesis

    Science.gov (United States)

    Choi, Sik-Won; Lee, Kwang-Sik; Lee, Jin Hwan; Kang, Hyeon Jung; Lee, Mi Ja; Kim, Hyun Young; Park, Kie-In; Kim, Sun-Lim; Shin, Hye Kyoung; Seo, Woo Duck

    2016-01-01

    Hypoxia-inducible factor (HIF)-1α is a key regulator associated with tumorigenesis, angiogenesis, and metastasis. HIF-1α regulation under hypoxia has been highlighted as a promising therapeutic target in angiogenesis-related diseases. Here, we demonstrate that diacetyl atractylodiol (DAA) from Atractylodes japonica (A. japonica) is a potent HIF-1α inhibitor that inhibits the Akt signaling pathway. DAA dose-dependently inhibited hypoxia-induced HIF-1α and downregulated Akt signaling without affecting the stability of HIF-1α protein. Furthermore, DAA prevented hypoxia-mediated angiogenesis based on in vitro tube formation and in vivo chorioallantoic membrane (CAM) assays. Therefore, DAA might be useful for treatment of hypoxia-related tumorigenesis, including angiogenesis. [BMB Reports 2016; 49(9): 508-513] PMID:27439603

  7. The expression of HIF-1 α in liver tissues in the rat model of paraquat poisoning

    Institute of Scientific and Technical Information of China (English)

    熊莺

    2014-01-01

    Objective To observe the levels of HIF-αand TGF-βin the liver tissue,change of serum transaminase in different phases after paraquat(PQ)toxicity and liver histopathology change in PQ-induced liver toxicity of rat models in order to analyze the relationship between HIF-αand hepatic toxicity induced by PQ.Methods A total of 48 healthy SD rats were randomly(random number)assigned into 2 groups:PQ poisoning group(n=42,

  8. Development drivers for waste management.

    Science.gov (United States)

    Wilson, David C

    2007-06-01

    This paper identifies six broad groups of drivers for development in waste management. Public health led to the emergence of formalized waste collection systems in the nineteenth century, and remains a key driver in developing countries. Environmental protection came to the forefront in the 1970s, with an initial focus on eliminating uncontrolled disposal, followed by the systematic increasing of technical standards. Today, developing countries seem still to be struggling with these first steps; while climate change is also emerging as a key driver. The resource value of waste, which allows people to make a living from discarded materials, was an important driver historically, and remains so in developing countries today. A current trend in developed countries is closing the loop, moving from the concept of 'end-of-pipe' waste management towards a more holistic resource management. Two underpinning groups of drivers are institutional and responsibility issues, and public awareness. There is no, one single driver for development in waste management: the balance between these six groups of drivers has varied over time, and will vary between countries depending on local circumstances, and between stakeholders depending on their perspective. The next appropriate steps towards developing a sustainable, integrated waste management system will also vary in each local situation.

  9. HIF2A and IGF2 Expression Correlates in Human Neuroblastoma Cells and Normal Immature Sympathetic Neuroblasts

    Directory of Open Access Journals (Sweden)

    Sofie Mohlin

    2013-03-01

    Full Text Available During normal sympathetic nervous system (SNS development, cells of the ganglionic lineage can malignantly transform and develop into the childhood tumor neuroblastoma. Hypoxia-inducible transcription factors (HIFs mediate cellular responses during normal development and are central in the adaptation to oxygen shortage. HIFs are also implicated in the progression of several cancer forms, and high HIF-2α expression correlates with disseminated disease and poor outcome in neuroblastoma. During normal SNS development, HIF2A is transiently expressed in neuroblasts and chromaffin cells. SNS cells can, during development, be distinguished by distinct gene expression patterns, and insulin-like growth factor 2 (IGF2 is a marker of sympathetic chromaffin cells, whereas sympathetic neuroblasts lack IGF2 expression. Despite the neuronal derivation of neuroblastomas, we show that neuroblastoma cell lines and specimens express IGF2 and that expression of HIF2A and IGF2 correlates, with the strongest correlation in high-stage tumors. In neuroblastoma, both IGF2 and HIF2A are hypoxia-driven and knocking down IGF2 at hypoxia resulted in downregulated HIF2A levels. HIF-2α and IGF2 were strongly expressed in subsets of immature neuroblastoma cells, suggesting that these two genes could be co-expressed also at early stages of SNS development. We show that IGF2 is indeed expressed in sympathetic chain ganglia at embryonic week 6.5, a developmental stage when HIF-2α is present. These findings provide a rationale for the unexpected IGF2 expression in neuroblastomas and might suggest that IGF2 and HIF2A positive neuroblastoma cells are arrested at an embryonic differentiation stage corresponding to the stage when sympathetic chain ganglia begins to coalesce.

  10. Bilirubin activates transcription of HIF-1α in human proximal tubular cells cultured in the physiologic oxygen content.

    Science.gov (United States)

    Kim, Sung Gyun; Ahn, Shin-Young; Lee, Eun Seong; Kim, Sejoong; Na, Ki Young; Chae, Dong-Wan; Chin, Ho Jun

    2014-09-01

    The expression of hypoxia-inducible factor (HIF) is influenced by reactive oxygen species (ROS). Effect of bilirubin on HIF-1 expression in proximal tubular cells was investigated under physiological oxygen concentration, which is relative hypoxic condition mimicking oxygen content in the medulla of renal tissue. The human kidney (HK2) cells were cultured in 5% oxygen with or without bilirubin. HIF-1α protein expression was increased by bilirubin treatment at 0.01-0.2 mg/dL concentration. The messenger RNA expression of HIF-1α was increased by 1.69±0.05 folds in the cells cultured with 0.1 mg/dL bilirubin, compared to the control cells. The inhibitors of PI3K/mTOR, PI3K/AKT, and ERK 1/2 pathways did not attenuate increased HIF-1α expression by bilirubin. HIF-1α expression decreased by 10 µM exogenous hydrogen peroxide (H2O2); scavenger of ROS with or without bilirubin in the HK2 cells increased HIF-1α concentration more than that in the cells without bilirubin. Exogenous H2O2 decreased the phosphorylation of P70S6 kinase, which was completely reversed by bilirubin treatment. Knockdown of NOX4 gene by small interfering RNA (siRNA) increased HIF-1α mRNA expression. In coonclusion, bilirubin enhances HIF-1α transcription as well as the up-regulation of HIF-1α protein translation through the attenuation of ROS and subunits of NADPH oxidase.

  11. NF-kappaB links innate immunity to the hypoxic response through transcriptional regulation of HIF-1alpha.

    Science.gov (United States)

    Rius, Jordi; Guma, Monica; Schachtrup, Christian; Akassoglou, Katerina; Zinkernagel, Annelies S; Nizet, Victor; Johnson, Randall S; Haddad, Gabriel G; Karin, Michael

    2008-06-05

    The hypoxic response is an ancient stress response triggered by low ambient oxygen (O2) (ref. 1) and controlled by hypoxia-inducible transcription factor-1 (HIF-1), whose alpha subunit is rapidly degraded under normoxia but stabilized when O2-dependent prolyl hydroxylases (PHDs) that target its O2-dependent degradation domain are inhibited. Thus, the amount of HIF-1alpha, which controls genes involved in energy metabolism and angiogenesis, is regulated post-translationally. Another ancient stress response is the innate immune response, regulated by several transcription factors, among which NF-kappaB plays a central role. NF-kappaB activation is controlled by IkappaB kinases (IKK), mainly IKK-beta, needed for phosphorylation-induced degradation of IkappaB inhibitors in response to infection and inflammation. IKK-beta is modestly activated in hypoxic cell cultures when PHDs that attenuate its activation are inhibited. However, defining the relationship between NF-kappaB and HIF-1alpha has proven elusive. Using in vitro systems, it was reported that HIF-1alpha activates NF-kappaB, that NF-kappaB controls HIF-1alpha transcription and that HIF-1alpha activation may be concurrent with inhibition of NF-kappaB. Here we show, with the use of mice lacking IKK-beta in different cell types, that NF-kappaB is a critical transcriptional activator of HIF-1alpha and that basal NF-kappaB activity is required for HIF-1alpha protein accumulation under hypoxia in cultured cells and in the liver and brain of hypoxic animals. IKK-beta deficiency results in defective induction of HIF-1alpha target genes including vascular endothelial growth factor. IKK-beta is also essential for HIF-1alpha accumulation in macrophages experiencing a bacterial infection. Hence, IKK-beta is an important physiological contributor to the hypoxic response, linking it to innate immunity and inflammation.

  12. Regular endurance training reduces the exercise induced HIF-1alpha and HIF-2alpha mRNA expression in human skeletal muscle in normoxic conditions

    DEFF Research Database (Denmark)

    Lundby, Carsten; Gassmann, Max; Pilegaard, Henriette

    2005-01-01

    Regular exercise induces a variety of adaptive responses that enhance the oxidative and metabolic capacity of human skeletal muscle. Although the physiological adjustments of regular exercise have been known for decades, the underlying mechanisms are still unclear. The hypoxia inducible factors 1...... with a single exercise bout, and that this response is blunted with training. We obtained muscle biopsies from a trained (5 days/week during 4 weeks) and untrained leg from the same human subject before, immediately after, and during the recovery from a 3 h two-legged knee extensor exercise bout, where the two......alpha and HIF-2alpha mRNA levels are transiently increased in untrained human skeletal muscle in response to an acute exercise bout, but this response is blunted after exercise training. We propose that HIFs expression is upregulated with exercise and that it may be an important transcription factor...

  13. Rosalind Driver studentships

    Science.gov (United States)

    1999-11-01

    The School of Education at King's College London can now offer funded studentships to those wishing to undertake research in science education. These studentships, which are funded through the generous benefaction of the late Rosalind Driver, can be for a full-time student (over a maximum of three years) or several part-time students (a maximum of six years). Applications from anyone working in science education are welcome but preference will be given to those originating from practising science teachers. Applicants will be expected to register for the award of a MPhil/PhD or EdD and are normally expected to have a first degree. Preliminary ideas about a topic for investigation would also be helpful. Further details and application forms are obtainable from Chiz Dube, School of Education, King's College London, Franklin - Wilkins Building, Waterloo Road, London SE1 8WA (tel: 020-7848-3089, e-mail: chiz.dube@kcl.ac.uk). The deadline for the first round of applications was the middle of October, but preliminary informal enquiries may be made to Dr Jonathan Osborne at the School of Education (tel: 020-7848-3094, e-mail: jonathan.osborne@kcl.ac.uk).

  14. Modulating Effects of HIF-1α on the Promoter Activity of Snail and the Interaction Site Evaluation%HIF-1α调控Snail基因启动子活性及作用位点的鉴定

    Institute of Scientific and Technical Information of China (English)

    朱光辉; 周联明; 王时光; 刘海军; 单远洲

    2016-01-01

    [目的]探讨缺氧诱导因子-1α(HIF-1α)对Snail启动子活性的作用,并鉴定其作用位点.[方法]应用transfac软件分析Snail启动子区域潜在的HIF-1α结合位点.通过荧光素酶报告基因载体、电泳迁移率实验(EMSA)和染色质免疫沉淀(ChIP)实验研究HIF-1α是否能够与Snail启动子区域潜在的位点结合,直接调控Snail的表达.[结果]生物信息学分析发现Snail启动子区域存在2个HIF-1α可能的结合位点(-653,-649)和(-543,-538).双荧光素酶报告基因检测发现(-543,-538)位点是HIF-1α特异性结合于Snail的位点,HIF-1α蛋白与之结合后调节Snail基因转录活性.EMSA和ChIP实验证实了HIF-1α蛋白和Snail启动子区(-543,-538)结合,直接上调Snail的表达.[结论]Snail启动子区域存在HIF-1α的结合位点,HIF-1α与之结合后直接调控Snail的表达.

  15. HIF3A DNA Methylation Is Associated with Childhood Obesity and ALT.

    Science.gov (United States)

    Wang, Shuo; Song, Jieyun; Yang, Yide; Zhang, Yining; Wang, Haijun; Ma, Jun

    2015-01-01

    Gene polymorphisms associated so far with body mass index (BMI) can explain only 1.18-1.45% of observed variation in BMI. Recent studies suggest that epigenetic modifications, especially DNA methylation, could contribute to explain part of the missing heritability, and two epigenetic genome-wide analysis studies (EWAS) have reported that Hypoxia Inducible Factor 3 Alpha Subunit (HIF3A) methylation was associated with BMI or BMI change. We therefore assessed whether the HIF3A methylation is associated with obesity and other obesity-related phenotypes in Chinese children. The subjects included 110 severe obese cases aged 7-17y and 110 normal-weight controls matched by age and gender for measurement of blood DNA methylation levels at the HIF3A gene locus using the Sequenom's MassARRAY system. We observed significantly higher methylation levels in obese children than in controls at positions 46801642 and 46801699 in HIF3A gene (Pobesity, and has a BMI-independent association with ALT. The results provide evidence for identifying epigenetic factors of elivated ALT and may be useful for risk assessment and personalized medicine of liver diseases such as non-alcoholic fatty liver disease (NAFLD).

  16. Reactivating HIF prolyl hydroxylases under hypoxia results in metabolic catastrophe and cell death.

    Science.gov (United States)

    Tennant, D A; Frezza, C; MacKenzie, E D; Nguyen, Q D; Zheng, L; Selak, M A; Roberts, D L; Dive, C; Watson, D G; Aboagye, E O; Gottlieb, E

    2009-11-12

    Cells exposed to low-oxygen conditions (hypoxia) alter their metabolism to survive. This response, although vital during development and high-altitude survival, is now known to be a major factor in the selection of cells with a transformed metabolic phenotype during tumorigenesis. It is thought that hypoxia-selected cells have increased invasive capacity and resistance to both chemo- and radiotherapies, and therefore represent an attractive target for antitumor therapy. Hypoxia inducible factors (HIFs) are responsible for the majority of gene expression changes under hypoxia, and are themselves controlled by the oxygen-sensing HIF prolyl hydroxylases (PHDs). It was previously shown that mutations in succinate dehydrogenase lead to the inactivation PHDs under normoxic conditions, which can be overcome by treatment with alpha-ketoglutarate derivatives. Given that solid tumors contain large regions of hypoxia, the reactivation of PHDs in these conditions could induce metabolic catastrophe and therefore prove an effective antitumor therapy. In this report we demonstrate that derivatized alpha-ketoglutarate can be used as a strategy for maintaining PHD activity under hypoxia. By increasing intracellular alpha-ketoglutarate and activating PHDs we trigger PHD-dependent reversal of HIF1 activation, and PHD-dependent hypoxic cell death. We also show that derivatized alpha-ketoglutarate can permeate multiple layers of cells, reducing HIF1alpha levels and its target genes in vivo.

  17. Hypoxia and HIFs in regulating the development of the hematopoietic system

    NARCIS (Netherlands)

    P. Imanirad (Parisa); E.A. Dzierzak (Elaine)

    2013-01-01

    textabstractMany physiologic processes during the early stages of mammalian ontogeny, particularly placental and vascular development, take place in the low oxygen environment of the uterus. Organogenesis is affected by hypoxia inducible factor (HIF) transcription factors that are sensors of hypoxia

  18. HIF-1α activation by intermittent hypoxia requires NADPH oxidase stimulation by xanthine oxidase.

    Science.gov (United States)

    Nanduri, Jayasri; Vaddi, Damodara Reddy; Khan, Shakil A; Wang, Ning; Makarenko, Vladislav; Semenza, Gregg L; Prabhakar, Nanduri R

    2015-01-01

    Hypoxia-inducible factor 1 (HIF-1) mediates many of the systemic and cellular responses to intermittent hypoxia (IH), which is an experimental model that simulates O2 saturation profiles occurring with recurrent apnea. IH-evoked HIF-1α synthesis and stability are due to increased reactive oxygen species (ROS) generated by NADPH oxidases, especially Nox2. However, the mechanisms by which IH activates Nox2 are not known. We recently reported that IH activates xanthine oxidase (XO) and the resulting increase in ROS elevates intracellular calcium levels. Since Nox2 activation requires increased intracellular calcium levels, we hypothesized XO-mediated calcium signaling contributes to Nox activation by IH. We tested this possibility in rat pheochromocytoma PC12 cells subjected to IH consisting alternating cycles of hypoxia (1.5% O2 for 30 sec) and normoxia (21% O2 for 5 min). Kinetic analysis revealed that IH-induced XO preceded Nox activation. Inhibition of XO activity either by allopurinol or by siRNA prevented IH-induced Nox activation, translocation of the cytosolic subunits p47phox and p67phox to the plasma membrane and their interaction with gp91phox. ROS generated by XO also contribute to IH-evoked Nox activation via calcium-dependent protein kinase C stimulation. More importantly, silencing XO blocked IH-induced upregulation of HIF-1α demonstrating that HIF-1α activation by IH requires Nox2 activation by XO.

  19. HIF-1[alpha] effects on angiogenic potential in human small cell lung carcinoma

    National Research Council Canada - National Science Library

    Wan, Jun; Chai, Huiping; Yu, Zaicheng; Ge, Wei; Kang, Ningning; Xia, Wanli; Che, Yun

    2011-01-01

    ...] in vitro and in vivo. In vivo we used an alternative method to study the effect of HIF-1a on angiogenic potential of SCLC by buliding NCI-H446 cell transplantation tumor on the chick embryo chorioallantoic membrane (CAM) surface...

  20. Antihelminthic benzimidazoles are novel HIF activators that prevent oxidative neuronal death via binding to tubulin.

    Science.gov (United States)

    Aleyasin, Hossein; Karuppagounder, Saravanan S; Kumar, Amit; Sleiman, Sama; Basso, Manuela; Ma, Thong; Siddiq, Ambreena; Chinta, Shankar J; Brochier, Camille; Langley, Brett; Haskew-Layton, Renee; Bane, Susan L; Riggins, Gregory J; Gazaryan, Irina; Starkov, Anatoly A; Andersen, Julie K; Ratan, Rajiv R

    2015-01-10

    Pharmacological activation of the adaptive response to hypoxia is a therapeutic strategy of growing interest for neurological conditions, including stroke, Huntington's disease, and Parkinson's disease. We screened a drug library with known safety in humans using a hippocampal neuroblast line expressing a reporter of hypoxia-inducible factor (HIF)-dependent transcription. Our screen identified more than 40 compounds with the ability to induce hypoxia response element-driven luciferase activity as well or better than deferoxamine, a canonical activator of hypoxic adaptation. Among the chemical entities identified, the antihelminthic benzimidazoles represented one pharmacophore that appeared multiple times in our screen. Secondary assays confirmed that antihelminthics stabilized the transcriptional activator HIF-1α and induced expression of a known HIF target gene, p21(cip1/waf1), in post-mitotic cortical neurons. The on-target effect of these agents in stimulating hypoxic signaling was binding to free tubulin. Moreover, antihelminthic benzimidazoles also abrogated oxidative stress-induced death in vitro, and this on-target effect also involves binding to free tubulin. These studies demonstrate that tubulin-binding drugs can activate a component of the hypoxic adaptive response, specifically the stabilization of HIF-1α and its downstream targets. Tubulin-binding drugs, including antihelminthic benzimidazoles, also abrogate oxidative neuronal death in primary neurons. Given their safety in humans and known ability to penetrate into the central nervous system, antihelminthic benzimidazoles may be considered viable candidates for treating diseases associated with oxidative neuronal death, including stroke.

  1. HIF-mediated innate immune responses: cell signaling and therapeutic implications

    Directory of Open Access Journals (Sweden)

    Harris AJ

    2014-05-01

    Full Text Available Alison J Harris, AA Roger Thompson, Moira KB Whyte, Sarah R Walmsley Academic Unit of Respiratory Medicine, Department of Infection and Immunity, University of Sheffield, Sheffield, UK Abstract: Leukocytes recruited to infected, damaged, or inflamed tissues during an immune response must adapt to oxygen levels much lower than those in the circulation. Hypoxia inducible factors (HIFs are key mediators of cellular responses to hypoxia and, as in other cell types, HIFs are critical for the upregulation of glycolysis, which enables innate immune cells to produce adenosine triphosphate anaerobically. An increasing body of evidence demonstrates that hypoxia also regulates many other innate immunological functions, including cell migration, apoptosis, phagocytosis of pathogens, antigen presentation and production of cytokines, chemokines, and angiogenic and antimicrobial factors. Many of these functions are mediated by HIFs, which are not only stabilized posttranslationally by hypoxia, but also transcriptionally upregulated by inflammatory signals. Here, we review the role of HIFs in the responses of innate immune cells to hypoxia, both in vitro and in vivo, with a particular focus on myeloid cells, on which the majority of studies have so far been carried out. Keywords: hypoxia, neutrophils, monocytes, macrophages

  2. Copper activates HIF-1α/GPER/VEGF signalling in cancer cells.

    Science.gov (United States)

    Rigiracciolo, Damiano Cosimo; Scarpelli, Andrea; Lappano, Rosamaria; Pisano, Assunta; Santolla, Maria Francesca; De Marco, Paola; Cirillo, Francesca; Cappello, Anna Rita; Dolce, Vincenza; Belfiore, Antonino; Maggiolini, Marcello; De Francesco, Ernestina Marianna

    2015-10-27

    Copper promotes tumor angiogenesis, nevertheless the mechanisms involved remain to be fully understood. We have recently demonstrated that the G-protein estrogen receptor (GPER) cooperates with hypoxia inducible factor-1α (HIF-1α) toward the regulation of the pro-angiogenic factor VEGF. Here, we show that copper sulfate (CuSO4) induces the expression of HIF-1α as well as GPER and VEGF in breast and hepatic cancer cells through the activation of the EGFR/ERK/c-fos transduction pathway. Worthy, the copper chelating agent TEPA and the ROS scavenger NAC prevented the aforementioned stimulatory effects. We also ascertained that HIF-1α and GPER are required for the transcriptional activation of VEGF induced by CuSO4. In addition, in human endothelial cells, the conditioned medium from breast cancer cells treated with CuSO4 promoted cell migration and tube formation through HIF-1α and GPER. The present results provide novel insights into the molecular mechanisms involved by copper in triggering angiogenesis and tumor progression. Our data broaden the therapeutic potential of copper chelating agents against tumor angiogenesis and progression.

  3. An association study between hypoxia inducible factor-1alpha (HIF-1α polymorphisms and osteonecrosis.

    Directory of Open Access Journals (Sweden)

    Georgia Chachami

    Full Text Available Bone hypoxia resulting from impaired blood flow is the final pathway for the development of osteonecrosis (ON. The aim of this study was to evaluate if HIF-1α, the major transcription factor triggered by hypoxia, is genetically implicated in susceptibility to ON. For this we analyzed frequencies of three known HIF-1α polymorphisms: one in exon 2 (C111A and two in exon 12 (C1772T and G1790A and their association with ON in a Greek population. Genotype analysis was performed using PCR-RFLP and rare alleles were further confirmed with sequencing. We found that genotype and allele frequency of C1772T and G1790A SNP of HIF-1α (SNPs found in our cohort were not significantly different in ON patients compared to control patients. Furthermore these SNPs could not be associated with the different subgroups of ON. At the protein level we observed that the corresponding mutations (P582S and A588T, respectively are not significant for protein function since the activity, expression and localization of the mutant proteins is practically indistinguishable from wt in HEK293 and Saos-2 cells. These results suggest that these missense mutations in the HIF-1α gene are not important for the risk of developing ON.

  4. Stanniocalcin-2 is a HIF-1 target gene that promotes cell proliferation in hypoxia

    Energy Technology Data Exchange (ETDEWEB)

    Law, Alice Y.S. [Department of Biology, Hong Kong Baptist University, Kowloon Tong (Hong Kong); Wong, Chris K.C., E-mail: ckcwong@hkbu.edu.hk [Department of Biology, Hong Kong Baptist University, Kowloon Tong (Hong Kong)

    2010-02-01

    Stanniocalcin-2 (STC2), the paralog of STC1, has been suggested as a novel target of oxidative stress response to protect cells from apoptosis. The expression of STC2 has been reported to be highly correlated with human cancer development. In this study, we reported that STC2 is a HIF-1 target gene and is involved in the regulation of cell proliferation. STC2 was shown to be up-regulated in different breast and ovarian cancer cells, following exposure to hypoxia. Using ovarian cancer cells (SKOV3), the underlying mechanism of HIF-1 mediated STC2 gene transactivation was characterized. Hypoxia-induced STC2 expression was found to be HIF-1{alpha} dependent and required the recruitment of p300 and HDAC7. Using STC2 promoter deletion constructs and site-directed mutagenesis, two authentic consensus HIF-1 binding sites were identified. Under hypoxic condition, the silencing of STC2 reduced while the overexpression of STC2 increased the levels of phosphorylated retinoblastoma and cyclin D in both SKOV3 and MCF7 cells. The change in cell cycle proteins correlated with the data of the serial cell counts. The results indicated that cell proliferation was reduced in STC2-silenced cells but was increased in STC2-overexpressing hypoxic cells. Solid tumor progression is usually associated with hypoxia. The identification and functional analysis of STC2 up-regulation by hypoxia, a feature of the tumor microenvironment, sheds light on a possible role for STC2 in tumors.

  5. HIF-1α mediates tumor hypoxia to confer a perpetual mesenchymal phenotype for malignant progression.

    Science.gov (United States)

    Yoo, Young-Gun; Christensen, Jared; Gu, Jie; Huang, L Eric

    2011-06-21

    Although tumor progression involves genetic and epigenetic alterations to normal cellular biology, the underlying mechanisms of these changes remain obscure. Numerous studies have shown that hypoxia-inducible factor 1α (HIF-1α) is overexpressed in many human cancers and up-regulates a host of hypoxia-responsive genes for cancer growth and survival. We recently identified an alternative mechanism of HIF-1α function that induces genetic alterations by suppressing DNA repair. Here, we show that long-term hypoxia, which mimics the tumor microenvironment, drives a perpetual epithelial-mesenchymal transition (EMT) through up-regulation of the zinc finger E-box binding homeobox protein ZEB2, whereas short-term hypoxia induces a reversible EMT that requires the transcription factor Twist1. Moreover, we show that the perpetual EMT driven by chronic hypoxia depends on HIF-1α induction of genetic alterations rather than its canonical transcriptional activator function. These mesenchymal tumor cells not only acquire tumorigenicity but also display characteristics of advanced cancers, including necrosis, aggressive invasion, and metastasis. Hence, these results reveal a mechanism by which HIF-1α promotes a perpetual mesenchymal phenotype, thereby advancing tumor progression.

  6. Transcription factor HIF-1 is a necessary mediator of the pasteur effect in mammalian cells.

    Science.gov (United States)

    Seagroves, T N; Ryan, H E; Lu, H; Wouters, B G; Knapp, M; Thibault, P; Laderoute, K; Johnson, R S

    2001-05-01

    The ability to respond to differential levels of oxygen is important to all respiring cells. The response to oxygen deficiency, or hypoxia, takes many forms and ranges from systemic adaptations to those that are cell autonomous. Perhaps the most ancient of the cell-autonomous adaptations to hypoxia is a metabolic one: the Pasteur effect, which includes decreased oxidative phosphorylation and an increase in anaerobic fermentation. Because anaerobic fermentation produces far less ATP than oxidative phosphorylation per molecule of glucose, increased activity of the glycolytic pathway is necessary to maintain free ATP levels in the hypoxic cell. Here, we present genetic and biochemical evidence that, in mammalian cells, this metabolic switch is regulated by the transcription factor HIF-1. As a result, cells lacking HIF-1alpha exhibit decreased growth rates during hypoxia, as well as decreased levels of lactic acid production and decreased acidosis. We show that this decrease in glycolytic capacity results in dramatically lowered free ATP levels in HIF-1alpha-deficient hypoxic cells. Thus, HIF-1 activation is an essential control element of the metabolic state during hypoxia; this requirement has important implications for the regulation of cell growth during development, angiogenesis, and vascular injury.

  7. miR-190 Enhances HIF-Dependent Responses to Hypoxia in Drosophila by Inhibiting the Prolyl-4-hydroxylase Fatiga.

    Science.gov (United States)

    De Lella Ezcurra, Ana Laura; Bertolin, Agustina Paola; Kim, Kevin; Katz, Maximiliano Javier; Gándara, Lautaro; Misra, Tvisha; Luschnig, Stefan; Perrimon, Norbert; Melani, Mariana; Wappner, Pablo

    2016-05-01

    Cellular and systemic responses to low oxygen levels are principally mediated by Hypoxia Inducible Factors (HIFs), a family of evolutionary conserved heterodimeric transcription factors, whose alpha- and beta-subunits belong to the bHLH-PAS family. In normoxia, HIFα is hydroxylated by specific prolyl-4-hydroxylases, targeting it for proteasomal degradation, while in hypoxia the activity of these hydroxylases decreases due to low oxygen availability, leading to HIFα accumulation and expression of HIF target genes. To identify microRNAs required for maximal HIF activity, we conducted an overexpression screen in Drosophila melanogaster, evaluating the induction of a HIF transcriptional reporter. miR-190 overexpression enhanced HIF-dependent biological responses, including terminal sprouting of the tracheal system, while in miR-190 loss of function embryos the hypoxic response was impaired. In hypoxic conditions, miR-190 expression was upregulated and required for induction of HIF target genes by directly inhibiting the HIF prolyl-4-hydroxylase Fatiga. Thus, miR-190 is a novel regulator of the hypoxia response that represses the oxygen sensor Fatiga, leading to HIFα stabilization and enhancement of hypoxic responses.

  8. Intestine-specific Disruption of Hypoxia-inducible Factor (HIF)-2α Improves Anemia in Sickle Cell Disease*

    Science.gov (United States)

    Das, Nupur; Xie, Liwei; Ramakrishnan, Sadeesh K.; Campbell, Andrew; Rivella, Stefano; Shah, Yatrik M.

    2015-01-01

    Sickle cell disease (SCD) is caused by genetic defects in the β-globin chain. SCD is a frequently inherited blood disorder, and sickle cell anemia is a common type of hemoglobinopathy. During anemia, the hypoxic response via the transcription factor hypoxia-inducible factor (HIF)-2α is highly activated in the intestine and is essential in iron absorption. Intestinal disruption of HIF-2α protects against tissue iron accumulation in iron overload anemias. However, the role of intestinal HIF-2α in regulating anemia in SCD is currently not known. Here we show that in mouse models of SCD, disruption of intestinal HIF-2α significantly decreased tissue iron accumulation. This was attributed to a decrease in intestinal iron absorptive genes, which were highly induced in a mouse model of SCD. Interestingly, disruption of intestinal HIF-2α led to a robust improvement in anemia with an increase in RBC, hemoglobin, and hematocrit. This was attributed to improvement in RBC survival, hemolysis, and insufficient erythropoiesis, which is evident from a significant decrease in serum bilirubin, reticulocyte counts, and serum erythropoietin following intestinal HIF-2α disruption. These data suggest that targeting intestinal HIF-2α has a significant therapeutic potential in SCD pathophysiology. PMID:26296885

  9. Mechanism Underlying Post-menopausal Osteoporosis: HIF1α is Required for Osteoclast Activation by Estrogen Deficiency.

    Science.gov (United States)

    Miyamoto, Takeshi

    2015-01-01

    The aging of the population worldwide has sharply increased the number of post-menopausal osteoporosis patients. Bone fragility caused by osteoporosis often results in fractures; therefore, controlling osteoporosis is crucial to prevent such injuries. To date, various drugs to treat osteoporosis have been developed and launched; however, the molecular mechanisms underlying post-menopausal osteoporosis have not been fully elucidated, and additional factors that could be targeted to treat patients remain to be characterized. Recently, hypoxia inducible factor 1 alpha (HIF1α) was identified as essential for osteoclast activation, an activity that promotes bone loss following menopausal estrogen deficiency. Although osteoclasts, which are located in hypoxic regions of the bone surface, express HIF1α mRNA, in pre-menopausal conditions the presence of estrogen decreases HIF1α protein levels in these cells. In menopausal conditions, however, estrogen deficiency allows HIF1α protein to accumulate in osteoclasts, leading to osteoclast activation and bone loss. Osteoclast-specific conditional HIF1α inactivation protects mice from estrogen deficiency-induced osteoclast activation and bone loss, as does systemic administration of a HIF1α inhibitor. Therefore, HIF1α represents a potential therapeutic target to prevent osteoclast activation and bone loss in post-menopausal patients.

  10. [Effect of flumatinib mesylate on C-MYC, HIF-1α and VEGF in U226 line].

    Science.gov (United States)

    Chang, Ming-Xing; Ma, Yan-Ping

    2013-12-01

    The objective of this study was to investigate the effect of the new generation of tyrosine kinase inhibitor flumatinib mesylate on C-MYC, HIF-1α and VEGF in multiple myeloma (MM) cell line U266. Different concentrations (1, 5, 10 µmol/L) of flumatinib mesylate were used to act on U266 cell line for 8, 16 and 24 h, and the expression of C-MYC, and HIF-1α genes was detected by real-time fluorescence-quantitative PCR, the expression of C-MYC, HIF-1α and VEGF was measured by Western blot. The results showed that the gene expression of C-MYC and HIF-1 genes decreased gradually with the increasing of flumatinib mesylate concentration (P C-MYC and HIF-1α gene decreased gradually with prolonging of treatment time with the flumatinib mesylate (P C-MYC, HIF-1α and VEGF decreased gradually with the increasing of flumatinib mesylate concentration (P C-MYC, HIF-1 α and VEGF in U266 cell line in a time- and dose-dependent manners, so flumatinib mesylate may become a new drug for MM therapy.

  11. HIF-1 regulates iron homeostasis in Caenorhabditis elegans by activation and inhibition of genes involved in iron uptake and storage.

    Directory of Open Access Journals (Sweden)

    Steven Joshua Romney

    2011-12-01

    Full Text Available Caenorhabditis elegans ftn-1 and ftn-2, which encode the iron-storage protein ferritin, are transcriptionally inhibited during iron deficiency in intestine. Intestinal specific transcription is dependent on binding of ELT-2 to GATA binding sites in an iron-dependent enhancer (IDE located in ftn-1 and ftn-2 promoters, but the mechanism for iron regulation is unknown. Here, we identify HIF-1 (hypoxia-inducible factor -1 as a negative regulator of ferritin transcription. HIF-1 binds to hypoxia-response elements (HREs in the IDE in vitro and in vivo. Depletion of hif-1 by RNA interference blocks transcriptional inhibition of ftn-1 and ftn-2 reporters, and ftn-1 and ftn-2 mRNAs are not regulated in a hif-1 null strain during iron deficiency. An IDE is also present in smf-3 encoding a protein homologous to mammalian divalent metal transporter-1. Unlike the ftn-1 IDE, the smf-3 IDE is required for HIF-1-dependent transcriptional activation of smf-3 during iron deficiency. We show that hif-1 null worms grown under iron limiting conditions are developmentally delayed and that depletion of FTN-1 and FTN-2 rescues this phenotype. These data show that HIF-1 regulates intestinal iron homeostasis during iron deficiency by activating and inhibiting genes involved in iron uptake and storage.

  12. Effect of proline analogues on activity of human prolyl hydroxylase and the regulation of HIF signal transduction pathway.

    Directory of Open Access Journals (Sweden)

    Xiaoyan Ma

    Full Text Available Hypoxia inducible factor 1 (HIF-1 plays a pivotal role in cellular responses to hypoxia. Prolyl hydroxylase 3 (PHD3 degrades HIF-1α under normoxic conditions through the hydroxylation of HIF-1α for proteolysis. Inhibiting PHD3 activity is crucial for up-regulating HIF-1α, thereby acting as a potential target for treating hypoxia-related diseases. In this study, two proline analogues (PA1 and PA2 were screened as PHD3 inhibitors with apparent EC50 values of 1.53 and 3.17 µM respectively, indicating good inhibition potency. Nine proteins, significantly regulated by PA1, were identified using 2-DE coupled with MALDI-TOF/TOF MS. Pyruvate kinase isozymes M1/M2 (PKM and alpha-enolase 1 (ENO1, which are key modulators of glycolysis, are directly regulated by HIF-1α. Moreover, VEGF, a signal protein stimulating angiogenesis, was strongly promoted by PA1. Our findings suggest that PA1 stabilized HIF-1α as well as up-regulated glycolysis and angiogenesis proteins. Herein, for the first time, we systematically studied proline analogue PA1 as a PHD3 inhibitor, which provides innovative evidence for the treatment of HIF-related diseases.

  13. Relative Contribution of Prolyl Hydroxylase-Dependent and -Independent Degradation of HIF-1alpha by Proteasomal Pathways in Cerebral Ischemia

    Directory of Open Access Journals (Sweden)

    Yomna Badawi

    2017-05-01

    Full Text Available Hypoxia inducible factor-1 (HIF-1 is a key regulator in hypoxia and can determine the fate of brain cells during ischemia. However, the mechanism of HIF-1 regulation is still not fully understood in ischemic brains. We tested a hypothesis that both the 26S and the 20S proteasomal pathways were involved in HIF-1α degradation under ischemic conditions. Using in vitro ischemic model (oxygen and glucose deprivation and a mouse model of middle cerebral artery occlusion, we tested effects of inhibitors of proteasomes and prolyl hydroxylase (PHD on HIF-1α stability and brain injury in cerebral ischemia. We observed that 30 and 60 min of oxygen-glucose deprivation significantly increased the 20S proteasomal activity. We demonstrated that proteasome inhibitors increased HIF-1α stabilization and cell viability and were more effective than PHD inhibitors in primary cultured cortical neurons exposed to oxygen and glucose deprivation. Furthermore, the administration of the proteasome inhibitor, epoxomicin, to mice resulted in smaller infarct size and brain edema than a PHD inhibitor. Our results indicate that 20S proteasomes are involved in HIF-1α degradation in ischemic neurons and that proteasomal inhibition provides more HIF-1α stabilization and neuroprotection than PHD inhibition in cerebral ischemia.

  14. Experimental Study on the Expression of HIF-1α and Its Relationship to Apoptosis in Tissues around Cerebral Bleeding Loci

    Institute of Scientific and Technical Information of China (English)

    朱遂强; 唐洲平; 郭守刚; 彭岚; 方思羽; 张苏明

    2004-01-01

    The expression of hypoxia inducible factor-1 alpha (HIF-1α) and its relationship to apoptosis in tissues around cerebral bleeding loci was studied. The expression of HIF-1α and apoptosis in 37 samples of tissues around cerebral bleeding loci and 9 samples of normal cerebral tissues was assessed by immunohistochemical straining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling methods. In 37 tissue samples around cerebral bleeding loci, the positive rate of the HIF-1α expression was 40.6 %. Especially in the patients with amount of bleeding>60 ml, the positive rate (88.9 %) of the HIF-1α expression was significantly higher than those with the amount of bleeding ranging from 30-45 ml or 45-60 ml (P<0.05). The expression of HIF-1α was increased as the amount of bleeding and operative time increased (P<0.05). There existed a positive correlation between HIF-1α labeling index and apoptosis index (n= 12, r= 0.56, P<0.01). These results suggested that the expression of HIF-1α was closely related with the time of hemorrhage and the amount of bleeding, and could induce the apoptosis of neurons.

  15. Intestine-specific Disruption of Hypoxia-inducible Factor (HIF)-2α Improves Anemia in Sickle Cell Disease.

    Science.gov (United States)

    Das, Nupur; Xie, Liwei; Ramakrishnan, Sadeesh K; Campbell, Andrew; Rivella, Stefano; Shah, Yatrik M

    2015-09-25

    Sickle cell disease (SCD) is caused by genetic defects in the β-globin chain. SCD is a frequently inherited blood disorder, and sickle cell anemia is a common type of hemoglobinopathy. During anemia, the hypoxic response via the transcription factor hypoxia-inducible factor (HIF)-2α is highly activated in the intestine and is essential in iron absorption. Intestinal disruption of HIF-2α protects against tissue iron accumulation in iron overload anemias. However, the role of intestinal HIF-2α in regulating anemia in SCD is currently not known. Here we show that in mouse models of SCD, disruption of intestinal HIF-2α significantly decreased tissue iron accumulation. This was attributed to a decrease in intestinal iron absorptive genes, which were highly induced in a mouse model of SCD. Interestingly, disruption of intestinal HIF-2α led to a robust improvement in anemia with an increase in RBC, hemoglobin, and hematocrit. This was attributed to improvement in RBC survival, hemolysis, and insufficient erythropoiesis, which is evident from a significant decrease in serum bilirubin, reticulocyte counts, and serum erythropoietin following intestinal HIF-2α disruption. These data suggest that targeting intestinal HIF-2α has a significant therapeutic potential in SCD pathophysiology.

  16. Sulforaphane inhibits hypoxia-induced HIF-1α and VEGF expression and migration of human colon cancer cells.

    Science.gov (United States)

    Kim, Dong Hwan; Sung, Bokyung; Kang, Yong Jung; Hwang, Seong Yeon; Kim, Min Jeong; Yoon, Jeong-Hyun; Im, Eunok; Kim, Nam Deuk

    2015-12-01

    The effects of sulforaphane (a natural product commonly found in broccoli) was investigated on hypoxia inducible factor-1α (HIF-1α) expression in HCT116 human colon cancer cells and AGS human gastric cancer cells. We found that hypoxia-induced HIF-1α protein expression in HCT116 and AGS cells, while treatment with sulforaphane markedly and concentration-dependently inhibited HIF-1α expression in both cell lines. Treatment with sulforaphane inhibited hypoxia-induced vascular endothelial growth factor (VEGF) expression in HCT116 cells. Treatment with sulforaphane modulated the effect of hypoxia on HIF-1α stability. However, degradation of HIF-1α by sulforaphane was not mediated through the 26S proteasome pathway. We also found that the inhibition of HIF-1α by sulforaphane was not mediated through AKT and extracellular signal-regulated kinase phosphorylation under hypoxic conditions. Finally, hypoxia-induced HCT116 cell migration was inhibited by sulforaphane. These data suggest that sulforaphane may inhibit human colon cancer progression and cancer cell angiogenesis by inhibiting HIF-1α and VEGF expression. Taken together, these results indicate that sulforaphane is a new and potent chemopreventive drug candidate for treating patients with human colon cancer.

  17. Driver style and driver skills – clustering drivers differing in their potential danger in traffic

    DEFF Research Database (Denmark)

    Martinussen, Laila Marianne; Møller, Mette; Prato, Carlo Giacomo

    and the DSI. Moreover, the joint use of the two instruments was applied to identify sub-groups of drivers that differ in their potential danger in traffic, as well as to test for heterogeneity across the population, namely whether the sub-groups of drivers differed in characteristics such as age, gender......, annual mileage and accident involvement. 3908 drivers aged 18–84 participated in the survey. The results suggested that the drivers have good insight into their own driving ability, as the driving skill level mirrored the frequency of aberrant driving behaviors. K-means cluster analysis revealed four...... distinct clusters that differed in the frequency of aberrant driving behavior and driving skills, as well as individual characteristics and driving related factors such as annual mileage, accident frequency and number of tickets and fines. Thus, two sub-groups were identified as more unsafe than the two...

  18. Regulation of IL-12p40 by HIF controls Th1/Th17 responses to prevent mucosal inflammation.

    Science.gov (United States)

    Marks, E; Naudin, C; Nolan, G; Goggins, B J; Burns, G; Mateer, S W; Latimore, J K; Minahan, K; Plank, M; Foster, P S; Callister, R; Veysey, M; Walker, M M; Talley, N J; Radford-Smith, G; Keely, S

    2017-01-25

    Intestinal inflammatory lesions are inherently hypoxic, due to increased metabolic demands created by cellular infiltration and proliferation, and reduced oxygen supply due to vascular damage. Hypoxia stabilizes the transcription factor hypoxia-inducible factor-1α (HIF) leading to a coordinated induction of endogenously protective pathways. We identified IL12B as a HIF-regulated gene and aimed to define how the HIF-IL-12p40 axis influenced intestinal inflammation. Intestinal lamina propria lymphocytes (LPL) were characterized in wild-type and IL-12p40(-/-) murine colitis treated with vehicle or HIF-stabilizing prolyl-hydroxylase inhibitors (PHDi). IL12B promoter analysis was performed to examine hypoxia-responsive elements. Immunoblot analysis of murine and human LPL supernatants was performed to characterize the HIF/IL-12p40 signaling axis. We observed selective induction of IL-12p40 following PHDi-treatment, concurrent with suppression of Th1 and Th17 responses in murine colitis models. In the absence of IL-12p40, PHDi-treatment was ineffective. Analysis of the IL12B promoter identified canonical HIF-binding sites. HIF stabilization in LPLs resulted in production of IL-12p40 homodimer which was protective against colitis. The selective induction of IL-12p40 by HIF-1α leads to a suppression of mucosal Th1 and Th17 responses. This HIF-IL12p40 axis may represent an endogenously protective mechanism to limit the progression of chronic inflammation, shifting from pro-inflammatory IL-12p70 to an antagonistic IL-12p40 homodimer.Mucosal Immunology advance online publication, 25 January 2017; doi:10.1038/mi.2016.135.

  19. Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia

    Directory of Open Access Journals (Sweden)

    Syarifah Dewi

    2009-11-01

    Full Text Available Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1 is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β, the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of this study was to analyze the adaptive tissue responses during induced systemic hypoxia by comparation of relative expression of mRNA HIF-1α in rat heart, brain and blood. Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia by placing them in the hypoxic chamber supplied by 8-10% of O2 for 0, 1, 7, 14 and 21 days, respectively. The relative expression level of HIF-1α mRNA in brain, heart and leucocyte cells were analyzed using quantitative RT-PCR assay (Real Time PCR based on Pfaff's formula. This study demonstrates that the increased of relative expression of HIF-1α mRNA during induced systemic hypoxia reached its maximum level at day 7 (in heart or at day 14 (in brain, whereas in leucocyte cells the stimulation of HIF-1α expression was intensively maintained up to 21 days although the expression has reached the remarkably high level. We could conclude that HIF-1α as an oxygen sensing during systemic hypoxia has different capacity and sensitivity in brain, heart and blood tissues, due to the importance of oxygen homeostasis in each tissue.

  20. Adaptation to HIF-1 deficiency by upregulation of the AMP/ATP ratio and phosphofructokinase activation in hepatomas

    Directory of Open Access Journals (Sweden)

    Airley Rachel E

    2011-05-01

    Full Text Available Abstract Background HIF-1 deficiency has marked effects on tumour glycolysis and growth. We therefore investigated the consequences of HIF-1 deficiency in mice, using the well established Hepa-1 wild-type (WT and HIF-1β-deficient (c4 model. These mechanisms could be clinically relevant, since HIF-1 is now a therapeutic target. Methods Hepa-1 WT and c4 tumours grown in vivo were analysed by 18FDG-PET and 19FDG Magnetic Resonance Spectroscopy for glucose uptake; by HPLC for adenine nucleotides; by immunohistochemistry for GLUTs; by immunoblotting and by DIGE followed by tandem mass spectrometry for protein expression; and by classical enzymatic methods for enzyme activity. Results HIF-1β deficient Hepa-1 c4 tumours grew significantly more slowly than WT tumours, and (as expected showed significantly lower expression of many glycolytic enzymes. However, HIF-1β deficiency caused no significant change in the rate of glucose uptake in c4 tumours compared to WT when assessed in vivo by measuring fluoro-deoxyglucose (FDG uptake. Immunohistochemistry demonstrated less GLUT-1 in c4 tumours, whereas GLUT-2 (liver type was similar to WT. Factors that might upregulate glucose uptake independently of HIF-1 (phospho-Akt, c-Myc were shown to have either lower or similar expression in c4 compared to WT tumours. However the AMP/ATP ratio was 4.5 fold higher (p Conclusions Despite their defective HIF-1 and consequent down-regulation of glycolytic enzyme expression, Hepa-1 c4 tumours maintain glucose uptake and glycolysis because the resulting low [ATP] high [AMP] allosterically activate PFK-1. This mechanism of resistance would keep glycolysis functioning and also result in activation of AMP-Kinase and growth inhibition; it may have major implications for the therapeutic activity of HIF inhibitors in vivo. Interestingly, this control mechanism does not involve transcriptional control or proteomics, but rather the classical activation and inhibition mechanisms

  1. ATR controls cellular adaptation to hypoxia through positive regulation of hypoxia-inducible factor 1 (HIF-1) expression.

    Science.gov (United States)

    Fallone, F; Britton, S; Nieto, L; Salles, B; Muller, C

    2013-09-12

    Tumor cells adaptation to severe oxygen deprivation (hypoxia) plays a major role in tumor progression. The transcription factor HIF-1 (hypoxia-inducible factor 1), whose α-subunit is stabilized under hypoxic conditions is a key component of this process. Recent studies showed that two members of the phosphoinositide 3-kinase-related kinases (PIKKs) family, ATM (ataxia telangiectasia mutated) and DNA-PK (DNA-dependent protein kinase), regulate the hypoxic-dependent accumulation of HIF-1. These proteins initiate cellular stress responses when DNA damage occurs. In addition, it has been demonstrated that extreme hypoxia induces a replicative stress resulting in regions of single-stranded DNA at stalled replication forks and the activation of ATR (ataxia telangiectasia and Rad3 related protein), another member of the PIKKs family. Here, we show that even less severe hypoxia (0.1% O2) also induces activation of ATR through replicative stress. Importantly, in using either transiently silenced ATR cells, cells expressing an inactive form of ATR or cells exposed to an ATR inhibitor (CGK733), we demonstrate that hypoxic ATR activation positively regulates the key transcription factor HIF-1 independently of the checkpoint kinase Chk1. We show that ATR kinase activity regulates HIF-1α at the translational level and we find that the elements necessary for the regulation of HIF-1α translation are located within the coding region of HIF-1α mRNA. Finally, by using three independent cellular models, we clearly show that the loss of ATR expression and/or kinase activity results in the decrease of HIF-1 DNA binding under hypoxia and consequently affects protein expression levels of two HIF-1 target genes, GLUT-1 and CAIX. Taken together, our data show a new function for ATR in cellular adaptation to hypoxia through regulation of HIF-1α translation. Our work offers new prospect for cancer therapy using ATR inhibitors with the potential to decrease cellular adaptation in hypoxic

  2. Expression of HIF-1{alpha} in irradiated tissue is altered by topical negative-pressure therapy

    Energy Technology Data Exchange (ETDEWEB)

    Grimm, A.; Stange, S.; Labanaris, A.; Horch, R.E. [Erlangen-Nuernberg Univ., Erlangen (Germany). Dept. of Plastic and Hand Surgery; Dimmler, A. [Erlangen-Nuernberg Univ., Erlangen (Germany). Dept. of Pathology; Sauer, R.; Grabenbauer, G. [Erlangen-Nuernberg Univ. (Germany). Dept. of Radiation Oncology

    2007-03-15

    Background and Purpose: Despite the enormous therapeutic potential of modern radiotherapy, common side effects such as radiation-induced wound healing disorders remain a well-known clinical phenomenon. Topical negative pressure therapy (TNP) is a novel tool to alleviate intraoperative, percutaneous irradiation or brachytherapy. Since TNP has been shown to positively influence the perfusion of chronic, poorly vascularized wounds, the authors applied this therapeutic method to irradiated wounds and investigated the effect on tissue oxygenation in irradiated tissue in five patients. Material and Methods: With informed patients' consent, samples prior to and 4 and 8 days after continuous TNP with -125 mmHg were obtained during routine wound debridements. Granulation tissue was stained with hematoxylin-eosin, and additionally with CD31, HIF-1{alpha} (hypoxia-inducible factor-1{alpha}), and D2-40 to detect blood vessels, measure indirect signs of hypoxia, and lymph vessel distribution within the pre- and post-TNP samples. Results: In this first series of experiments, a positive influence of TNP onto tissue oxygenation in radiation-induced wounds could be demonstrated. TNP led to a significant decrease of 53% HIF-1{alpha}-positive cell nuclei. At the same time, a slight reduction of CD31-stained capillaries was seen in comparison to samples before TNP. Immunostaining with D2-40 revealed an increased number of lymphatic vessels with distended lumina and an alteration of the parallel orientation within the post-TNP samples. Conclusion: This study is, to the authors' knowledge, the first report on a novel previously not described histological marker to demonstrate the effects of TNP on HIF-1{alpha} expression as an indirect marker of tissue oxygenation in irradiated wounds, as demonstrated by a reduction of HIF-1{alpha} concentration after TNP. Since this observation may be of significant value to develop possible new strategies to treat radiation-induced tissue

  3. Raised HIF1α during normoxia in high altitude pulmonary edema susceptible non-mountaineers.

    Science.gov (United States)

    Soree, Poonam; Gupta, Rajinder K; Singh, Krishan; Desiraju, Koundinya; Agrawal, Anurag; Vats, Praveen; Bharadwaj, Abhishek; Baburaj, T P; Chaudhary, Pooja; Singh, Vijay K; Verma, Saroj; Bajaj, Amir Chand; Singh, Shashi Bala

    2016-05-23

    High altitude pulmonary edema (HAPE) susceptibility is associated with EGLN1 polymorphisms, we hypothesized that HAPE-susceptible (HAPE-S, had HAPE episode in past) subjects may exhibit abnormal HIF1α levels in normoxic conditions. We measured HIF1α levels in HAPE-S and HAPE resistant (HAPE-R, no HAPE episode) individuals with similar pulmonary functions. Hemodynamic responses were also measured before and after normobaric hypoxia (Fi02 = 0.12 for 30 min duration at sea level) in both groups. . HIF1α was higher in HAPE-S (320.3 ± 267.5 vs 58.75 ± 33.88 pg/ml, P < 0.05) than HAPE-R, at baseline, despite no significant difference in baseline oxygen saturations (97.7 ± 1.7% and 98.8 ± 0.7). As expected, HAPE-S showed an exaggerated increase in pulmonary artery pressure (27.9 ± 6 vs 19.3 ± 3.7 mm Hg, P < 0.05) and a fall in peripheral oxygen saturation (66.9 ± 11.7 vs 78.7 ± 3.8%, P < 0.05), when exposed to hypoxia. HIF1α levels at baseline could accurately classify members of the two groups (AUC = 0.87). In a subset of the groups where hemoglobin fractions were additionally measured to understand the cause of elevated hypoxic response at baseline, two of four HAPE-S subjects showed reduced HbA. In conclusion, HIF 1 α levels during normoxia may represent an important marker for determination of HAPE susceptibility.

  4. Locomotor diseases among male long-haul truck drivers and other professional drivers

    DEFF Research Database (Denmark)

    Jensen, Anker; Kaerlev, Linda; Tüchsen, Finn

    2007-01-01

    -249) and for other truck drivers (SHR: 130, 95% CI: 108-156) compared to bus drivers (SHR: 110, 95% CI: 79-149). All drivers had high SHR for lesions of the ulnar nerve (SHR: 159, 95% CI: 119-207), especially bus drivers (SHR: 197, 95% CI: 116-311). Long-haul truck drivers had high SHRs for synovitis and bursitis...

  5. Locomotor diseases among male long-haul truck drivers and other professional drivers

    DEFF Research Database (Denmark)

    Jensen, Anker; Kaerlev, Linda; Tüchsen, Finn

    2007-01-01

    -249) and for other truck drivers (SHR: 130, 95% CI: 108-156) compared to bus drivers (SHR: 110, 95% CI: 79-149). All drivers had high SHR for lesions of the ulnar nerve (SHR: 159, 95% CI: 119-207), especially bus drivers (SHR: 197, 95% CI: 116-311). Long-haul truck drivers had high SHRs for synovitis and bursitis...

  6. Aminoflavone, a ligand of the Aryl Hydrocarbon Receptor (AhR), inhibits HIF-1α expression in an AhR-independent fashion

    Science.gov (United States)

    Terzuoli, Erika; Puppo, Maura; Rapisarda, Annamaria; Uranchimeg, Badarch; Cao, Liang; Burger, Angelika M.; Ziche, Marina; Melillo, Giovanni

    2010-01-01

    Aminoflavone (AF), the active component of a novel anticancer agent (AFP464) in phase I clinical trials, is a ligand of the aryl hydrocarbon receptor (AhR). AhR dimerizes with HIF-1β/ARNT, which is shared with HIF-1α, a transcription factor critical for the response of cells to oxygen deprivation. To address whether pharmacological activation of the AhR pathway might be a potential mechanism for inhibition of HIF-1, we tested the effects of AF on HIF-1 expression. AF inhibited HIF-1α transcriptional activity and protein accumulation in MCF-7 cells. However, inhibition of HIF-1α by AF was independent from a functional AhR pathway. Indeed, AF inhibited HIF-1α expression in AhR100 cells, in which the AhR pathway is functionally impaired, yet did not induce cytotoxicity, providing evidence that these effects are mediated by distinct signaling pathways. Moreover, AF was inactive in MDA-MB-231 cells, yet inhibited HIF-1α in MDA-MB-231 cells transfected with the SULT1A1 gene. AF inhibited HIF-1α mRNA expression by approximately 50%. Notably, actinomycin-D completely abrogated the ability of AF to down-regulate HIF-1α mRNA, indicating that active transcription was required for the inhibition of HIF-1α expression. Finally, AF inhibited HIF-1α protein accumulation and the expression of HIF-1-target genes in MCF-7 xenografts. These results demonstrate that AF inhibits HIF-1α in an AhR-independent fashion and they unveil additional activities of AF that may be relevant for its further clinical development. PMID:20736373

  7. HIF-1α inhibition reverses multidrug resistance in colon cancer cells via downregulation of MDR1/P-glycoprotein.

    Directory of Open Access Journals (Sweden)

    Jianfang Chen

    Full Text Available BACKGROUND: Multidrug resistance (MDR is one of the major reasons chemotherapy-based treatments fail. Hypoxia is generally associated with tumor chemoresistance. However, the correlation between the heterodimeric hypoxia-inducible factor-1 (HIF-1 and the multidrug resistance (MDR1 gene/transporter P-glycoprotein (P-gp remains unclear. This study aims to explore the molecular mechanisms of reversing colon cancer MDR by focusing on the target gene HIF-1α. METHODS: A chemotherapeutic sensitivity assay was used to observe the efficiency of MDR reversal in LoVo multicellular spheroids (MCS. The apoptotic level induced by different drugs was examined by flow cytometry (FCM. Binding of HIF-1α to the MDR1 gene promoter was evaluated by Chromatin immunoprecipitation (ChIP. The relationship between HIF-1α/P-gp expression and sensitivity to chemotherapy was analyzed. RESULTS: The sensitivity of LoVo MCS to all four chemotherapy drugs was decreased to varying degrees under hypoxic conditions. After silencing the HIF-1α gene, the sensitivities of LoVo MCS to all four chemotherapy drugs were restored. The apoptotic levels that all the drugs induced were all decreased to various extents in the hypoxic group. After silencing HIF-1α, the apoptosis level induced by all four chemotherapy drugs increased. The expression of HIF-1α and P-gp was significantly enhanced in LoVo MCS after treatment with hypoxia. Inhibiting HIF-1α significantly decreased the expression of MDR1/P-gp mRNA or protein in both the LoVo monolayers and LoVo MCS. The ChIP assay showed that HIF-1α was bound to the MDR1 gene promoter. Advanced colon carcinoma patients with expression of both HIF-1α and P-gp were more resistant to chemotherapy than that with non expression. CONCLUSIONS: HIF-1α inhibition reverses multidrug resistance in colon cancer cells via downregulation of MDR1/P-gp. The expression of HIF-1α and MDR1/P-gp can be used as a predictive marker for chemotherapy resistance

  8. Sexual behavior among truck drivers.

    Science.gov (United States)

    Singh, Rajiv Kumar; Joshi, Hari Shankar

    2012-01-01

    A cross-sectional study was conducted on Lucknow highway in Bareilly district of Uttar Pradesh to study the knowledge of truck drivers about HIV transmission and prevention and to study the sexual behaviour of these drivers with reference to HIV/AIDS. Age, marital status, education, income, drinking alcohol, length of stay away from home, knowledge about transmission and prevention of HIV, and HIV-prone behavior of truck drivers were studied. Chi-square, mean, and SD were calculated. In all, 289 (97.6%) drivers had heard about HIV/AIDS. Only 242 (81.8%) were aware of HIV transmission by heterosexual route. Misconceptions such as HIV transmission by mosquito bites, living in same room, shaking hands, and sharing food were found. Out of 174 (58.8%) who visited Commercial Sex Workers (CSW), 146 (83.9%) used a condom. 38 (12.8%) visited more than 5 CSW in the last 3 months. Time away from home on the road, marital status, alcohol use, and income class were associated with visiting CSW. High-risk behavior was established in the study population. Safe sex and use of condoms need to be promoted among the truck drivers and better condom availability needs to be assured on highways.

  9. The accumulations of HIF-1α and HIF-2α by JNK and ERK are involved in biphasic effects induced by different levels of arsenite in human bronchial epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Yuan; Li, Yuan [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Li, Huiqiao [Qujing Center for Disease Control and Prevention, Qujing 655000, Yunnan (China); Pang, Ying; Zhao, Yue; Jiang, Rongrong; Shen, Lu [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Zhou, Jianwei; Wang, Xinru [The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Liu, Qizhan, E-mail: drqzliu@hotmail.com [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China)

    2013-01-15

    The biphasic effects of arsenite, in which low levels of arsenite induce cell proliferation and high levels of arsenite induce DNA damage and apoptosis, apparently contribute to arsenite-induced carcinogenesis. However, the mechanisms underlying this phenomenon are not well understood. In this study, we investigated the effects of different levels of arsenite on cell proliferation, DNA damage and apoptosis as well as on signal transduction pathways in human bronchial epithelial (HBE) cells. Our results show that a low level of arsenite activates extracellular signal-regulated kinases (ERK), which probably mediate arsenite-inhibited degradation of ubiquitinated hypoxia-inducible factor-2α (HIF-2α) in HBE cells. ERK inhibition blocks cell proliferation induced by a low level of arsenite, in part via HIF-2α. In contrast, a high level of arsenite activates c-Jun N-terminal kinases (JNK), which provoke a response to suppress ubiquitinated HIF-1α degradation. Down-regulation of HIF-1α by inhibiting JNK, however, increases the DNA damage but decreases the apoptosis induced by a high level of arsenite. Thus, data in the present study suggest that the accumulations of HIF-1α and HIF-2α by JNK and ERK are involved in different levels of arsenite-induced biphasic effects, with low levels of arsenite inducing cell proliferation and high levels of arsenite inducing DNA damage and apoptosis in HBE cells. -- Highlights: ► Biphasic effects induced by different concentrations of arsenite. ► Different regulation of ERK or JNK signal pathway by arsenite. ► Different regulation of HIF1α or HIF 2α by arsenite.

  10. Construction and indentification of CRISPR/Cas9 plasmid targeting HIF-1α gene%靶向HIF-1α基因的CRISPR/Cas9基因敲除质粒的构建与鉴定

    Institute of Scientific and Technical Information of China (English)

    何玉婷; 李娟; 孙冉冉; 陈晓龙; 申珅; 阚全程; 余祖江

    2016-01-01

    目的:构建靶向低氧诱导因子(HIF)-1α基因的CRISPR/Cas9基因敲除质粒,并体外鉴定其敲除效果.方法:设计靶向HIF-1α基因的gRNA序列,将其插入到CRISPR/Cas9质粒骨架载体pCAG-T7中,转化后挑取克隆,进行测序验证.将构建好的重组质粒体外转染人肝癌细胞HepG2和SK-Hep-I,利用嘌呤霉素进行筛选并扩大培养获得HIF-1α基因敲除混合克隆细胞,应用Western blot技术检测未转染细胞和混合克隆细胞中HIF-1α蛋白的表达情况.结果:经测序验证,成功构建了靶向HIF-1α的重组质粒pCAG-T7-HIF-1α-gRNA,经Western blot验证,转染该重组质粒后人肝癌细胞株HepG2和SK-Hep-1经CoCl2诱导HIF-1α蛋白表达明显减弱.结论:成功构建了靶向HIF-1α基因的CRISPR/Cas9基因敲除质粒载体.

  11. Key drivers of airline loyalty

    Science.gov (United States)

    Dolnicar, Sara; Grabler, Klaus; Grün, Bettina; Kulnig, Anna

    2011-01-01

    This study investigates drivers of airline loyalty. It contributes to the body of knowledge in the area by investigating loyalty for a number of a priori market segments identified by airline management and by using a method which accounts for the multi-step nature of the airline choice process. The study is based on responses from 687 passengers. Results indicate that, at aggregate level, frequent flyer membership, price, the status of being a national carrier and the reputation of the airline as perceived by friends are the variables which best discriminate between travellers loyal to the airline and those who are not. Differences in drivers of airline loyalty for a number of segments were identified. For example, loyalty programs play a key role for business travellers whereas airline loyalty of leisure travellers is difficult to trace back to single factors. For none of the calculated models satisfaction emerged as a key driver of airline loyalty. PMID:27064618

  12. Key drivers of airline loyalty.

    Science.gov (United States)

    Dolnicar, Sara; Grabler, Klaus; Grün, Bettina; Kulnig, Anna

    2011-10-01

    This study investigates drivers of airline loyalty. It contributes to the body of knowledge in the area by investigating loyalty for a number of a priori market segments identified by airline management and by using a method which accounts for the multi-step nature of the airline choice process. The study is based on responses from 687 passengers. Results indicate that, at aggregate level, frequent flyer membership, price, the status of being a national carrier and the reputation of the airline as perceived by friends are the variables which best discriminate between travellers loyal to the airline and those who are not. Differences in drivers of airline loyalty for a number of segments were identified. For example, loyalty programs play a key role for business travellers whereas airline loyalty of leisure travellers is difficult to trace back to single factors. For none of the calculated models satisfaction emerged as a key driver of airline loyalty.

  13. [Effect of HIF-1α Gene Silence on Biological Characteristics of Human Colon Cancer Cells SW480].

    Science.gov (United States)

    Wang, Qiang; Hao, Lang-song; Shi, Jia; Huang, Jian

    2015-07-01

    To investigate changes in proliferation and apoptosis of human colon cancer SW480 cells after silencing hypoxia inducible factor-lα (HIF-1α) expression. siRNA interference technology was performed to silence the expression of HIF-1α using lipofectamine mediation to transfect siRNA into human colon cancer SW480 cells. The siRNA interfered SW480 cells were compared with a negative control group, an empty vector group, and a blank control group. Real-time PCR and Western blot were used to measure the expressions of HIF-lα protein and mRNA. MTT and flow cytometry (FCM) were used to evaluate the apoptosis and proliferation of SW480 cells. The interfered SW480 cells had a higher level of silence of HIF-lα mRNA (> 80%) compared with those of in the three control groups (P silencing promotes apoptosis and inhibits the proliferation of SW480 cells in vitro.

  14. Predictive values of H.I.F.-1 alpha, H.I.F.-2 alpha and C.A. 9 expressions by prostate adenocarcinomas treated by exclusive irradiation. Ancillary study of the G.E.T.U.G. 06 protocol; Valeurs predictives des expressions de HIF-1 alpha, HIF-2 alpha et CA 9 par les adenocarcinomes de la prostate traites par irradiation exclusive. Etude ancillaire du protocole GETUG 06

    Energy Technology Data Exchange (ETDEWEB)

    Simon, J.M.; Mazeron, J.J. [Groupe Hospitalier de la Pitie-Salpetriere, APHP, Service de Radiotherapie, 75 - Paris (France); Comperat, E. [Groupe Hospitalier de la Pitie-Salpetriere, APHP, Lab. d' Anatomie Pathologique, 75 - Paris (France); Beckendorf, V. [Centre Alexis-Vautrin, Dept. de Radiotherapie, 54 - Vandoeuvre-les-Nancy (France); Bey, P. [Institut Curie, 75 - Paris (France); Jaillon, P. [Hopital Saint-Antoine, APHP, Service de Pharmacologie, 75 - Paris (France)

    2007-11-15

    The adenocarcinomas of the prostate are potentially hypoxic tumors. The strong expression of markers of hypoxia H.I.F.-2 alpha and C.A. 9 are independent predictor factors of biochemical relapse after exclusive radiotherapy. (N.C.)

  15. Effect of overexpression of HIF-2 alpha on movement and invasion of hepatocellular carcinoma cells%过表达HIF-2α对肝癌细胞运动和侵袭的影响

    Institute of Scientific and Technical Information of China (English)

    孙海香; 杨柳晓

    2016-01-01

    目的:探讨缺氧诱导因子(hypoxia inducible factor-2α,HIF-2α)对肝癌细胞运动和侵袭的影响。方法:构建 HIF-2α过表达载体,转染肝癌细胞,构建过表达稳定表达细胞株。采用实时荧光定量多聚酶联反应(quantitative real-time polymerase chain reaction ,qRT-PCR)和蛋白质印迹法检测HIF-2α表达水平,通过Transwell运动实验、划痕实验体外分析过表达HIF-2α对肝癌细胞运动的影响;将过表达HIF-2α细胞原位接种裸鼠,观察小鼠肺脏的转移情况。采用蛋白质印迹法检测EM T相关指标分析 HIF-2α促进肝癌细胞运动、侵袭能力的作用机制。结果:将过表达载体转染肝癌细胞后,HIF-2αmRNA和蛋白表达水平均升高(P<0.05)。Transwell运动实验发现,高表达 HIF-2α后,穿膜细胞数目明显增加(P<0.01);划痕实验结果也表明高表达HIF-2α的细胞运动能力增强。体内侵袭实验发现高表达 HIF-2α组小鼠肺转移灶大小增加。蛋白质印迹结果显示HIF-2α可促进间质细胞相关标志物蛋白表达增加。结论:过表达 HIF-2α可能通过促进肝癌细胞的EM T转化提高肝癌细胞的运动、侵袭能力。%Objective:To investigate the effects of hypoxia inducible factor 2 alpha (HIF-2α ) on movement and invasion of hepatocellular carcinoma cells (HCC) .Methods:The plasmid of over-expression of HIF-2α was constructed and transfected into HCC cells ,then the stable expression cell strains were selected .HIF-2α expression levels were detected by quantitative real-time polymerase chain reaction (RT-PCR) and Western Bloting .The effects of HIF-2αover-expression on the movement ability of HCC were analyzed in vitro by Transwell assay and scrach assay .The over-expressed HIF-2αcells were planted in situ in nude mice ,and the lung metastasis were observed .we checked the EMT markers by Western Bloting .The mechanism of HIF-2α promoting cancer

  16. 妊娠期糖尿病胎盘组织HIF-1α及EPO表达及意义%EXPRESSIONS OF HIF-1α AND EPO AND THEIR SIGNIFICATION IN PLATENTA OF GESTATIONAL DIABTES MELLITUS

    Institute of Scientific and Technical Information of China (English)

    孙玲玲; 杨晓菊; 谭萍; 陈燕秋; 吴洪婧

    2014-01-01

    目的 探讨缺氧诱导因子1α(HIF-1α)和促红细胞生成素(EPO)在妊娠期糖尿病(GDM)胎盘组织的表达及临床意义.方法 采用RT-PCR方法检测44例GDM孕妇、37例正常产妇胎盘组织中HIF-1α mRNA和EPO mRNA的表达.结果 GDM组HIF-1α mRNA水平高于正常对照组,差异有显著意义(t=15.907,P<0.05);GDM组EPO mRNA表达水平高于正常对照组,差异有显著性(t=10.167,P<0.05).GDM巨大儿组新生儿出生体质量与HIF-1α及EPO表达均呈正相关(r=0.662、0.475,P<0.05).结论 胎盘组织中存在HIF-1α和EPO的基因表达,与GDM巨大儿的发生有关.HIF-1α的高表达可能是EPO表达上调的机制之一.

  17. Noscapine inhibits hypoxia-mediated HIF-1alpha expression andangiogenesis in vitro: a novel function for an old drug.

    Science.gov (United States)

    Newcomb, Elizabeth W; Lukyanov, Yevgeniy; Schnee, Tona; Ali, M Aktar; Lan, Li; Zagzag, David

    2006-05-01

    Overexpression of hypoxia-inducible factor-1 (HIF-1) is a common feature in solid malignancies related to oxygen deficiency. Since increased HIF-1 expression correlates with advanced disease stage, increased angiogenesis and poor prognosis, HIF-1 and its signaling pathway have become targets for cancer chemotherapy. In this study, we identified noscapine to be a novel small molecule inhibitor of the HIF-1 pathway based on its structure-function relation-ships with HIF-1 pathway inhibitors belonging to the benzylisoquinoline class of plant metabolites and/or to microtubule binding agents. We demonstrate that noscapine treatment of human glioma U87MG and T98G cell lines exposed to the hypoxic mimetic agent, CoCl2, inhibits hypoxia-mediated HIF-1alpha expression and transcriptional activity as measured by decreased secretion of VEGF, a HIF-1 target gene. Inhibition of hypoxia-mediated HIF-1alpha expression was due, in part, to its ability to inhibit accumulation of HIF-1alpha in the nucleus and target it for degradation via the proteasome. One mechanism of action of microtubule binding agents is their antiangiogenic activity associated with disruption of endothelial tubule formation. We show that noscapine has similar properties in vitro. Thus, noscapine may possess novel antiangiogenic activity associated with two broad mechanisms of action: first, by decreasing HIF-1alpha expression in hypoxic tumor cells, upregulation of target genes, such as VEGF, would be decreased concomitant with its associated angiogenic activity; second, by inhibiting endothelial cells from forming blood vessels in response to VEGF stimulation, it may limit the process of neo-vascularization, correlating with antitumor activity in vivo. For more than 75 years, noscapine has traditionally been used as an oral cough suppressant with no known toxic side effects in man. Thus, the studies reported here have found a novel function for an old drug. Given its low toxicity profile, its demonstrated

  18. The bone-forming effects of HIF-1α-transduced BMSCs promote osseointegration with dental implant in canine mandible.

    Directory of Open Access Journals (Sweden)

    Duohong Zou

    Full Text Available The presence of insufficient bone volume remains a major clinical problem for dental implant placement to restore the oral function. Gene-transduced stem cells provide a promising approach for inducing bone regeneration and enhancing osseointegration in dental implants with tissue engineering technology. Our previous studies have demonstrated that the hypoxia-inducible factor-1α (HIF-1α promotes osteogenesis in rat bone mesenchymal stem cells (BMSCs. In this study, the function of HIF-1α was validated for the first time in a preclinical large animal canine model in term of its ability to promote new bone formation in defects around implants as well as the osseointegration between tissue-engineered bone and dental implants. A lentiviral vector was constructed with the constitutively active form of HIF-1α (cHIF. The ectopic bone formation was evaluated in nude mice. The therapeutic potential of HIF-1α-overexpressing canine BMSCs in bone repair was evaluated in mesi-implant defects of immediate post-extraction implants in the canine mandible. HIF-1α mediated canine BMSCs significantly promoted new bone formation both subcutaneously and in mesi-implant defects, including increased bone volume, bone mineral density, trabecular thickness, and trabecular bone volume fraction. Furthermore, osseointegration was significantly enhanced by HIF-1α-overexpressing canine BMSCs. This study provides an important experimental evidence in a preclinical large animal model concerning to the potential applications of HIF-1α in promoting new bone formation as well as the osseointegration of immediate implantation for oral function restoration.

  19. Estrogen Receptor beta 2 Induces Hypoxia Signature of Gene Expression by Stabilizing HIF-1 alpha in Prostate Cancer

    OpenAIRE

    Prasenjit Dey; Velazquez-Villegas, Laura A.; Michelle Faria; Anthony Turner; Philp Jonsson; Paul Webb; Cecilia Williams; Jan-Åke Gustafsson; Ström, Anders M.

    2015-01-01

    The estrogen receptor (ER) beta variant ER beta 2 is expressed in aggressive castration-resistant prostate cancer and has been shown to correlate with decreased overall survival. Genome-wide expression analysis after ER beta 2 expression in prostate cancer cells revealed that hypoxia was an overrepresented theme. Here we show that ER beta 2 interacts with and stabilizes HIF-1 alpha protein in normoxia, thereby inducing a hypoxic gene expression signature. HIF-1 alpha is known to stimulate met...

  20. Early Diagnosis of Clear Cell Kidney Cancer via VHL/HIF Pathway Regulated-Circulating microRNA

    Science.gov (United States)

    2016-05-01

    Award Number: W81XWH-11-1-0715 TITLE: Early Diagnosis of Clear Cell Kidney Cancer via VHL/HIF Pathway -Regulated Circulating microRNA PRINCIPAL...TITLE AND SUBTITLE Sa. CONTRACT NUMBER Early Diagnosis of Clear Cell Kidney Cancer via VHL/HIF Pathway - Regulated Circulating microRNA Sb. GRANT NUMBER...panel of diagnostic miRNAs that are measurable in serum and will be able to identify kidney cancer in its earliest stages. We hypothesized that serum

  1. The dietary flavonoid kaempferol effectively inhibits HIF-1 activity and hepatoma cancer cell viability under hypoxic conditions

    Energy Technology Data Exchange (ETDEWEB)

    Mylonis, Ilias; Lakka, Achillia; Tsakalof, Andreas [Laboratory of Biochemistry, School of Medicine, University of Thessaly, BIOPOLIS, 41110 Larissa (Greece); Institute of Biomedical Research and Technology (BIOMED), 51 Papanastasiou str., 41222 Larissa (Greece); Simos, George, E-mail: simos@med.uth.gr [Laboratory of Biochemistry, School of Medicine, University of Thessaly, BIOPOLIS, 41110 Larissa (Greece); Institute of Biomedical Research and Technology (BIOMED), 51 Papanastasiou str., 41222 Larissa (Greece)

    2010-07-16

    Research highlights: {yields} Kaempferol inhibits HIF-1 activity in hepatocarcinoma cells; {yields} Kaempferol causes cytoplasmic mislocalization of HIF-1{alpha} by impairing the MAPK pathway. {yields} Viability of hepatocarcinoma cells under hypoxia is reduced by kaempferol. -- Abstract: Hepatocellular carcinoma (HCC) is characterized by high mortality rates and resistance to conventional treatment. HCC tumors usually develop local hypoxia, which stimulates proliferation of cancer cells and renders them resilient to chemotherapy. Adaptation of tumor cells to the hypoxic conditions depends on the hypoxia-inducible factor 1 (HIF-1). Over-expression of its regulated HIF-1{alpha} subunit, an important target of anti-cancer therapy, is observed in many cancers including HCC and is associated with severity of tumor growth and poor patient prognosis. In this report we investigate the effect of the dietary flavonoid kaempferol on activity, expression levels and localization of HIF-1{alpha} as well as viability of human hepatoma (Huh7) cancer cells. Treatment of Huh7 cells with kaempferol under hypoxic conditions (1% oxygen) effectively inhibited HIF-1 activity in a dose-dependent manner (IC{sub 50} = 5.16 {mu}M). The mechanism of this inhibition did not involve suppression of HIF-1{alpha} protein levels but rather its mislocalization into the cytoplasm due to inactivation of p44/42 MAPK by kaempferol (IC{sub 50} = 4.75 {mu}M). Exposure of Huh7 cells to 10 {mu}{Mu} kaempferol caused significant reduction of their viability, which was remarkably more evident under hypoxic conditions. In conclusion, kaempferol, a non-toxic natural food component, inhibits both MAPK and HIF-1 activity at physiologically relevant concentrations (5-10 {mu}M) and suppresses hepatocarcinoma cell survival more efficiently under hypoxia. It has, therefore, potential as a therapeutic or chemopreventive anti-HCC agent.

  2. Interactions between HIF-1α and AMPK in the regulation of cellular hypoxia adaptation in chronic kidney disease.

    Science.gov (United States)

    Li, Hui; Satriano, Joseph; Thomas, Joanna L; Miyamoto, Satoshi; Sharma, Kumar; Pastor-Soler, Núria M; Hallows, Kenneth R; Singh, Prabhleen

    2015-09-01

    Renal hypoxia contributes to chronic kidney disease (CKD) progression, as validated in experimental and human CKD. In the early stages, increased oxygen consumption causes oxygen demand/supply mismatch, leading to hypoxia. Hence, early targeting of the determinants and regulators of oxygen consumption in CKD may alter the disease course before permanent damage ensues. Here, we focus on hypoxia inducible factor-1α (HIF-1α) and AMP-activated protein kinase (AMPK) and on the mechanisms by which they may facilitate cellular hypoxia adaptation. We found that HIF-1α activation in the subtotal nephrectomy (STN) model of CKD limits protein synthesis, inhibits apoptosis, and activates autophagy, presumably for improved cell survival. AMPK activation was diminished in the STN kidney and was remarkably restored by HIF-1α activation, demonstrating a novel role for HIF-1α in the regulation of AMPK activity. We also investigated the independent and combined effects of HIF-1α and AMPK on cell survival and death pathways by utilizing pharmacological and knockdown approaches in cell culture models. We found that the effect of HIF-1α activation on autophagy is independent of AMPK, but on apoptosis it is partially AMPK dependent. The effects of HIF-1α and AMPK activation on inhibiting protein synthesis via the mTOR pathway appear to be additive. These various effects were also observed under hypoxic conditions. In conclusion, HIF-1α and AMPK appear to be linked at a molecular level and may act as components of a concerted cellular response to hypoxic stress in the pathophysiology of CKD.

  3. Role of Hypoxia Inducible Factor-1α (HIF-1α in Innate Defense against Uropathogenic Escherichia coli Infection.

    Directory of Open Access Journals (Sweden)

    Ann E Lin

    2015-04-01

    Full Text Available Uropathogenic E. coli (UPEC is the primary cause of urinary tract infections (UTI affecting approximately 150 million people worldwide. Here, we revealed the importance of transcriptional regulator hypoxia-inducible factor-1 α subunit (HIF-1α in innate defense against UPEC-mediated UTI. The effects of AKB-4924, a HIF-1α stabilizing agent, were studied using human uroepithelial cells (5637 and a murine UTI model. UPEC adherence and invasion were significantly reduced in 5637 cells when HIF-1α protein was allowed to accumulate. Uroepithelial cells treated with AKB-4924 also experienced reduced cell death and exfoliation upon UPEC challenge. In vivo, fewer UPEC were recovered from the urine, bladders and kidneys of mice treated transurethrally with AKB-4924, whereas increased bacteria were recovered from bladders of mice with a HIF-1α deletion. Bladders and kidneys of AKB-4924 treated mice developed less inflammation as evidenced by decreased pro-inflammatory cytokine release and neutrophil activity. AKB-4924 impairs infection in uroepithelial cells and bladders, and could be correlated with enhanced production of nitric oxide and antimicrobial peptides cathelicidin and β-defensin-2. We conclude that HIF-1α transcriptional regulation plays a key role in defense of the urinary tract against UPEC infection, and that pharmacological HIF-1α boosting could be explored further as an adjunctive therapy strategy for serious or recurrent UTI.

  4. Expression of HIF-1α and Its Target Genes in the Nanorana parkeri Heart:Implications for High Altitude Adaptation

    Institute of Scientific and Technical Information of China (English)

    Qiong ZHANG; Xingzhi HAN; Yinzi YE; Robert H S KRAUS; Liqing FAN; Le YANG; Yi TAO

    2016-01-01

    Hypoxia-inducible factor 1 alpha (HIF-1α) and its target genes vascular endothelial growth factor (VEGF) and transferrins (TF) play an important role in native endothermic animals’ adaptation to the high altitude environments. For ectothermic animals – especially frogs – it remains undetermined whether HIF-1α and its target genes (VEGF and TF) play an important role in high altitude adaptation, too. In this study, we compared the gene sequences and expression of HIF-1α and its target genes (VEGF and TF) between three Nanorana parkeri populations from different altitudes (3008 m a.s.l., 3440 m a.s.l. and 4312 m a.s.l.). We observed that the cDNA sequences of HIF-1A exhibited high sequence similarity (99.38%) among the three altitudinally separated populations; but with increasing altitude, the expression of HIF-1A and its target genes (VEGF and TF) increased significantly. These results indicate that HIF-1αplays an important role in N. parkeri adaptation to the high altitude, similar to its role in endothermic animals.

  5. Role of Hypoxia Inducible Factor-1α (HIF-1α) in Innate Defense against Uropathogenic Escherichia coli Infection.

    Science.gov (United States)

    Lin, Ann E; Beasley, Federico C; Olson, Joshua; Keller, Nadia; Shalwitz, Robert A; Hannan, Thomas J; Hultgren, Scott J; Nizet, Victor

    2015-04-01

    Uropathogenic E. coli (UPEC) is the primary cause of urinary tract infections (UTI) affecting approximately 150 million people worldwide. Here, we revealed the importance of transcriptional regulator hypoxia-inducible factor-1 α subunit (HIF-1α) in innate defense against UPEC-mediated UTI. The effects of AKB-4924, a HIF-1α stabilizing agent, were studied using human uroepithelial cells (5637) and a murine UTI model. UPEC adherence and invasion were significantly reduced in 5637 cells when HIF-1α protein was allowed to accumulate. Uroepithelial cells treated with AKB-4924 also experienced reduced cell death and exfoliation upon UPEC challenge. In vivo, fewer UPEC were recovered from the urine, bladders and kidneys of mice treated transurethrally with AKB-4924, whereas increased bacteria were recovered from bladders of mice with a HIF-1α deletion. Bladders and kidneys of AKB-4924 treated mice developed less inflammation as evidenced by decreased pro-inflammatory cytokine release and neutrophil activity. AKB-4924 impairs infection in uroepithelial cells and bladders, and could be correlated with enhanced production of nitric oxide and antimicrobial peptides cathelicidin and β-defensin-2. We conclude that HIF-1α transcriptional regulation plays a key role in defense of the urinary tract against UPEC infection, and that pharmacological HIF-1α boosting could be explored further as an adjunctive therapy strategy for serious or recurrent UTI.

  6. The LINK-A lncRNA Activates Normoxic HIF1α Signaling in Triple-negative Breast Cancer

    Science.gov (United States)

    Lin, Aifu; Li, Chunlai; Xing, Zhen; Hu, Qingsong; Liang, Ke; Han, Leng; Wang, Cheng; Hawke, David H.; Wang, Shouyu; Zhang, Yanyan; Wei, Yongkun; Ma, Guolin; Park, Peter K.; Zhou, Jianwei; Zhou, Yan; Hu, Zhibin; Zhou, Yubin; Marks, Jeffery R.; Liang, Han; Hung, Mien-Chie; Lin, Chunru; Yang, Liuqing

    2016-01-01

    Although long noncoding RNAs (lncRNAs) predominately reside in nuclear and exert their functions in many biological processes, their potential involvement in cytoplasmic signal transduction remains unexplored. Here, we identified a cytoplasmic lncRNA, Long-Intergenic Noncoding RNA for Kinase Activation (LINK-A), which mediates HB-EGF triggered, EGFR:GPNMB heterodimer-dependent HIF1α phosphorylation at Tyr565 and Ser797 by BRK and LRRK2 respectively. These events cause HIF1α stabilization, HIF1α-p300 interaction, and activation of HIF1α transcriptional programs under normoxic conditions. Mechanistically, LINK-A facilitates the recruitment of BRK to EGFR:GPNMB complex and BRK kinase activation. The BRK-dependent HIF1α Tyr565 phosphorylation interferes with Pro564 hydroxylation, leading to normoxic HIF1α stabilization. Both LINK-A and LINK-A-dependent signaling pathway activation correlate with TNBC, promoting breast cancer glycolysis reprogramming and tumorigenesis. Our findings illustrate the magnitude and diversity of cytoplasmic lncRNAs in signal transduction and highlight the important roles of lncRNAs in cancer. PMID:26751287

  7. HIF-1 attenuates Ref-1 expression in endothelial cells: reversal by siRNA and inhibition of geranylgeranylation.

    Science.gov (United States)

    Loboda, Agnieszka; Stachurska, Anna; Dorosz, Jerzy; Zurawski, Marek; Wegrzyn, Joanna; Kozakowska, Magdalena; Jozkowicz, Alicja; Dulak, Jozef

    2009-01-01

    Redox factor-1 (Ref-1), a multifunctional protein with DNA repairing activities, plays a cytoprotective function by post-translational redox modification of numerous transcription factors, including hypoxia inducible factor-1 (HIF-1). In the present study, activation of HIF-1 by hypoxia and dimethyloxaloylglycine (DMOG), a hypoxia mimic, diminished Ref-1 mRNA and protein expression in human microvascular endothelial cells (HMEC-1). Similarly, adenoviral delivery of the stabilized form of HIF-1alpha decreased Ref-1 mRNA and protein levels. Accordingly, HIF-1alpha siRNA abolished the hypoxia-induced inhibition of Ref-1 expression, indicating the role of HIF-1 in down-regulation of Ref-1. Also, translocation of Ref-1 from nucleus to cytoplasm after HIF-1 activation was noted. Interestingly, we observed the restoration of Ref-1 expression in hypoxia by pharmacologically relevant doses of atorvastatin. This effect was dependent on the inhibition of protein geranylgeranylation, but not farnesylation, as only the inhibitor of the former but not the latter prenylation step restored the Ref-1 expression. The regulation of Ref-1 by statins may be considered as a novel mechanism of their beneficial effects on endothelium.

  8. TwinDrivers: Semi-Automatic Derivation of Fast and Safe Hypervisor Network Drivers from Guest OS Drivers

    OpenAIRE

    2009-01-01

    In a virtualized environment, device drivers are often run inside a virtual machine (VM) rather than in the hypervisor. Doing so protects the hypervisor from bugs in the driver, and also allows the reuse of the device driver and its support infrastructure in the VM. Unfortunately, this approach results in poor performance for I/O intensive devices such as network cards. The alternative approach is to run device drivers directly in the hypervisor. Although this approach results in better perfo...

  9. Structural basis for oxygen degradation domain selectivity of the HIF prolyl hydroxylases

    Science.gov (United States)

    Chowdhury, Rasheduzzaman; Leung, Ivanhoe K. H.; Tian, Ya-Min; Abboud, Martine I.; Ge, Wei; Domene, Carmen; Cantrelle, François-Xavier; Landrieu, Isabelle; Hardy, Adam P.; Pugh, Christopher W.; Ratcliffe, Peter J.; Claridge, Timothy D. W.; Schofield, Christopher J.

    2016-08-01

    The response to hypoxia in animals involves the expression of multiple genes regulated by the αβ-hypoxia-inducible transcription factors (HIFs). The hypoxia-sensing mechanism involves oxygen limited hydroxylation of prolyl residues in the N- and C-terminal oxygen-dependent degradation domains (NODD and CODD) of HIFα isoforms, as catalysed by prolyl hydroxylases (PHD 1-3). Prolyl hydroxylation promotes binding of HIFα to the von Hippel-Lindau protein (VHL)-elongin B/C complex, thus signalling for proteosomal degradation of HIFα. We reveal that certain PHD2 variants linked to familial erythrocytosis and cancer are highly selective for CODD or NODD. Crystalline and solution state studies coupled to kinetic and cellular analyses reveal how wild-type and variant PHDs achieve ODD selectivity via different dynamic interactions involving loop and C-terminal regions. The results inform on how HIF target gene selectivity is achieved and will be of use in developing selective PHD inhibitors.

  10. Fuel-Cell Drivers Wanted

    Science.gov (United States)

    Clark, Todd; Jones, Rick

    2004-01-01

    While the political climate seems favorable for the development of fuel-cell vehicles for personal transportation, the market's demand may not be so favorable. Nonetheless, middle level students will be the next generation of drivers and voters, and they need to be able to make informed decisions regarding the nation's energy and transportation…

  11. Drivers and Limits for Transport

    DEFF Research Database (Denmark)

    Kristensen, Niels Buus; Nielsen, Thomas A. Sick; Gudmundsson, Henrik;

    This report summarizes key outcomes of the study ’Drivers and Limits’ that was supported for the period 2009-2013 by a research grant from the Danish Strategic Research Council. The project investigated - for the empirical context of Denmark - key driving forces behind transport growth, as well...

  12. SLEEPINESS AMONG IRANIAN LORRY DRIVERS

    Directory of Open Access Journals (Sweden)

    K. Sadeghniiat Y. Labbafinejad

    2007-06-01

    Full Text Available Excessive daytime sleepiness (EDS denotes a propensity to doze off or fall asleep unintentionally during the day, particularly in passive situations. There is cumulative evidence pointing to an association between sleepiness and probability of involvement in motor vehicle crashes. This study was designed to determine the prevalence of sleepiness in a group of Iranian lorry drivers and its association with accidents. A cross-sectional study was carried out in lorry drivers of Tehran goods transportation terminal in 2005. This study used a questionnaire and the Epworth Sleepiness Scale (ESS. The questionnaire included questions regarding demographic features, professional data, sleep habits and excessive daytime sleepiness. A total of 386 male drivers, aged 43.23 ± 9.72 years were included in the study. ESS was higher than 10 points in 9.1% of the interviewees; 50.8% never have driven drowsy, although 36% rarely, 7.3% half of the times, 4.9% almost always and 1% always have driven drowsy. Logistic regression analysis indicated that EDS, age and job satisfaction were associated with an increased risk of accidents. Sleepiness is a prevailing symptom in lorry drivers and is probably related to accidents.

  13. HIF-1α和AMPK在前列腺癌中的表达及临床意义%Expression and clinical significance of HIF-1α and AMPK in prostate cancer

    Institute of Scientific and Technical Information of China (English)

    江绍钦; 李梦强; 王玉兵; 王辉丽; 许恩赐

    2016-01-01

    目的 探讨缺氧诱导因子1α(HIF-1α)及腺苷酸活化蛋白激酶(AMPK)在前列腺癌中的表达及临床意义.方法 收集我院2011年3月至2013年5月手术切除的前列腺癌标本34例,采用免疫组化SABC法染色观察癌组织中HIF-1α、AMPK的表达情况.结果 前列腺癌组织免疫组化显示:年龄分组中,HIF-1α在高龄组阳性表达率为77.27%,显著高于低年龄组的41.67%(P<0.05).Gleason评分中,高分组HIF-1α的阳性表达率为74.07%,显著高于低分组的28.57% (P<0.05);高分组AMPK的阳性表达率为22.22%,显著低于低分组的71.43%(P<0.05).在TNM分组中,HIF-1α在高危组阳性表达率为84.21%,显著高于低危组的40.00%(P<0.05).HIF-1α在前列腺癌组织的高表达与AMPK的低表达呈负相关,具有统计学(P<0.05).AMPK的阳性表达率在不同年龄、血清PSA、TNM分组比较中均无显著差异(P>0.05);HIF-1α在血清PSA分组比较无显著差异(P>0.05).结论 HIF-1α和AMPK表达均同前列腺癌的病理及临床预后相关.前列腺癌组织HIF-1α的高表达同AMPK低表达呈负相关,而AMPK有可能是前列腺癌细胞中HIF-1α活化调控的关键组分.

  14. 缺氧诱导因子HIF-1在肿瘤Warburg效应中作用机制的研究进展%The Progress of Hypoxia-Inducible Factor HIF-1 Functions in Glycolysis in Tumor Cells

    Institute of Scientific and Technical Information of China (English)

    周扬梅; 肖嵘; 吴元强; 唐玲; 陈尧磊; 殷俊; 曾庆海

    2012-01-01

    Usually human cells use aerobic phosphorylation as a major way to gain energy. However, in tumor cells, energy that cells need mainly come from glycolysis even in the presence of enough oxygen, this phenomenon is known as the Warburg effect. In cancer cells, HIF-1 is interrelated with glycolysis. By up regulating a series of genes that related with glyeolysis's metabolism, angiogenesis, survival of cancer cells and erythropoiesis-related genes, HIF-1 promote Warburg effect' happen remarkably. During the cancer cell metabolism reprogramming process, pyruvate kinase M2 and HIF-1 form a positive feedback, while FIH suppress HIF-l's activity by inhibiting HIF-1 recruit CBP/p300, an important transcript factor.%正常状态下人体细胞的能量主要来源于有氧磷酸化,而在肿瘤细胞,其能量主要来源于糖酵解,即使在含有充足氧气的环境中肿瘤细胞依然进行糖酵解,这种现象被称为Warburg效应.在肿瘤细胞中,缺氧诱导因子HIF-1水平的升高与糖酵解活动的增强密切相关,HIF-1上调一系列与糖酵解能量代谢、血管新生、肿瘤细胞存活和红细胞生成相关的基因,从而促进了肿瘤细胞Warburg效应的发生.在肿瘤细胞代谢重编程过程中,丙酮酸激酶M2(PKM2)与HIF-1之间构成一个正反馈过程,而缺氧诱导因子抑制因子1 (FIH-1)能通过抑制HIF-1对重要基因转录因子CPB/p300的招募,来抑制HIF-1的活性.

  15. 部分胃癌PET-CT显像与HIF-1α等指标的相关分析%Analysis of relationship among HIF-1 α, COX-2, VEGF and SUVmax in gastric cancer

    Institute of Scientific and Technical Information of China (English)

    林夏雯; 李爱梅; 施鸣; 许守林; 郭万华

    2012-01-01

    Objective To study the relationship among the expression of hypoxia inducible factor-la (HIF-la) ,cyclooxygenase-2 (COX-2) ,vascular endothelial growth factor (VEGF) and 18F-fluorodeoxyglucose (' F-FDG) uptake of positron emission tomography ( PET-CT) imaging in patients with gastric cancer. Methods From August 2010 to August 2011 ,14 patients with gastric cancer were examined by PET-CT. The maximum standard uptake values ( SUVmax) were measured. Resected tumor samples were pathologically examined. The expression of HIF-la, COX-2 and VEGF of 14 cases were graded semi-quantitatively by immunohistochem-istry. Kendall correlation was analysed for HIF-la,COX-2,VEGF with SUVmax. Results The correlation coefficient (r) for the SUVmax and HIF-la,COX-2,VEGF expression were 0. 499,0. 686,0. 279,were all positively related (P <0.05). Conclusion 18F-FDG uptake correlates with expression of HIF-la,COX-2 and VEGF in patients with gastric cancer.%目的 探讨胃癌正电子发射体层显像(PET-CT)中18F-氟代脱氧葡萄糖(18F-FDG)摄取水平与相应组织中缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)、环氧合酶-2(cyclooxygenase-2,COX-2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的关系.方法 对14例胃癌患者术前进行PET-CT检查,测定肿瘤最大标准摄取值(standard uptake value,SUVmax),并对手术切除标本进行病理检查,应用免疫组织化学法检测肿瘤组织HIF-1α、COX-2和VEGF的表达,分析PET-CT的SUVmax与HIF-1α、COX-2和VEGF表达的相关性.结果 SUVmax与HIF-1α、COX-2和VEGF表达的相关系数(r)分别为0.499、0.686、0.279,均呈正相关(P<0.05).结论 胃癌组织HIF-1α、COX-2和VEGF表达量越大,PET-CT显像中18F-FDG摄取越多,浓聚越明显.

  16. XBP1 promotes triple-negative breast cancer by controlling the HIF1α pathway.

    Science.gov (United States)

    Chen, Xi; Iliopoulos, Dimitrios; Zhang, Qing; Tang, Qianzi; Greenblatt, Matthew B; Hatziapostolou, Maria; Lim, Elgene; Tam, Wai Leong; Ni, Min; Chen, Yiwen; Mai, Junhua; Shen, Haifa; Hu, Dorothy Z; Adoro, Stanley; Hu, Bella; Song, Minkyung; Tan, Chen; Landis, Melissa D; Ferrari, Mauro; Shin, Sandra J; Brown, Myles; Chang, Jenny C; Liu, X Shirley; Glimcher, Laurie H

    2014-04-01

    Cancer cells induce a set of adaptive response pathways to survive in the face of stressors due to inadequate vascularization. One such adaptive pathway is the unfolded protein (UPR) or endoplasmic reticulum (ER) stress response mediated in part by the ER-localized transmembrane sensor IRE1 (ref. 2) and its substrate XBP1 (ref. 3). Previous studies report UPR activation in various human tumours, but the role of XBP1 in cancer progression in mammary epithelial cells is largely unknown. Triple-negative breast cancer (TNBC)--a form of breast cancer in which tumour cells do not express the genes for oestrogen receptor, progesterone receptor and HER2 (also called ERBB2 or NEU)--is a highly aggressive malignancy with limited treatment options. Here we report that XBP1 is activated in TNBC and has a pivotal role in the tumorigenicity and progression of this human breast cancer subtype. In breast cancer cell line models, depletion of XBP1 inhibited tumour growth and tumour relapse and reduced the CD44(high)CD24(low) population. Hypoxia-inducing factor 1α (HIF1α) is known to be hyperactivated in TNBCs. Genome-wide mapping of the XBP1 transcriptional regulatory network revealed that XBP1 drives TNBC tumorigenicity by assembling a transcriptional complex with HIF1α that regulates the expression of HIF1α targets via the recruitment of RNA polymerase II. Analysis of independent cohorts of patients with TNBC revealed a specific XBP1 gene expression signature that was highly correlated with HIF1α and hypoxia-driven signatures and that strongly associated with poor prognosis. Our findings reveal a key function for the XBP1 branch of the UPR in TNBC and indicate that targeting this pathway may offer alternative treatment strategies for this aggressive subtype of breast cancer.

  17. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia

    Science.gov (United States)

    Holden, Victoria I.; Breen, Paul; Houle, Sébastien; Dozois, Charles M.

    2016-01-01

    ABSTRACT Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α) protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6), CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia. PMID:27624128

  18. Intestinal HIF2α promotes tissue-iron accumulation in disorders of iron overload with anemia.

    Science.gov (United States)

    Anderson, Erik R; Taylor, Matthew; Xue, Xiang; Ramakrishnan, Sadeesh K; Martin, Angelical; Xie, Liwei; Bredell, Bryce X; Gardenghi, Sara; Rivella, Stefano; Shah, Yatrik M

    2013-12-10

    Several distinct congenital disorders can lead to tissue-iron overload with anemia. Repeated blood transfusions are one of the major causes of iron overload in several of these disorders, including β-thalassemia major, which is characterized by a defective β-globin gene. In this state, hyperabsorption of iron is also observed and can significantly contribute to iron overload. In β-thalassemia intermedia, which does not require blood transfusion for survival, hyperabsorption of iron is the leading cause of iron overload. The mechanism of increased iron absorption in β-thalassemia is unclear. We definitively demonstrate, using genetic mouse models, that intestinal hypoxia-inducible factor-2α (HIF2α) and divalent metal transporter-1 (DMT1) are activated early in the pathogenesis of β-thalassemia and are essential for excess iron accumulation in mouse models of β-thalassemia. Moreover, thalassemic mice with established iron overload had significant improvement in tissue-iron levels and anemia following disruption of intestinal HIF2α. In addition to repeated blood transfusions and increased iron absorption, chronic hemolysis is the major cause of tissue-iron accumulation in anemic iron-overload disorders caused by hemolytic anemia. Mechanistic studies in a hemolytic anemia mouse model demonstrated that loss of intestinal HIF2α/DMT1 signaling led to decreased tissue-iron accumulation in the liver without worsening the anemia. These data demonstrate that dysregulation of intestinal hypoxia and HIF2α signaling is critical for progressive iron overload in β-thalassemia and may be a novel therapeutic target in several anemic iron-overload disorders.

  19. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1α Stabilization during Pneumonia

    Directory of Open Access Journals (Sweden)

    Victoria I. Holden

    2016-09-01

    Full Text Available Klebsiella pneumoniae is a Gram-negative pathogen responsible for a wide range of infections, including pneumonia and bacteremia, and is rapidly acquiring antibiotic resistance. K. pneumoniae requires secretion of siderophores, low-molecular-weight, high-affinity iron chelators, for bacterial replication and full virulence. The specific combination of siderophores secreted by K. pneumoniae during infection can impact tissue localization, systemic dissemination, and host survival. However, the effect of these potent iron chelators on the host during infection is unknown. In vitro, siderophores deplete epithelial cell iron, induce cytokine secretion, and activate the master transcription factor hypoxia inducible factor-1α (HIF-1α protein that controls vascular permeability and inflammatory gene expression. Therefore, we hypothesized that siderophore secretion by K. pneumoniae directly contributes to inflammation and bacterial dissemination during pneumonia. To examine the effects of siderophore secretion independently of bacterial growth, we performed infections with tonB mutants that persist in vivo but are deficient in siderophore import. Using a murine model of pneumonia, we found that siderophore secretion by K. pneumoniae induces the secretion of interleukin-6 (IL-6, CXCL1, and CXCL2, as well as bacterial dissemination to the spleen, compared to siderophore-negative mutants at an equivalent bacterial number. Furthermore, we determined that siderophore-secreting K. pneumoniae stabilized HIF-1α in vivo and that bacterial dissemination to the spleen required alveolar epithelial HIF-1α. Our results indicate that siderophores act directly on the host to induce inflammatory cytokines and bacterial dissemination and that HIF-1α is a susceptibility factor for bacterial invasion during pneumonia.

  20. The association between let-7, RAS and HIF-1α in Ewing Sarcoma tumor growth

    Science.gov (United States)

    Yaniv, Isaac; Ash, Shifra; Cohen, Ian J.; Kodman, Yona; Haklai, Ronit; Elad-Sfadia, Galit; Kloog, Yoel; Chepurko, Elena; Feinmesser, Meora; Issakov, Josephine; Sher, Osnat; Luria, Drorit; Kollender, Yehuda; Weizman, Avraham; Avigad, Smadar

    2015-01-01

    Ewing Sarcoma (ES) is the second most common primary malignant bone tumor in children and adolescents. microRNAs (miRNAs) are involved in cancer as tumor suppressors or oncogenes. We studied the involvement of miRNAs located on chromosomes 11q and 22q that participate in the most common translocation in ES. Of these, we focused on 3 that belong to the let-7 family. We studied the expression levels of let-7a, and let-7b and detected a significant correlation between low expression of let-7b and increased risk of relapse. let-7 is known to be a negative regulator of the RAS oncogene. Indeed, we detected an inverse association between the expression of let-7 and RAS protein levels and its downstream target p-ERK, following transfection of let-7 mimics and inhibitors. Furthermore, we identified let-7 as a negative regulator of HIF-1α and EWS-FLI-1. Moreover, we were able to show that HIF-1α directly binds to the EWS-FLI-1 promoter. Salirasib treatment in-vitro resulted in the reduction of cell viability, migration ability, and in the decrease of cells in S-phase. A significant reduction in tumor burden and in the expression levels of both HIF-1α and EWS-FLI-1 proteins were observed in mice after treatment. Our results support the hypothesis that let-7 is a tumor suppressor that negatively regulates RAS, also in ES, and that HIF-1α may contribute to the aggressive metastatic behavior of ES. Moreover, the reduction in the tumor burden in a mouse model of ES following Salirasib treatment, suggests therapeutic potential for this RAS inhibitor in ES. PMID:26393682

  1. Fumaric acid esters stimulate astrocytic VEGF expression through HIF-1α and Nrf2.

    Directory of Open Access Journals (Sweden)

    Diana Wiesner

    Full Text Available Fumaric acid esters (FAE are oral analogs of fumarate that have recently been shown to decrease relapse rate and disease progression in multiple sclerosis (MS, prompting to investigate their protective potential in other neurological diseases such as amyotrophic lateral sclerosis (ALS. Despite efficacy in MS, mechanisms of action of FAEs are still largely unknown. FAEs are known to activate the transcription factor Nrf2 and downstream anti-oxidant responses through the succination of Nrf2 inhibitor KEAP1. However, fumarate is also a known inhibitor of prolyl-hydroxylases domain enzymes (PhD, and PhD inhibition might lead to stabilization of the HIF-1α transcription factor under normoxic conditions and subsequent activation of a pseudo hypoxic response. Whether Nrf2 activation is associated with HIF-1α stabilization in response to FAEs in cell types relevant to MS or ALS remains unknown. Here, we show that FAEs elicit HIF-1α accumulation, and VEGF release as its expected consequence, in astrocytes but not in other cell types of the central nervous system. Reporter assays demonstrated that increased astrocytic VEGF release in response to FAEs was dependent upon both HIF-1α and Nrf2 activation. Last, astrocytes of transgenic mice expressing SOD1(G93A, an animal model of ALS, displayed reduced VEGF release in response to FAEs. These studies show that FAEs elicit different signaling pathways in cell types from the central nervous system, in particular a pseudo-hypoxic response in astrocytes. Disease relevant mutations might affect this response.

  2. Stabilization of HIF-1α and HIF-2α, up-regulation of MYCC and accumulation of stabilized p53 constitute hallmarks of CNS-PNET animal model

    Science.gov (United States)

    Malchenko, Sergey; Sredni, Simone Treiger; Bi, Yingtao; Margaryan, Naira V.; Boyineni, Jerusha; Mohanam, Indra; Tomita, Tadanori; Davuluri, Ramana V.; Soares, Marcelo B.

    2017-01-01

    Recently, we described a new animal model of CNS primitive neuroectodermal tumors (CNS-PNET), which was generated by orthotopic transplantation of human Radial Glial (RG) cells into NOD-SCID mice’s brain sub-ventricular zone. In the current study we conducted comprehensive RNA-Seq analyses to gain insights on the mechanisms underlying tumorigenesis in this mouse model of CNS-PNET. Here we show that the RNA-Seq profiles derived from these tumors cluster with those reported for patients’ PNETs. Moreover, we found that (i) stabilization of HIF-1α and HIF-2α, which are involved in mediation of the hypoxic responses in the majority of cell types, (ii) up-regulation of MYCC, a key onco-protein whose dysregulation occurs in ~70% of human tumors, and (iii) accumulation of stabilized p53, which is commonly altered in human cancers, constitute hallmarks of our tumor model, and might represent the basis for CNS-PNET tumorigenesis in this model. We discuss the possibility that these three events might be interconnected. These results indicate that our model may prove invaluable to uncover the molecular events leading to MYCC and TP53 alterations, which would be of broader interest considering their relevance to many human malignancies. Lastly, this mouse model might prove useful for drug screening targeting MYCC and related members of its protein interaction network. PMID:28249000

  3. Driver ASICs for Advanced Deformable Mirrors Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The overall goal of the SBIR program is to develop a new Application Specified Integrated Circuit (ASIC) driver to be used in driver electronics of a deformable...

  4. Anthropogenic Drivers of Ecosystem Change: an Overview

    Directory of Open Access Journals (Sweden)

    Gerald C. Nelson

    2006-12-01

    Full Text Available This paper provides an overview of what the Millennium Ecosystem Assessment (MA calls "indirect and direct drivers" of change in ecosystem services at a global level. The MA definition of a driver is any natural or human-induced factor that directly or indirectly causes a change in an ecosystem. A direct driver unequivocally influences ecosystem processes. An indirect driver operates more diffusely by altering one or more direct drivers. Global driving forces are categorized as demographic, economic, sociopolitical, cultural and religious, scientific and technological, and physical and biological. Drivers in all categories other than physical and biological are considered indirect. Important direct drivers include changes in climate, plant nutrient use, land conversion, and diseases and invasive species. This paper does not discuss natural drivers such as climate variability, extreme weather events, or volcanic eruptions.

  5. Inhibition of HIF-1{alpha} activity by BP-1 ameliorates adjuvant induced arthritis in rats

    Energy Technology Data Exchange (ETDEWEB)

    Shankar, J. [Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago (United States); Thippegowda, P.B., E-mail: btprabha@uic.edu [Department of Pharmacology, (M/C 868), College of Medicine, University of Illinois at Chicago, 835 S. Wolcott Ave., Chicago, IL 60612 (United States); Kanum, S.A. [Department of Chemistry, Yuvaraj' s College, University of Mysore, Mysore (India)

    2009-09-18

    Rheumatoid arthritis (RA) is a chronic inflammatory, angiogenic disease. Inflamed synovitis is a hallmark of RA which is hypoxic in nature. Vascular endothelial growth factor (VEGF), one of the key regulators of angiogenesis, is overexpressed in the pathogenesis of RA. VEGF expression is regulated by hypoxia-inducible factor-1{alpha} (HIF-1{alpha}), a master regulator of homeostasis which plays a pivotal role in hypoxia-induced angiogenesis. In this study we show that synthetic benzophenone analogue, 2-benzoyl-phenoxy acetamide (BP-1) can act as a novel anti-arthritic agent in an experimental adjuvant induced arthritis (AIA) rat model by targeting VEGF and HIF-1{alpha}. BP-1 administered hypoxic endothelial cells and arthritic animals clearly showed down regulation of VEGF expression. Further, BP-1 inhibits nuclear translocation of HIF-1{alpha}, which in turn suppresses transcription of the VEGF gene. These results suggest a further possible clinical application of the BP-1 derivative as an anti-arthritic agent in association with conventional chemotherapeutic agents.

  6. Lipid-Albumin Nanoparticles (LAN) for Therapeutic Delivery of Antisense Oligonucleotide against HIF-1α.

    Science.gov (United States)

    Li, Hong; Quan, Jishan; Zhang, Mengzi; Yung, Bryant C; Cheng, Xinwei; Liu, Yang; Lee, Young B; Ahn, Chang-Ho; Kim, Deog Joong; Lee, Robert J

    2016-07-01

    Lipid-albumin nanoparticles (LAN) were synthesized for delivery of RX-0047, an antisense oligonucleotide (ASO) against the hypoxia inducible factor-1 alpha (HIF-1α) to solid tumor. These lipid nanoparticles (LNs) incorporated a human serum albumin-pentaethylenehexamine (HSA-PEHA) conjugate, which is cationic and can form electrostatic complexes with negatively charged oligonucleotides. The delivery efficiency of LAN-RX-0047 was investigated in KB cells and a KB murine xenograft model. When KB cells were treated with LAN-RX-0047, significant HIF-1α downregulation and enhanced cellular uptake were observed compared to LN-RX-0047. LN-RX-0047 and LAN-RX-0047 showed similar cytotoxicity against KB cells with IC50 values of 19.3 ± 3.8 and 20.1 ± 4.2 μM, respectively. LAN-RX-0047 was shown to be taken up by the cells via the macropinocytosis and caveolae-mediated endocytosis pathways while LN-RX-0047 was taken up by cells via caveolae-mediated endocytosis. In the KB xenograft tumor model, LAN-RX-0047 exhibited tumor suppressive activity and significantly reduced intratumoral HIF-1α expression compared to LN-RX-0047. Furthermore, LAN-RX-0047 greatly increased survival time of mice bearing KB-1 xenograft tumors at doses of either 3 mg/kg or 16 mg/kg. These results indicated that LAN-RX-0047 is a highly effective vehicle for therapeutic delivery of antisense agents to tumor.

  7. Cell autonomy of HIF effects in Drosophila: tracheal cells sense hypoxia and induce terminal branch sprouting.

    Science.gov (United States)

    Centanin, Lázaro; Dekanty, Andrés; Romero, Nuria; Irisarri, Maximiliano; Gorr, Thomas A; Wappner, Pablo

    2008-04-01

    Drosophila tracheal terminal branches are plastic and have the capacity to sprout out projections toward oxygen-starved areas, in a process analogous to mammalian angiogenesis. This response involves the upregulation of FGF/Branchless in hypoxic tissues, which binds its receptor Breathless on tracheal cells. Here, we show that extra sprouting depends on the Hypoxia-Inducible Factor (HIF)-alpha homolog Sima and on the HIF-prolyl hydroxylase Fatiga that operates as an oxygen sensor. In mild hypoxia, Sima accumulates in tracheal cells, where it induces breathless, and this induction is sufficient to provoke tracheal extra sprouting. In nontracheal cells, Sima contributes to branchless induction, whereas overexpression of Sima fails to attract terminal branch outgrowth, suggesting that HIF-independent components are also required for full induction of the ligand. We propose that the autonomous response to hypoxia that occurs in tracheal cells enhances tracheal sensitivity to increasing Branchless levels, and that this mechanism is a cardinal step in hypoxia-dependent tracheal sprouting.

  8. Lights on for HIF-1α: genetically enhanced mouse cardiomyocytes for heart tissue imaging.

    Science.gov (United States)

    Hesse, Amke R; Levent, Elif; Zieseniss, Anke; Tiburcy, Malte; Zimmermann, Wolfram H; Katschinski, Dörthe M

    2014-01-01

    The hypoxia inducible factor-1 (HIF-1) is a suitable marker for tissue oxygenation. We intended to develop cardiomyocytes (CMs) expressing the oxygen-dependent degradation domain of HIF-1α fused to the firefly luciferase (ODD-Luc) followed by proof-of-concept for its applicability in the assessment of heart muscle oxygenation. We first generated embryonic stem cell (ESC) lines (ODD-Luc ESCs) from a Tg ROSA26 ODD-Luc/+ mouse. Subsequent CMs selection was facilitated by stable integration of an antibiotic resistance expressed under the control of the αMHC promoter. ODD-Luc ESCs showed a strong Luc-signal within 1 h of hypoxia (1% oxygen), which coincided with endogenous HIF-1α. Engineered heart muscle (EHM) constructed with ODD-Luc CMs confirmed the utility of the model to sense hypoxia, and monitor reoxygenation also in a multicellular heart muscle model. Pharmacologically induced inotropy/chronotropy under isoprenaline resulted in enhanced Luc-signal suggesting enhanced oxygen consumption, leading to notable myocardial hypoxia. ODD-Luc-CMs can be used to monitor dynamic changes of cardiomyocyte oxygenation in living heart muscle samples. We provide proof-of-concept for pharmacologically induced myocardial interventions and envision applications of the developed model in drug screens and fundamental studies of ischemia/reperfusion injury. © 2014 S. Karger AG, Basel.

  9. Lights on for HIF-1α: Genetically Enhanced Mouse Cardiomyocytes for Heart Tissue Imaging

    Directory of Open Access Journals (Sweden)

    Amke R. Hesse

    2014-07-01

    Full Text Available Background/Aims: The hypoxia inducible factor-1 (HIF-1 is a suitable marker for tissue oxygenation. We intended to develop cardiomyocytes (CMs expressing the oxygen-dependent degradation domain of HIF-1α fused to the firefly luciferase (ODD-Luc followed by proof-of-concept for its applicability in the assessment of heart muscle oxygenation. Methods and Results: We first generated embryonic stem cell (ESC lines (ODD-Luc ESCs from a Tg ROSA26 ODD-Luc/+ mouse. Subsequent CMs selection was facilitated by stable integration of an antibiotic resistance expressed under the control of the αMHC promoter. ODD-Luc ESCs showed a strong Luc-signal within 1 h of hypoxia (1% oxygen, which coincided with endogenous HIF-1α. Engineered heart muscle (EHM constructed with ODD-Luc CMs confirmed the utility of the model to sense hypoxia, and monitor reoxygenation also in a multicellular heart muscle model. Pharmacologically induced inotropy/chronotropy under isoprenaline resulted in enhanced Luc-signal suggesting enhanced oxygen consumption, leading to notable myocardial hypoxia. Conclusions: ODD-Luc-CMs can be used to monitor dynamic changes of cardiomyocyte oxygenation in living heart muscle samples. We provide proof-of-concept for pharmacologically induced myocardial interventions and envision applications of the developed model in drug screens and fundamental studies of ischemia/reperfusion injury.

  10. HIF-1α/MDR1 pathway confers chemoresistance to cisplatin in bladder cancer.

    Science.gov (United States)

    Sun, Yi; Guan, Zhenfeng; Liang, Liang; Cheng, Yongyi; Zhou, Jiancheng; Li, Jing; Xu, Yonggang

    2016-03-01

    Bladder cancer (BCa) is the 9th most common malignant tumor and the 13th leading cause of death due to cancer. The development of surgery and target drugs bring new challenges for the traditional concept for BCa therapy, and chemotherapy is still the final option for many BCa patients, and cisplatin-containing regimen the most effective one. However, the ubiquitous application of cisplatin-containing regimen in BCa results in the cisplatin-resistance, in addition, the cisplatin‑resistant BCa manifests enhanced malignant behavior, the mechanism of which is unclear. In the present study, we used BCa cell lines to to clarify this point. BCa cell lines T24/J82 were pretreated with cisplatin >3 months to construct stable cisplatin-resistant cell lines (tagged T24(Cis-R) and J82(Cis-R)), which manifested as enhanced capacity of proliferation and malignant behavior in vivo and in vitro, accompanied by cisplatin, and even doxorubicin resistance. The following mechanism dissection revealed that prolonged treatment time of T24/J82 cells led to elevated expression of HIF-1α, which targeted the increased expression of MDR1 on the one hand, and contributed to BCa cell proliferation, migration/invasion on the other hand. Finally, IHC staining of human BCa tissue supported our conclusion that the expression of HIF-1α and MDR1 was higher in chemoresistant tissue vs. chemosensitive tissue. Our results provided a new view of HIF-1α in chemotherapy.

  11. The relationships between hypoxia-dependent markers: HIF-1alpha, EPO and EPOR in colorectal cancer.

    Science.gov (United States)

    Baltaziak, Marek; Wincewicz, Andrzej; Kanczuga-Koda, Luiza; Lotowska, Joanna M; Koda, Mariusz; Sulkowska, Urszula; Baltaziak, Marcin; Podbielski, Monika; Sobaniec-Lotowska, Maria E; Sulkowski, Stanislaw

    2013-01-01

    Hypoxia triggers production of several cytoprotective proteins. Hypoxia-inducible factor 1alpha (HIF-1α) is a powerful stimulator of transcription of many genes, including erythropoietin (EPO) in hypoxia-affected cells. Recent data have also implicated signaling by EPO receptor (EPOR) as a new factor influencing tumor progression. The aim of the study was to detect by immunohistochemistry the presence of HIF-1α, EPO and EPOR in colorectal cancer (CRC) in reference to clinicopathological variables. We found the presence of the studied proteins in specimens of all 125 CRC patients which is suggestive of the occurrence of hypoxia in colorectal cancer tissues. The expression of HIF-1α correlated significantly with the presence of EPO and EPOR in all samples (P < 0.001, r = 0.549 and P < 0.001, r = 0.536, respectively). Significant correlations (from P < 0.024 to P < 0.001) were found in the analyses of CRC subgroups such as histopathological type tumor, tumor grade, tumor stage and patients with lymph nodes metastases. The same high significant correlations (P < 0.001) were observed in group of sex, age and tumor location. However, the values of the correlation coefficients (r) which usually ranged from 0.5 to 0.6 suggest the existence of independent or concurrent mechanism stimulating generation of these proteins in colorectal cancer.

  12. HIF1alpha synergizes with glucocorticoids to promote BFU-E progenitor self-renewal.

    Science.gov (United States)

    Flygare, Johan; Rayon Estrada, Violeta; Shin, Chanseok; Gupta, Sumeet; Lodish, Harvey F

    2011-03-24

    With the aim of finding small molecules that stimulate erythropoiesis earlier than erythropoietin and that enhance erythroid colony-forming unit (CFU-E) production, we studied the mechanism by which glucocorticoids increase CFU-E formation. Using erythroid burst-forming unit (BFU-E) and CFU-E progenitors purified by a new technique, we demonstrate that glucocorticoids stimulate the earliest (BFU-E) progenitors to undergo limited self-renewal, which increases formation of CFU-E cells > 20-fold. Interestingly, glucocorticoids induce expression of genes in BFU-E cells that contain promoter regions highly enriched for hypoxia-induced factor 1α (HIF1α) binding sites. This suggests activation of HIF1α may enhance or replace the effect of glucocorticoids on BFU-E self-renewal. Indeed, HIF1α activation by a prolyl hydroxylase inhibitor (PHI) synergizes with glucocorticoids and enhances production of CFU-Es 170-fold. Because PHIs are able to increase erythroblast production at very low concentrations of glucocorticoids, PHI-induced stimulation of BFU-E progenitors thus represents a conceptually new therapeutic window for treating erythropoietin-resistant anemia.

  13. The mitochondrial calcium uniporter regulates breast cancer progression via HIF-1α.

    Science.gov (United States)

    Tosatto, Anna; Sommaggio, Roberta; Kummerow, Carsten; Bentham, Robert B; Blacker, Thomas S; Berecz, Tunde; Duchen, Michael R; Rosato, Antonio; Bogeski, Ivan; Szabadkai, Gyorgy; Rizzuto, Rosario; Mammucari, Cristina

    2016-05-02

    Triple-negative breast cancer (TNBC) represents the most aggressive breast tumor subtype. However, the molecular determinants responsible for the metastatic TNBC phenotype are only partially understood. We here show that expression of the mitochondrial calcium uniporter (MCU), the selective channel responsible for mitochondrial Ca(2+) uptake, correlates with tumor size and lymph node infiltration, suggesting that mitochondrial Ca(2+) uptake might be instrumental for tumor growth and metastatic formation. Accordingly, MCU downregulation hampered cell motility and invasiveness and reduced tumor growth, lymph node infiltration, and lung metastasis in TNBC xenografts. In MCU-silenced cells, production of mitochondrial reactive oxygen species (mROS) is blunted and expression of the hypoxia-inducible factor-1α (HIF-1α) is reduced, suggesting a signaling role for mROS and HIF-1α, downstream of mitochondrial Ca(2+) Finally, in breast cancer mRNA samples, a positive correlation of MCU expression with HIF-1α signaling route is present. Our results indicate that MCU plays a central role in TNBC growth and metastasis formation and suggest that mitochondrial Ca(2+) uptake is a potential novel therapeutic target for clinical intervention.

  14. Expression of HIF-1α in breast cancer and precancerous lesions and the relationship to clinicopathological features

    Institute of Scientific and Technical Information of China (English)

    Yun’ai Liang; Zengxin Li; Gangping Wang

    2014-01-01

    Objective: The aim of this study was to observe the expressions and clinical significance of HIF-1α in breast cancer and precancerous lesions, and analyze the relationship between the expressions and clinicopathological features in breast cancer. Methods: We analyzed the HIF-1α expression in 128 cases of invasive ductal carcinomas, 146 precancerous lesions patients including 89 cases of ductal carcinoma in situ and 57 cases of atypical ductal hyperplasia. 53 cases of usual ductal hyperplasia breast tissues were selected as a control group. The specimens were evaluated for HIF-1α, estrogen re-ceptor (ER) & progesterone receptor (PR), epidermal growth factor receptor type 2 (HER2/neu) and Ki-67. Immunoreactivity was semi-quantitatively evaluated in at least 1000 cells examined under the microscope at 40 × magnification and recorded as the percentage of positive tumor cells over the total number of cells examined in the same area. The percentage scores were subsequently categorized. The express of HIF-1α and their relationship with multiple biological parameters including ER& PR, HER2/neu and Ki-67, the biomarkers levels of CA153, CA125 TSGF, and CEA in blood serum and nipple discharge, histological grade, region lymph node metastasis, distant metastasis and recurrence on files were also assessed. Results:Compared with usual ductal hyperplasia, the positive expression rate of HIF-1α in atypical ductal hyperplasia, ductal carci-noma in situ and invasive ductal carcinomas group was significantly increased (P 14% groups, histological grade (I + II) and grade III invasive ductal carcinomas groups, with lymph node metastasis, distant metastasis and recurrence groups (P50 years), tumor diameter (≤ 2 cm vs > 2 cm; P > 0.05). The nipple discharge and serum levels of CA153, TSGF, CA125 and CEA in invasive ductal carcinomas HIF-1α positive patients were significantly higher than those in the negative patients (P <0.05). Conclusion: In breast cancer, HIF-1α expression

  15. HIF-1α Activation Attenuates IL-6 and TNF-α Pathways in Hippocampus of Rats Following Transient Global Ischemia

    Directory of Open Access Journals (Sweden)

    Jihong Xing

    2016-07-01

    Full Text Available Background/Aims: This study was to examine the role played by hypoxia inducible factor-1 (HIF-1α in regulating pro-inflammatory cytokines (PICs pathway in the rat hippocampus after cardiac arrest (CA induced-transient global ischemia followed by cardiopulmonary resuscitation (CPR. Those PICs include interleukin-1β (IL-1β, interleukin-6 (IL-6 and tumor necrosis factor-α (TNF-α. Methods: A rat model of CA induced by asphyxia was used in the current study. Following CPR, the hippocampus CA1 region was obtained for ELISA to determine the levels of HIF-1α and PICs; and Western Blot analysis to determine the protein levels of PIC receptors. Results: Our data show that IL-1β, IL-6 and TNF-α were significant elevated in the hippocampus after CPR as compared with control group. This was companied with increasing of HIF-1α and the time courses for HIF-1α and PICs were similar. In addition, PIC receptors, namely IL-1R, IL-6R and TNFR1 were upregulated in CA rats. Also, stimulation of HIF-1α by systemic administration of ML228, HIF-1α activator, significantly attenuated the amplified IL-6/IL-6R and TNF-α /TNFR1 pathway in the hippocampus of CA rats, but did not modify IL-1β and its receptor. Moreover, ML228 attenuated upregulated expression of Caspase-3 indicating cell apoptosis evoked by CA. Conclusion: Transient global ischemia induced by CA increases the levels of IL-1β, IL-6 and TNF-α and thereby leads to enhancement in their respective receptor in the rat hippocampus. Stabilization of HIF-1α plays a role in attenuating amplified expression IL-6R, TNFR1 and Caspase-3 in the processing of transient global ischemia. Results of our study suggest that PICs contribute to cerebral injuries evoked by transient global ischemia and in this pathophysiological process activation of HIF-1α improves tissues against ischemic injuries. Our data revealed specific signaling pathways in alleviating CA-evoked global cerebral ischemia by elucidating that

  16. HIF-1 and c-Src mediate increased glucose uptake induced by endothelin-1 and connexin43 in astrocytes.

    Directory of Open Access Journals (Sweden)

    José Carlos Valle-Casuso

    Full Text Available In previous work we showed that endothelin-1 (ET-1 increases the rate of glucose uptake in astrocytes, an important aspect of brain function since glucose taken up by astrocytes is used to supply the neurons with metabolic substrates. In the present work we sought to identify the signalling pathway responsible for this process in primary culture of rat astrocytes. Our results show that ET-1 promoted an increase in the transcription factor hypoxia-inducible factor-1α (HIF-1α in astrocytes, as shown in other cell types. Furthermore, HIF-1α-siRNA experiments revealed that HIF-1α participates in the effects of ET-1 on glucose uptake and on the expression of GLUT-1, GLUT-3, type I and type II hexokinase. We previously reported that these effects of ET-1 are mediated by connexin43 (Cx43, the major gap junction protein in astrocytes. Indeed, our results show that silencing Cx43 increased HIF-1α and reduced the effect of ET-1 on HIF-1α, indicating that the effect of ET-1 on HIF-1α is mediated by Cx43. The activity of oncogenes such as c-Src can up-regulate HIF-1α. Since Cx43 interacts with c-Src, we investigated the participation of c-Src in this pathway. Interestingly, both the treatment with ET-1 and with Cx43-siRNA increased c-Src activity. In addition, when c-Src activity was inhibited neither ET-1 nor silencing Cx43 were able to up-regulate HIF-1α. In conclusion, our results suggest that ET-1 by down-regulating Cx43 activates c-Src, which in turn increases HIF-1α leading to the up-regulation of the machinery required to take up glucose in astrocytes. Cx43 expression can be reduced in response not only to ET-1 but also to various physiological and pathological stimuli. This study contributes to the identification of the signalling pathway evoked after Cx43 down-regulation that results in increased glucose uptake in astrocytes. Interestingly, this is the first evidence linking Cx43 to HIF-1, which is a master regulator of glucose metabolism.

  17. A holistic perspective on corporate sustainability drivers

    NARCIS (Netherlands)

    Lozano, R.

    2013-01-01

    Since company boards are increasingly discussing 'sustainability', it becomes necessary to examine the nature of sustainability drivers. Most approaches to corporate sustainability drivers have focused either on internal or external drivers. This paper is aimed at providing a more holistic perspecti

  18. A holistic perspective on corporate sustainability drivers

    NARCIS (Netherlands)

    Lozano, Rodrigo

    2015-01-01

    Since company boards are increasingly discussing 'sustainability', it becomes necessary to examine the nature of sustainability drivers. Most approaches to corporate sustainability drivers have focused either on internal or external drivers. This paper is aimed at providing a more holistic perspecti

  19. Brief anoxia preconditioning and HIF prolyl-hydroxylase inhibition enhances neuronal resistance in organotypic hippocampal slices on model of ischemic damage.

    Science.gov (United States)

    Lushnikova, Iryna; Orlovsky, Maxim; Dosenko, Victor; Maistrenko, Anastasiia; Skibo, Galina

    2011-04-22

    It is well known that a brief anoxia or hypoxia episodes can render brain resistant to a subsequent ischemia. Recent investigations indicate that mechanisms of such stimulated endogenous neuroprotection are related to the family of hypoxia-inducible factors (HIF), however there are still little data available on the role of HIF family members in hippocampus-a brain structure, highly sensitive to oxygen deficiency. We have used the model of cultured hippocampal slices and single-cell quantitative RT-PCR to study HIF-1α and HIF-3α mRNA expression following triple 5-min mild anoxia, 30-min oxygen-glucose deprivation and their combination. We also tested the effects of HIF prolyl-hydroxylase inhibition with 2,4-pyridinedicarboxylic acid diethyl ester pre-treatment followed by a 30-min oxygen-glucose deprivation. It was found that neuronal damage induced by oxygen-glucose deprivation was accompanied by a significant decrease in both HIF-1α and HIF-3α mRNA levels in CA1 but not CA3 neurons. Anoxia preconditioning did not affect cell viability and HIF mRNA levels but applied before oxygen-glucose deprivation prevented neuronal damage and suppression of HIF-1α and HIF-3α mRNA expression. It was also found that effects of the prolyl-hydroxylase inhibitor were similar to anoxia preconditioning. These results suggest that anoxia preconditioning increases anti-ischemic neuronal resistance which to a certain extent correlates with the changes of HIF-1α and HIF-3α expression.

  20. Interaction of HIF and USF signaling pathways at human genes flanked by hypoxia-response elements and E-box palindromes

    OpenAIRE

    2011-01-01

    Rampant activity of the hypoxia-inducible factor (HIF)-1 in cancer is frequently associated with the malignant progression into a harder-to-treat, increasingly aggressive phenotype. Clearly, anti-HIF strategies in cancer cells are of considerable clinical interest. One way to fine-tune, or inhibit, HIF's transcriptional outflow independently of hydroxylase activities could be through competing transcription factors. A CACGTG-binding activity in human hepatoma cells was previously found to res...

  1. Genome-wide analysis reveals NRP1 as a direct HIF1α-E2F7 target in the regulation of motorneuron guidance in vivo

    NARCIS (Netherlands)

    de Bruin, Alain; Cornelissen, Peter W A; Kirchmaier, Bettina C; Mokry, Michal; Iich, Elhadi; Nirmala, Ella; Liang, Kuo-Hsuan; Végh, Anna M D; Scholman, Koen T; Groot Koerkamp, Marian J; Holstege, Frank C; Cuppen, Edwin; Schulte-Merker, Stefan; Bakker, Walbert J

    2016-01-01

    In this study, we explored the existence of a transcriptional network co-regulated by E2F7 and HIF1α, as we show that expression of E2F7, like HIF1α, is induced in hypoxia, and because of the previously reported ability of E2F7 to interact with HIF1α. Our genome-wide analysis uncovers a transcriptio

  2. VEGF, HIF-1α expression and MVD as an angiogenic network in familial breast cancer.

    Science.gov (United States)

    Saponaro, Concetta; Malfettone, Andrea; Ranieri, Girolamo; Danza, Katia; Simone, Giovanni; Paradiso, Angelo; Mangia, Anita

    2013-01-01

    Angiogenesis, which plays an important role in tumor growth and progression of breast cancer, is regulated by a balance between pro- and anti-angiogenic factors. Expression of vascular endothelial growth factor (VEGF) is up-regulated during hypoxia by hypoxia-inducible factor-1α (HIF-1α). It is known that there is an interaction between HIF-1α and BRCA1 carrier cancers, but little has been reported about angiogenesis in BRCA1-2 carrier and BRCAX breast cancers. In this study, we investigated the expression of VEGF and HIF-1α and microvessel density (MVD) in 26 BRCA1-2 carriers and 58 BRCAX compared to 77 sporadic breast cancers, by immunohistochemistry. VEGF expression in BRCA1-2 carriers was higher than in BRCAX cancer tissues (p = 0.0001). Furthermore, VEGF expression was higher in both BRCA1-2 carriers and BRCAX than the sporadic group (p<0.0001). VEGF immunoreactivity was correlated with poor tumor grade (p = 0.0074), hormone receptors negativity (p = 0.0206, p = 0.0002 respectively), and MIB-1-labeling index (p = 0.0044) in familial cancers (BRCA1-2 and BRCAX). The percentage of nuclear HIF-1α expression was higher in the BRCA1-2 carriers than in BRCAX cancers (p<0.05), and in all familial than in sporadic tumor tissues (p = 0.0045). A higher MVD was observed in BRCA1-2 carrier than in BRCAX and sporadic cancer tissues (p = 0.002, p = 0.0001 respectively), and in all familial tumors than in sporadic tumors (p = 0.01). MVD was positively related to HIF-1α expression in BRCA1-2 carriers (r = 0.521, p = 0.006), and, in particular, we observed a highly significant correlation in the familial group (r = 0.421, p<0.0001). Our findings suggest that angiogenesis plays a crucial role in BRCA1-2 carrier breast cancers. Prospective studies in larger BRCA1-2 carrier series are needed to improve the best therapeutic strategies for this subgroup of breast cancer patients.

  3. The epigenetic regulation of HIF-1α by SIRT1 in MPP{sup +} treated SH-SY5Y cells

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Su-Yan; Guo, Yan-Jie; Feng, Ya; Cui, Xin-Xin [Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080 (China); Kuo, Sheng-Han [Department of Neurology, College of Physicians and Surgeons, Columbia University, New York (United States); Liu, Te, E-mail: liute1979@126.com [Shanghai Geriatric Institute of Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200031 (China); Wu, Yun-Cheng, E-mail: yunchw@medmail.com.cn [Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080 (China)

    2016-02-05

    Both silent information regulator 1 (SIRT1) and hypoxia inducible factor 1 (HIF-1) have been found to play important roles in the pathophysiology of Parkinson's disease (PD). However, their mechanisms and their relationship still require further study. In the present study, we focused on the change and relationship of SIRT1 and HIF-1α in PD. PD cell models were established by using methyl-4-phenylpyridinium (MPP{sup +}), which induced inhibition of cell proliferation, cell cycle arrest and apoptosis. We found that the expression of HIF-1α and its target genes VEGFA and LDHA increased and that SIRT1 expression was inhibited in MPP{sup +} treated cells. With further analysis, we found that the acetylation of H3K14 combined with the HIF-1α promoter was dramatically increased in cells treated with MPP{sup +}, which resulted in the transcriptional activation of HIF-1α. Moreover, the acetylation of H3K14 and the expression of HIF-1α increased when SIRT1 was knocked down, suggesting that SIRT1 was involved in the epigenetic regulation of HIF-1α. At last, phenformin, another mitochondrial complex1 inhibitor, was used to testify that the increased HIF-1a was not due to off target effects of MPP{sup +}. Therefore, our results support a link between PD and SIRT1/HIF-1α signaling, which may serve as a clue for understanding PD.

  4. Molecular-targeted antitumor agents: the Saururus cernuus dineolignans manassantin B and 4-O-demethylmanassantin B are potent inhibitors of hypoxia-activated HIF-1.

    Science.gov (United States)

    Hodges, Tyler W; Hossain, Chowdhury Faiz; Kim, Yong-Pil; Zhou, Yu-Dong; Nagle, Dale G

    2004-05-01

    The transcription factor hypoxia-inducible factor-1 (HIF-1) is a key regulator of tumor cell adaptation and survival under hypoxic conditions. Selective HIF-1 inhibitors represent an important new class of potential molecular-targeted antitumor therapeutic agents. Extracts of plants and marine organisms were evaluated using a T47D human breast tumor cell-based reporter assay for HIF-1 inhibitors. Bioassay-guided fractionation of the lipid extract of Saururus cernuus resulted in the isolation of manassantin B (1) and a new compound, 4-O-demethylmanassantin B (2). The structure of 2 was determined spectroscopically. The absolute configurations of manassantin-type dineolignans have not been previously reported. Therefore, the absolute configurations of the chiral centers in each side chain were deduced from spectroscopic analysis of the Mosher MTPA ester derivatives of 1. Both 1 and 2 are among the most potent small molecule HIF-1 inhibitors discovered, to date, with IC(50) values of 3 and 30 nM, respectively. Compounds 1 and 2 selectively inhibited hypoxia-activated HIF-1 in contrast to iron chelator-activated HIF-1. Compounds 1 and 2 also inhibited hypoxic induction of the angiogenic factor VEGF. Further study revealed that 1 selectively blocked the induction of HIF-1alpha protein, the oxygen regulated HIF-1 subunit that determines HIF-1 activity.

  5. Pneumococcal infection of respiratory cells exposed to welding fumes; Role of oxidative stress and HIF-1 alpha

    Science.gov (United States)

    Grigg, Jonathan; Miyashita, Lisa

    2017-01-01

    Welders are more susceptible to pneumococcal pneumonia. The mechanisms are yet unclear. Pneumococci co-opt the platelet activating factor receptor (PAFR) to infect respiratory epithelial cells. We previously reported that exposure of respiratory cells to welding fumes (WF), upregulates PAFR–dependent pneumococcal infection. The signaling pathway for this response is unknown, however, in intestinal cells, hypoxia-inducible factor-1 α (HIF 1α) is reported to mediate PAFR-dependent infection. We sought to assess whether oxidative stress plays a role in susceptibility to pneumococcal infection via the platelet activating factor receptor. We also sought to evaluate the suitability of nasal epithelial PAFR expression in welders as a biomarker of susceptibility to infection. Finally, we investigated the generalisability of the effect of welding fumes on pneumococcal infection and growth using a variety of different welding fume samples. Nasal epithelial PAFR expression in welders and controls was analysed by flow cytometry. WF were collected using standard methodology. The effect of WF on respiratory cell reactive oxygen species production, HIF-1α expression, and pneumococcal infection was determined using flow cytometry, HIF-1α knockdown and overexpression, and pneumococcal infection assays. We found that nasal PAFR expression is significantly increased in welders compared with controls and that WF significantly increased reactive oxygen species production, HIF-1α and PAFR expression, and pneumococcal infection of respiratory cells. In unstimulated cells, HIF-1α knockdown decreased PAFR expression and HIF-1α overexpression increased PAFR expression. However, in knockdown cells pneumococcal infection was paradoxically increased and in overexpressing cells infection was unaffected. Nasal epithelial PAFR expression may be used as a biomarker of susceptibility to pneumococcal infection in order to target individuals, particularly those at high risk such as welders

  6. The biphasic redox sensing of SENP3 accounts for the HIF-1 transcriptional activity shift by oxidative stress

    Institute of Scientific and Technical Information of China (English)

    Ying WANG; Jie YANG; Kai YANG; Hui CANG; Xin-zhi HUANG; Hui LI; Jing YI

    2012-01-01

    Aim:To investigate the mechanisms underlying the biphasic redox regulation of hypoxia-inducible factor-1 (HIF-1) transcriptional activity under different levels of oxidative stress caused by reactive oxidative species (ROS).Methods:HeLa cells were exposed to different concentrations of H2O2 as a simple model for mild and severe oxidative stress.Luciferase reporter assay and/or quantitative real-time PCR were used to investigate the transcriptional activity.Immunoblot was used to detect protein expression.Chromatin immunoprecipitation assay was used to detect HIF-1/DNA binding.The interaction of p300with HIF-1α or with SENP3,and the SUMO2/3 conjugation states of p300 were examined by coimmunoprecipitation.Results:HIF-1 transcriptional activity in HeLa cells was enhanced by low doses (0.05-0.5 mmol/L) of H202,but suppressed by high doses (0.75-8.0 mmol/L) of H2O2.The amount of co-activator p300 bound to HIF-1α in HeLa cells was increased under mild oxidative stress,but decreased under severe oxidative stress.The ROS levels differentially modified cysteines 243 and 532 in the cysteine protease SENP3,regulating the interaction of SENP3 with p300 to cause different SUMOylation of p300,thus shifting HIF-1 transcriptional activity.Conclusion:The shift of HIF-1 transactivation by ROS is correlated with and dependent on the biphasic redox sensing of SENP3 that leads to the differential SENP3/p300 interaction and the consequent fluctuation in the p300 SUMOylation status.

  7. Propranolol represses infantile hemangioma cell growth through the β2-adrenergic receptor in a HIF-1α-dependent manner.

    Science.gov (United States)

    Li, Peng; Guo, Zhengtuan; Gao, Ya; Pan, Weikang

    2015-06-01

    Propranolol, as a non-selective blocker of the β-adrenergic receptor (AR), is utilised as the first-line treatment for infantile hemangiomas. However, the underlying mechanism remains poorly understood. The present study was designed to investigate the molecular basis of propranolol on the regression of infantile hemangiomas using a proliferating infantile hemangioma-derived endothelial cell line. In infantile hemangioma patients, we found that propranolol significantly decreased the expression levels of the hypoxia inducible factor (HIF)-1α in serum and urine, as well as in hemangioma tissues. In vitro analysis revealed that propranolol reduces the expression of HIF-1α in hemangioma cells in a dose- and time-dependent manner, mainly by acting on β2-AR. Interestingly, it was observed that overexpression of HIF-1α apparently abrogated the inhibitory effects of propranolol on vascular endothelial growth factor (VEGF) expression and cell growth. Our data further demonstrated that propranolol inhibited the signal transducer and activator of transcription 3 (STAT3), a critical oncogenic signaling molecule, and the anti-apoptotic protein Bcl-2. Additionally, overexpression of HIF-1α significantly reversed the inhibitory effects of propranolol on STAT3 signaling. In a mouse xenograft hemangioma model, overexpression of HIF-1α significantly attenuated the therapeutic effects of propranolol and inhibited propranolol-induced hemangioma cell apoptosis. Moreover, the protein levels of VEGF, phosphorylated STAT3, total STAT3 and Bcl-2 were significantly upregulated by HIF-1α overexpression in propranolol-treated nude mice bearing hemangiomas. Collectively, our data provide evidence that propranolol may regress infantile hemangiomas by suppressing VEGF and STAT3 signaling pathways in an HIF-1α-dependent manner.

  8. Cobalt Chloride Upregulates Impaired HIF-1α Expression to Restore Sevoflurane Post-conditioning-Dependent Myocardial Protection in Diabetic Rats

    Science.gov (United States)

    Wu, Jianjiang; Yang, Long; Xie, Peng; Yu, Jin; Yu, Tian; Wang, Haiying; Maimaitili, Yiliyaer; Wang, Jiang; Ma, Haiping; Yang, Yining; Zheng, Hong

    2017-01-01

    Previous studies from our group have demonstrated that sevoflurane post-conditioning (SPC) protects against myocardial ischemia reperfusion injury via elevating the intranuclear expression of hypoxia inducible factor-1 alpha (HIF-1α). However, diabetic SPC is associated with decreased myocardial protection and disruption of the HIF-1 signaling pathway. Previous studies have demonstrated that cobalt chloride (CoCl2) can upregulate HIF-1α expression under diabetic conditions, but whether myocardial protection by SPC can be restored afterward remains unclear. We established a rat model of type 2 diabetes and a Langendorff isolated heart model of ischemia-reperfusion injury. Prior to reperfusion, 2.4% sevoflurane was used as a post-conditioning treatment. The diabetic rats were treated with CoCl2 24 h before the experiment. At the end of reperfusion, tests were performed to assess myocardial function, infarct size, mitochondrial morphology, nitric oxide (NO), Mitochondrial reactive oxygen species (ROS), mitochondrial respiratory function and enzyme activity, HIF-1α, vascular endothelial growth factor (VEGF) and endothelial NO synthase (eNOS) protein levels. In addition, myocardial protection by SPC was monitored after the blood glucose levels were lowered by insulin. The diabetic state was associated with deficient SPC protection and decreased HIF-1α expression. After treating the diabetic rats with CoCl2, SPC significantly upregulated the expression of HIF-1α, VEGF and eNOS, which markedly improved cardiac function, NO, mitochondrial respiratory function, and enzyme activity and decreased the infarction areas and ROS. In addition, these effects were not influenced by blood glucose levels. This study proved that CoCl2activates the HIF-1α signaling pathway, which restores SPC-dependent myocardial protection under diabetic conditions, and the protective effects of SPC were independent of blood glucose levels. PMID:28659817

  9. Cardiac glycosides suppress the maintenance of stemness and malignancy via inhibiting HIF-1α in human glioma stem cells

    Science.gov (United States)

    Lee, Dae-Hee; Oh, Sang Cheul; Giles, Amber J.; Jung, Jinkyu; Gilbert, Mark R.; Park, Deric M.

    2017-01-01

    Tissue hypoxia contributes to solid tumor pathogenesis by activating a series of adaptive programs. We previously showed that hypoxia promotes the preferential expansion and maintenance of CD133 positive human glioma stem cells (GSC) in a hypoxia inducible factor 1 alpha (HIF-1α)-dependent mechanism. Here, we examined the activity of digitoxin (DT), a cardiac glycoside and a putative inhibitor of HIF-1α, on human GSC in vitro and in vivo. During hypoxic conditions (1% O2), we observed the effect of DT on the intracellular level of HIF-1α and the extracellular level of vascular endothelial growth factor (VEGF) in human GSC. We found that DT at clinically achievable concentrations, suppressed HIF-1α accumulation during hypoxic conditions in human GSC and established glioma cell lines. DT treatment also significantly attenuated hypoxia-induced expression of VEGF, a downstream target of HIF-1α. Exposure to DT also reduced hypoxia-induced activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. Furthermore, DT potently inhibited neurosphere formation, and decreased CD133 expression even at concentrations that were not overtly cytotoxic. Lastly, treatment with DT reduced GSC engraftment in an in vivo xenograft model of glioblastoma. Intraperitoneal injections of DT significantly inhibited the growth of established glioblastoma xenografts, and suppressed expression of HIF-1α and carbonic anhydrase (CA9), a surrogate marker of hypoxia. Taken together, these results suggest that DT at clinically achievable concentration functions as an inhibitor of HIF-1α, worthy of further investigations in the therapy of glioblastoma. PMID:28410215

  10. Factors Contributing to Crashes among Young Drivers

    Directory of Open Access Journals (Sweden)

    Lyndel J. Bates

    2014-08-01

    Full Text Available Young drivers are the group of drivers most likely to crash. There are a number of factors that contribute to the high crash risk experienced by these drivers. While some of these factors are intrinsic to the young driver, such as their age, gender or driving skill, others relate to social factors and when and how often they drive. This article reviews the factors that affect the risk of young drivers crashing to enable a fuller understanding of why this risk is so high in order to assist in developing effective countermeasures.

  11. Factors Contributing to Crashes among Young Drivers.

    Science.gov (United States)

    Bates, Lyndel J; Davey, Jeremy; Watson, Barry; King, Mark J; Armstrong, Kerry

    2014-08-01

    Young drivers are the group of drivers most likely to crash. There are a number of factors that contribute to the high crash risk experienced by these drivers. While some of these factors are intrinsic to the young driver, such as their age, gender or driving skill, others relate to social factors and when and how often they drive. This article reviews the factors that affect the risk of young drivers crashing to enable a fuller understanding of why this risk is so high in order to assist in developing effective countermeasures.

  12. Visualization drivers for Geant4

    Energy Technology Data Exchange (ETDEWEB)

    Beretvas, Andy; /Fermilab

    2005-10-01

    This document is on Geant4 visualization tools (drivers), evaluating pros and cons of each option, including recommendations on which tools to support at Fermilab for different applications. Four visualization drivers are evaluated. They are OpenGL, HepRep, DAWN and VRML. They all have good features, OpenGL provides graphic output without an intermediate file. HepRep provides menus to assist the user. DAWN provides high quality plots and even for large files produces output quickly. VRML uses the smallest disk space for intermediate files. Large experiments at Fermilab will want to write their own display. They should proceed to make this display graphics independent. Medium experiment will probably want to use HepRep because of it's menu support. Smaller scale experiments will want to use OpenGL in the spirit of having immediate response, good quality output and keeping things simple.

  13. Food consumption trends and drivers

    OpenAIRE

    Kearney, John

    2010-01-01

    A picture of food consumption (availability) trends and projections to 2050, both globally and for different regions of the world, along with the drivers largely responsible for these observed consumption trends are the subject of this review. Throughout the world, major shifts in dietary patterns are occurring, even in the consumption of basic staples towards more diversified diets. Accompanying these changes in food consumption at a global and regional level have been considerable health co...

  14. Global desertification: Drivers and feedbacks

    Science.gov (United States)

    D'Odorico, Paolo; Bhattachan, Abinash; Davis, Kyle F.; Ravi, Sujith; Runyan, Christiane W.

    2013-01-01

    Desertification is a change in soil properties, vegetation or climate, which results in a persistent loss of ecosystem services that are fundamental to sustaining life. Desertification affects large dryland areas around the world and is a major cause of stress in human societies. Here we review recent research on the drivers, feedbacks, and impacts of desertification. A multidisciplinary approach to understanding the drivers and feedbacks of global desertification is motivated by our increasing need to improve global food production and to sustainably manage ecosystems in the context of climate change. Classic desertification theories look at this process as a transition between stable states in bistable ecosystem dynamics. Climate change (i.e., aridification) and land use dynamics are the major drivers of an ecosystem shift to a “desertified” (or “degraded”) state. This shift is typically sustained by positive feedbacks, which stabilize the system in the new state. Desertification feedbacks may involve land degradation processes (e.g., nutrient loss or salinization), changes in rainfall regime resulting from land-atmosphere interactions (e.g., precipitation recycling, dust emissions), or changes in plant community composition (e.g., shrub encroachment, decrease in vegetation cover). We analyze each of these feedback mechanisms and discuss their possible enhancement by interactions with socio-economic drivers. Large scale effects of desertification include the emigration of “environmental refugees” displaced from degraded areas, climatic changes, and the alteration of global biogeochemical cycles resulting from the emission and long-range transport of fine mineral dust. Recent research has identified some possible early warning signs of desertification, which can be used as indicators of resilience loss and imminent shift to desert-like conditions. We conclude with a brief discussion on some desertification control strategies implemented in different

  15. The drivers of tropical speciation.

    Science.gov (United States)

    Smith, Brian Tilston; McCormack, John E; Cuervo, Andrés M; Hickerson, Michael J; Aleixo, Alexandre; Cadena, Carlos Daniel; Pérez-Emán, Jorge; Burney, Curtis W; Xie, Xiaoou; Harvey, Michael G; Faircloth, Brant C; Glenn, Travis C; Derryberry, Elizabeth P; Prejean, Jesse; Fields, Samantha; Brumfield, Robb T

    2014-11-20

    Since the recognition that allopatric speciation can be induced by large-scale reconfigurations of the landscape that isolate formerly continuous populations, such as the separation of continents by plate tectonics, the uplift of mountains or the formation of large rivers, landscape change has been viewed as a primary driver of biological diversification. This process is referred to in biogeography as vicariance. In the most species-rich region of the world, the Neotropics, the sundering of populations associated with the Andean uplift is ascribed this principal role in speciation. An alternative model posits that rather than being directly linked to landscape change, allopatric speciation is initiated to a greater extent by dispersal events, with the principal drivers of speciation being organism-specific abilities to persist and disperse in the landscape. Landscape change is not a necessity for speciation in this model. Here we show that spatial and temporal patterns of genetic differentiation in Neotropical birds are highly discordant across lineages and are not reconcilable with a model linking speciation solely to landscape change. Instead, the strongest predictors of speciation are the amount of time a lineage has persisted in the landscape and the ability of birds to move through the landscape matrix. These results, augmented by the observation that most species-level diversity originated after episodes of major Andean uplift in the Neogene period, suggest that dispersal and differentiation on a matrix previously shaped by large-scale landscape events was a major driver of avian speciation in lowland Neotropical rainforests.

  16. Driving fatigue in professional drivers: a survey of truck and taxi drivers.

    Science.gov (United States)

    Meng, Fanxing; Li, Shuling; Cao, Lingzhi; Li, Musen; Peng, Qijia; Wang, Chunhui; Zhang, Wei

    2015-01-01

    Fatigue among truck drivers has been studied extensively; however, less is known regarding the fatigue experience of taxi drivers in heavily populated metropolitan areas. This study aimed to compare the differences and similarities between truck and taxi driver fatigue to provide implications for the fatigue management and education of professional drivers. A sample of 274 truck drivers and 286 taxi drivers in Beijing was surveyed via a questionnaire, which included items regarding work characteristics, fatigue experience, accident information, attitude toward fatigue, and methods of counteracting fatigue. Driver fatigue was prevalent among professional drivers, and it was even more serious for taxi drivers. Taxi drivers reported more frequent fatigue experiences and were involved in more accidents. Among the contributing factors to fatigue, prolonged driving time was the most important factor identified by both driver groups. Importantly, the reason for the engagement in prolonged driving was neither due to the lack of awareness concerning the serious outcome of fatigue driving nor because of their poor detection of fatigue. The most probable reason was the optimism bias, as a result of which these professional drivers thought that fatigue was more serious for other drivers than for themselves, and they thought that they were effective in counteracting the effect of fatigue on their driving performance. Moreover, truck drivers tended to employ methods that require stopping to counteract fatigue, whereas taxi drivers preferred methods that were simultaneous with driving. Although both driver groups considered taking a nap as one of the most effective means to address fatigue, this method was not commonly used. Interestingly, these drivers were aware that the methods they frequently used were not the most effective means to counteract fatigue. This study provides knowledge on truck and taxi drivers' characteristics in fatigue experience, fatigue attitude, and

  17. Expression and clinical significance of the HIF-1a/ET-2 signaling pathway during the development and treatment of polycystic ovary syndrome.

    Science.gov (United States)

    Wang, Fan; Zhang, Zhenghong; Wang, Zhaokai; Xiao, Kaizhuan; Wang, Qing; Su, Jingqian; Wang, Zhengchao

    2015-04-01

    Polycystic ovary syndrome (PCOS) is a major health problem in reproductive-aged women worldwide, but the precise pathogenesis of PCOS remains unclear. Our previous study revealed that hypoxia-inducible factor (HIF)-1a mediated endothelin (ET)-2 signaling plays an important role in ovulation in rats. Therefore, the present study used a PCOS rat model to test the hypotheses that HIF-1a signaling is expressed and inhibited in ovaries during PCOS formation and that the HIF-1a/ET-2 signaling pathway is a target of dimethyldiguanide (DMBG) in the clinical treatment of PCOS. First, the development of a PCOS model and the effect of DMBG treatment were examined through ovarian histology and serum hormone levels, which were consistent with previous reports. Second, HIF-1a and ET-2 expression were detected by immunohistochemistry and western blot. The results showed decreased HIF-1a/ET-2 expression in the ovaries of PCOS rats, whereas DMBG treatment reversed the protein decreases and improved the PCOS symptoms. Third, to understand the molecular mechanism, HIF-1a/ET-2 mRNA expression was also examined. Interestingly, HIF-1a mRNA increased in the ovaries of PCOS rats, while ET-2 mRNA decreased, indicating that HIF-1a protein degradation may be involved in POCS development and treatment. Finally, HIF prolyl hydroxylase (PHD) activity was examined to further clarify the contribution of HIF-1a signaling to the development and treatment of PCOS. The results suggested that the inhibition of HIF-1a/ET-2 signaling may be caused by increased PHD activity in PCOS. DMBG-treated PCOS may further activate HIF-1a signaling at least partly through inhibiting PHD activity. Taken together, these results indicate that HIF-1a signaling is inhibited in a PCOS rat model through increasing PHD activity. DMBG treatment improved PCOS by rescuing this pathway, suggesting that HIF-1a signaling plays an important role in the development and treatment of PCOS. This HIF-1a-mediated ET-2 signaling pathway

  18. Kaempferol inhibits angiogenesis and VEGF expression through both HIF dependent and independent pathways in human ovarian cancer cells.

    Science.gov (United States)

    Luo, Haitao; Rankin, Gary O; Liu, Lingzhi; Daddysman, Matthew K; Jiang, Bing-Hua; Chen, Yi Charlie

    2009-01-01

    Ovarian cancer is 1 of the most significant malignancies in the Western world, and the antiangiogenesis strategy has been postulated for prevention and treatment of ovarian cancers. Kaempferol is a natural flavonoid present in many fruits and vegetables. The antiangiogenesis potential of kaempferol and its underlying mechanisms were investigated in two ovarian cancer cell lines, OVCAR-3 and A2780/CP70. Kaempferol mildly inhibits cell viability but significantly reduces VEGF gene expression at mRNA and protein levels in both ovarian cancer cell lines. In chorioallantoic membranes of chicken embryos, kaempferol significantly inhibits OVCAR-3-induced angiogenesis and tumor growth. HIF-1alpha, a regulator of VEGF, is downregulated by kaempferol treatment in both ovarian cancer cell lines. Kaempferol also represses AKT phosphorylation dose dependently at 5 to 20 muM concentrations. ESRRA is a HIF-independent VEGF regulator, and it is also downregulated by kaempferol in a dose-dependent manner. Overall, this study demonstrated that kaempferol is low in cytotoxicity but inhibits angiogenesis and VEGF expression in human ovarian cancer cells through both HIF-dependent (Akt/HIF) and HIF-independent (ESRRA) pathways and deserves further studies for possible application in angio prevention and treatment of ovarian cancers.

  19. The HIF-1α/CXCR4 pathway supports hypoxia-induced metastasis of human osteosarcoma cells.

    Science.gov (United States)

    Guan, Guofeng; Zhang, Yinglong; Lu, Yao; Liu, Lijuan; Shi, Doufei; Wen, Yanhua; Yang, Lianjia; Ma, Qiong; Liu, Tao; Zhu, Xiaodong; Qiu, Xiuchun; Zhou, Yong

    2015-02-01

    HIF-1α mediates hypoxia-induced expression of the chemokine receptor CXCR4 and contributes to metastasis in many different cancers. We have previously shown that hypoxia promotes migration of human osteosarcoma cells by activating the HIF-1α/CXCR4 pathway. Here, immunohistochemical analysis showed that unlike control osteochondroma samples, osteosarcoma specimens were characterized by elevated expression levels of HIF-1α and CXCR4. Moreover, we found that hypoxia-induced invasiveness was more pronounced in high metastatic potential F5M2 osteosarcoma cells than in low metastatic potential F4 cells, and that this induction was sensitive to treatment with the CXCR4 antagonist AMD3100 and the HIF-1α inhibitor KC7F2. Interestingly, hypoxia-induced CXCR4 expression persisted after cultured osteosarcoma cells were returned to normoxic conditions. These observations were confirmed by experiments in a mouse model of osteosarcoma lung metastasis showing that hypoxia stimulation of pulmonary metastasis was greater in F5M2 than in F4 cells, and was sensitive to treatment with AMD3100. Our study provides further evidence of the contributions of hypoxia and the HIF-1α/CXCR4 pathway to the progression of osteosarcoma, and suggests that this axis might be efficiently leveraged in the development of novel osteosarcoma therapeutics. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  20. Association of Dll4/notch and HIF-1a -VEGF signaling in the angiogenesis of missed abortion.

    Directory of Open Access Journals (Sweden)

    Yan Fang

    Full Text Available BACKGROUND: Dll4/Notch and HIF-1a-VEGF have been shown to play an important role during angiogenesis, but there are no data about their roles and association in missed abortion. In this study, we investigated the association of Dll4/Notch and HIF-1a-VEGF signaling in missed abortion. METHODS: Women with missed abortion (n=27 and healthy controls (n=26 were included in the study. Real-time Reverse Transcription-PCR Analyses (RT-PCR was used to analyze the mRNA levels of Dll4/Notch and HIF-1a-VEGF signaling molecules. The protein level for Dll4 was measured by immunohistochemistry. RESULTS: Compared with induced abortion, the expression of VEGF was statistically reduced while the level of VEGFR1 and Notch1 was significantly up-regulated in missed abortion. Though other molecules (VEGFR2 and Dll4 were marginally higher in missed abortion, no statistical difference was observed. The expression of HIF-1a was significantly up-regulated, and close negatively correlated with VEGF in missed abortion. Both in induced abortion and missed abortion, Dll4 was positively correlated with Notch1. CONCLUSIONS: The early pregnancy is in a hypoxic environment, this may encourage the angiogenesis, but severe hypoxic may inhibit the angiogenesis. Aberrant Dll4/Notch and HIF-1a-VEGF signaling may have a role in missed abortion.

  1. ELR510444 inhibits tumor growth and angiogenesis by abrogating HIF activity and disrupting microtubules in renal cell carcinoma.

    Directory of Open Access Journals (Sweden)

    Jennifer S Carew

    Full Text Available BACKGROUND: Hypoxia-inducible factor (HIF is an attractive therapeutic target for renal cell carcinoma (RCC as its high expression due to the loss of von Hippel-Lindau (VHL promotes RCC progression. Considering this, we hypothesized that ELR510444, a novel orally available small molecule inhibitor of HIF activity, would reduce angiogenesis and possess significant activity in RCC. The mechanism of action and therapeutic efficacy of ELR510444 were investigated in in vitro and in vivo models of RCC. PRINCIPAL FINDINGS: ELR510444 decreased HIF-1α and HIF-2α levels, reduced RCC cell viability and clonogenic survival, and induced apoptosis. VHL-deficient RCC cells were more sensitive to ELR510444-mediated apoptosis and restoration of VHL promoted drug resistance. Higher concentrations of ELR51044 promoted apoptosis independently of VHL status, possibly due to the microtubule destabilizing properties of this agent. ELR510444 significantly reduced tumor burden in the 786-O and A498 RCC xenograft models. These effects were associated with increased necrosis and apoptosis and inhibition of angiogenesis. CONCLUSIONS: ELR510444 is a promising new HIF inhibitor that reduced RCC cell viability, induced apoptosis, and diminished tumor burden in RCC xenograft models. ELR510444 also destabilized microtubules suggesting that it possesses vascular disrupting and anti-angiogenic properties. Further investigation of ELR510444 for the therapy of RCC is warranted.

  2. The role of hypoxia and HIF1α in the regulation of STAR-mediated steroidogenesis in granulosa cells.

    Science.gov (United States)

    Kowalewski, Mariusz Pawel; Gram, Aykut; Boos, Alois

    2015-02-05

    The adaptive responses to hypoxia are mediated by hypoxia-inducible factor 1 alpha (HIF1α). Its role, however, in regulating steroidogenesis remains poorly understood. We examined the role of hypoxia and HIF1α in regulating steroid acute regulatory protein (STAR) expression and steroidogenesis in immortalized (KK1) mouse granulosa cells under progressively lowering O2 concentrations (20%, 15%, 10%, 5%, 1%). Basal and dbcAMP-stimulated progesterone synthesis was decreased under severe hypoxia (1% and 5% O2). The partial hypoxia revealed opposing effects, with a significant increase in steroidogenic response at 10% O2 in dbcAMP-treated cells: Star-promoter activity, mRNA and protein expression were increased. The hypoxia-stimulated STAR expression was PKA-dependent. Binding of HIF1α to the Star-promoter was potentiated under partial hypoxia. Inhibition of the transcriptional activity or expression of HIF1α suppressed STAR-expression. HIF1α appears to be a positive regulator of basal and stimulated STAR-expression, which under partial hypoxia is capable of increasing the steroidogenic capacity of granulosa cells.

  3. CD133 Modulate HIF-1α Expression under Hypoxia in EMT Phenotype Pancreatic Cancer Stem-Like Cells

    Directory of Open Access Journals (Sweden)

    Koki Maeda

    2016-06-01

    Full Text Available Although CD133 is a known representative cancer stem cell marker, its function in tumor aggressiveness under hypoxia is not fully known. The aim of this study is to demonstrate that CD133 regulates hypoxia inducible factor (HIF-1α expression with tumor migration. The CD133+ pancreatic cancer cell line, Capan1M9, was compared with the CD133− cell line, shCD133M9, under hypoxia. HIF-1α expression levels were compared by Western blot, HIF-1α nucleus translocation assay and real-time (RT-PCR. The hypoxia responsive element (HRE was observed by luciferase assay. The migration ability was analyzed by migration and wound healing assays. Epithelial mesenchymal transition (EMT related genes were analyzed by real-time RT-PCR. HIF-1α was highly expressed in Capan1M9 compared to shCD133M9 under hypoxia because of the high activation of HRE. Furthermore, the migration ability of Capan1M9 was higher than that of shCD133M9 under hypoxia, suggesting higher expression of EMT related genes in Capan1M9 compared to shCD133M9. Conclusion: HIF-1α expression under hypoxia in CD133+ pancreatic cancer cells correlated with tumor cell migration through EMT gene expression. Understanding the function of CD133 in cancer aggressiveness provides a novel therapeutic approach to eradicate pancreatic cancer stem cells.

  4. HIF-1α inhibition by siRNA or chetomin in human malignant glioma cells: effects on hypoxic radioresistance and monitoring via CA9 expression

    Directory of Open Access Journals (Sweden)

    Bache Matthias

    2010-11-01

    Full Text Available Abstract Background Hypoxia induces activation of the HIF-1 pathway and is an essential characteristic of malignant gliomas. Hypoxia has been linked to tumor progression, therapy resistance and poor prognosis. However, little is known about the impact of HIF-1α inhibition on radioresistance of malignant glioma. Methods In this study, we investigated the effects of the inhibition of HIF-1α on cell survival and radiosensitivity in U251MG and U343MG glioma cells, using two different strategies. HIF-1α inhibition was achieved by siRNA targeting of HIF-1α or via chetomin, a disruptor of interactions between HIF-1α and p300. The inhibition of the HIF-1 pathway was monitored by quantitative real-time PCR and Western blot analyses of the expression levels of HIF-1α and CA9. CA9 expression was investigated as a potential indicator of the efficacy of HIF-1 inhibition and the resulting radiosensitivity of malignant glioma cell lines was determined by clonogenic assay after irradiation under normoxic (2-10 Gy or hypoxic (2-15 Gy conditions. Results Although siRNA and chetomin show distinct modes of action, both attenuated the hypoxia-induced radioresistance of malignant glioma cell lines U251MG (DMF10: 1.35 and 1.18 and U343MG (DMF10: 1.78 and 1.48. However, siRNA and chetomin showed diverse effects on radiosensitivity under normoxic conditions in U251MG (DMF10: 0.86 and 1.35 and U343MG (DMF10: 1.33 and 1.02 cells. Conclusions Results from this in vitro study suggest that inhibition of HIF-1α is a promising strategy to sensitize human malignant gliomas to radiotherapy and that CA9 could serve as an indicator of effective HIF-1-related radiosensitization.

  5. The acquired radioresistance in HeLa cells under conditions mimicking hypoxia was attenuated by a decreased expression of HIF subunit genes induced by RNA interference

    Energy Technology Data Exchange (ETDEWEB)

    Doi, Nobutaka [Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194 (Japan); New Products Research & Development, Gene Engineering Division, NIPPON GENE Co., Ltd. (Japan); Ogawa, Ryohei, E-mail: ogawa@med.u-toyama.ac.jp [Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194 (Japan); Cui, Zheng-Guo [Department of Public Health, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama (Japan); Morii, Akihiro; Watanabe, Akihiko [Department of Urology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama (Japan); Kanayama, Shinji; Yoneda, Yuko [New Products Research & Development, Gene Engineering Division, NIPPON GENE Co., Ltd. (Japan); Kondo, Takashi [Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194 (Japan)

    2015-05-01

    The cancer cells residing in the hypoxic layer are resistant to radiation and these are ones responsible for cancer recurrence after radiation therapy. One of the reasons why hypoxic cancer cells acquire radioresistance may be attributable to changes in the gene expression profile by the activation of hypoxia inducible factors (HIFs). However, the details underlying this process remain unknown. In this study, we investigated the effects of knockdown of HIF subunit genes to elucidate how HIF subunit genes may be involved in the radioresistance acquired by HeLa cells following exposure to a hypoxia mimic. Interestingly, HIF-1α and HIF-2α seemed mutually complementary for each other when either of them was suppressed. We thus suppressed the expression of both genes simultaneously. To do this, we developed a short hairpin RNA (shRNA) targeting a high homology region between HIF-1α and HIF-2α. It was shown that the expression of the shRNA effectively suppressed the acquisition of radioresistance following the hypoxia mimic. Moreover, it was confirmed that suppression of both subunits resulted in the downregulation of stem cell markers and the suppression of spheroid formation during the hypoxia mimicking-conditions. This shRNA-mediated knockdown method targeting a common region shared by a family of genes may offer a new candidate cancer treatment. - Highlights: • Incubation with CoCl{sub 2} confers radioresistance to HeLa cells. • Both HIF-1α and HIF-2α are involved in the acquisition of radioresistance. • An shRNA to a homology region of HIF-1α and HIF-2α suppressed the radioresistance. • The shRNA decreased cells with stem cell markers and a stem cell phenotype.

  6. Grill kak stil zhizni / Hilda Vall

    Index Scriptorium Estoniae

    Vall, Hilda

    2007-01-01

    Eesti grillvorstide tootmise turuliider ettevõte Wõro on sel aastal müünud juba üle 300 tonni suvetooteid. Jaanipäevaga seoses prognoositakse müügiarvude suurenemist kahekordseks. Rakvere Lihakombinaat plaanib suurendada grilltoodete müüki 26 %

  7. Grill kak stil zhizni / Hilda Vall

    Index Scriptorium Estoniae

    Vall, Hilda

    2007-01-01

    Eesti grillvorstide tootmise turuliider ettevõte Wõro on sel aastal müünud juba üle 300 tonni suvetooteid. Jaanipäevaga seoses prognoositakse müügiarvude suurenemist kahekordseks. Rakvere Lihakombinaat plaanib suurendada grilltoodete müüki 26 %

  8. HIF-1α和VEGF-C在前列腺癌中的表达及意义%Expression and Clinical Significance of HIF-1α and VEGF-C in Prostate Cancer

    Institute of Scientific and Technical Information of China (English)

    王晋龙; 白铁男; 胡海龙; 韩瑞发; 赵朋

    2012-01-01

    Objective: To study the expression and clinical significance of hypoxia- inducible factor la (HIF-1α) and vascular endothelial growth factor C (VEGF-C) in the prostate carcinoma (PC). Methods: The expression levels of HIF-1α and VEGF-C were detected by SABC immunohistochemical analysis in 59 cases of PC and 13 cases of benign prostatic hyper-plasia (BPH). The expression and relationship of HIF-1α and VEGF-C with clinic-pathological features were analyzed. Re-Sults: The positive expression rates of HIF-1α and VEGF-C were significantly higher in PC (66.1% and 76.3% respectively) compared with those of BPH (7.7% and 0 respectively, P 0.05). There was a positive correlation between the expressions of HIF-1α and VEGF-C in prostatic carcinoma (r, = 0.261, P < 0.05). Conclusion: The over-expression of HIF-1α induced tumor invasion by activating the transcription of VEGF-C, which may play a great role in the genesis and development of prostate tumor.%目的:探讨缺氧诱导因子(HIF)-1α和血管内皮生长因子(VEGF)-C在前列腺癌中的表达及其临床意义.方法:应用免疫组织化学SABC法检测59例前列腺癌(PC)组织及13例良性前列腺增生(BPH)组织中HIF-1α和VEGF-C表达情况,分析两者相关性及其表达与临床病理特征的关系.结果:HIF-1α和VEGF-C在PC组中的阳性表达率分别为66.1%和76.3%,明显高于BPH组(7.7%和0),差异有统计学意义(P<0.01).HIF-1α和VEGF-C阳性表达率在不同Gleason评分、TNM分期和有无淋巴结转移的患者间差异有统计学意义(P< 0.05或P<0.01),而不同年龄和前列腺特异性抗原(PSA)患者间差异无统计学意义(P> 0.05).在PC中HIF-1α与VEGF-C的表达呈正相关(rs=0.261,P<0.05).结论:HIF-1α的过表达可能通过激活下游基因VEGF-C的表达协同参与前列腺癌的发生、发展,促进肿瘤的转移.

  9. PKM2和HIF-1α在食管癌中的表达及意义%Expression and significance of PKM2 and HIF-1αin esophageal carcinoma

    Institute of Scientific and Technical Information of China (English)

    顾国青; 费素娟

    2016-01-01

    Objective:To explore the expression and significance of M2-pyruvate kinase (PKM2) and hypoxia inducible factor-1α(HIF-1α) in esophageal carcinoma. Methods:The carcinoma tissues of 100 cases of esophageal carcinoma confirmed as squamous carcinoma clinically and pathologically were collected ( esophageal carcinoma group) and detected by immunohistochemistry for the expression of PKM2 and HIF-1α, and they were compared with 80 cases of esophagitis ( control group) . Results:The rate of positive expression of PKM2 and HIF-1αin esophageal carcinoma group were higher than that in control group( P <0. 05); the positive expression of PKM2 and HIF-1α was related to TNM stage, lymph node metastasis, chemotherapy resistance and 3-year survival rate;PKM2 and HIF-1αexpression were positively correlated ( r =0. 47, P <0. 05). Conclusion:The expression of PKM2 and HIF-1α is closely related to TNM stage, lymph node metastasis and chemotherapy resist-ance, over-expression of which can negatively affect the prognosis of patients with esophageal carcinoma.%目的::探讨M2型丙酮酸激酶(M2-pyruvate kinase,PKM2)与缺氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1α)在食管癌中的表达及意义。方法:取临床及病理证实为鳞癌的100例食管癌患者的癌组织(食管癌组),通过免疫组化方法检测其PKM2与HIF-1α的表达,并与食管炎患者80例(对照组)进行比较。结果:食管癌组PKM2与HIF-1α的阳性表达率高于对照组( P <0.05);PKM2与HIF-1α的阳性表达与TNM分期、淋巴结转移、化疗抵抗及3年生存率相关;PKM2与HIF-1α的表达呈正性相关( r =0.47, P <0.05)。结论:PKM2与HIF-1α的表达与食管癌的分期、远处转移、化疗抵抗相关,两者表达过度可负性影响食管癌患者预后。

  10. Cognitive characteristics of older Japanese drivers

    Directory of Open Access Journals (Sweden)

    Susilowati Indri H

    2012-02-01

    Full Text Available Abstract Background Some causes of accidents among older drivers are: not paying attention to traffic signals; missing stop lines; and having to deal with and misjudging emergency situations. These causes of accidents reveal problems with attention and cognition. Such incidents are also related to driver perception and stress-coping mechanisms. It is important to examine the relation of stress reactions to attention and cognition as a factor influencing the causes of accidents commonly involving older drivers. Finding Subjects were 10 young drivers (23.3 ± 3.33 years and 25 older drivers divided into two groups (older1 [60 to 65 years] and older2 [> 65 years]. This study revealed the correlation within driver stress inventory and driver coping questionnaires parameters was observed only in older drivers. They also needed a longer response time for Trail Making Test A and B. The factors affected the attention and cognition of older drivers by age but not driving experience itself, and coping parameters such as emotion focus, reappraisal, and avoidance were not included as stress inventory parameters. Being prone to fatigue was less for younger drivers than older drivers. Because they have shorter distances, shorter drive times, and no need for expressways, older drivers also had a significantly lower risk of thrill-seeking behaviour and more patience. Conclusion The intervention addressing their attention skills, aggressive feelings, and emotion focus should be considered. The technological improvements in cars will make older drivers feel safer and make driving easier which might lower the attention paid to the road, and regular driving training might be needed to assess and enhance their safety.

  11. Cognitive characteristics of older Japanese drivers.

    Science.gov (United States)

    Susilowati, Indri H; Yasukouchi, Akira

    2012-02-29

    Some causes of accidents among older drivers are: not paying attention to traffic signals; missing stop lines; and having to deal with and misjudging emergency situations. These causes of accidents reveal problems with attention and cognition. Such incidents are also related to driver perception and stress-coping mechanisms. It is important to examine the relation of stress reactions to attention and cognition as a factor influencing the causes of accidents commonly involving older drivers. Subjects were 10 young drivers (23.3 ± 3.33 years) and 25 older drivers divided into two groups (older1 [60 to 65 years] and older2 [> 65 years]). This study revealed the correlation within driver stress inventory and driver coping questionnaires parameters was observed only in older drivers. They also needed a longer response time for Trail Making Test A and B. The factors affected the attention and cognition of older drivers by age but not driving experience itself, and coping parameters such as emotion focus, reappraisal, and avoidance were not included as stress inventory parameters. Being prone to fatigue was less for younger drivers than older drivers. Because they have shorter distances, shorter drive times, and no need for expressways, older drivers also had a significantly lower risk of thrill-seeking behaviour and more patience. The intervention addressing their attention skills, aggressive feelings, and emotion focus should be considered. The technological improvements in cars will make older drivers feel safer and make driving easier which might lower the attention paid to the road, and regular driving training might be needed to assess and enhance their safety.

  12. Insufficient radiofrequency ablation promotes angiogenesis of residual hepatocellular carcinoma via HIF-1α/VEGFA.

    Directory of Open Access Journals (Sweden)

    Jian Kong

    Full Text Available BACKGROUND: The mechanism of rapid growth of the residual tumor after radiofrequency (RF ablation is poorly understood. In this study, we investigated the effect of hyperthermia on HepG2 cells and generated a subline with enhanced viability and dys-regulated angiogenesis in vivo, which was used as a model to further determine the molecular mechanism of the rapid growth of residual HCC after RF ablation. METHODOLOGY/PRINCIPAL FINDINGS: Heat treatment was used to establish sublines of HepG2 cells. A subline (HepG2 k with a relatively higher viability and significant heat tolerance was selected. The cellular protein levels of VEGFA, HIF-1α and p-Akt, VEGFA mRNA and secreted VEGFA were measured, and all of these were up-regulated in this subline compared to parental HepG2 cells. HIF-1α inhibitor YC-1 and VEGFA siRNA inhibited the high viability of the subline. The conditioned media from the subline exerted stronger pro-angiogenic effects. Bevacizumab, VEGFA siRNA and YC-1 inhibited proangiogenic effects of the conditioned media of HepG2 k cells and abolished the difference between parental HepG2 cells and HepG2 k cells. For in vivo studies, a nude mouse model was used, and the efficacy of bavacizumab was determined. HepG2 k tumor had stronger pro-angiogenic effects than parental HepG2 tumor. Bevacizumab could inhibit the tumor growth and angiogenesis, and also eliminate the difference in tumor growth and angiogenesis between parental HepG2 tumor and HepG2 k tumor in vivo. CONCLUSIONS/SIGNIFICANCE: The angiogenesis induced by HIF1α/VEGFA produced by altered cells after hyperthermia treatment may play an important role in the rapid growth of residual HCC after RF ablation. Bevacizumab may be a good candidate drug for preventing and treating the process.

  13. Dynamic Expression of HIF -1α and COX -2 in Perihematoma after Intracerebral Hemorrhage in Rats%大鼠脑出血灶周HIF-1α和COX-2的动态变化

    Institute of Scientific and Technical Information of China (English)

    李婷婷; 牛小媛

    2012-01-01

    目的 观察大鼠脑出血后血肿灶周脑组织中HIF -1α、COX -2蛋白的动态表达,探讨二者与脑水肿的关系,以及HIF -1α和COX -2蛋白的表达在血肿灶周脑组织损伤中的作用机制.方法 SD大鼠35只,随机分为假手术组、脑出血6h、1、3、7、14、21天组,每组5只.采用自体血体内注射法建立脑出血动物模型,用免疫组化法检测脑出血后血肿灶周脑组织中HIF -1α和COX -2蛋白的动态表达,并用干/湿比重法测定脑含水量.结果 脑出血后6h HIF - 1α、COX -2蛋白的表达和脑水含量明显升高,3~7天达到高峰,第14天明显降低,COX -2及脑水含量在21天基本降至正常.HIF - 1α在21天仍有较高水平的表达.造模后6h,脑出血组与假手术组相比,HIF -1 α及COX -2蛋白的表达和脑水含量明显增高,其差异有统计学意义(P<0.05).脑出血组的各时间点间比较,其差异均有统计学意义(P<0.05).HIF - 1α蛋白的表达与脑水含量呈正相关(r =0.636,P<0.01);COX -2蛋白的表达与脑水含量呈正相关(r=0.927,P<0.01).结论 大鼠脑出血后血肿灶周HIF -1α、COX -2蛋白的表达呈一定的变化规律.随着脑出血时间的延长HIF -1α蛋白、COX -2蛋白的表达程度和脑组织水肿程度呈正相关.%Objective To investigate the dynamic changes of HIF-lot and COX-2 protein in perihematoma after Intracerebral hemorrhage in rats, to analysis their correlation with the time of ICH and brain edema, and try to explore the mechanism of action of the expression HIF-lα and COX-2 protein on the brain tissue injury. Methods Thirty-five healthy SD rats were randomly divided into sham operation group (n=5) and ICH model groups (n =30). Rats were sacrificed at 6h,1days,3days,7days, 14days and 21 days after operation, respectively. The expressions of HIF-1α and COX-2 were investigated using immunohistochemistry, Brain water content was measured by the wet/dry specific gravity method. Results HIF-1

  14. Effect of TMP on the Expression of VEGF and HIF-1α of PG CSC Like Cells%川芎嗪对PG干细胞样细胞VEGF、HIF-1α蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    杨栋; 张培彤; 王耀焓; 郭秀伟; 马雪曼

    2015-01-01

    [目的]研究活血药川芎有效成分川芎嗪对高转移性人巨细胞肺癌细胞PG-BE1干细胞样细胞VEGF、HIF-1α蛋白表达的影响.[方法]利用无血清成球培养法分选PG-BE1中的干细胞亚群并验证其干性.用含不同浓度川芎嗪的培养液干预PG-BE1干细胞样细胞,同时设立空白对照组及顺铂(DDP)组,采用ELISA法检测PG-BE1亲代细胞、干细胞样细胞(常氧环境和低氧环境)以及不同浓度药物干预后的PG-BE1干细胞样细胞的VEGF表达,采用Wester blot法检测各组HIF-1α蛋白表达.[结果]PG-BE1干细胞样细胞VEGF表达强于其亲代细胞(P<0.001),VEGF在低氧环境中的表达高于常氧环境(P<0.001),不同浓度川芎嗪均可下调VEGF表达(P<0.001),且具有剂量依赖性;PG-BE1干细胞样细胞HIF-1α表达强于其亲代细胞(P<0.001),HIF-1α在低氧环境中的表达高于常氧环境(P=0.001),不同浓度川芎嗪均可下调HIF-1α表达(P<0.001),但不同剂量之间差异无统计学意义(P=0.477).[结论]PG-BE1干细胞样细胞VEGF、HIF-1α表达高于其亲代细胞,低氧环境下VEGF、HIF-1α表达上调,川芎嗪对VEGF、HIF-1α表达有一定的抑制作用.

  15. Definition of a Novel Feed-Forward Mechanism for Glycolysis-HIF1α Signaling in Hypoxic Tumors Highlights Aldolase A as a Therapeutic Target.

    Science.gov (United States)

    Grandjean, Geoffrey; de Jong, Petrus R; James, Brian P; Koh, Mei Yee; Lemos, Robert; Kingston, John; Aleshin, Alexander; Bankston, Laurie A; Miller, Claudia P; Cho, Eun Jeong; Edupuganti, Ramakrishna; Devkota, Ashwini; Stancu, Gabriel; Liddington, Robert C; Dalby, Kevin N; Powis, Garth

    2016-07-15

    The hypoxia-inducible transcription factor HIF1α drives expression of many glycolytic enzymes. Here, we show that hypoxic glycolysis, in turn, increases HIF1α transcriptional activity and stimulates tumor growth, revealing a novel feed-forward mechanism of glycolysis-HIF1α signaling. Negative regulation of HIF1α by AMPK1 is bypassed in hypoxic cells, due to ATP elevation by increased glycolysis, thereby preventing phosphorylation and inactivation of the HIF1α transcriptional coactivator p300. Notably, of the HIF1α-activated glycolytic enzymes we evaluated by gene silencing, aldolase A (ALDOA) blockade produced the most robust decrease in glycolysis, HIF-1 activity, and cancer cell proliferation. Furthermore, either RNAi-mediated silencing of ALDOA or systemic treatment with a specific small-molecule inhibitor of aldolase A was sufficient to increase overall survival in a xenograft model of metastatic breast cancer. In establishing a novel glycolysis-HIF-1α feed-forward mechanism in hypoxic tumor cells, our results also provide a preclinical rationale to develop aldolase A inhibitors as a generalized strategy to treat intractable hypoxic cancer cells found widely in most solid tumors. Cancer Res; 76(14); 4259-69. ©2016 AACR.

  16. Immunohistological expression of HIF-1α, GLUT-1, Bcl-2 and Ki-67 in consecutive biopsies during chemoradiotherapy in patients with rectal cancer

    DEFF Research Database (Denmark)

    Havelund, Birgitte Mayland; Sørensen, Flemming Brandt; Pløen, John;

    2013-01-01

    receiving preoperative CRT (>50.4 Gy and Uracil/Tegafur). Immunohistological expressions of HIF-1α, GLUT-1, Bcl-2 and Ki-67 were investigated in biopsies taken before treatment, after 2, 4 and 6 weeks of CRT and in specimens from the operation. Decreasing expressions of HIF-1α, Bcl-2 and Ki-67 were observed...

  17. Digoxin Downregulates NDRG1 and VEGF through the Inhibition of HIF-1α under Hypoxic Conditions in Human Lung Adenocarcinoma A549 Cells

    Directory of Open Access Journals (Sweden)

    Dong Wei

    2013-04-01

    Full Text Available Digoxin, an inhibitor of Na+/K+ ATPase, has been used in the treatment of heart-related diseases (such as congestive heart failure and atrial arrhythmia for decades. Recently, it was reported that digoxin is also an effective HIF-1α inhibitor. We investigated whether digoxin could suppress tumor cell growth through HIF-1α in non-small cell lung cancer cells (A549 cells under hypoxic conditions. An MTT assay was used to measure cell viability. RT-PCR and western blotting were performed to analyze the mRNA and protein expression of VEGF, NDRG1, and HIF-1α. HIF-1α nuclear translocation was then determined by EMSA. Digoxin was found to inhibit the proliferation of A549 cells under hypoxic conditions. Our results showed that hypoxia led to the upregulation of VEGF, NDRG1, and HIF-1α both at the mRNA and protein levels. We also found that the hypoxia-induced overexpression of VEGF, NDRG1, and HIF-1α was suppressed by digoxin in a concentration-dependent manner. As expected, our EMSA results demonstrated that under hypoxic conditions HIF-1α nuclear translocation was also markedly reduced by digoxin in a concentration-dependent manner. Our results suggest that digoxin downregulated hypoxia-induced overexpression of VEGF and NDRG1 at the transcriptional level probably through the inhibition of HIF-1α synthesis in A549 cells.

  18. Design, synthesis and insight into the structure-activity relationship of 1,3-disubstituted indazoles as novel HIF-1 inhibitors.

    Science.gov (United States)

    An, Hongchan; Kim, Nam-Jung; Jung, Jong-Wha; Jang, Hannah; Park, Jong-Wan; Suh, Young-Ger

    2011-11-01

    Design, synthesis and insight into the structure-activity relationship (SAR) of 1,3-disubstituted indazoles as novel HIF-1 inhibitors are described. In particular, the substituted furan moiety on indazole skeleton as well as its substitution pattern turns out crucial for the high HIF-1 inhibition.

  19. 18F-FDG imaging of human atherosclerotic carotid plaques reflects gene expression of the key hypoxia marker HIF-1α

    DEFF Research Database (Denmark)

    Pedersen, Sune Folke; Græbe, Martin; Hag, Anne Mette F;

    2013-01-01

    and CD68 gene expression co-variated and accordingly when entering the variables into multivariate linear regression models with SUV-values as dependent variables, HIF-1α was eliminated in the final models. (18)F-FDG-uptake (SUVmax) is correlated with HIF-1α gene expression indicating an association...

  20. Precise Characterization of a Laser Current Driver

    Science.gov (United States)

    Troxel, Daylin

    2009-10-01

    I will be presenting a characterization of our unique low-noise laser current driver. Our current driver improves on the typical model used in laboratories, giving extra current stability and lower noise. I will discuss our techniques for measuring the noise and drift and the results we obtained. The current driver has a lower noise and drift than any other current driver with a published value, so it has value in making precision measurements. Many other labs have expressed interest in our design as there is a need for this type of current driver in many applications. The current driver demonstrates some interesting applications of electronics principles and uses of electric components, as well as practical considerations in designing circuitry.

  1. Driver models for personalised driving assistance

    Science.gov (United States)

    Lefèvre, Stéphanie; Carvalho, Ashwin; Gao, Yiqi; Tseng, H. Eric; Borrelli, Francesco

    2015-12-01

    We propose a learning-based driver modelling approach which can identify manoeuvres performed by drivers on the highway and predict the future driver inputs. We show how this approach can be applied to provide personalised driving assistance. In a first example, the driver model is used to predict unintentional lane departures and a model predictive controller is used to keep the car in the lane. In a second example, the driver model estimates the preferred acceleration of the driver during lane keeping, and a model predictive controller is implemented to provide a personalised adaptive cruise control. For both applications, we use a combination of real data and simulation to evaluate the proposed approaches.

  2. Restoring physiological levels of ascorbate slows tumor growth and moderates HIF-1 pathway activity in Gulo(-/-) mice.

    Science.gov (United States)

    Campbell, Elizabeth J; Vissers, Margreet C M; Bozonet, Stephanie; Dyer, Arron; Robinson, Bridget A; Dachs, Gabi U

    2015-02-01

    Hypoxia-inducible factor-1 (HIF-1) governs cellular adaption to the hypoxic microenvironment and is associated with a proliferative, metastatic, and treatment-resistant tumor phenotype. HIF-1 levels and transcriptional activity are regulated by proline and asparagine hydroxylases, which require ascorbate as cofactor. Ascorbate supplementation reduced HIF-1 activation in vitro, but only limited data are available in relevant animal models. There is no information of the effect of physiological levels of ascorbate on HIF activity and tumor growth, which was measured in this study. C57BL/6 Gulo(-/-) mice (a model of the human ascorbate dependency condition) were supplemented with 3300 mg/L, 330 mg/L, or 33 mg/L of ascorbate in their drinking water before and during subcutaneous tumor growth of B16-F10 melanoma or Lewis lung carcinoma (LL/2). Ascorbate levels in tumors increased significantly with elevated ascorbate intake and restoration of wild-type ascorbate levels led to a reduction in growth of B16-F10 (log phase P ascorbate supplementation increased for both tumor models (P ascorbate was inversely correlated with levels of the HIF-1 target proteins CA-IX, GLUT-1, and VEGF in both B16-F10 and LL/2 tumors (P ascorbate groups but varied between models (30% for B16-F10 and 21% for LL/2), indicating that ascorbate did not affect tumor hypoxia. Our data support the hypothesis that restoration of optimal intracellular ascorbate levels reduces tumor growth via moderation of HIF-1 pathway activity.

  3. NADPH oxidase 4 mediates insulin-stimulated HIF-1α and VEGF expression, and angiogenesis in vitro.

    Directory of Open Access Journals (Sweden)

    Dan Meng

    Full Text Available Acute intensive insulin therapy causes a transient worsening of diabetic retinopathy in type 1 diabetes patients and is related to VEGF expression. Reactive oxygen species (ROS have been shown to be involved in HIF-1α and VEGF expression induced by insulin, but the role of specific ROS sources has not been fully elucidated. In this study we examined the role of NADPH oxidase subunit 4 (Nox4 in insulin-stimulated HIF-1α and VEGF expression, and angiogenic responses in human microvascular endothelial cells (HMVECs. Here we demonstrate that knockdown of Nox4 by siRNA reduced insulin-stimulated ROS generation, the tyrosine phosphorylation of IR-β and IRS-1, but did not change the serine phosphorylation of IRS-1. Nox4 gene silencing had a much greater inhibitory effect on insulin-induced AKT activation than ERK1/2 activation, whereas it had little effect on the expression of the phosphatases such as MKP-1 and SHIP. Inhibition of Nox4 expression inhibited the transcriptional activity of VEGF through HIF-1. Overexpression of wild-type Nox4 was sufficient to increase VEGF transcriptional activity, and further enhanced insulin-stimulated the activation of VEGF. Downregulation of Nox4 expression decreased insulin-stimulated mRNA and protein expression of HIF-1α, but did not change the rate of HIF-1α degradation. Inhibition of Nox4 impaired insulin-stimulated VEGF expression, cell migration, cell proliferation, and tube formation in HMVECs. Our data indicate that Nox4-derived ROS are essential for HIF-1α-dependent VEGF expression, and angiogenesis in vitro induced by insulin. Nox4 may be an attractive therapeutic target for diabetic retinopathy caused by intensive insulin treatment.

  4. Salidroside Inhibits Inflammation Through PI3K/Akt/HIF Signaling After Focal Cerebral Ischemia in Rats.

    Science.gov (United States)

    Wei, Yicong; Hong, Haimian; Zhang, Xiaoqin; Lai, Wenfang; Wang, Yingzheng; Chu, Kedan; Brown, John; Hong, Guizhu; Chen, Lidian

    2017-08-01

    Salidroside is being investigated for its therapeutic potential in stroke because it is neuroprotective over an extended therapeutic window of time. In the present study, we investigated the mechanisms underlying the anti-inflammatory effects of salidroside (50 mg/kg intraperitoneally) in rats, given 1 h after reperfusion of a middle cerebral artery that had been occluded for 2 h. After 24 h, we found that salidroside increased the neuronal nuclear protein NeuN and reduced the marker of microglia and macrophages CD11b in the peri-infarct area of the brain. Salidroside also decreased IL-6, IL-1β, TNF-α, CD14, CD44, and iNOs mRNAs. At the same time, salidroside increased the ratio of phosphorylated protein kinase B (p-Akt) to total Akt. The phosphoinositide 3-kinase (PI3K) inhibitor LY294002 prevented this increase in p-Akt and reversed the inhibitory effects of salidroside on CD11b and inflammatory mediators. Salidroside also elevated the protein levels of hypoxia-inducible factor (HIF) subunits HIF1α, HIF2α, HIF3α, and of erythropoietin (EPO). The stimulatory effects of salidroside on HIFα subunits were blocked by LY294002. Moreover, YC-1, a HIF inhibitor, abolished salidroside-mediated increase of HIF1α and prevented the inhibitory effects of salidroside on CD11b and inflammatory mediators. Taken together, our results provide evidence for the first time that all three HIFα subunits and EPO can be regulated by PI3K/Akt in cerebral tissue, and that salidroside entrains this signaling pathway to induce production of HIFα subunits and EPO, one or more of which mediate the anti-inflammatory effects of salidroside after cerebral IRI.

  5. Kelch-like ECT2-interacting protein KLEIP regulates late-stage pulmonary maturation via Hif-2α in mice

    Directory of Open Access Journals (Sweden)

    Nicole Woik

    2014-06-01

    Full Text Available Respiratory distress syndrome (RDS caused by preterm delivery is a major clinical problem with limited mechanistic insight. Late-stage embryonic lung development is driven by hypoxia and the hypoxia-inducible transcription factors Hif-1α and Hif-2α, which act as important regulators for lung development. Expression of the BTB-and kelch-domain-containing (BTB-kelch protein KLEIP (Kelch-like ECT2-interacting protein; also named Klhl20 is controlled by two hypoxia response elements, and KLEIP regulates stabilization and transcriptional activation of Hif-2α. Based on the available data, we hypothesized an essential role for KLEIP in murine lung development and function. Therefore, we have performed a functional, histological, mechanistic and interventional study in embryonic and neonatal KLEIP−/− mice. Here, we show that about half of the KLEIP−/− neonates die due to respiratory failure that is caused by insufficient aeration, reduced septal thinning, reduced glycogenolysis, type II pneumocyte immaturity and reduced surfactant production. Expression analyses in embryonic day (E 18.5 lungs identified KLEIP in lung capillaries, and showed strongly reduced mRNA and protein levels for Hif-2α and VEGF; such reduced levels are associated with embryonic endothelial cell apoptosis and lung bleedings. Betamethasone injection in pregnant females prevented respiratory failure in KLEIP−/− neonates, normalized lung maturation, vascularization, aeration and function, and increased neonatal Hif-2α expression. Thus, the experimental study shows that respiratory failure in KLEIP−/− neonates is determined by insufficient angiocrine Hif-2α–VEGF signaling and that betamethasone activates this newly identified signaling cascade in late-stage embryonic lung development.

  6. Modelling Driver Assitance Systems by Optimal Control

    OpenAIRE

    Wang, M.; Daamen, W.; Hoogendoorn, S.P.; Van Arem, B.

    2012-01-01

    Driver assistance systems support drivers in operating vehicles in a safe, comfortable and efficient way, and thus may induce changes in traffic flow characteristics. This paper put forward a receding horizon control framework to model driver assistance systems. The accelerations of automated vehicles are determined to optimise a cost function, assuming other vehicles driving at stationary conditions over a prediction horizon. The flexibility of the framework is demonstrated with controller d...

  7. Pretreatment HIF-1α and GLUT-1 expressions do not correlate with outcome after preoperative chemoradiotherapy in rectal cancer

    DEFF Research Database (Denmark)

    Havelund, Birgitte Mayland; Sørensen, Flemming Brandt; Lindebjerg, Jan

    2011-01-01

    The aim of the present study was to investigate hypoxia-inducible factor 1α (HIF-1α) and glucose transporter-1 (GLUT-1) expressions as predictors of response and survival after chemoradiotherapy in pretreatment biopsy specimens from patients with rectal cancer.......The aim of the present study was to investigate hypoxia-inducible factor 1α (HIF-1α) and glucose transporter-1 (GLUT-1) expressions as predictors of response and survival after chemoradiotherapy in pretreatment biopsy specimens from patients with rectal cancer....

  8. Expression of HIF-1α and VEGF in human dental pulp cells under mechanical stretch%牵张力作用下人牙髓细胞HIF-1α和VEGF的表达

    Institute of Scientific and Technical Information of China (English)

    魏福兰; 耿杰; 王春玲; 王惠; 张本君; 张凡

    2012-01-01

    目的:研究牵张力作用下人牙髓细胞HIF-1α和VEGF基因的表达变化,探讨 HIF-1α和VEGF与正畸牙髓组织维持内环境稳定的关系.方法:通过细胞牵张应力加载系统,对细胞施加频率为1.0 Hz、大小为15%形变率的牵张应力,分别加力6h、12h、24h、48 h和72 h实时定量PCR检测不同时间点HIF-1α和VEGF mRNA的表达变化采用SPSS 12.0软件包对数据进行统计学分析 结果:未加力前,HIF-1 αmRNA表达极其微弱;加力6 h时,表达开始升高(P<0.05);持续增加至24 h时,表达最显著(P<0.05);加力48 h时,表达缓慢下降(P<0.01);至加力72 h时,表达下降至未加力前(P> 0.05).未加力前,VEGFmRNA表达微弱,加力6h表达开始升高,但无显著差异(P>0.05);加力12 h 时,VEGF mRNA表达开始明显增强(P<0.05),持续增加至72 h加力结束时(P<0.05).结论:牵张力可诱导人牙髓细胞HIF-1α和VEGF mRNA的表达变化,两者可能在维持正畸受力后牙髓内环境稳定中发挥重要作用.%To examine the expression of HIF-la and VEGF after application of mechanical stretch on human dental pulp cells (HDPCs), and to investigate the role of them in maintaining homeostasis of dental pulp during orthodontic movement. METHODS: HDPCs were subjected to 15% elongation by 1.0 Hz stretching frequency for 6, 12. 24, 48 and 72 h. The expression of HIF-la and VEGF mRNA was measured by real-time polymerase chain reaction (PCR). SPSS12.0 software package was applied for statistical analysis. RESULTS: At the initial time point, HIF-la mRNA had a weak expression. The mRNA level of HIF-la increased gradually and then decreased towards its pre-pressure levels. The mRNA level of VEGF was upregulated in a time梔ependent manner. CONCLUSIONS: The mRNA expression of HIF-la and VEGF was enhanced by mechanical stress in HDPCs, which indicates that HIF-la and VEGF may play an important role in retaining homeostasis of dental pulp during orthodontic movement. Supported

  9. Economic drivers of mineral supply

    Science.gov (United States)

    Wagner, Lorie A.; Sullivan, Daniel E.; Sznopek, John L.

    2003-01-01

    The debate over the adequacy of future supplies of mineral resources continues in light of the growing use of mineral-based materials in the United States. According to the U.S. Geological Survey, the quantity of new materials utilized each year has dramatically increased from 161 million tons2 in 1900 to 3.2 billion tons in 2000. Of all the materials used during the 20th century in the United States, more than half were used in the last 25 years. With the Earth?s endowment of natural resources remaining constant, and increased demand for resources, economic theory states that as depletion approaches, prices rise. This study shows that many economic drivers (conditions that create an economic incentive for producers to act in a particular way) such as the impact of globalization, technological improvements, productivity increases, and efficient materials usage are at work simultaneously to impact minerals markets and supply. As a result of these economic drivers, the historical price trend of mineral prices3 in constant dollars has declined as demand has risen. When price is measured by the cost in human effort, the price trend also has been almost steadily downward. Although the United States economy continues its increasing mineral consumption trend, the supply of minerals has been able to keep pace. This study shows that in general supply has grown faster than demand, causing a declining trend in mineral prices.

  10. Pin1, a new player in the fate of HIF-1α degradation: an hypothetical mechanism inside vascular damage as Alzheimer’s disease risk factor.

    Directory of Open Access Journals (Sweden)

    Elena eLonati

    2014-01-01

    Full Text Available Aetiology of neurodegenerative mechanisms underlying Alzheimer's disease (AD are still under elucidation. The contribution of cerebrovascular deficiencies (such as cerebral ischemia/stroke has been strongly endorsed in recent years. Reduction of blood supply leading to hypoxic condition is known to activate cellular responses mainly controlled by hypoxia-inducible transcription factor-1 (HIF-1. Thus alterations of oxygen responsive HIF-1α subunit in the central nervous system may contribute to the cognitive decline, especially influencing mechanisms associated to APP (amyloid precursor protein amyloidogenic metabolism. Although HIF-1α protein level is known to be regulated by von Hippel-Lindau (VHL ubiquitin-proteasome system, it has been recently suggested that Gsk-3β (glycogen synthase kinase-3β promotes a VHL-independent HIF-1α degradation. Here we provide evidences that in rat primary hippocampal cell cultures, HIF-1α degradation might be mediated by a synergic action of Gsk-3β and Pin1 (peptidyl-prolyl cis/trans isomerase. In post-ischemic conditions, such as those mimicked with oxygen glucose deprivation (OGD, HIF-1α protein level increases remaining unexpectedly high for long time after normal condition restoration jointly with the increase of LDH (lactate dehydrogenase and BACE1 (β-secretase 1 protein expression (70% and 140% respectively. Interestingly the Pin1 activity decreases about 40%-60% and Pin1S16 inhibitory phosphorylation significantly increases, indicating that Pin1 binding to its substrate and enzymatic activity are reduced by treatment. Co-immunoprecipitation experiments demonstrate that HIF-1α/Pin1 in normoxia are associated, and that in presence of specific Pin1 and Gsk-3β inhibitors their interaction is reduced in parallel to an increase of HIF-1α protein level. Thus we suggest that in post-OGD neurons the high level of HIF-1α might be due to Pin1 binding ability and activity reduction which affects HIF-1

  11. Macrophages transmit potent proangiogenic effects of oxLDL in vitro and in vivo involving HIF-1α activation: a novel aspect of angiogenesis in atherosclerosis.

    Science.gov (United States)

    Hutter, Randolph; Speidl, Walter S; Valdiviezo, Carolina; Sauter, Bernhard; Corti, Roberto; Fuster, Valentin; Badimon, Juan J

    2013-08-01

    Neovascularization has been linked to the progression and vulnerability of atherosclerotic lesions. Angiogenesis is increased in lipid-rich plaque. Hypoxia-inducible factor alpha (HIF-1α) is a key transcriptional regulator responding to hypoxia and activating genes, which promote angiogenesis, among them vascular endothelial growth factor (VEGF). Oxidized low-density lipoprotein (oxLDL) is generated in lipid-rich plaque by oxidative stress. It triggers an inflammatory response and was traditionally thought to inhibit endothelial cells. New data, however, suggest that oxLDL can activate HIF-1α in monocytes in a hypoxia-independent fashion. We hypothesized that HIF-1α activation in monocyte-macrophages could transmit proangiogenic effects of oxLDL linking hyperlipidemia, inflammation, and angiogenesis in atherosclerosis. First, we examined the effect of oxLDL on HIF-1α and VEGF expression in monocyte-macrophages and on their proangiogenic effect on endothelial cells in vitro in a monocyte-macrophage/endothelial co-culture model. OxLDL strongly induced HIF-1α and VEGF in monocyte-macrophages and significantly increased tube formation in co-cultured endothelial cells. HIF-1α inhibition reversed this effect. Second, we demonstrated a direct proangiogenic effect of oxLDL in an in vivo angiogenesis assay. Again, HIF-1α inhibition abrogated the proangiogenic effect of oxLDL. Third, in a rabbit atherosclerosis model, we studied the effect of dietary lipid lowering on arterial HIF-1α and VEGF expression. The administration of low-lipid diet significantly reduced the expression of both HIF-1α and VEGF, resulting in decreased plaque neovascularization. Our data point to oxLDL as a proangiogenic agent linking hyperlipidemia, inflammation, and angiogenesis in atherosclerosis. This effect is dependent on macrophages and, at least in part, on the induction of the HIF-1α pathway.

  12. High concentrations of L-ascorbic acid specifically inhibit the growth of human leukemic cells via downregulation of HIF-1α transcription.

    Directory of Open Access Journals (Sweden)

    Hiroshi Kawada

    Full Text Available We examined the antileukemic effects of high concentrations of L-ascorbic acid (high AA on human leukemic cells. In vitro, high AA markedly induced apoptosis in various leukemic cell lines by generating hydrogen peroxide (H2O2 but not in normal hematopoietic stem/progenitor cells. High AA significantly repressed leukemic cell proliferation as well as neoangiogenesis in immunodeficient mice. We then noted that in leukemic cells, HIF-1α transcription was strongly suppressed by high AA and correlated with the transcription of VEGF. Our data indicate that exposure to high AA markedly increased the intracellular AA content of leukemic cells and inhibited the nuclear translocation of NF-κB, which mediates expression of HIF-1α. We next generated K562 cells that overexpressed HIF-1α (K562-HIF1α cells and assessed the mechanistic relationship between inhibition of HIF-1α transcription and the antileukemic effect of high AA. The ability of high AA to induce apoptosis was significantly lower in K562-HIF1α cells than in K562 cells in vitro. We found that expression of HIF-1α-regulated antiapoptotic proteins of the Bcl-2 family, such as Mcl-1, Bcl-xL, and Bcl-2, was significantly suppressed by high AA in K562 cells, but was sustained at higher levels in K562-HIF1α cells, regardless of high AA exposure. Moreover, repression of cell proliferation and neoangiogenesis by high AA was completely abrogated in mice receiving transplants of K562-HIF1α cells. These results indicate that, along with H2O2 generation, downregulation of HIF-1α transcription plays a crucial role in growth inhibition of human leukemic cells by high AA.

  13. Knock down of HIF-1α in glioma cells reduces migration in vitro and invasion in vivo and impairs their ability to form tumor spheres

    Directory of Open Access Journals (Sweden)

    Esencay Mine

    2010-06-01

    Full Text Available Abstract Background Glioblastoma (GBM is the most common and malignant primary intracranial human neoplasm. GBMs are characterized by the presence of extensive areas of necrosis and hypoxia. Hypoxia and its master regulator, hypoxia inducible factor 1 (HIF-1 play a key role in glioma invasion. Results To further elucidate the functional role of HIF-1α in glioma cell migration in vitro and in invasion in vivo, we used a shRNA approach to knock down HIF-1α expression complemented with genome-wide expression profiling, performed in both normoxic and hypoxic conditions. Our data show that knock down of HIF-1α in glioma cells significantly impairs their migration in vitro as well as their ability to invade into the brain parenchyma in vivo. Next, we assessed the role that HIF-1α plays in maintaining the characteristics of cancer stem cells (CSCs. By using the tumor sphere forming assay, we demonstrate that HIF-1α plays a role in the survival and self-renewal potential of CSCs. Finally, expression profiling experiments in glioma cells provided detailed insight into a broad range of specific biological pathways and processes downstream of HIF-1α. We discuss the role of these processes in the migratory and invasive properties, as well as the stem cell biology of glioblastomas Conclusions Our data show that knock down of HIF-1α in human and murine glioma cells impairs their migration in vitro and their invasion in vivo. In addition, our data suggest that HIF-1α plays a role in the survival and self-renewal potential of CSCs and identify genes that might further elucidate the role of HIF-1α in tumor migration, invasion and stem cell biology.

  14. Kinome-Wide Functional Genomics Screen Reveals a Novel Mechanism of TNFα-Induced Nuclear Accumulation of the HIF-1α Transcription Factor in Cancer Cells

    Science.gov (United States)

    Schoolmeesters, Angela; Brown, Daniel D.; Fedorov, Yuriy

    2012-01-01

    Hypoxia-inducible factor-1 (HIF-1) and its most important subunit, HIF-1α, plays a central role in tumor progression by regulating genes involved in cancer cell survival, proliferation and metastasis. HIF-1α activity is associated with nuclear accumulation of the transcription factor and regulated by several mechanisms including modulation of protein stability and degradation. Among recent advances are the discoveries that inflammation-induced cytokines and growth factors affect protein accumulation of HIF-1α under normoxia conditions. TNFα, a major pro-inflammatory cytokine that promotes tumorigenesis is known as a stimulator of HIF-1α activity. To improve our understanding of TNFα-mediated regulation of HIF-1α nuclear accumulation we screened a kinase-specific siRNA library using a cell imaging–based HIF-1α-eGFP chimera reporter assay. Interestingly, this systematic analysis determined that depletion of kinases involved in conventional TNFα signaling (IKK/NFκB and JNK pathways) has no detrimental effect on HIF-1α accumulation. On the other hand, depletion of PRKAR2B, ADCK2, TRPM7, and TRIB2 significantly decreases the effect of TNFα on HIF-1α stability in osteosarcoma and prostate cancer cell lines. These newly discovered regulators conveyed their activity through a non-conventional RELB-depended NFκB signaling pathway and regulation of superoxide activity. Taken together our data allow us to conclude that TNFα uses a distinct and complex signaling mechanism to induce accumulation of HIF-1α in cancer cells. In summary, our results illuminate a novel mechanism through which cancer initiation and progression may be promoted by inflammatory cytokines, highlighting new potential avenues for fighting this disease. PMID:22355351

  15. Enhanced hypoxia-inducible factor (HIF)-1α stability induced by 5-hydroxymethyl-2-furfural (5-HMF) contributes to protection against hypoxia.

    Science.gov (United States)

    He, Yun-Ling; Li, Ming-Ming; Wu, Li-Ying; Zhao, Tong; Di, Yao; Huang, Xin; Ding, Xue-Feng; Wu, Kui-Wu; Fan, Ming; Zhu, Ling-Ling

    2014-01-01

    We first reported the role of 5-hydroxymethyl-2-furfural (5-HMF) against hypoxia. Here, we studied the mechanism by using oxygen-dependent degradation domain (ODD)-Luc mice, which are a useful model to probe the stabilization of hypoxia-inducible factor 1α (HIF-1α). Compared with three other compounds that have been reported to have a role in stabilizing HIF-1α, 5-HMF caused stronger bioluminescence, which is indicative of HIF-1α stability in the brain and kidney of ODD-Luc mice. We further demonstrated that the HIF-1α protein accumulated in response to 5-HMF in the brains and kidneys of these mice, as well as in PC12 cells. Additionally, 5-HMF promoted the nuclear translocation of HIF-1α and the transcriptional activity of HIF-1, which was evaluated by detecting vascular endothelial growth factor (VEGF ) mRNA expression. These results suggest that 5-HMF stabilized HIF-1α and increased its activity. Considering the role of proline hydroxylases (PHDs) in negatively regulating HIF-1α stability, we explored whether 5-HMF interacts with the substrates and cofactors of PHDs, such as 2-oxoglutarate (2-OG), Fe(2+) and vitamin C (VC), which affects the activity of PHDs. The result revealed that 5-HMF did not interact with Fe(2+) or 2-OG but interacted with VC. This interaction was confirmed by subsequent experiments, in which 5-HMF entered into cells and reduced the VC content. The enhanced stability of HIF-1α by 5-HMF was reversed by VC supplementation, and the improved survival of mice caused by 5-HMF under hypoxia was abrogated by VC supplementation. Thus, we demonstrated for the first time that 5-HMF increases HIF-1α stability by reducing the VC content, which mediates the protection against hypoxia.

  16. Is the interaction between HIF1A P582S and ACTN3 R577X determinant for power/sprint performance?

    Science.gov (United States)

    Eynon, Nir; Alves, Alberto Jorge; Meckel, Yoav; Yamin, Chen; Ayalon, Moshe; Sagiv, Michael; Sagiv, Moran

    2010-06-01

    Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that regulates gene expression in response to hypoxia and has been associated with athletic performance. The aims of this study were (1) to determine the frequency distribution of HIF1A Pro582Ser (rs11549465) polymorphism among 155 Israeli athletes (sprinters and endurance athletes) and 240 healthy controls and (2) to analyze the influence of the interaction between HIF1A Pro582Ser and ACTN3 R577X (rs1815739) genotypes on sprint performance. There were no differences across the HIF1A genotype and allele frequencies among endurance athletes, sprinters, and controls. Similarly, no differences were found between the subgroups of top-level and national-level endurance athletes, or between top-level and national-level sprinters. Conversely, interaction effects were found between HIF1A Pro582Ser and ACTN3 R577X polymorphisms and sprinters. The proportion of HIF1A Pro/Pro + ACTN3 R/R genotypes was significantly higher in sprinters than in endurance athletes and healthy controls (P = .002). In addition, the odds ratio for HIF1A Pro/Pro + ACTN3 R/R genotype carriers being a sprinter was 2.25 (95% confidence interval, 1.24-4.1); and that for HIF1A Pro/Pro + ACTN3 R/R genotype carriers being an endurance athlete was 0.5 (95% confidence interval, 0.2-1.24). We conclude that HIF1A Pro582Ser polymorphism by itself is not critical in determining sprint performance. However, sprinter performance is determined by the interaction between the wild-type HIF1A Pro/Pro genotype and ACTN3 RR genotype.

  17. Structural basis for oxygen degradation domain selectivity of the HIF prolyl hydroxylases

    Science.gov (United States)

    Chowdhury, Rasheduzzaman; Leung, Ivanhoe K. H.; Tian, Ya-Min; Abboud, Martine I.; Ge, Wei; Domene, Carmen; Cantrelle, François-Xavier; Landrieu, Isabelle; Hardy, Adam P.; Pugh, Christopher W.; Ratcliffe, Peter J.; Claridge, Timothy D. W.; Schofield, Christopher J.

    2016-01-01

    The response to hypoxia in animals involves the expression of multiple genes regulated by the αβ-hypoxia-inducible transcription factors (HIFs). The hypoxia-sensing mechanism involves oxygen limited hydroxylation of prolyl residues in the N- and C-terminal oxygen-dependent degradation domains (NODD and CODD) of HIFα isoforms, as catalysed by prolyl hydroxylases (PHD 1–3). Prolyl hydroxylation promotes binding of HIFα to the von Hippel–Lindau protein (VHL)–elongin B/C complex, thus signalling for proteosomal degradation of HIFα. We reveal that certain PHD2 variants linked to familial erythrocytosis and cancer are highly selective for CODD or NODD. Crystalline and solution state studies coupled to kinetic and cellular analyses reveal how wild-type and variant PHDs achieve ODD selectivity via different dynamic interactions involving loop and C-terminal regions. The results inform on how HIF target gene selectivity is achieved and will be of use in developing selective PHD inhibitors. PMID:27561929

  18. Axotomy-induced HIF-serotonin signalling axis promotes axon regeneration in C. elegans

    Science.gov (United States)

    Alam, Tanimul; Maruyama, Hiroki; Li, Chun; Pastuhov, Strahil Iv.; Nix, Paola; Bastiani, Michael; Hisamoto, Naoki; Matsumoto, Kunihiro

    2016-01-01

    The molecular mechanisms underlying the ability of axons to regenerate after injury remain poorly understood. Here we show that in Caenorhabditis elegans, axotomy induces ectopic expression of serotonin (5-HT) in axotomized non-serotonergic neurons via HIF-1, a hypoxia-inducible transcription factor, and that 5-HT subsequently promotes axon regeneration by autocrine signalling through the SER-7 5-HT receptor. Furthermore, we identify the rhgf-1 and rga-5 genes, encoding homologues of RhoGEF and RhoGAP, respectively, as regulators of axon regeneration. We demonstrate that one pathway initiated by SER-7 acts upstream of the C. elegans RhoA homolog RHO-1 in neuron regeneration, which functions via G12α and RHGF-1. In this pathway, RHO-1 inhibits diacylglycerol kinase, resulting in an increase in diacylglycerol. SER-7 also promotes axon regeneration by activating the cyclic AMP (cAMP) signalling pathway. Thus, HIF-1-mediated activation of 5-HT signalling promotes axon regeneration by activating both the RhoA and cAMP pathways. PMID:26790951

  19. mTOR signaling pathway regulates HIF-1α and VEGF%mTOR信号通路调节HIF-1α及VEGF

    Institute of Scientific and Technical Information of China (English)

    陈洪菊; 唐彬秩; 屈艺; 母得志

    2011-01-01

    哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)是一种非典型丝氨酸/苏氨酸蛋白激酶,可整合营养、能量及生长因子等多种细胞外信号,参与基因转录、蛋白质翻译、核糖体合成等生物过程,在细胞生长和细胞程序性死亡中发挥极为重要的作用.研究表明,mTOR信号通路可能通过调节下游靶蛋白来调控细胞生物过程,如缺氧诱导因子(hypoxiainducible factor,HIF-1α),血管内皮生长因子(vascular endothelial growth factor,VEGF)等.mTOR通过调节这些细胞因子,可能影响蛋白质合成、血管生成和细胞凋亡自噬等,从而影响疾病发展和结局,这为mTOR成为疾病治疗靶点提供证据.%The mammalian target of rapamycin (mTOR) is an evolutionarily conserved atypical Ser/Thr protein kinase, functioning as a sensor of extracellular signals from nutrients, oxygen, energy and growth factors. Through modulating its downstream targets in the cellular nucleus, mTOR regulates transcription, cell growth, protein translation, proliferation, survival and programmed cell death. It is proved that mTOR can regulate the cellular biological processes through some cytokines, such as hypoxia-inducible factor (HIF-la), vascular endothelial growth factor (VEGF), etc. These processes include protein synthesis, angiogenesis, apoptosis or autophagy, which participate in the pathological or survival mechanisms of diseases. Therefore, mTOR could become a candidate target for treatment of diseases.

  20. The predictive values of HIF-1 and VEGF in early diabetic nephropathy%HIF-1α、VEGF在预测早期糖尿病肾脏病中的价值

    Institute of Scientific and Technical Information of China (English)

    宋秋艳; 余玲; 董瑞鸿

    2016-01-01

    Objective To investigate the predictive values of hypoxia-induced factor(HIF)- 1α and vascular endothelial growth factor (VEGF) in patients with type 2 diabetic kidney disease, and its relationship between HIF-1α, VEGF and glomerular filtration rate (eGFR). Methods Seventy-six patients with type 2 diabetic kidney disease were divided into two groups:DKD1 group [eGFR>90 mL/(min·1.73 m2),n=48] and DKD2 group [60 mL/(min·1.73 m2)0.05). Serum levels of HIF-1α and VEGF were significantly higher in DKD2 group than those of DKD1 group (P<0.05). There were negative correlation between VEGF and HIF-1αlevels with eGFR level (r=-0.59, P<0.01;r=-0.55, P<0.01). ROC curve showed that serum VEGF and HIF-1αlevels were above the opportunity diagonal. The diagnostic sensitivities of HIF-1 to DKD1 and DKD2 were 92.1%and 87.4%. The diagnostic specificities of HIF-1 to DKD1 and DKD2 were 91.7%and 85.4%. The diagnostic sensitivities of VEGF to DKD1 and DKD2 were 96.1%and 86.2%, and specificities were 93.3%and 89.1%. Conclusion Serum levels of VEGF and HIF-1αchange with the severity of diabetic nephropathy are likely to be an early predictor for the progression of diabetic kidney disease.%目的:通过分析缺氧诱导因子(HIF)-1α和血管内皮生长因子(VEGF)水平与肾小球滤过率(eGFR)的关系,探讨HIF-1α、VEGF在预测早期糖尿病肾脏病(DKD)中的价值。方法76例糖尿病肾脏病患者根据eGFR水平分为DKD1组[eGFR>90 mL(min·1.73 m2),48例]和DKD2组[60 mL/(min·1.73 m2)<eGFR≤90 mL/(min·1.73 m2),28例],另择同期健康体检者23例为对照组。采用液相免疫沉淀透射比浊法检测24 h尿微量白蛋白,计算尿白蛋白排泄率(UAE);酶联免疫吸附试验(ELISA)检测HIF-1α、VEGF。结果 DKD1组、DKD2组UAE、HIF-1α、VEGF水平均明显高于对照组(均P<0.05);DKD2组UAE水平与DKD1组差异无统计学意义,HIF-1α、VEGF水平高于DKD1组(均P<0

  1. Intelligent Speed Adaptation for involuntary drivers

    DEFF Research Database (Denmark)

    Agerholm, Niels; Tradisauskas, Nerius; Juhl, Jens

    2012-01-01

    The Danish Intelligent Speed Adaptation (ISA) trial ISA C included 26 commercial cars and 51 drivers a number of whom were involuntary. After a baseline period, ISA was activated for one year. The drivers should identify themselves with a personal key ID before driving. As well as being informative...

  2. Switched mode piezo-panel driver

    NARCIS (Netherlands)

    Slakhorst, R.J.

    2007-01-01

    Abstract The subject of this thesis is the design of a system which can drive piezo-panels. This system is called the piezo driver. The piezo-panels are used for an Active Noise Cancelling (ANC) system which is being developed to be used inside the cabin of airplanes. The piezo driver fills the gap

  3. Developing Linux kernel space device driver

    Institute of Scientific and Technical Information of China (English)

    Zheng Wei; Wang Qinruo; Wu Naiyou

    2003-01-01

    This thesis introduces how to develop kernel level device drivers on Linux platform in detail. On the basis of comparing proc file system with dev file system, we choose PCI devices and USB devices as instances to introduce the method of writing device drivers for character devices by using these two file systems.

  4. Should Passengers Be Responsible For Drunk Drivers?

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    In late September, China’s Ministry of Public Security expanded its nationwide campaign against drunk driving by releasing a document suggesting that passengers sharing a car with a drunk driver be punished together with the driver and that passengers who do not prevent drunk driving be fined.

  5. Frictional Dermatosis in a Courier Driver

    Directory of Open Access Journals (Sweden)

    Uwe Wollina

    2017-07-01

    Full Text Available Frictional hypermelanosis is an uncommon finding in Caucasians. We report the unusual case of 56-year-old male courier driver who developed linear and patchy hypermelanosis of the back caused by the driver's seat. Histology has included other pathologies. Treatment of the asymptomatic hyper pigmentation was not warranted.

  6. Physics at an upgraded Fermilab proton driver

    Energy Technology Data Exchange (ETDEWEB)

    Geer, S.; /Fermilab

    2005-07-01

    In 2004 the Fermilab Long Range Planning Committee identified a new high intensity Proton Driver as an attractive option for the future, primarily motivated by the recent exciting developments in neutrino physics. Over the last few months a physics study has developed the physics case for the Fermilab Proton Driver. The potential physics opportunities are discussed.

  7. Physics at an upgraded Fermilab proton driver

    Energy Technology Data Exchange (ETDEWEB)

    Geer, S.; /Fermilab

    2005-07-01

    In 2004 the Fermilab Long Range Planning Committee identified a new high intensity Proton Driver as an attractive option for the future, primarily motivated by the recent exciting developments in neutrino physics. Over the last few months a physics study has developed the physics case for the Fermilab Proton Driver. The potential physics opportunities are discussed.

  8. 49 CFR 177.816 - Driver training.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Driver training. 177.816 Section 177.816... Information and Regulations § 177.816 Driver training. (a) In addition to the training requirements of § 177... employee who will operate a motor vehicle has been trained in the applicable requirements of 49 CFR...

  9. Displaceable Gear Torque Controlled Driver

    Science.gov (United States)

    Cook, Joseph S., Jr. (Inventor)

    1997-01-01

    Methods and apparatus are provided for a torque driver including a displaceable gear to limit torque transfer to a fastener at a precisely controlled torque limit. A biasing assembly biases a first gear into engagement with a second gear for torque transfer between the first and second gear. The biasing assembly includes a pressurized cylinder controlled at a constant pressure that corresponds to a torque limit. A calibrated gage and valve is used to set the desired torque limit. One or more coiled output linkages connect the first gear with the fastener adaptor which may be a socket for a nut. A gear tooth profile provides a separation force that overcomes the bias to limit torque at the desired torque limit. Multiple fasteners may be rotated simultaneously to a desired torque limit if additional output spur gears are provided. The torque limit is adjustable and may be different for fasteners within the same fastener configuration.

  10. The drivers of plant diversity

    DEFF Research Database (Denmark)

    Jensen, Kristine Engemann

    In this thesis we use a “big data” approach to describe and explain large-scale patterns of plant diversity. The botanical data used for the six papers come from three different databases covering the New World, North America, and Europe respectively. The data on plant distributions were combined...... and beta diversity over time for woody forest communities in North America, using a 20 year forest plot dataset from the United States Department of Agriculture Forest Inventory and Analysis program. To assess functional diversity, we combined the plot data with data on four functional traits. Over time...... with environmental data on climate, soil, topography, and disturbance to identify the drivers of macroecological plant diversity patterns. Unless otherwise stated, the botanical data used in the papers come from the Botanical Information and Ecology Network. Paper I describes how we compiled a new plant growth form...

  11. BDC 500 branch driver controller

    CERN Document Server

    Dijksman, A

    1981-01-01

    This processor has been designed for very fast data acquisition and date pre-processing. The dataway and branch highway speeds have been optimized for approximately 1.5 mu sec. The internal processor cycle is approximately 0.8 mu sec. The standard version contains the following functions (slots): crate controller type A1; branch highway driver including terminator; serial I/O port (TTY, VDU); 24 bit ALU and 24 bit program counter; 16 bit memory address counter and 4 word stack; 4k bit memory for program and/or data; battery backup for the memory; CNAFD and crate LAM display; request/grant logic for time- sharing operation of several BDCs. The free slots can be equipped with e.g. extra RAM, computer interfaces, hardware multiplier/dividers, etc. (0 refs).

  12. TCDD induces the hypoxia-inducible factor (HIF)-1α regulatory pathway in human trophoblastic JAR cells.

    Science.gov (United States)

    Liao, Tien-Ling; Chen, Su-Chee; Tzeng, Chii-Reuy; Kao, Shu-Huei

    2014-09-30

    The exposure to dioxin can compromise pregnancy outcomes and increase the risk of preterm births. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) has been demonstrated to induce placental hypoxia at the end of pregnancy in a rat model, and hypoxia has been suggested to be the cause of abnormal trophoblast differentiation and placental insufficiency syndromes. In this study, we demonstrate that the non-hypoxic stimulation of human trophoblastic cells by TCDD strongly increased hypoxia inducible factor-1 alpha (HIF-1α) stabilization. TCDD exposure induced the generation of reactive oxygen species (ROS) and nitric oxide. TCDD-induced HIF-1α stabilization and Akt phosphorylation was inhibited by pretreatment with wortmannin (a phosphatidylinositol 3-kinase (PI3K) inhibitor) or N-acetylcysteine (a ROS scavenger). The augmented HIF-1α stabilization by TCDD occurred via the ROS-dependent activation of the PI3K/Akt pathway. Additionally, a significant increase in invasion and metallomatrix protease-9 activity was found in TCDD-treated cells. The gene expression of vascular endothelial growth factor and placental growth factor was induced upon TCDD stimulation, whereas the protein levels of peroxisome proliferator-activated receptor γ (PPARγ), PPARγ coactivator-1α, mitochondrial transcription factor, and uncoupling protein 2 were decreased. Our results indicate that an activated HIF-1α pathway, elicited oxidative stress, and induced metabolic stress contribute to TCDD-induced trophoblastic toxicity. These findings may provide molecular insight into the TCDD-induced impairment of trophoblast function and placental development.

  13. Pien Tze Huang Inhibits Hypoxia-Induced Angiogenesis via HIF-1α/VEGF-A Pathway in Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Hongwei Chen

    2015-01-01

    Full Text Available Hypoxia-induced angiogenesis plays an important role in the development and metastasis of solid tumors and is highly regulated by HIF-1α/VEGF-A pathway. Therefore, inhibiting tumor angiogenesis via suppression of HIF-1α/VEGF-A signaling represents a promising strategy for anticancer treatment. As a traditional Chinese medicine formula, Pien Tze Huang (PZH has long been used as a folk remedy for cancer in China and Southeast Asia. Previously, we reported that PZH inhibits colorectal cancer (CRC growth both in vivo and in vitro. To elucidate the antitumor mechanisms of PZH, in the present study we used human umbilical vein endothelial cells (HUVEC and colorectal carcinoma HCT-8 cells to evaluate the effects of PZH on hypoxia-induced angiogenesis and investigated the underlying molecular mechanisms. We found that PZH could inhibit hypoxia-induced migration and tube formation of HUVEC cells in a dose-dependent manner, although the low concentrations of PZH had no effect on HUVEC viability. Moreover, PZH inhibited hypoxia-induced activation of HIF-1α signaling and the expression of VEGF-A and/or VEGFR2 in both HCT-8 and HUVEC cells. Collectively, our findings suggest that PZH can inhibit hypoxia-induced tumor angiogenesis via suppression of HIF-1α/VEGF-A pathway.

  14. PML promotes metastasis of triple-negative breast cancer through transcriptional regulation of HIF1A target genes.

    Science.gov (United States)

    Ponente, Manfredi; Campanini, Letizia; Cuttano, Roberto; Piunti, Andrea; Delledonne, Giacomo A; Coltella, Nadia; Valsecchi, Roberta; Villa, Alessandra; Cavallaro, Ugo; Pattini, Linda; Doglioni, Claudio; Bernardi, Rosa

    2017-02-23

    Elucidating the molecular basis of tumor metastasis is pivotal for eradicating cancer-related mortality. Triple-negative breast cancer (TNBC) encompasses a class of aggressive tumors characterized by high rates of recurrence and metastasis, as well as poor overall survival. Here, we find that the promyelocytic leukemia protein PML exerts a prometastatic function in TNBC that can be targeted by arsenic trioxide. We found that, in TNBC patients, constitutive HIF1A activity induces high expression of PML, along with a number of HIF1A target genes that promote metastasis at multiple levels. Intriguingly, PML controls the expression of these genes by binding to their regulatory regions along with HIF1A. This mechanism is specific to TNBC cells and does not occur in other subtypes of breast cancer where PML and prometastatic HIF1A target genes are underexpressed. As a consequence, PML promotes cell migration, invasion, and metastasis in TNBC cell and mouse models. Notably, pharmacological inhibition of PML with arsenic trioxide, a PML-degrading agent used to treat promyelocytic leukemia patients, delays tumor growth, impairs TNBC metastasis, and cooperates with chemotherapy by preventing metastatic dissemination. In conclusion, we report identification of a prometastatic pathway in TNBC and suggest clinical development toward the use of arsenic trioxide for TNBC patients.

  15. Effects of HIF-1 inhibition by chetomin on hypoxia-related transcription and radiosensitivity in HT 1080 human fibrosarcoma cells

    Directory of Open Access Journals (Sweden)

    Baier Kurt

    2007-11-01

    Full Text Available Abstract Background Hypoxia-inducible factor-1 (HIF-1 overexpression has been linked to tumor progression and poor prognosis. We investigated whether targeting of HIF-1 using chetomin, a disrupter of the interaction of HIF-1 with the transcriptional coactivator p300, influences the radiosensitivity of hypoxic HT 1080 human fibrosarcoma cells. Methods Optimal dose of chetomin was determined by EGFP-HRE gene reporter assay in stably transfected HT 1080 cells. Cells were assayed for expression of the hypoxia-inducible genes carbonic anhydrase 9 (CA9 and vascular endothelial growth factor (VEGF by RT-PCR and for clonogenic survival after irradiation with 2, 5 or 10 Gy, under normoxic or hypoxic (0.1% O2, 12 h conditions in the presence or absence of chetomin (150 nM, 12 h, pre-treatment of 4 h. Results Chetomin treatment significantly reduced CA9 and VEGF mRNA expression in hypoxic cells to 44.4 ± 7.2% and 39.6 ± 16.0%, respectively, of untreated hypoxic controls. Chetomin clearly reduced the modified oxygen enhancement ratio (OER' compared to untreated cells, from 2.02 to 1.27, from 1.86 to 1.22 and from 1.49 to 1.06 at the 50%, 37% and 10% clonogenic survival levels, respectively. Conclusion HIF-1 inhibition by chetomin effectively reduces hypoxia-dependent transcription and radiosensitizes hypoxic HT 1080 human fibrosarcoma cells in vitro.

  16. Ursolic acid sensitized colon cancer cells to chemotherapy under hypoxia by inhibiting MDR1 through HIF-1α*

    Science.gov (United States)

    Shan, Jian-zhen; Xuan, Yan-yan; Zhang, Qi; Huang, Jian-jin

    2016-01-01

    Objective: To explore the efficacy of ursolic acid in sensitizing colon cancer cells to chemotherapy under hypoxia and its underlying mechanisms. Methods: Three colon cancer cell lines (RKO, LoVo, and SW480) were used as in vitro models. 5-Fluorouracil (5-FU) and oxaliplatin were used as chemotherapeutic drugs. Cell viability and apoptosis were tested to evaluate the sensitivity of colon cancer cells to chemotherapy. The transcription and expression levels of hypoxia-inducible factor-1α (HIF-1α), multidrug resistance gene 1 (MDR1), and vascular endothelial growth factors (VEGF) were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) and immunoblotting. Cycloheximide and MG132 were used to inhibit protein synthesis and degradation, respectively. In vitro tube formation assay was used to evaluate angiogenesis. Results: We demonstrated the chemosensitizing effects of ursolic acid with 5-FU and oxaliplatin in three colon cancer cell lines under hypoxia. This effect was correlated to its inhibition of MDR1 through HIF-1α. Moreover, ursolic acid was capable of inhibiting HIF-1α accumulation with little effects on its constitutional expression in normoxia. In addition, ursolic acid also down-regulated VEGF and inhibited tumor angiogenesis. Conclusions: Ursolic acid exerted chemosensitizing effects in colon cancer cells under hypoxia by inhibiting HIF-1α accumulation and the subsequent expression of the MDR1 and VEGF. PMID:27604859

  17. Proinflammatory signal suppresses proliferation and shifts macrophage metabolism from Myc-dependent to HIF1α-dependent.

    Science.gov (United States)

    Liu, Lingling; Lu, Yun; Martinez, Jennifer; Bi, Yujing; Lian, Gaojian; Wang, Tingting; Milasta, Sandra; Wang, Jian; Yang, Mao; Liu, Guangwei; Green, Douglas R; Wang, Ruoning

    2016-02-01

    As a phenotypically plastic cellular population, macrophages change their physiology in response to environmental signals. Emerging evidence suggests that macrophages are capable of tightly coordinating their metabolic programs to adjust their immunological and bioenergetic functional properties, as needed. Upon mitogenic stimulation, quiescent macrophages enter the cell cycle, increasing their bioenergetic and biosynthetic activity to meet the demands of cell growth. Proinflammatory stimulation, however, suppresses cell proliferation, while maintaining a heightened metabolic activity imposed by the production of bactericidal factors. Here, we report that the mitogenic stimulus, colony-stimulating factor 1 (CSF-1), engages a myelocytomatosis viral oncogen (Myc)-dependent transcriptional program that is responsible for cell cycle entry and the up-regulation of glucose and glutamine catabolism in bone marrow-derived macrophages (BMDMs). However, the proinflammatory stimulus, lipopolysaccharide (LPS), suppresses Myc expression and cell proliferation and engages a hypoxia-inducible factor alpha (HIF1α)-dependent transcriptional program that is responsible for heightened glycolysis. The acute deletion of Myc or HIF1α selectively impaired the CSF-1- or LPS-driven metabolic activities in BMDM, respectively. Finally, inhibition of glycolysis by 2-deoxyglucose (2-DG) or genetic deletion of HIF1α suppressed LPS-induced inflammation in vivo. Our studies indicate that a switch from a Myc-dependent to a HIF1α-dependent transcriptional program may regulate the robust bioenergetic support for an inflammatory response, while sparing Myc-dependent proliferation.

  18. Gamma rays induce a p53-independent mitochondrial biogenesis that is counter-regulated by HIF1α.

    Science.gov (United States)

    Bartoletti-Stella, A; Mariani, E; Kurelac, I; Maresca, A; Caratozzolo, M F; Iommarini, L; Carelli, V; Eusebi, L H; Guido, A; Cenacchi, G; Fuccio, L; Rugolo, M; Tullo, A; Porcelli, A M; Gasparre, G

    2013-06-13

    Mitochondrial biogenesis is an orchestrated process that presides to the regulation of the organelles homeostasis within a cell. We show that γ-rays, at doses commonly used in the radiation therapy for cancer treatment, induce an increase in mitochondrial mass and function, in response to a genotoxic stress that pushes cells into senescence, in the presence of a functional p53. Although the main effector of the response to γ-rays is the p53-p21 axis, we demonstrated that mitochondrial biogenesis is only indirectly regulated by p53, whose activation triggers a murine double minute 2 (MDM2)-mediated hypoxia-inducible factor 1α (HIF1α) degradation, leading to the release of peroxisome-proliferator activated receptor gamma co-activator 1β inhibition by HIF1α, thus promoting mitochondrial biogenesis. Mimicking hypoxia by HIF1α stabilization, in fact, blunts the mitochondrial response to γ-rays as well as the induction of p21-mediated cell senescence, indicating prevalence of the hypoxic over the genotoxic response. Finally, we also show in vivo that post-radiotherapy mitochondrial DNA copy number increase well correlates with lack of HIF1α increase in the tissue, concluding this may be a useful molecular tool to infer the trigger of a hypoxic response during radiotherapy, which may lead to failure of activation of cell senescence.

  19. HIF-1α induces the epithelial-mesenchymal transition in gastric cancer stem cells through the Snail pathway.

    Science.gov (United States)

    Yang, Shi-Wei; Zhang, Zhi-Gang; Hao, Ying-Xue; Zhao, Yong-Liang; Qian, Feng; Shi, Yan; Li, Ping-Ang; Liu, Chun-Yang; Yu, Pei-Wu

    2017-02-07

    Substantial evidence suggests that the epithelial-mesenchymal transition (EMT) phenotype is associated with the invasive characteristics of cancer stem cells (CSCs),which possess an EMT phenotype that may predominate in tumor invasion and metastasis. However, the mechanisms for the generation and regulation of these CSCs have not been clearly defined. As hypoxia and EMT-related factors may have important functions in EMT-like CSCs, the aim of this study was to investigate the effects of hypoxia on these cells. CSCs were established from the gastric cancer cell lines MGC-803 and SGC7901, and the relationship between hypoxia and EMT-like CSCs was investigated in gastric cancer. After hypoxia treatment, some gastric CSCs exhibited a marked increase in hypoxia-inducible factor-1α (HIF-1α)expression and increased migration and invasion capabilities compared with the normoxic control. These CSCs were defined by activation of the mesenchymal cell marker Vimentin and by inhibition of the epithelial cell marker E-cadherin. Our analyses also show that HIF-1α was responsible for activating EMT via increased expression of the transcription factor Snail in gastric CSCs. Moreover, inhibition of Snail by shRNA reduced HIF-1α-induced EMT in gastric CSCs. The results demonstrated that hypoxia-induced EMT-like CSCs rely on HIF-1αto activate Snail, which may result in recurrence and metastasis of gastric cancer.

  20. PML promotes metastasis of triple-negative breast cancer through transcriptional regulation of HIF1A target genes

    Science.gov (United States)

    Ponente, Manfredi; Campanini, Letizia; Cuttano, Roberto; Piunti, Andrea; Delledonne, Giacomo A.; Coltella, Nadia; Valsecchi, Roberta; Villa, Alessandra

    2017-01-01

    Elucidating the molecular basis of tumor metastasis is pivotal for eradicating cancer-related mortality. Triple-negative breast cancer (TNBC) encompasses a class of aggressive tumors characterized by high rates of recurrence and metastasis, as well as poor overall survival. Here, we find that the promyelocytic leukemia protein PML exerts a prometastatic function in TNBC that can be targeted by arsenic trioxide. We found that, in TNBC patients, constitutive HIF1A activity induces high expression of PML, along with a number of HIF1A target genes that promote metastasis at multiple levels. Intriguingly, PML controls the expression of these genes by binding to their regulatory regions along with HIF1A. This mechanism is specific to TNBC cells and does not occur in other subtypes of breast cancer where PML and prometastatic HIF1A target genes are underexpressed. As a consequence, PML promotes cell migration, invasion, and metastasis in TNBC cell and mouse models. Notably, pharmacological inhibition of PML with arsenic trioxide, a PML-degrading agent used to treat promyelocytic leukemia patients, delays tumor growth, impairs TNBC metastasis, and cooperates with chemotherapy by preventing metastatic dissemination. In conclusion, we report identification of a prometastatic pathway in TNBC and suggest clinical development toward the use of arsenic trioxide for TNBC patients. PMID:28239645

  1. HIF1α is a regulator of hematopoietic progenitor and stem cell development in hypoxic sites of the mouse embryo

    Directory of Open Access Journals (Sweden)

    Parisa Imanirad

    2014-01-01

    Full Text Available Hypoxia affects many physiologic processes during early stages of mammalian ontogeny, particularly placental and vascular development. In the adult, the hypoxic bone marrow microenvironment plays a role in regulating hematopoietic stem cell (HSC function. HSCs are generated from the major vasculature of the embryo, but whether the hypoxic response affects the generation of these HSCs is as yet unknown. Here we examined whether Hypoxia Inducible Factor1-alpha (HIF1α, a key modulator of the response to hypoxia, is essential for HSC development. We found hypoxic cells in embryonic tissues that generate and expand hematopoietic cells (aorta, placenta and fetal liver, and specifically aortic endothelial and hematopoietic cluster cells. A Cre/loxP conditional knockout (cKO approach was taken to delete HIF1α in Vascular Endothelial-Cadherin expressing endothelial cells, the precursors to definitive hematopoietic cells. Functional assays show that HSC and hematopoietic progenitor cells (HPCs are significantly reduced in cKO aorta and placenta. Moreover, decreases in phenotypic aortic hematopoietic cluster cells in cKO embryos indicate that HIF1α is necessary for generation and/or expansion of HPCs and HSCs. cKO adult BM HSCs are also affected under transplantation conditions. Thus, HIF1α is a regulator of HSC generation and function beginning at the earliest embryonic stages.

  2. Visual behaviour analysis and driver cognitive model

    Energy Technology Data Exchange (ETDEWEB)

    Baujon, J.; Basset, M.; Gissinger, G.L. [Mulhouse Univ., (France). MIPS/MIAM Lab.

    2001-07-01

    Recent studies on driver behaviour have shown that perception - mainly visual but also proprioceptive perception - plays a key role in the ''driver-vehicle-road'' system and so considerably affects the driver's decision making. Within the framework of the behaviour analysis and studies low-cost system (BASIL), this paper presents a correlative, qualitative and quantitative study, comparing the information given by visual perception and by the trajectory followed. This information will help to obtain a cognitive model of the Rasmussen type according to different driver classes. Many experiments in real driving situations have been carried out for different driver classes and for a given trajectory profile, using a test vehicle and innovative, specially designed, real-time tools, such as the vision system or the positioning module. (orig.)

  3. Stability analysis of automobile driver steering control

    Science.gov (United States)

    Allen, R. W.

    1981-01-01

    In steering an automobile, the driver must basically control the direction of the car's trajectory (heading angle) and the lateral deviation of the car relative to a delineated pathway. A previously published linear control model of driver steering behavior which is analyzed from a stability point of view is considered. A simple approximate expression for a stability parameter, phase margin, is derived in terms of various driver and vehicle control parameters, and boundaries for stability are discussed. A field test study is reviewed that includes the measurement of driver steering control parameters. Phase margins derived for a range of vehicle characteristics are found to be generally consistent with known adaptive properties of the human operator. The implications of these results are discussed in terms of driver adaptive behavior.

  4. Reducing risky driver behaviour through the implementation of a driver risk management system

    Directory of Open Access Journals (Sweden)

    Rose Luke

    2014-03-01

    Full Text Available South Africa has one of the highest incidences of road accidents in the world. Most accidents are avoidable and are caused by driver behaviour and errors. The purpose of this article was to identify the riskiest driver behaviours in commercial fleets in South Africa, to determine the business impact of such behaviour, to establish a framework for the management of risky driver behaviour and to test the framework by applying a leading commercial driver behaviour management system as a case study. The case study comprised three South African commercial fleets. Using data from these fleets, critical incident triangles were used to determine the ratio data of risky driver behaviour to near-collisions and collisions. Based on managing the riskiest driver behaviours as causes of more serious incidents and accidents, the results indicated that through the implementation of an effective driver risk management system, risky incidents were significantly reduced.

  5. Modeling aggressive driver behavior at unsignalized intersections.

    Science.gov (United States)

    Kaysi, Isam A; Abbany, Ali S

    2007-07-01

    The processing of vehicles at unsignalized intersections is a complex and highly interactive process, whereby each driver makes individual decisions about when, where, and how to complete the required maneuver, subject to his perceptions of distances, velocities, and own car's performance. Typically, the performance of priority-unsignalized intersections has been modeled with probabilistic approaches that consider the distribution of gaps in the major-traffic stream and their acceptance by the drivers of minor street vehicles based on the driver's "critical gap". This paper investigates the aggressive behavior of minor street vehicles at intersections that are priority-unsignalized but operate with little respect of control measures. The objective is to formulate a behavioral model that predicts the probability that a driver performs an aggressive maneuver as a function of a set of driver and traffic attributes. Parameters that were tested and modeled include driver characteristics (gender and age), car characteristics (performance and model year), and traffic attributes (number of rejected gaps, total waiting time at head of queue, and major-traffic speed). Binary probit models are developed and tested, based on a collected data set from an unsignalized intersection in the city of Beirut, to determine which of the studied variables are statistically significant in determining the aggressiveness of a specific driver. Primary conclusions reveal that age, car performance, and average speed on the major road are the major determinants of aggressive behavior. Another striking conclusion is that the total waiting time of the driver while waiting for an acceptable gap is of little significance in incurring the "forcing" behavior. The obtained model is incorporated in a simple simulation framework that reflects driver behavior and traffic stream interactions in estimating delay and conflict measures at unsignalized intersections. The simulation results were then compared

  6. Driver drowsiness detection using multimodal sensor fusion

    Science.gov (United States)

    Andreeva, Elena O.; Aarabi, Parham; Philiastides, Marios G.; Mohajer, Keyvan; Emami, Majid

    2004-04-01

    This paper proposes a multi-modal sensor fusion algorithm for the estimation of driver drowsiness. Driver sleepiness is believed to be responsible for more than 30% of passenger car accidents and for 4% of all accident fatalities. In commercial vehicles, drowsiness is blamed for 58% of single truck accidents and 31% of commercial truck driver fatalities. This work proposes an innovative automatic sleep-onset detection system. Using multiple sensors, the driver"s body is studied as a mechanical structure of springs and dampeners. The sleep-detection system consists of highly sensitive triple-axial accelerometers to monitor the driver"s upper body in 3-D. The subject is modeled as a linear time-variant (LTV) system. An LMS adaptive filter estimation algorithm generates the transfer function (i.e. weight coefficients) for this LTV system. Separate coefficients are generated for the awake and asleep states of the subject. These coefficients are then used to train a neural network. Once trained, the neural network classifies the condition of the driver as either awake or asleep. The system has been tested on a total of 8 subjects. The tests were conducted on sleep-deprived individuals for the sleep state and on fully awake individuals for the awake state. When trained and tested on the same subject, the system detected sleep and awake states of the driver with a success rate of 95%. When the system was trained on three subjects and then retested on a fourth "unseen" subject, the classification rate dropped to 90%. Furthermore, it was attempted to correlate driver posture and sleepiness by observing how car vibrations propagate through a person"s body. Eight additional subjects were studied for this purpose. The results obtained in this experiment proved inconclusive which was attributed to significant differences in the individual habitual postures.

  7. The angiogenic response to PLL-g-PEG-mediated HIF-1α plasmid DNA delivery in healthy and diabetic rats.

    Science.gov (United States)

    Thiersch, Markus; Rimann, Markus; Panagiotopoulou, Vasiliki; Öztürk, Ece; Biedermann, Thomas; Textor, Marcus; Lühmann, Tessa C; Hall, Heike

    2013-05-01

    Impaired angiogenesis is a major clinical problem and affects wound healing especially in diabetic patients. Improving angiogenesis is a reasonable strategy to increase diabetes-impaired wound healing. Recently, our lab described a system of transient gene expression due to pegylated poly-l-lysine (PLL-g-PEG) polymer-mediated plasmid DNA delivery in vitro. Here we synthesized peptide-modified PLL-g-PEG polymers with two functionalities, characterized them in vitro and utilized them in vivo via a fibrin-based delivery matrix to induce dermal wound angiogenesis in diabetic rats. The two peptides were 1) a TG-peptide to covalently bind these nanocondensates to the fibrin matrix (TG-peptide) for a sustained release and 2) a polyR peptide to improve cellular uptake of these nanocondensates. In order to induce angiogenesis in vivo we condensed modified and non-modified polymers with plasmid DNA encoding a truncated form of the therapeutic candidate gene hypoxia-inducible transcription factor 1α (HIF-1α). HIF-1α is the primarily oxygen-dependent regulated subunit of the heterodimeric transcription factor HIF-1, which controls angiogenesis among other physiological pathways. The truncated form of HIF-1α lacks the oxygen-dependent degradation domain (ODD) and therefore escapes degradation under normoxic conditions. PLL-g-PEG polymer-mediated HIF-1α-ΔODD plasmid DNA delivery was found to lead to a transiently induced gene expression of angiogenesis-related genes Acta2 and Pecam1 as well as the HIF-1α target gene Vegf in vivo. Furthermore, HIF-1α gene delivery was shown to enhance the number endothelial cells and smooth muscle cells - precursors for mature blood vessels - during wound healing. We show that - depending on the selection of the therapeutic target gene - PLL-g-PEG nanocondensates are a promising alternative to viral DNA delivery approaches, which might pose a risk to health. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Decursin reduce radio-resistance of hypoxic regions under the proton beam therapy by induced HIF-1α degradation

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Myung Hwan; Kim, Kye Ryung [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2013-10-15

    Protons induce cancer-cell apoptosis in vitro and block blood vessel formation in vivo through the generation of reactive oxygen species (ROS). The fact that proton severely inhibits blood vessel development in zebrafish embryos suggests a higher sensitivity of vascular endothelial cells to proton beam. Decursin, a coumarin compound, was originally isolated from Angelica gigas Nakai (Dang Gui). A. gigas root has been traditionally used in Korean folk medicine for the treatment of anemia and other common diseases. In previous reports, decursin was reported to exhibit anti-tumor activity against various cancer cells and to inhibit the activities of the androgen and androgen-receptor (AR) signaling pathway in prostate cancer, induction of cell cycle arrest and apoptosis in various cancer cells, such as prostate, breast, bladder, and colon cancer cells. Decursin also inhibits VEGF-induced angiogenesis through the suppression of the VEGFR-2-signaling pathway. However, the mechanism of decursin mediates change of HIF-1α activities is not clear. In this research, we identified regulations of the HIF-1α and the anti-angiogenesis effects of decursin in proton-beam-irradiated human lung cancer, prostate cancer and Hepatic cancer cells. We investigated the underlying mechanisms of positive effects of protonbeam-induced anti-angiogenesis. Our data indicate that the groups co-treated with decursin and a proton-beam had significant reduced HIF-1α activity compared with the groups treated with only a proton beam under the hypoxic condition caused by DFX(desferrioxamine). Decursin was found to induced HIF-1α degradation. Therefore, we suggest that decursin may be a potential candidate for use as a sensitizer for proton-beaminduced cell apoptosis. Here we have shown that decursin successfully reduced HIF-1α stability under hypoxic condition by induced desferrioxamine. We showed novel candidates for anti-angiogenic compound, decursin, leading to complete inhibition of radio

  9. Transcriptional activation of hypoxia-inducible factor-1 (HIF-1) in myeloid cells promotes angiogenesis through VEGF and S100A8.

    Science.gov (United States)

    Ahn, G-One; Seita, Jun; Hong, Beom-Ju; Kim, Young-Eun; Bok, Seoyeon; Lee, Chan-Ju; Kim, Kwang Soon; Lee, Jerry C; Leeper, Nicholas J; Cooke, John P; Kim, Hak Jae; Kim, Il Han; Weissman, Irving L; Brown, J Martin

    2014-02-18

    Emerging evidence indicates that myeloid cells are essential for promoting new blood vessel formation by secreting various angiogenic factors. Given that hypoxia-inducible factor (HIF) is a critical regulator for angiogenesis, we questioned whether HIF in myeloid cells also plays a role in promoting angiogenesis. To address this question, we generated a unique strain of myeloid-specific knockout mice targeting HIF pathways using human S100A8 as a myeloid-specific promoter. We observed that mutant mice where HIF-1 is transcriptionally activated in myeloid cells (by deletion of the von Hippel-Lindau gene) resulted in erythema, enhanced neovascularization in matrigel plugs, and increased production of vascular endothelial growth factor (VEGF) in the bone marrow, all of which were completely abrogated by either genetic or pharmacological inactivation of HIF-1. We further found that monocytes were the major effector producing VEGF and S100A8 proteins driving neovascularization in matrigel. Moreover, by using a mouse model of hindlimb ischemia we observed significantly improved blood flow in mice intramuscularly injected with HIF-1-activated monocytes. This study therefore demonstrates that HIF-1 activation in myeloid cells promotes angiogenesis through VEGF and S100A8 and that this may become an attractive therapeutic strategy to treat diseases with vascular defects.

  10. Structural Insights into the Association of Hif1 with Histones H2A-H2B Dimer and H3-H4 Tetramer.

    Science.gov (United States)

    Zhang, Mengying; Liu, Hejun; Gao, Yongxiang; Zhu, Zhongliang; Chen, Zijun; Zheng, Peiyi; Xue, Lu; Li, Jixi; Teng, Maikun; Niu, Liwen

    2016-10-04

    Histone chaperones are critical for guiding specific post-transcriptional modifications of histones, safeguarding the histone deposition (or disassociation) of nucleosome (dis)assembly, and regulating chromatin structures to change gene activities. HAT1-interacting factor 1 (Hif1) has been reported to be an H3-H4 chaperone and to be involved in telomeric silencing and nucleosome (dis)assembly. However, the structural basis for the interaction of Hif1 with histones remains unknown. Here, we report the complex structure of Hif1 binding to H2A-H2B for uncovering the chaperone specificities of Hif1 on binding to both the H2A-H2B dimer and the H3-H4 tetramer. Our findings reveal that Hif1 interacts with the H2A-H2B dimer and the H3-H4 tetramer via distinct mechanisms, suggesting that Hif1 is a pivotal scaffold on alternate binding of H2A-H2B and H3-H4. These specificities are conserved features of the Sim3-Hif1-NASP interrupted tetratricopeptide repeat proteins, which provide clues for investigating their potential roles in nucleosome (dis)assembly. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. HIF-1α transgenic bone marrow cells can promote tissue repair in cases of corticosteroid-induced osteonecrosis of the femoral head in rabbits.

    Directory of Open Access Journals (Sweden)

    Hao Ding

    Full Text Available Although corticosteroid-induced osteonecrosis of the femoral head (ONFH is common, the treatment for it remains limited and largely ineffective. We examined whether implantation of hypoxia inducible factor-1α (HIF-1α transgenic bone marrow cells (BMCs can promote the repair of the necrotic area of corticosteroid-induced ONFH. In this study, we confirmed that HIF-1α gene transfection could enhance mRNA expression of osteogenic genes in BMCs in vitro. Alkaline phosphatase activity assay and alizarin red-S staining indicated HIF-1α transgenic BMCs had enhanced osteogenic differentiation capacity in vitro. Furthermore, enzyme linked immunosorbent assay (ELISA for VEGF revealed HIF-1α transgenic BMCs secreted more VEGF as compared to normal BMCs. An experimental rabbit model of early-stage corticosteroid-induced ONFH was established and used for an evaluation of cytotherapy. Transplantation of HIF-1α transgenic BMCs dramatically improved the bone regeneration of the necrotic area of the femoral head. The number and volume of blood vessel were significantly increased in the necrotic area of the femoral head compared to the control groups. These results support HIF-1α transgenic BMCs have enhanced osteogenic and angiogenic activity in vitro and in vivo. Transplantation of HIF-1α transgenic BMCs can potentially promote the repair of the necrotic area of corticosteroid-induced ONFH.

  12. 15-Deoxy-Delta(12,14)-prostaglandin-J(2) reveals a new pVHL-independent, lysosomal-dependent mechanism of HIF-1alpha degradation.

    Science.gov (United States)

    Olmos, Gemma; Arenas, María I; Bienes, Raquel; Calzada, María Jose; Aragonés, Julián; Garcia-Bermejo, Maria Laura; Landazuri, Manuel O; Lucio-Cazaña, Javier

    2009-07-01

    Hypoxia-inducible factor-1alpha (HIF-1alpha) protein is degraded under normoxia by its association to von Hippel-Lindau protein (pVHL) and further proteasomal digestion. However, human renal cells HK-2 treated with 15-deoxy-Delta(12,14)-prostaglandin-J(2) (15d-PGJ(2)) accumulate HIF-1alpha in normoxic conditions. Thus, we aimed to investigate the mechanism involved in this accumulation. We found that 15d-PGJ(2) induced an over-accumulation of HIF-1alpha in RCC4 cells, which lack pVHL and in HK-2 cells treated with inhibitors of the pVHL-proteasome pathway. These results indicated that pVHL-proteasome-independent mechanisms are involved, and therefore we aimed to ascertain them. We have identified a new lysosomal-dependent mechanism of HIF-1alpha degradation as a target for 15d-PGJ(2) based on: (1) HIF-1alpha colocalized with the specific lysosomal marker Lamp-2a, (2) 15d-PGJ(2) inhibited the activity of cathepsin B, a lysosomal protease, and (3) inhibition of lysosomal activity did not result in over-accumulation of HIF-1alpha in 15d-PGJ(2)-treated cells. Therefore, expression of HIF-1alpha is also modulated by lysosomal degradation.

  13. HIF-1α inhibition by 2-methoxyestradiol induces cell death via activation of the mitochondrial apoptotic pathway in acute myeloid leukemia.

    Science.gov (United States)

    Zhe, Nana; Chen, Shuya; Zhou, Zhen; Liu, Ping; Lin, Xiaojing; Yu, Meisheng; Cheng, Bingqing; Zhang, Yaming; Wang, Jishi

    2016-06-02

    The bone marrow microenvironment plays an important role in the development and progression of AML. Leukemia stem cells are in a hypoxic condition, which induces the expression of HIF-1α. Aberrant activation of HIF-1α is implicated in the poor prognosis of patients with acute myeloid leukemia (AML). Herein, we investigated the expression of HIF-1α in AML and tested 2-methoxyestradiol (2ME2) as a candidate HIF-1α inhibitor for the treatment of AML. We found that HIF-1α was overexpressed in AML. HIF-1α suppression by 2ME2 significantly induced apoptosis of AML cells, and it outperformed traditional chemotherapy drugs such as cytarabine. At the same time, 2ME2 downregulated the transcriptional levels of VEGF, GLUT1 and HO-1 in cellular assays. Additionally, 2ME2 displayed antileukemia activity in bone marrow blasts from AML patients, but showed little effect on normal cells. 2ME2-induced activation of mitochondrial apoptotic pathway is mediated by reactive oxygen species (ROS), which decreased the slight effect of drug on normal cells. Our data show that supression of HIF-1α expression significantly reduced the survival of AML cell lines, suggesting that 2ME2 may represent a powerful therapeutic approach for patients with AML.

  14. Nanoparticle delivery of HIF1α siRNA combined with photodynamic therapy as a potential treatment strategy for head-and-neck cancer.

    Science.gov (United States)

    Chen, Wei-Hua; Lecaros, Rumwald Leo G; Tseng, Yu-Cheng; Huang, Leaf; Hsu, Yih-Chih

    2015-04-01

    Combination therapy has become a major strategy in cancer treatment. We used anisamide-targeted lipid-calcium-phosphate (LCP) nanoparticles to efficiently deliver HIF1α siRNA to the cytoplasm of sigma receptor-expressing SCC4 and SAS cells that were also subjected to photodynamic therapy (PDT). HIF1α siRNA nanoparticles effectively reduced HIF1α expression, increased cell death, and significantly inhibited cell growth following photosan-mediated photodynamic therapy in cultured cells. Intravenous injection of the same nanoparticles into human SCC4 or SAS xenografted mice likewise resulted in concentrated siRNA accumulation and reduced HIF1α expression in tumor tissues. When combined with photodynamic therapy, HIF1α siRNA nanoparticles enhanced the regression in tumor size resulting in a ~40% decrease in volume after 10 days. Combination therapy was found to be substantially more effective than either HIF1α siRNA or photodynamic therapy alone. Results from caspase-3, TUNEL, and CD31 marker studies support this conclusion. Our results show the potential use of LCP nanoparticles for efficient delivery of HIF1α siRNA into tumors as part of combination therapy along with PDT in the treatment of oral squamous cell carcinoma. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  15. Development of a Novel Anti-HIF-1α Screening System Coupled with Biochemical and Biological Validation for Rapidly Selecting Potent Anti-Cancer Compounds.

    Science.gov (United States)

    Lu, Yi; Madu, Chikezie; Masters, Jordan; Lu, Andrew; Li, Liyuan

    2014-01-01

    Breast cancer (BCa) is the most diagnosed cancer and the second leading cause of cancer death in the American women. Adaptation to the hypoxic environment seen in solid tumors is critical for tumor cell survival and growth. The activation of hypoxia inducible factor-1 alpha (HIF-1α), an important master transcriptional factor that is induced and stabilized by intratumoral hypoxia, stimulates a group of HIF-1α-regulated genes including vascular endothelial growth factor (VEGF), leading tumor cells towards malignant progression. Therefore, a promising therapeutic approach to cancer treatment is to target HIF-1α. The goal of this project was to develop and validate a screening system coupled with secondary screen/validation process that has the capability to screen large numbers of potential anti-cancer small-molecule compounds based on their anti-HIF-1α activities. Breast cancer MDA-231 cells were used as the model to select potent anti-HIF-1α compounds by their abilities to inhibit transactivation of a VEGF promoter fused to a luciferase reporter gene under hypoxia. Positive compounds were then validated by a series of assays that confirm compounds' anti-HIF-1α activities including measurement of HIF-1α downstream VEGF gene expression and angiogenic ability of BCa cells. Results of our pilot screening demonstrate that this prototype screening coupled with validation system can effectively select highly potent anti-HIF-1α agents from the compound library, suggesting that this prototype screen system has the potential to be developed into a high-throughput screen (HTS) coupled with automated validation process for the screening and identification of novel and effective anti-cancer drugs based on anti-HIF-1α mechanism.

  16. Erythropoietin inhibits HIF-1α expression via upregulation of PHD-2 transcription and translation in an in vitro model of hypoxia-ischemia.

    Science.gov (United States)

    Souvenir, Rhonda; Flores, Jerry J; Ostrowski, Robert P; Manaenko, Anatol; Duris, Kamil; Tang, Jiping

    2014-02-01

    Hypoxia inducible factor (HIF)-1α is the central transcriptional factor for the regulation of oxygen-associated genes in response to hypoxia. Erythropoietin (EPO), a hematopoietic growth factor, increases oxygen availability during hypoxia/ischemia and is associated with neuroprotection following hypoxia-ischemia in laboratory models of stroke. However, EPO has failed to translate in a clinical setting. Thus, it is critical to elucidate the key players in EPO-induced neuroprotection. Our preliminary studies have shown that EPO, as a downstream gene of HIF, inhibits HIF-1α in a dose-dependent manner in an in vitro model of hypoxia-ischemia. This study is designed to elucidate the primary mediator of EPO-induced HIF-1α inhibition and subsequent cell survival/neuroprotection. Oxygen and glucose deprivation (OGD) of nerve growth factor-differentiated rat pheochromocytoma (PC-12) cells were used to model hypoxia-ischemia in an in vitro environment. The profile of HIF-1α, HIF-2α and prolyl hydroxylase domain 2 (PHD-2) expression; HIF-1α and prolyl hydroxylase (PHD-2) mRNA levels; matrix metalloproteinase (MMP)-9; and cell death was evaluated in the presence and absence of either EPO or PHD-2 inhibitor during OGD. Our findings showed that EPO treatment resulted in an increase in PHD-2 transcription and translation, inhibition of HIF-1α expression, reactive oxygen species formation, and MMP-9 activity, resulting in increased cell survival after OGD. We also observed that EPO-induced cell survival/neuroprotection was reversed by siRNA silencing of PHD-2. This led to the conclusion that PHD-2 is a key mediator of EPO-induced HIF-1α inhibition and subsequent neuroprotection in an in vitro model of hypoxia-ischemia.

  17. Pilot trial of EZN-2968, an antisense oligonucleotide inhibitor of hypoxia-inducible factor-1 alpha (HIF-1α), in patients with refractory solid tumors.

    Science.gov (United States)

    Jeong, Woondong; Rapisarda, Annamaria; Park, Sook Ryun; Kinders, Robert J; Chen, Alice; Melillo, Giovanni; Turkbey, Baris; Steinberg, Seth M; Choyke, Peter; Doroshow, James H; Kummar, Shivaani

    2014-02-01

    Hypoxia-inducible factor-1 (HIF-1) facilitates the adaptation of normal and tumor tissues to oxygen deprivation. HIF-1 is frequently overexpressed in cancer cells, where it is involved in the upregulation of many genes necessary for survival. EZN-2968 is an antisense oligodeoxynucleotide that specifically targets HIF-1α, one of the subunits of HIF-1. We conducted a trial of EZN-2968 in patients with refractory solid tumors to evaluate antitumor response and to measure modulation of HIF-1α mRNA and protein levels as well as HIF-1 target genes. Adult patients with refractory advanced solid tumors were administered EZN-2968 as a 2-h IV infusion at a dose of 18 mg/kg once a week for three consecutive weeks followed by 3-week off; in a 6-week cycle. Tumor biopsies and dynamic contrast enhanced MRI (DCE-MRI) were performed at baseline and after the third dose. Ten patients were enrolled, of whom all were evaluable for response; one patient with a duodenal neuroendocrine tumor had prolonged stabilization of disease (24 weeks). Reduction in HIF-1α mRNA levels compared to baseline was demonstrated in 4 of 6 patients with paired tumor biopsies. Reductions in levels of HIF-1α protein and mRNA levels of some target genes were observed in two patients. Quantitative analysis of DCE-MRI from two patients revealed changes in K (trans) and k ep. The trial was closed prematurely when the sponsor suspended development of this agent. This trial provides preliminary proof of concept for modulation of HIF-1α mRNA and protein expression and target genes in tumor biopsies following the administration of EZN-2968.

  18. Strong Expression of Hypoxia-Inducible Factor-1α (HIF-1α Is Associated with Axl Expression and Features of Aggressive Tumors in African Breast Cancer.

    Directory of Open Access Journals (Sweden)

    Hawa Nalwoga

    Full Text Available Inhibition of hypoxia-inducible factor (HIF and Axl receptor tyrosine kinase is being evaluated for targeted therapy in solid tumors. Both HIF-1α and Axl influence tumor growth and metastatic potential, and they have been linked to treatment failure in many cancers. However, there is a lack of reports on HIF-1α expression in African breast cancer, which has a poor prognosis, and novel treatment targets must therefore be established. Here, we aimed to evaluate HIF-1α in relation to Axl expression, angiogenesis markers, and other tumor characteristics in a series of African breast cancer.Using immunohistochemistry, we examined 261 invasive breast cancers on tissue microarrays for HIF-1α and Axl as well as several other markers, and a subset of 185 cases had information on VEGF (vascular endothelial growth factor expression, microvessel density (MVD, proliferating microvessel density (pMVD and vascular proliferation index (VPI for important comparisons.Strong HIF-1α expression was associated with increased Axl (p = 0.007, VEGF (p<0.0005, and p53 (p = 0.032 expression, as well as high tumor cell proliferation by Ki-67 (p = 0.006, and high tumor grade (p = 0.003. Tumors with strong HIF-1α expression had significantly higher MVD (p = 0.019 and higher pMVD (p = 0.027 than tumors with weak expression.High HIF-1α expression is significantly associated with Axl and VEGF expression, and with markers of poor prognosis in this series of breast cancer, suggesting HIF-1α and Axl as potential therapeutic targets in African breast cancer.

  19. Neuronal damage and functional deficits are ameliorated by inhibition of aquaporin and HIF1α after traumatic brain injury (TBI).

    Science.gov (United States)

    Shenaq, Mohammed; Kassem, Hassan; Peng, Changya; Schafer, Steven; Ding, Jamie Y; Fredrickson, Vance; Guthikonda, Murali; Kreipke, Christian W; Rafols, José A; Ding, Yuchuan

    2012-12-15

    The present study, using a rodent model of closed-head diffuse traumatic brain injury (TBI), investigated the role of dysregulated aquaporins (AQP) 4 and 9, as well as hypoxia inducible factor -1α(HIF-1α) on brain edema formation, neuronal injury, and functional deficits. TBI was induced in adult (400-425 g), male Sprague-Dawley rats using a modified Marmarou's head impact-acceleration device (450 g weight dropped from 2m height). Animals in each treatment group were administered intravenous anti-AQP4 or -AQP9 antibodies or 2-Methoxyestradiol (2ME2, an inhibitor of HIF-1α) 30 min after injury. At 24h post-TBI, animals (n=6 each group) were sacrificed to examine the extent of brain edema by water content, as well as protein expression of AQP and HIF-1α by Western immune-blotting. At 48-hours post-TBI, neuronal injury (n=8 each group) was assessed by FluoroJade (FJ) histochemistry. Spatial learning and memory deficits were evaluated by radial arm maze (n=8 each group) up to 21 days post-TBI. Compared to non-injured controls, significant (pTBI was associated with increases (p TBI animals, AQP or HIF-1α inhibition significantly (pTBI. Taken together, the present data supports a causal relation between HIF-AQP mediated cerebral edema, secondary neuronal injury, and tertiary behavioral deficits post-TBI. The data further suggests that upstream modulation of the molecular patho-trajectory effectively ameliorates both neuronal injury and behavioral deficits post-TBI.

  20. Mimicking hypoxia to treat anemia: HIF-stabilizer BAY 85-3934 (Molidustat stimulates erythropoietin production without hypertensive effects.

    Directory of Open Access Journals (Sweden)

    Ingo Flamme

    Full Text Available Oxygen sensing by hypoxia-inducible factor prolyl hydroxylases (HIF-PHs is the dominant regulatory mechanism of erythropoietin (EPO expression. In chronic kidney disease (CKD, impaired EPO expression causes anemia, which can be treated by supplementation with recombinant human EPO (rhEPO. However, treatment can result in rhEPO levels greatly exceeding the normal physiological range for endogenous EPO, and there is evidence that this contributes to hypertension in patients with CKD. Mimicking hypoxia by inhibiting HIF-PHs, thereby stabilizing HIF, is a novel treatment concept for restoring endogenous EPO production. HIF stabilization by oral administration of the HIF-PH inhibitor BAY 85-3934 (molidustat resulted in dose-dependent production of EPO in healthy Wistar rats and cynomolgus monkeys. In repeat oral dosing of BAY 85-3934, hemoglobin levels were increased compared with animals that received vehicle, while endogenous EPO remained within the normal physiological range. BAY 85-3934 therapy was also effective in the treatment of renal anemia in rats with impaired kidney function and, unlike treatment with rhEPO, resulted in normalization of hypertensive blood pressure in a rat model of CKD. Notably, unlike treatment with the antihypertensive enalapril, the blood pressure normalization was achieved without a compensatory activation of the renin-angiotensin system. Thus, BAY 85-3934 may provide an approach to the treatment of anemia in patients with CKD, without the increased risk of adverse cardiovascular effects seen for patients treated with rhEPO. Clinical studies are ongoing to investigate the effects of BAY 85-3934 therapy in patients with renal anemia.

  1. Correlation of HIF-2α, ABCG2 and OCT-4 with chemotherapy resistance in human gastric cancer

    Directory of Open Access Journals (Sweden)

    Hong-mei ZHANG

    2015-11-01

    Full Text Available Objective To investigate the correlation of HIF-2α, ABCG2 and OCT-4 with chemotherapy resistant gastric cancer in humans. Methods Fifty-two patients who were confirmed to have advanced gastric cancer with the aid of electronic endoscopy and pathology in the Department of Gastroenterology, Affiliated Hospital of Weifang Medical College, were enrolled in the study. According to the effect of FOL-FOX4 chemotherapy that these patients had experienced, they were divided into three groups: CR+PR (complete remission+partial remission group, SD (stable disease group and PD (progressive disease group. The expression levels of HIF-2α, ABCG2, and OCT-4 mRNA and protein were assessed in different groups by using RT-PCR and immunocytochemistry. Results Two patients achieved CR , 19 achieved PR , 25 showed SD, and 6 showed PD. In other words, CR+PR were seen in 21 patients (40.4%, SD in 25(48.1%, PD in 6(11.5%. In CR+PR group, the expression levels of HIF-2α, ABCG2 and OCT4 mRNA and protein were low, but the above mentioned expressions were significantly increased in SD group and PD group. The expression levels of HIF-2α, ABCG2 and Oct-4 mRNA and protein were highest in the PD group, lower in the SD group, and lowest in the CR + PR groups (all P<0.05. Conclusions The expression of the markers HIF-2α, ABCG2 and OCT4 in human tumor tissues is related to the effect of chemotherapy for gastric cancer. A high expression of tumor markers is perhaps the main reason for low efficacy of chemotherapy due to drug resistance. DOI: 10.11855/j.issn.0577-7402.2015.10.09

  2. Pilocarpine protects cobalt chloride-induced apoptosis of RGC-5 cells: involvement of muscarinic receptors and HIF-1 alpha pathway.

    Science.gov (United States)

    Zhu, Xu; Zhou, Wei; Cui, Yongyao; Zhu, Liang; Li, Juan; Feng, Xuemei; Shao, Biyun; Qi, Hong; Zheng, Jun; Wang, Hao; Chen, Hongzhuan

    2010-04-01

    The retina is the most metabolically active tissue in the human body and hypoxia-induced retinal ganglion cell (RGC) death has been implicated in glaucomatous optic neuropathy. The aim of this study is to determine whether muscarinic receptor agonist pilocarpine, a classic antiglaucoma drug, possesses neuroprotection against cobalt chloride (CoCl(2))-mimetic hypoxia-induced apoptosis of rat retinal ganglion cells (RGC-5 cells) and its underlying mechanisms. Cell viability was determined by Cell Counting Kit-8 assay and apoptosis was examined by annexin V and mitochondrial membrane potential (MMP) assays. Expressions of hypoxia-induced factor-1 alpha (HIF-1 alpha), p53, and BNIP3 were investigated by quantitative real-time PCR and western blot analysis. After treatment of 200 microM CoCl(2) for 24 h, RGC-5 cells showed a marked decrease of cell viability by approximately 30%, increased apoptosis rate and obvious decline in MMP, which could largely be reversed by the pretreatment of 1 microM pilocarpine mainly via the activation of muscarinic receptors. Meanwhile, pretreatment of 1 microM pilocarpine could significantly prevent CoCl(2)-induced HIF-1 alpha translocation from cytoplasm to nucleus and down-regulate the expression of HIF-1 alpha, p53, and BNIP3. These studies demonstrated that pilocarpine had effective protection against hypoxia-induced apoptosis in RGCs via muscarinic receptors and HIF-1 alpha pathway. The findings suggest that HIF-1 alpha pathway as a "master switch" may be used as a therapeutic target in the cholinergic treatment of glaucoma.

  3. Dual inhibition of plasminogen kringle 5 on angiogenesis and chemotaxis suppresses tumor metastasis by targeting HIF-1α pathway.

    Directory of Open Access Journals (Sweden)

    Wei-Bin Cai

    Full Text Available We had demonstrated that plasminogen kringle 5 (K5, a potent angiogenic inhibitor, inhibited retinal neovascularization and hepatocellular carcinoma growth by anti-angiogenesis. The current study investigated the effects and the underlying mechanisms of K5 on both tumor growth and spontaneous pulmonary metastasis in Lewis lung carcinoma (LLC implanted mouse model. Similarly, K5 could decrease expression of VEGF in LLC cells and grafted tissues and suppress tumor angiogenesis and growth. K5 had no direct effect on proliferation and apoptosis of LLC. However, K5 could significantly inhibit SDF-1α-induced chemotaxis movement of LLC cells and resulted in a great reduction of surface metastatic nodules and micrometastases in the lungs of LLC tumor-bearing mice. K5 also decreased expression of chemokine (C-X-C motif receptor 4 (CXCR4 in LLC cells and grafted tissues. Furthermore, K5 down-regulated SDF-1α expression in metastatic lung tissues of LLC-bearing mice. Therefore, K5 may suppress tumor pulmonary metastasis through inhibiting SDF-1α-CXCR4 chemotaxis movement and down-regulation of VEGF. Moreover, the role of hypoxia inducible factor-1α (HIF-1α, a crucial transcriptional factor for both VEGF and CXCR4 expression, was evaluated. The siRNA of HIF-1α attenuated expression of VEGF and CXCR4 and inhibited LLC migration. K5 decreased HIF-1α protein level and impaired nuclear HIF-1α accumulation. These results showed for the first time that K5 inhibits LLC growth and metastasis via the dual effects of anti-angiogenesis and suppression of tumor cell motility by targeting the pivotal molecule, HIF-1α.

  4. Variants of the Low Oxygen Sensors EGLN1 and HIF-1AN Associated with Acute Mountain Sickness

    Directory of Open Access Journals (Sweden)

    Enhao Zhang

    2014-11-01

    Full Text Available Two low oxygen sensors, Egl nine homolog 1 (EGLN1 and hypoxia-inducible factor 1-α inhibitor (HIF-1AN, play pivotal roles in the regulation of HIF-1α, and high altitude adaption may be involved in the pathology of acute mountain sickness (AMS. Here, we aimed to analyze single nucleotide polymorphisms (SNPs in the untranslated regions of the EGLN1 and HIF-1AN genes and SNPs chosen from a genome-wide adaptation study of the Han Chinese population. To assess the association between EGLN1 and HIF-1AN SNPs and AMS in a Han Chinese population, a case–control study was performed including 190 patients and 190 controls. In total, thirteen SNPs were genotyped using the MassARRAY® MALDI-TOF system. Multiple genetic models were tested; The Akaike’s information criterion (AIC and Bayesian information criterion (BIC values indicated that the dominant model may serve as the best-fit model for rs12406290 and rs2153364 of significant difference. However, these data were not significant after Bonferroni correction. No significant association was noted between AMS and rs12757362, rs1339894, rs1361384, rs2009873, rs2739513 or rs2486729 before and after Bonferroni correction. Further haplotype analyses indicated the presence of two blocks in EGLN1; one block consists of rs12406290-rs2153364, located upstream of the EGLN1 gene. Carriers of the “GG” haplotype of rs12406290-rs2153364 exhibited an increased risk of AMS after adjustments for age and smoking status. However, no significant association was observed among HIF-1AN 3'-untranslated region (3'-UTR polymorphisms, haplotype and AMS. Our study indicates that variants in the EGLN1 5'-UTR influence the susceptibility to AMS in a Han Chinese population.

  5. Targeting the lactate transporter MCT1 in endothelial cells inhibits lactate-induced HIF-1 activation and tumor angiogenesis.

    Directory of Open Access Journals (Sweden)

    Pierre Sonveaux

    Full Text Available Switching to a glycolytic metabolism is a rapid adaptation of tumor cells to hypoxia. Although this metabolic conversion may primarily represent a rescue pathway to meet the bioenergetic and biosynthetic demands of proliferating tumor cells, it also creates a gradient of lactate that mirrors the gradient of oxygen in tumors. More than a metabolic waste, the lactate anion is known to participate to cancer aggressiveness, in part through activation of the hypoxia-inducible factor-1 (HIF-1 pathway in tumor cells. Whether lactate may also directly favor HIF-1 activation in endothelial cells (ECs thereby offering a new druggable option to block angiogenesis is however an unanswered question. In this study, we therefore focused on the role in ECs of monocarboxylate transporter 1 (MCT1 that we previously identified to be the main facilitator of lactate uptake in cancer cells. We found that blockade of lactate influx into ECs led to inhibition of HIF-1-dependent angiogenesis. Our demonstration is based on the unprecedented characterization of lactate-induced HIF-1 activation in normoxic ECs and the consecutive increase in vascular endothelial growth factor receptor 2 (VEGFR2 and basic fibroblast growth factor (bFGF expression. Furthermore, using a variety of functional assays including endothelial cell migration and tubulogenesis together with in vivo imaging of tumor angiogenesis through intravital microscopy and immunohistochemistry, we documented that MCT1 blockers could act as bona fide HIF-1 inhibitors leading to anti-angiogenic effects. Together with the previous demonstration of MCT1 being a key regulator of lactate exchange between tumor cells, the current study identifies MCT1 inhibition as a therapeutic modality combining antimetabolic and anti-angiogenic activities.

  6. SU-C-303-02: Correlating Metabolic Response to Radiation Therapy with HIF-1alpha Expression

    Energy Technology Data Exchange (ETDEWEB)

    Campos, D [University of Wisconsin Madison, Madison, WI (United States); Peeters, W [Radboud University Medical Center, Nijmegen, GA (United States); Nickel, K [University of Wisconsin, Madison, WI (United States); Eliceiri, K; Kimple, R; Van Der Kogel, A; Kissick, M [University of Wisconsin, Madison, Wisconsin (United States)

    2015-06-15

    Purpose: To understand radiation induced alterations in cellular metabolism which could be used to assess treatment or normal tissue response to aid in patient-specific adaptive radiotherapy. This work aims to compare the metabolic response of two head and neck cell lines, one malignant (UM-SCC-22B) and one benign (Normal Oral Keratinocyte), to ionizing radiation. Responses are compared to alterations in HIF-1alpha expression. These dynamics can potentially serve as biomarkers in assessing treatment response allowing for patient-specific adaptive radiotherapy. Methods: Measurements of metabolism and HIF-1alpha expression were taken before and X minutes after a 10 Gy dose of radiation delivered via an orthovoltage x-ray source. In vitro changes in metabolic activity were measured via fluorescence lifetime imaging (FLIM) to assess the mean lifetime of NADH autofluorescence following a dose of 10 Gy. HIF-1alpha expression was measured via immunohistochemical staining of in vitro treated cells and expression was quantified using the FIJI software package. Results: FLIM demonstrated a decrease in the mean fluorescence lifetime of NADH by 100 ps following 10 Gy indicating a shift towards glycolytic pathways for malignant cells; whereas this benign cell line showed little change in metabolic signature. Immunohistochemical analysis showed significant changes in HIF-1alpha expression in response to 10 Gy of radiation that correlate to metabolic profiles. Conclusion: Radiation induces significant changes in metabolic activity and HIF-1alpha expression. These alterations occur on time scales approximating the duration of common radiation treatments (approximately tens of minutes). Further understanding these dynamics has important implications with regard to improvement of therapy and biomarkers of treatment response.

  7. Transcriptional activation of HIF-1 by a ROS-ERK axis underlies the resistance to photodynamic therapy

    Science.gov (United States)

    Pansa, María Florencia; Vera, Renzo Emanuel; Fernández-Zapico, Martín Ernesto; Rumie Vittar, Natalia Belén; Rivarola, Viviana Alicia

    2017-01-01

    Photodynamic therapy (PDT), a promising treatment option for cancer, involves the activation of a photosensitizer (PS) by local irradiation with visible light. Excitation of the PS leads to a series of photochemical reactions and consequently the local generation of harmful reactive oxygen species (ROS) causing limited or none systemic defects. However, the development of resistance to this promising therapy has slowed down its translation into the clinical practice. Thus, there is an increase need in understanding of the molecular mechanism underlying resistance to PDT. Here, we aimed to examine whether a relationship exists between PDT outcome and ROS-involvement in the resistance mechanism in photosensitized cancer cells. In order to recapitulate tumor architecture of the respective original tumor, we developed a multicellular three-dimensional spheroid system comprising a normoxic periphery, surrounding a hypoxic core. Using Me-ALA, a prodrug of the PS PpIX, in human colorectal spheroids we demonstrate that HIF-1 transcriptional activity was strongly up-regulated and mediates PDT resistant phenotype. RNAi knockdown of HIF-1 impairs resistance to PDT. Oxidative stress-mediated activation of ERK1/2 followed PDT was involved on positive modulation of HIF-1 transcriptional activity after photodynamic treatment. ROS scavenging and MEK/ERK pathway inhibition abrogated the PDT-mediated HIF-1 upregulation. Together our data demonstrate that resistance to PDT is in part mediated by the activation of a ROS-ERK1/2-HIF-1 axis, thus, identifying novel therapeutic targets that could be used in combination with PDT. PMID:28545088

  8. Effect of trifluoperazine on toxicity, HIF-1α induction and hepatocyte regeneration in acetaminophen toxicity in mice

    Energy Technology Data Exchange (ETDEWEB)

    Chaudhuri, Shubhra, E-mail: SCHAUDHURI@uams.edu [Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas (United States); Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); McCullough, Sandra S., E-mail: mcculloughsandras@uams.edu [Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas (United States); Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); Hennings, Leah, E-mail: lhennings@uams.edu [Department of Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas (United States); Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); Brown, Aliza T., E-mail: brownalizat@uams.edu [Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas (United States); Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); Li, Shun-Hwa [Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI (United States); Simpson, Pippa M., E-mail: psimpson@mcw.edu [Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI (United States); Hinson, Jack A., E-mail: hinsonjacka@uams.edu [Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); James, Laura P., E-mail: jameslaurap@uams.edu [Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas (United States); Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States); Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, Arkansas Children' s Hospital Research Institute, Little Rock, AR (United States)

    2012-10-15

    Oxidative stress and mitochondrial permeability transition (MPT) are important mechanisms in acetaminophen (APAP) toxicity. The MPT inhibitor trifluoperazine (TFP) reduced MPT, oxidative stress, and toxicity in freshly isolated hepatocytes treated with APAP. Since hypoxia inducible factor-one alpha (HIF-1α) is induced very early in APAP toxicity, a role for oxidative stress in the induction has been postulated. In the present study, the effect of TFP on toxicity and HIF-1α induction in B6C3F1 male mice treated with APAP was examined. Mice received TFP (10 mg/kg, oral gavage) prior to APAP (200 mg/kg IP) and at 7 and 36 h after APAP. Measures of metabolism (hepatic glutathione and APAP protein adducts) were comparable in the two groups of mice. Toxicity was decreased in the APAP/TFP mice at 2, 4, and 8 h, compared to the APAP mice. At 24 and 48 h, there were no significant differences in toxicity between the two groups. TFP lowered HIF-1α induction but also reduced the expression of proliferating cell nuclear antigen, a marker of hepatocyte regeneration. TFP can also inhibit phospholipase A{sub 2}, and cytosolic and secretory PLA{sub 2} activity levels were reduced in the APAP/TFP mice compared to the APAP mice. TFP also lowered prostaglandin E{sub 2} expression, a known mechanism of cytoprotection. In summary, the MPT inhibitor TFP delayed the onset of toxicity and lowered HIF-1α induction in APAP treated mice. TFP also reduced PGE{sub 2} expression and hepatocyte regeneration, likely through a mechanism involving PLA{sub 2}. -- Highlights: ► Trifluoperazine reduced acetaminophen toxicity and lowered HIF-1α induction. ► Trifluoperazine had no effect on the metabolism of acetaminophen. ► Trifluoperazine reduced hepatocyte regeneration. ► Trifluoperazine reduced phospholipase A{sub 2} activity and prostaglandin E{sub 2} levels.

  9. Assessing the relationship between the Driver Behavior Questionnaire and the Driver Skill Inventory: Revealing sub-groups of drivers

    DEFF Research Database (Denmark)

    Martinussen, Laila Marianne; Møller, Mette; Prato, Carlo Giacomo

    2014-01-01

    The Driver Behavior Questionnaire and the Driver Skill Inventory are two of the most frequently used measures of self-reported driving style and driving skill. The motivation behind the present study was to identify sub-groups of drivers that potentially act dangerously in traffic (as measured by...... driving behaviors, and vice versa. The present findings highlight the need to look into driver’s attitudes towards safety, and to devise differential interventions targeting specific problematic groups of the population in the attempt to improve road safety nationwide....

  10. Gene transcripts encoding hypoxia-inducible factor (HIF) exhibit tissue- and muscle fiber type-dependent responses to hypoxia and hypercapnic hypoxia in the Atlantic blue crab, Callinectes sapidus.

    Science.gov (United States)

    Hardy, Kristin M; Follett, Chandler R; Burnett, Louis E; Lema, Sean C

    2012-09-01

    Hypoxia inducible factor (HIF) is a transcription factor that under low environmental oxygen regulates the expression of suites of genes involved in metabolism, angiogenesis, erythropoiesis, immune function, and growth. Here, we isolated and sequenced partial cDNAs encoding hif-α and arnt/hif-β from the Atlantic blue crab, Callinectes sapidus, an estuarine species that frequently encounters concurrent hypoxia (low O(2)) and hypercapnia (elevated CO(2)). We then examined the effects of acute exposure (1h) to hypoxia (H) and hypercapnic hypoxia (HH) on relative transcript abundance for hif-α and arnt/hif-β in different tissues (glycolytic muscle, oxidative muscle, hepatopancreas, gill, and gonads) using quantitative real-time RT-PCR. Our results indicate that hif-α and arnt/hif-β mRNAs were constitutively present under well-aerated normoxia (N) conditions in all tissues examined. Further, H and HH exposure resulted in both tissue-specific and muscle fiber type-specific effects on relative hif-α transcript abundance. In the gill and glycolytic muscle, relative hif-α mRNA levels were significantly lower under H and HH, compared to N, while no change (or a slight increase) was detected in oxidative muscle, hepatopancreas and gonadal tissues. H and HH did not affect relative transcript abundance for arnt/hif-β in any tissue or muscle fiber type. Thus, in crustaceans the HIF response to H and HH appears to involve changes in hif transcript abundance, with variation in hif-α and arnt/hif-β transcriptional dynamics occurring in both a tissue- and muscle fiber type-dependent manner. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. Expression of HIF-1α and GLUT-1 in endometrial carcinoma and its clinical significance%HIF-1α、GLUT-1在子宫内膜癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    王丽; 刘欣; 孙乃英

    2009-01-01

    目的 探讨缺氧诱导因子-1α(HIF-1α)、葡萄糖转运蛋白-1(GLUT-1)在子宫内膜癌中的表达及意义.方法 采用免疫组织化学PV-9000二步法检测36例子宫内膜癌、18例不典型增生、12例子宫内膜复杂性增生、15例子宫内膜单纯性增生组织中HIF-1α、GLUT-1的表达情况.结果 HIF-1α,在子宫内膜单纯性增生、复杂性增生、不典型增生和子宫内膜癌组织中的阳性表达率分别为6.7%、25.0%、38.9%和77.8%.内膜癌组分别与单纯性增生、复杂性增生及不典型增生组比较,差异均有统计学意义(P<0.05).GLUT-1在子宫内膜单纯性增生、复杂性增生、不典型增生和子宫内膜癌组织中的阳性表达率分别为0、8.3%、44.4%和83.3%.子宫内膜癌组分别与单纯性增生、复杂性增生及不典型增生组比较,差异均有统计学意义(P<0.05).子宫内膜病变组织中HIF-1α与GLUT-1表达水平间呈正相关(r=0.559,P<0.01).结论 HIF-1α、GLUT-1在子宫内膜单纯性增生、复杂性增生、不典型增生及子宫内膜腺癌中的阳性表达率逐渐增高,说明HIF-1α及其靶基因GLUT-1可能在子宫内膜病变由良性到恶性转变过程及恶性肿瘤的进展中起重要作用.HIF-1α能提高GLUT-1的表达.%Objective To investigate the expression and significance of hypoxia inducible factor-1 alpha(HIF-1α)and glucose transporter protein 1(GLUT-1)in endometrial carcinoma.Methods Immunohistochemical(PV-9000 two-step)method was applied to detect the expression of HIF-1α and GLUT-1 in 36 cases of endometrial carcinoma,18 cases of atypical hyperplasia,12 cases of complex hyperplasia,15 cases of simple hyperplsia,atypical hyperplasia and endometrial carcinoma were 6.7%,25.0%、38.9%and 77.8%.Compared expression of HIF-1αin endomelrial adenocarcinoma with those in endometrial simple hyperplasia,complex hyperplasia and atypical hyperplasia tissue,differences were statistically significant(P<0.05).The positive rates

  12. 前列腺癌中HIF-1α和COX-2的表达及与血管生成的关系%Expression of HIF-1αand COX-2 in prostate cancer and its relationship with angiogenesis

    Institute of Scientific and Technical Information of China (English)

    闫红丽; 张伟; 安丰; 张金立; 梁春威; 刘学凯; 孙莉

    2012-01-01

      目的探讨缺氧诱导因子-1α(HIF-1α)和环氧合酶-2(COX-2)在前列腺癌组织中的表达情况及与肿瘤血管形成的关系。方法应用免疫组化(SP)法检测72例不同病理级别前列腺癌组织中HIF-1α、COX-2的表达,用八因子相关抗原(VIII-R-Ag)标记微血管密度(MVD)。结果 HIF-1α、COX-2在不同病理级别前列腺癌中表达差异有统计学意义;HIF-1α、COX-2与前列腺癌病理分级存在显著相关性;MVD在不同病理级别前列腺癌组织中差异有统计学意义;HIF-1α、COX-2的表达与MVD存在显著相关性,HIF-1α与COX-2的表达呈正相关。结论 HIF-1α和COX-2与前列腺癌的恶性程度有关,并可能在促进肿瘤血管形成过程中起协同作用。%  Objective To study the expression of HIF-1αand COX-2 in prostate cancer and its relationship with tumor angiogenesis. Methods The expression of HIF-1αand COX-2 was examined by immunohistochemistry Streptavidin-Peroxidase (SP) method in 72 cases of prostatic carcinoma with different pathological grades, and the MVD was marked out by the VIII-R-Ag. Results There was a statistically significance in the expression differences of HIF-1α and COX-2 in prostatic carcinoma with different pathological grades; significant correlation was showed between HIF-1α and COX-2 and the pathological grading of prostate cancer; the differences of MVD in prostate cancer with different grades hade a statistically significance; the expression of HIF-1α and COX-2 had a significant correlation with MVD, and positive correlation was showed in the expression of HIF-1αand COX-2. conclusion HIF-1αand COX-2 relate to malignancy of prostate cancer, and have a synergistic effect in tumor angiogenesis process.

  13. Square pulse linear transformer driver

    Directory of Open Access Journals (Sweden)

    A. A. Kim

    2012-04-01

    Full Text Available The linear transformer driver (LTD technological approach can result in relatively compact devices that can deliver fast, high current, and high-voltage pulses straight out of the LTD cavity without any complicated pulse forming and pulse compression network. Through multistage inductively insulated voltage adders, the output pulse, increased in voltage amplitude, can be applied directly to the load. The usual LTD architecture [A. A. Kim, M. G. Mazarakis, V. A. Sinebryukhov, B. M. Kovalchuk, V. A. Vizir, S. N Volkov, F. Bayol, A. N. Bastrikov, V. G. Durakov, S. V. Frolov, V. M. Alexeenko, D. H. McDaniel, W. E. Fowler, K. LeCheen, C. Olson, W. A. Stygar, K. W. Struve, J. Porter, and R. M. Gilgenbach, Phys. Rev. ST Accel. Beams 12, 050402 (2009PRABFM1098-440210.1103/PhysRevSTAB.12.050402; M. G. Mazarakis, W. E. Fowler, A. A. Kim, V. A. Sinebryukhov, S. T. Rogowski, R. A. Sharpe, D. H. McDaniel, C. L. Olson, J. L. Porter, K. W. Struve, W. A. Stygar, and J. R. Woodworth, Phys. Rev. ST Accel. Beams 12, 050401 (2009PRABFM1098-440210.1103/PhysRevSTAB.12.050401] provides sine shaped output pulses that may not be well suited for some applications like z-pinch drivers, flash radiography, high power microwaves, etc. A more suitable power pulse would have a flat or trapezoidal (rising or falling top. In this paper, we present the design and first test results of an LTD cavity that generates such a type of output pulse by including within its circular array a number of third harmonic bricks in addition to the main bricks. A voltage adder made out of a square pulse cavity linear array will produce the same shape output pulses provided that the timing of each cavity is synchronized with the propagation of the electromagnetic pulse.

  14. Risk drivers pose to themselves and other drivers by violating traffic rules.

    Science.gov (United States)

    Penmetsa, Praveena; Pulugurtha, Srinivas S

    2017-01-02

    Violation of traffic rules is a major contributing factor in both crashes and fatalities in the United States. This study aims at quantifying risk that drivers pose to themselves and other drivers by violating traffic rules. Crash data from 2010 to 2013 were gathered for the state of North Carolina. Descriptive analysis was carried out to identify frequent traffic violations and who were committing the traffic violations that resulted in crashes. A multinomial logit model was then developed to examine the relation between different traffic violations and driver injury severity. Additionally, odds ratios were estimated to identify the likelihood (probability) of severe or moderate injury to the driver and other drivers due to a driver violating a traffic rule that led to a crash. Exceeding the speed limit is more likely to result in severe injury compared to disregarding traffic signals. However, going the wrong way is more likely to result in severe injury to other drivers when compared to any other traffic violation. Driving under the influence of alcohol is 2 times more likely to result in severe injury than driving under the influence of drugs. These 2 traffic violations by a driver are almost equally likely to result in severe injury to other drivers. Drivers often perceive that violating traffic rules will not result in a crash or severe injury. However, the results from this study show that a majority of the traffic violations lead to severe injury to the violator as well as to other drivers. The findings from this study serve as documented evidence to educate drivers about the risk they pose to themselves and to other drivers by violating traffic rules and encourage the adaptation of safe driving behavior in order to contribute toward reaching the "zero traffic deaths" vision. They also help make policy changes pertaining to penalty points and fines for violating a traffic rule.

  15. Approaches of truck drivers and non-truck drivers toward reckless on-road behavior.

    Science.gov (United States)

    Rosenbloom, Tova; Eldror, Ehud; Shahar, Amit

    2009-07-01

    The purpose of the study was to compare the reported approaches of truck drivers to those of non-truck drivers toward reckless on-road behaviors. One hundred and sixty-seven adult males, including 70 non-truck drivers, completed the questionnaires voluntarily. The truck drivers were employees of a concrete manufacturing company working at various company plants throughout Israel. Seventy were professional mixer truckers and 27 were tip-truckers. The participants completed the Reckless Driving Self-Report Scale based on Taubman Ben-Ari et al. [Taubman Ben-Ari, O., Florian, V., Mikulincer, M., 1999. The impact of mortality salience on reckless driving: a test of terror management mechanisms. Journal of Personality and Social Psychology 76, 35-45], adapted for truck drivers for this study. It was expected that non-professional, as compared to professional (truck) drivers, would be more permissive regarding reckless driving, since driving risks are less prominent in their daily driving experience. An ANOVA performed on mean reckless-driving scores yielded significant results. The post hoc Schéffe test indicated significantly higher reckless-driving scores for automobile drivers as compared to both mixer-truck driver scores and tip-truck driver scores. In addition, the reckless-driving scores for mixer-truck drivers were significantly higher than the tip-truck driver scores. We discuss various explanations for the findings and consider possible implications for training strategies in organizations as well as for media campaigns focused on mutual safe road use of truck drivers and private vehicle drivers.

  16. Tarantula: Killing driver bugs before they hatch

    DEFF Research Database (Denmark)

    Lawall, Julia Laetitia; Muller, Gilles; Urunuela, Richard

    2005-01-01

    The Linux operating system is undergoing continual evolution. Evolution in the kernel and generic driver modules often triggers the need for corresponding evolutions in specific device drivers. Such collateral evolutions are tedious, because of the large number of device drivers, and error......-prone, because of the complexity of the code modifications involved. We propose an automatic tool, Tarantula, to aid in this process. In this paper, we examine some recent evolutions in Linux and the collateral evolutions they trigger, and assess the corresponding requirements on Tarantula....

  17. CMOS Law-jitter Clock Driver Design

    OpenAIRE

    2012-01-01

    [ANGLÈS] Design of a low-jitter, low-phase noise clock driver in 40 nm CMOS technology. The work is in the field of analog integrated circuit (IC) design in nanometer CMOS technologies. [CASTELLÀ] Diseño de un circuito integrado "clock driver" de bajo jitter y bajo ruido de fase en tecnología CMOS 40 nm. El trabajo se contextualiza en el campo del diseño de circuitos integrados analógicos en tecnologías CMOS nanométricas. [CATALÀ] Disseny d'un circuit "clock driver" de baix jitter i bai...

  18. LC Oscillator Driver for Safety Critical Applications

    CERN Document Server

    Horsky, Pavel

    2011-01-01

    A CMOS harmonic signal LC oscillator driver for automotive applications working in a harsh environment with high safety critical requirements is described. The driver can be used with a wide range of external components parameters (LC resonance network of a sensor). Quality factor of the external LC network can vary two decades. Amplitude regulation of the driver is digitally controlled and the DAC is constructed as exponential with piece-wise-linear (PWL) approximation. Low current consumption for high quality resonance networks is achieved. Realized oscillator is robust, used in safety critical application and has low EMC emissions.

  19. Tarantula: Killing driver bugs before they hatch

    DEFF Research Database (Denmark)

    Lawall, Julia Laetitia; Muller, Gilles; Urunuela, Richard

    2005-01-01

    The Linux operating system is undergoing continual evolution. Evolution in the kernel and generic driver modules often triggers the need for corresponding evolutions in specific device drivers. Such collateral evolutions are tedious, because of the large number of device drivers, and error......-prone, because of the complexity of the code modifications involved. We propose an automatic tool, Tarantula, to aid in this process. In this paper, we examine some recent evolutions in Linux and the collateral evolutions they trigger, and assess the corresponding requirements on Tarantula....

  20. Optimizing the Universal Robots ROS driver

    DEFF Research Database (Denmark)

    Andersen, Thomas Timm

    In this report I will examine both the current and the possible performance of one of the most popular robotics platforms in research, the Universal Robot manipulator. I will solely focus on the ROS based approaches and show how the current driver can be improved. I will look at performance...... improvement both in terms of faster reaction as well as making it possible to control the robot using either ros_control or ordinary joint speed commands, which is required for many types of sensory based control like visual servoing. The developed driver is compared to the drivers already existing in the ROS...

  1. Driver style and driver skill – Clustering sub-groups of drivers differing in their potential danger in traffic

    DEFF Research Database (Denmark)

    Martinussen, Laila Marianne; Møller, Mette; Prato, Carlo Giacomo

    based on a combined use of the DBQ and the DSI. Moreover, the joint use of the two instruments was applied to identify sub-groups of drivers that differ in their potential danger in traffic (as measured by frequency of aberrant driving behaviors and level of driving skills), as well as to test whether...... the sub-groups of drivers differed in characteristics such as age, gender, annual mileage and accident involvement. 3908 drivers aged 18–84 participated in the survey. The results suggested that the drivers are consistent in their reporting of driving ability, as the self-reported driving skill level...... mirrored the self-reported frequency of aberrant driving behaviors. K-means cluster analysis revealed four distinct clusters that differed in the frequency of aberrant driving behavior and driving skills, as well as individual characteristics and driving related factors such as annual mileage, accident...

  2. Assessing the relationship between the Driver Behavior Questionnaire and the Driver Skill Inventory: Revealing sub-groups of drivers

    DEFF Research Database (Denmark)

    Martinussen, Laila Marianne; Møller, Mette; Prato, Carlo Giacomo

    2014-01-01

    The Driver Behavior Questionnaire and the Driver Skill Inventory are two of the most frequently used measures of self-reported driving style and driving skill. The motivation behind the present study was to identify sub-groups of drivers that potentially act dangerously in traffic (as measured...... by frequency of aberrant driving behaviors and level of driving skills), as well as to test whether the sub-groups differ in characteristics such as age, gender, annual mileage and accident involvement. Furthermore, the joint analysis of the two instruments was used to test drivers’ assessment of their own...... self-reported driving skills and whether the reported skill level was reflected in the reported aberrant driving behaviors. 3908 drivers aged 18–84 participated in the survey. K-means cluster analysis revealed four distinct sub-groups that differed in driving skills and frequency of aberrant driving...

  3. Food consumption trends and drivers.

    Science.gov (United States)

    Kearney, John

    2010-09-27

    A picture of food consumption (availability) trends and projections to 2050, both globally and for different regions of the world, along with the drivers largely responsible for these observed consumption trends are the subject of this review. Throughout the world, major shifts in dietary patterns are occurring, even in the consumption of basic staples towards more diversified diets. Accompanying these changes in food consumption at a global and regional level have been considerable health consequences. Populations in those countries undergoing rapid transition are experiencing nutritional transition. The diverse nature of this transition may be the result of differences in socio-demographic factors and other consumer characteristics. Among other factors including urbanization and food industry marketing, the policies of trade liberalization over the past two decades have implications for health by virtue of being a factor in facilitating the 'nutrition transition' that is associated with rising rates of obesity and chronic diseases such as cardiovascular disease and cancer. Future food policies must consider both agricultural and health sectors, thereby enabling the development of coherent and sustainable policies that will ultimately benefit agriculture, human health and the environment.

  4. Uncovering a new role for peroxidase enzymes as drivers of angiogenesis.

    Science.gov (United States)

    Panagopoulos, Vasilios; Zinonos, Irene; Leach, Damien A; Hay, Shelley J; Liapis, Vasilios; Zysk, Aneta; Ingman, Wendy V; DeNichilo, Mark O; Evdokiou, Andreas

    2015-11-01

    Peroxidases are heme-containing enzymes released by activated immune cells at sites of inflammation. To-date their functional role in human health has mainly been limited to providing a mechanism for oxidative defence against invading bacteria and other pathogenic microorganisms. Our laboratory has recently identified a new functional role for peroxidase enzymes in stimulating fibroblast migration and collagen biosynthesis, offering a new insight into the causative association between inflammation and the pro-fibrogenic events that mediate tissue repair and regeneration. Peroxidases are found at elevated levels within and near blood vessels however, their direct involvement in angiogenesis has never been reported. Here we report for the first time that myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are readily internalised by human umbilical vein endothelial cells (HUVEC) where they promote cellular proliferation, migration, invasion, and stimulate angiogenesis both in vitro and in vivo. These pro-angiogenic effects were attenuated using the specific peroxidase inhibitor 4-ABAH, indicating the enzyme's catalytic activity is essential in mediating this response. Mechanistically, we provide evidence that MPO and EPO regulate endothelial FAK, Akt, p38 MAPK, ERK1/2 phosphorylation and stabilisation of HIF-2α, culminating in transcriptional regulation of key angiogenesis pathways. These findings uncover for the first time an important and previously unsuspected role for peroxidases as drivers of angiogenesis, and suggest that peroxidase inhibitors may have therapeutic potential for the treatment of angiogenesis related diseases driven by inflammation.

  5. DriverNet: uncovering the impact of somatic driver mutations on transcriptional networks in cancer.

    Science.gov (United States)

    Bashashati, Ali; Haffari, Gholamreza; Ding, Jiarui; Ha, Gavin; Lui, Kenneth; Rosner, Jamie; Huntsman, David G; Caldas, Carlos; Aparicio, Samuel A; Shah, Sohrab P

    2012-12-22

    Simultaneous interrogation of tumor genomes and transcriptomes is underway in unprecedented global efforts. Yet, despite the essential need to separate driver mutations modulating gene expression networks from transcriptionally inert passenger mutations, robust computational methods to ascertain the impact of individual mutations on transcriptional networks are underdeveloped. We introduce a novel computational framework, DriverNet, to identify likely driver mutations by virtue of their effect on mRNA expression networks. Application to four cancer datasets reveals the prevalence of rare candidate driver mutations associated with disrupted transcriptional networks and a simultaneous modulation of oncogenic and metabolic networks, induced by copy number co-modification of adjacent oncogenic and metabolic drivers. DriverNet is available on Bioconductor or at http://compbio.bccrc.ca/software/drivernet/.

  6. Driver ASICs for Advanced Deformable Mirrors Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The program leverages on our extensive expertise in developing high-performance driver ASICs for deformable mirror systems and seeks to expand the capacities of the...

  7. Physics at a new Fermilab proton driver

    Energy Technology Data Exchange (ETDEWEB)

    Geer, Steve; /Fermilab

    2006-04-01

    In 2004, motivated by the recent exciting developments in neutrino physics, the Fermilab Long Range Planning Committee identified a new high intensity Proton Driver as an attractive option for the future. At the end of 2004 the APS ''Study on the Physics of Neutrinos'' concluded that the future US neutrino program should have, as one of its components, ''A proton driver in the megawatt class or above and neutrino superbeam with an appropriate very large detector capable of observing Cp violation and measuring the neutrino mass-squared differences and mixing parameters with high precision''. The presently proposed Fermilab Proton Driver is designed to accomplish these goals, and is based on, and would help develop, Linear Collider technology. In this paper the Proton Driver parameters are summarized, and the potential physics program is described.

  8. The importance of sight for drivers

    Directory of Open Access Journals (Sweden)

    Alicja Pas-Wyroślak

    2013-06-01

    Full Text Available Sight is the basic sense for drivers. Condition of the eye determines correct, comfortable and safe performance of the work as drivers. This article presents various factors influencing the sight condition. There are two groups of factors, external (environment, the kind and time of work, stress caused by work and internal (systemic and local disorders. All these factors can reduce significantly visual functions, such as visual acuity, field of vision, color vision, strereoscopic vision, twilight vision and glare sensitivity. There are also presented actual requirements for drivers and causes of the car accidents in various age groups. Impairments in vision functions can be dangerous for both the driver and other road users. Med Pr 2013;64(3:419–425

  9. Driver circuit for solid state light sources

    Energy Technology Data Exchange (ETDEWEB)

    Palmer, Fred; Denvir, Kerry; Allen, Steven

    2016-02-16

    A driver circuit for a light source including one or more solid state light sources, a luminaire including the same, and a method of so driving the solid state light sources are provided. The driver circuit includes a rectifier circuit that receives an alternating current (AC) input voltage and provides a rectified AC voltage. The driver circuit also includes a switching converter circuit coupled to the light source. The switching converter circuit provides a direct current (DC) output to the light source in response to the rectified AC voltage. The driver circuit also includes a mixing circuit, coupled to the light source, to switch current through at least one solid state light source of the light source in response to each of a plurality of consecutive half-waves of the rectified AC voltage.

  10. Hypoxia in Tumor Angiogenesis and Metastasis: Evaluation of VEGF and MMP Over-expression and Down-Regulation of HIF-1alpha with RNAi in Hypoxic Tumor Cells

    Science.gov (United States)

    Shah, Shruti

    Background: As tumor mass grows beyond a few millimeters in diameter, the angiogenic "switch" is turned on leading to recruitment of blood vessels from surrounding artery and veins. However, the tumor mass is poorly perfused and there are pockets of hypoxia or lower oxygen concentrations relative to normal tissue. Hypoxia-inducing factor-1a (HIF-1a), a transcription factor, is activated when the oxygen concentration is low. Upon activation of HIF-1a, a number of other genes also turn on that allows the tumor to become more aggressive and resistant to therapy. Purpose: The main objectives of this study were to evaluate the effect of hypoxia-induced HIF-1a followed by over-expression of angiogenic and metastatic markers in tumor cells and down-regulation of HIF-1a using nanoparticle-delivered RNA interference therapy. Methods: Human ovarian (SKOV3) and breast (MDA-MB-231) adenocarcinoma cells were incubated under normoxic and hypoxic conditions. Following hypoxia treatment of the cells, HIF-1α, vascular endothelial growth factor (VEGF), matrix metalloproteinase 2 (MMP-2), and MMP-9 expression was analyzed qualitatively and quantitatively. For intracellular delivery of HIF-1a gene silencing small interfering RNA (siRNA), type B gelatin nanoparticles were fabricated using the solvent displacement method and the surface was modified with poly(ethylene glycol) (PEG, Mol. wt. 2kDa). Cellular uptake and distribution of the nanoparticles was observed with Cy3-siRNA loaded, FITC-conjugated gelatin nanoparticles. Cytotoxicity of the nanoparticle formulations was evaluated in both the cell lines. siRNA was transfected in the gelatin nanoparticles under hypoxic conditions. Total cellular protein and RNA were extracted for analysis of HIF1a, VEGF, MMP-2 and MMP-9 expression. Results: MDA-MB-231 and SKOV3 cells show increased expression of HIF1a under hypoxic conditions compared to baseline levels at normoxic conditions. ELISA and western blots of VEGF, MMP-2 and MMP-9 appear to

  11. HIF-1α基因RNA干扰质粒在胃癌细胞中的构建与鉴定%Construction of a vector carrying a small hairpin RNA targeting the HIF-1α gene and its expression in gastric cancer cells

    Institute of Scientific and Technical Information of China (English)

    龚青; 方淑环; 高国全

    2012-01-01

    AIM: To construct a plasmid carrying a small hairpin RNA (shRNA) targeting the HIF-lct gene and to observe its expression in gastric cancer SGC-7901 cells. METHODS: SHIF-la-specific shRNA sequence was designed, synthesized and cloned into the expression vector pSilencer 1.0-U6. The recombi-nants were identified by double digestion with EcoR I + Apa I and DNA sequencing, and then transfected into SGC-7901 cells under hypoxia. Forty-eight hours after transfection, HIF-la expression was detected by Western blotting. RESULTS: Restriction enzyme digestion and DNA sequencing confirmed that the shRNA was correctly inserted into the plasmid pSilenc-er 1.0-U6. The recombinant plasmid carrying the shRNA targeting HIF-la was successfully transfected into SGC-7901 cells. Western blot analysis suggested that HIF-la expression was knocked down in transfected cells compared to control cells. CONCLUSION: A plasmid carrying a shRNA targeting HIF-la has been constructed successfully, and SGC-7901 cells stably transfected with this recombinant vector have been obtained. These will offer favorable tools for studying HIF-la-targeted gene therapy.%目的:构建转录因子HIF-1α表达的RNA干扰(RNA interference,RNAi)质粒,为研究HIF-1α参与的细胞信号通路及以HIF-1α为靶点的基因治疗提供稳定转染细胞的RNAi.方法:选取HIF-1α基因的19nt特异性序列,设计针对HIF-1α的shRNA序列,应用基因重组技术克隆到pSilencer 1.0-U6表达载体中,利用EcoRⅠ和paⅠ双酶切和DNA测序鉴定重组克隆.低氧条件下,将包含HIF-1α基因RNAi序列的重组载体转染人胃癌细胞SGC-7901,转染48 h后,收集细胞裂解液,Western blotting分析低氧条件下对照组与转染组HIF-1α蛋白的表达水平.结果:双酶切证实shRNA插入pSilencer 1.0-U6质粒,DNA测序证实插入的序列正确;HIF-1α基因RNAi质粒转染胃癌细胞SGC-7901成功;Western blotting结果显示与空质粒对照组比转染组细胞HIF-1α表达下调.结论:成功构建人HIF

  12. Drivers of desertification and their impact

    Science.gov (United States)

    Mantel, S.; Van Lynden, G. V. L.; Karavitis, C. A.; Kosmas, C.; Van der Werff ten Bosch, M. J.

    2012-04-01

    Drivers of desertification and their impact An inventory was made of drivers of desertification and how they impact on the degradation process. The major drivers of desertification were analysed and compared between 16 sites around the globe. For each of these sites factors were scored with a perceived influence on desertification. Most of these factors, from the socio-cultural, environmental, and economic dimensions, appeared to be related to land management and planning and to (de-)population. They cause a number of temporary or permanent changes in the landscape, which, by themselves or in combination, lead to degradation of vegetation and soils. Most sites have several forms of land degradation occurring in and around their study area of which erosion by water is the dominant one. Other degradation types occurring in sites were: wind erosion, soil salinization, seawater intrusion in the groundwater, vegetation and biodiversity decline, groundwater depletion, decreased productivity/ carrying capacity, soil fertility decline, water logging and water pollution. As a first step, data and information was gathered on policies, desertification status and processes and on socio-economic conditions. The DPSIR framework (Driving force, Pressure, State, Impact, Response) provides a structure for assessment of the impact of past measures on the status of the environment or to formulate effective measures. In analysing the data, the different data items were structured in and formulated to elements fitting the DPSIR chain. Then possible connections between these different aspects were analysed. In our analysis nine major drivers were reported for the various sites, of which one was environmental, three drivers were related to land management, one driver was related to planning and policies, three drivers were related to socio-economic conditions, and one driver related to legal land status. Depending on the specific desertification process, factors may be positively or

  13. Identification of drivers for modular production

    DEFF Research Database (Denmark)

    Brunoe, Thomas Ditlev; Bossen, Jacob; Nielsen, Kjeld

    2015-01-01

    Todays competitive environment in industry creates a need for companies to enhance their ability to introduce new products faster. To increase rampup speed reconfigurable manufacturing systems is a promising concept, however to implement this production platforms and modular manufacturing...... is required. This paper presents an analysis whether and which module drivers from general product development can be applied to the development process of a modular manufacturing system. The result is a compiled list of modular drivers for manufacturing and examples of their use....