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Sample records for force induces gene

  1. Forced expression of Hnf4a induces hepatic gene activation through directed differentiation.

    Science.gov (United States)

    Yahoo, Neda; Pournasr, Behshad; Rostamzadeh, Jalal; Fathi, Fardin

    2016-08-05

    Embryonic stem (ES) cells are capable of unlimited self-renewal and have a diverse differentiation potential. These unique features make ES cells as an attractive source for developmental biology studies. Having the mature hepatocyte in the lab with functional activities is valuable in drug discovery studies. Overexpression of hepatocyte lineage-specific transcription factors (TFs) becomes a promising approach in pluripotent cell differentiation toward liver cells. Many studies generate transgenic ES cell lines to examine the effects of specific TFs overexpression in cell differentiation. In the present report, we have addressed whether a suspension or adherent model of differentiation is an appropriate way to study the role of Hnf4a overexpression. We generated ES cells that carried a doxycycline (Dox)-inducible Hnf4a using lentiviral vectors. The transduced cells were subjected to induced Hnf4a overexpression through both spontaneous and directed differentiation methods. Gene expression analysis showed substantially increased expression of hepatic gene markers, particularly Ttr and endogenous Hnf4a, in transduced cells differentiated by the directed approach. These results demonstrated that forced expression of TFs during directed differentiation would be an appropriate way to study relevant gene activation and the effects of overexpression in the context of hepatic differentiation.

  2. Expression of ACC Oxidase Gene from Sugarcane Induced by Hormones and Environmental Force

    Institute of Scientific and Technical Information of China (English)

    WANG Ai-qin; YANG Li-tao; WANG Zi-zhang; WEI Yu-tuo; HE Long-fei; LI Yang-rui

    2005-01-01

    In the present study, a full-length cDNA encoding 1-aminocyclopropane-1-carboxylic acid oxidase gene has been cloned from sugarcane (named GZ-ACO). Two primers were designed for coding the ORF in the full-length cDNA of GZ-ACO gene from sugarcane. PCR amplification was performed with sugarcane DNA template, and a fragment of 1 104 bp (GZ34)was obtained. GZ34 was labeled with [α-32p] dCTP as the probe and used for hybridization after cloning and sequencing.Southern blotting analysis indicated that there were at least three other sequences, which weakly hybridized with the GZ34. Northern analysis showed that GZ34 was strongly induced by treatment with IAA, BA, ethephon, LiC1 and cold stress, respectively. As a contrast, the mRNA for ACO gene was at lower levels for both the light-grown and dark-grown plants without additional treatment. There were two transcripts in the dark-grown plants and three transcripts in the treatments with IAA, BA and cold stress, but there was only one transcript in ethephon treatment. It showed that GZ-ACO might be a gene connected with ethylene formation and take part in response to the induction of plant hormone and environmental stress.

  3. Force induced DNA melting

    Energy Technology Data Exchange (ETDEWEB)

    Santosh, Mogurampelly; Maiti, Prabal K [Center for Condensed Matter Theory, Department of Physics, Indian Institute of Science, Bangalore-12 (India)], E-mail: santosh@physics.iisc.ernet.in, E-mail: maiti@physics.iisc.ernet.in

    2009-01-21

    When pulled along the axis, double-strand DNA undergoes a large conformational change and elongates by roughly twice its initial contour length at a pulling force of about 70 pN. The transition to this highly overstretched form of DNA is very cooperative. Applying a force perpendicular to the DNA axis (unzipping), double-strand DNA can also be separated into two single-stranded DNA, this being a fundamental process in DNA replication. We study the DNA overstretching and unzipping transition using fully atomistic molecular dynamics (MD) simulations and argue that the conformational changes of double-strand DNA associated with either of the above mentioned processes can be viewed as force induced DNA melting. As the force at one end of the DNA is increased the DNA starts melting abruptly/smoothly above a critical force depending on the pulling direction. The critical force f{sub m}, at which DNA melts completely decreases as the temperature of the system is increased. The melting force in the case of unzipping is smaller compared to the melting force when the DNA is pulled along the helical axis. In the case of melting through unzipping, the double-strand separation has jumps which correspond to the different energy minima arising due to sequence of different base pairs. The fraction of Watson-Crick base pair hydrogen bond breaking as a function of force does not show smooth and continuous behavior and consists of plateaus followed by sharp jumps.

  4. Sex-Specific Diurnal Immobility Induced by Forced Swim Test in Wild Type and Clock Gene Deficient Mice

    Directory of Open Access Journals (Sweden)

    Ningyue Li

    2015-03-01

    Full Text Available Objective: The link between alterations in circadian rhythms and depression are well established, but the underlying mechanisms are far less elucidated. We investigated the circadian characteristics of immobility behavior in wild type (WT mice and mice with mutations in core Clock genes. Methods: All mice were tested with forced swim test (FST at 4 h intervals. Results: These experiments revealed significant diurnal rhythms associated with immobility behavior in both male and female WT mice with sex-different circadian properties. In addition, male mice showed significantly less immobility during the night phase in comparison to female mice. Female Per1Brdm1 mice also showed significant rhythmicity. However, the timing of rhythmicity was very different from that observed in female wild type mice. Male Per1Brdm1 mice showed a pattern of rhythmicity similar to that of wild type mice. Furthermore, female Per1Brdm1 mice showed higher duration of immobility in comparison to male Per1Brdm1 mice in both daytime and early night phases. Neither Per2Brdm1 nor ClockΔ19 mice showed significant rhythmicity, but both female Per2Brdm1 and ClockΔ19 mice had lower levels of immobility, compared to males. Conclusions: This study highlights the differences in the circadian characteristics of immobility induced by FST in WT, ClockΔ19, Per1, and Per2 deficient mice.

  5. Sex-Specific Diurnal Immobility Induced by Forced Swim Test in Wild Type and Clock Gene Deficient Mice

    OpenAIRE

    Ningyue Li; Yanhua Xu; Xiaojuan Chen; Qing Duan; Mei Zhao

    2015-01-01

    Objective: The link between alterations in circadian rhythms and depression are well established, but the underlying mechanisms are far less elucidated. We investigated the circadian characteristics of immobility behavior in wild type (WT) mice and mice with mutations in core Clock genes. Methods: All mice were tested with forced swim test (FST) at 4 h intervals. Results: These experiments revealed significant diurnal rhythms associated with immobility behavior in both male and female WT mice...

  6. Pedestrian-induced lateral forces on footbridges

    DEFF Research Database (Denmark)

    Ingólfsson, Einar Thór; Georgakis, Christos T.; Jönsson, Jeppe

    2011-01-01

    This paper investigates the phenomenon of excessive pedestrian-induced lateral vibrations as observed on several high-profile footbridges. The vibrations are a consequence of human-structure interaction, in which the forces generated by the pedestrians depend strongly on the vibration of the unde...... amplitude lateral vibrations are the results of correlated pedestrian forces in the form of negative damping, with amplitudes that depend on the relationship between the step frequency and the frequency of the lateral movement.......This paper investigates the phenomenon of excessive pedestrian-induced lateral vibrations as observed on several high-profile footbridges. The vibrations are a consequence of human-structure interaction, in which the forces generated by the pedestrians depend strongly on the vibration...... motion at varying combinations of frequencies (0.33-1.07 Hz) and amplitudes (4.5-48 mm). The component of the pedestrian-induced force which is caused by the laterally moving surface is herewith quantified through equivalent velocity and acceleration proportional coefficients. It is shown that large...

  7. Pedestrian-induced lateral forces on footbridges

    DEFF Research Database (Denmark)

    Ingolfsson, Einar Thor; Georgakis, Christos T.; Jönsson, Jeppe

    2012-01-01

    This paper investigates the phenomenon of excessive pedestrian-induced lateral vibrations as observed on several high-profile footbridges. The vibrations are a consequence of human-structure interaction, in which the forces generated by the pedestrians depend strongly on the vibration of the unde...... amplitude lateral vibrations are the results of correlated pedestrian forces in the form of negative damping, with amplitudes that depend on the relationship between the step frequency and the frequency of the lateral movement.......This paper investigates the phenomenon of excessive pedestrian-induced lateral vibrations as observed on several high-profile footbridges. The vibrations are a consequence of human-structure interaction, in which the forces generated by the pedestrians depend strongly on the vibration...... motion at varying combinations of frequencies (0.33-1.07 Hz) and amplitudes (4.5-48 mm). The component of the pedestrian-induced force which is caused by the laterally moving surface is herewith quantified through equivalent velocity and acceleration proportional coefficients. It is shown that large...

  8. Regulating gene-expression by mechanical force

    Science.gov (United States)

    Visscher, Koen

    2008-10-01

    Initiation of transcription is an attractive target for controlling gene expression. Initiation typically involves binding of RNA polymerase to the DNA, followed by a rapid transition into a ``closed'' complex, and a subsequent transition into the ``open'' complex in which the DNA is locally melted. Nature makes good use of this target, for example in the form of repressor proteins that bind DNA and inhibit transcription. Here we will show that initiation of transcription is also dependent upon DNA tension and thus may be controlled by force alone, without the need for any accessory proteins. Using a three-bead assay in conjunction with optical tweezers we have shown that transient interactions of T7 RNA polymerase with the DNA promoter site shorten significantly, by up to a factor of ˜20, when DNA tension is increased. Experiments in the presence and absence of nucleotides have allowed us to conclude that force is likely to affect the rate constants into and/or out of the open complex, rather than the off-rate from the closed complex.

  9. Force-Induced Changes in Subnuclear Movement and Rheology

    Science.gov (United States)

    Booth-Gauthier, Elizabeth A.; Alcoser, Turi A.; Yang, Ge; Dahl, Kris N.

    2012-01-01

    Extracellular mechanical forces result in changes in gene expression, but it is unclear how cells are able to permanently adapt to new mechanical environments because chemical signaling pathways are short-lived. We visualize force-induced changes in nuclear rheology to examine short- and long-time genome organization and movements. Punctate labels in the nuclear interior of HeLa, human umbilical vein endothelial, and osteosarcoma (Saos-2) cells allow tracking of nuclear movements in cells under varying levels of shear and compressive force. Under adequate shear stress two distinct regimes develop in cells under mechanical stimulation: an initial event of increased intranuclear movement followed by a regime of intranuclear movements that reflect the dose of applied force. At early times there is a nondirectionally oriented response with a small increase in nuclear translocations. After 30 min, there is a significant increase in nuclear movements, which scales with the amount of shear or compressive stress. The similarities in the nuclear response to shear and compressive stress suggest that the nucleus is a mechanosensitive element within the cell. Thus, applied extracellular forces stimulate intranuclear movements, resulting in repositioning of nuclear bodies and the associated chromatin within the nucleus. PMID:23260044

  10. Reactivation of developmentally silenced globin genes by forced chromatin looping.

    Science.gov (United States)

    Deng, Wulan; Rupon, Jeremy W; Krivega, Ivan; Breda, Laura; Motta, Irene; Jahn, Kristen S; Reik, Andreas; Gregory, Philip D; Rivella, Stefano; Dean, Ann; Blobel, Gerd A

    2014-08-14

    Distal enhancers commonly contact target promoters via chromatin looping. In erythroid cells, the locus control region (LCR) contacts β-type globin genes in a developmental stage-specific manner to stimulate transcription. Previously, we induced LCR-promoter looping by tethering the self-association domain (SA) of Ldb1 to the β-globin promoter via artificial zinc fingers. Here, we show that targeting the SA to a developmentally silenced embryonic globin gene in adult murine erythroblasts triggers its transcriptional reactivation. This activity depends on the LCR, consistent with an LCR-promoter looping mechanism. Strikingly, targeting the SA to the fetal γ-globin promoter in primary adult human erythroblasts increases γ-globin promoter-LCR contacts, stimulating transcription to approximately 85% of total β-globin synthesis, with a reciprocal reduction in adult β-globin expression. Our findings demonstrate that forced chromatin looping can override a stringent developmental gene expression program and suggest a novel approach to control the balance of globin gene transcription for therapeutic applications.

  11. Forced expression of the Oct-4 gene influences differentiation of embryonic stem cells

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    By transfecting an Oct-4 expression plasmid into embryonic stem cells (ES cells), the ES-O cell line was constructed, which sustained the expression of Oct-4 gene when induced by retinoic acid. Forced expression of Oct-4 gene could not sustain the stem property of ES-O cells without the differentiation inhibiting factor LIF, but if LIF exists,forced expression of Oct-4 gene could enhance the ability to sustain the undifferentiation state and inhibit cell differentiation induced by retinoic acid. It was indicated that Oct-4 must cooperate with LIF to sustain the undifferentiation state of ES cells. During the cell differentiation, ES-O cells tend to differentiate into neural cells, suggesting that forced expression of Oct-4 gene may be in relation with the differentiation of neuroderm.

  12. Measurement of Laterally Induced Optical Forces at the Nanoscale

    CERN Document Server

    Huang, Fei; Wickramasinghe, Hemanta Kumar

    2016-01-01

    We demonstrate the measurement of laterally induced optical forces using an Atomic Force Microscope (AFM). The lateral electric field distribution between a gold coated AFM probe and a nano-aperture in a gold film is mapped by measuring the lateral optical force between the apex of the AFM probe and the nano-aperture. Torsional eigenmodes of an AFM cantilever probe were used to detect the laterally induced optical forces. We engineered the cantilever shape using a focused ion beam to enhance the torsional eigenmode resonance. The measured lateral optical force agrees well with simulations. This technique can be extended to simultaneously detect both lateral and longitudinal optical forces at the nanoscale by using an AFM cantilever as a multichannel detector. This will enable simultaneous Photon Induced Force Microscopy (PIFM) detection of molecular responses with different incident field polarizations. The technique can be implemented on both cantilever and tuning fork based AFMs.

  13. Retinal Changes Induced by Epiretinal Tangential Forces

    Directory of Open Access Journals (Sweden)

    Mario R. Romano

    2015-01-01

    Full Text Available Two kinds of forces are active in vitreoretinal traction diseases: tangential and anterior-posterior forces. However, tangential forces are less characterized and classified in literature compared to the anterior-posterior ones. Tangential epiretinal forces are mainly due to anomalous posterior vitreous detachment (PVD, vitreoschisis, vitreopapillary adhesion (VPA, and epiretinal membranes (ERMs. Anomalous PVD plays a key role in the formation of the tangential vectorial forces on the retinal surface as consequence of gel liquefaction (synchysis without sufficient and fast vitreous dehiscence at the vitreoretinal interface. The anomalous and persistent adherence of the posterior hyaloid to the retina can lead to vitreomacular/vitreopapillary adhesion or to a formation of avascular fibrocellular tissue (ERM resulting from the proliferation and transdifferentiation of hyalocytes resident in the cortical vitreous remnants after vitreoschisis. The right interpretation of the forces involved in the epiretinal tangential tractions helps in a better definition of diagnosis, progression, prognosis, and surgical outcomes of vitreomacular interfaces.

  14. Effects of Continuous and Interrupted Forces on Gene Transcription in Periodontal Ligament Cells in Vitro

    Directory of Open Access Journals (Sweden)

    Seyed Nasser Ostad

    2011-10-01

    Full Text Available The biological mechanisms of tooth movement are based on the response of periodontal tissues to mechanical forces. The final result of these responses is remodeling of the extracellular matrix. Tissue reactions may vary depending upon the type, magnitude and duration of the applied forces. The purpose of the present study was to analyze the effects of centrifugal force on the transcription of collagen type-I (Col-I, matrix metalloproteinase-1 (MMP-1, and tissue inhibitor of metalloproteinase- 1 (TIMP-1 genes in human periodontal ligament (PDL fibroblasts. Human fibroblasts obtained from the PDL were cultured and subjected to centrifugal forces (36.3 g/cm2 for 30, 60 and 90 min continuously. This was also carried out interruptedly, three times for 30 min and six times for 15 min. The mRNAs encoding for Col-I, MMP-1, and TIMP-1 were quantified using RT-PCR. The mRNA levels of Col-I and MMP-1 were increased when continuous force was applied for 30 min and 60 min respectively. The interrupted force had almost no effect on Col-I, MMP-1 and TIMP-1 genes. These results indicate that continuous forces may have a greater effect in inducing gene expression during the remodeling process of PDL compared to interrupted forces with short rest periods.

  15. Current-induced forces: a simple derivation

    DEFF Research Database (Denmark)

    Todorov, Tchavdar N.; Dundas, Daniel; Lü, Jing-Tao;

    2014-01-01

    We revisit the problem of forces on atoms under current in nanoscale conductors. We derive and discuss the five principal kinds of force under steady-state conditions from a simple standpoint that-with the help of background literature-should be accessible to physics undergraduates. The discussion...

  16. BRD4 regulates fructose-inducible lipid accumulation-related genes in the mouse liver.

    Science.gov (United States)

    Yamada, Aki; Honma, Kazue; Mochizuki, Kazuki; Goda, Toshinao

    2016-10-01

    Fructose intake induces hepatic steatosis by activating fat synthesis. In this study, we searched for genes that showed acute induction in the livers of mice force-fed with fructose, and examined how this induction is regulated. We identified genes induced at 6h after the fructose force-feeding using a microarray and quantitative real-time RT-PCR. Histone acetylation and an acetylated histone binding protein bromodomain containing (BRD)4 binding around the fructose-inducible genes were examined using a chromatin immunoprecipitation assay. We examined whether (+)-JQ1, an inhibitor of the binding between the BRD4 and acetylated histones, inhibited the expressions of fructose-inducible genes, histone acetylation and BRD4 binding around the genes. We identified upregulated genes related to lipid accumulation, such as Cyp8b1, Dak and Plin5, in mice force-fed with fructose compared with those force-fed with glucose. Acetylation of histones H3 and H4, and BRD4 binding around the transcribed region of those fructose-inducible genes, were enhanced by fructose force-feeding. Meanwhile, (+)-JQ1 treatment reduced expressions of fructose-inducible genes, histone acetylation and BRD4 binding around these genes. Acute induction of genes related to lipid accumulation in the livers of mice force-fed with fructose is associated with the induction of histone acetylation and BRD4 binding around these genes. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Activation-induced force enhancement in human adductor pollicis.

    Science.gov (United States)

    Oskouei, Ali E; Herzog, Walter

    2009-10-01

    It has been known for a long time that the steady-state isometric force after muscle stretch is bigger than the corresponding force obtained in a purely isometric contraction for electrically stimulated and maximal voluntary contractions (MVC). Recent studies using sub-maximal voluntary contractions showed that force enhancement only occurred in a sub-group of subjects suggesting that force enhancement for sub-maximal voluntary contractions has properties different from those of electrically-induced and maximal voluntary contractions. Specifically, force enhancement for sub-maximal voluntary contractions may contain an activation-dependent component that is independent of muscle stretching. To address this hypothesis, we tested for force enhancement using (i) sub-maximal electrically-induced contractions and stretch and (ii) using various activation levels preceding an isometric reference contraction at 30% of MVC (no stretch). All tests were performed on human adductor pollicis muscles. Force enhancement following stretching was found for all subjects (n=10) and all activation levels (10%, 30%, and 60% of MVC) for electrically-induced contractions. In contrast, force enhancement at 30% of MVC, preceded by 6s of 10%, 60%, and 100% of MVC was only found in a sub-set of the subjects and only for the 60% and 100% conditions. This result suggests that there is an activation-dependent force enhancement for some subjects for sub-maximal voluntary contractions. This activation-dependent force enhancement was always smaller than the stretch-induced force enhancement obtained at the corresponding activation levels. Active muscle stretching increased the force enhancement in all subjects, independent whether they showed activation dependence or not. It appears that post-activation potentiation, and the associated phosphorylation of the myosin light chains, might account for the stretch-independent force enhancement observed here.

  18. Superlens induced loss-insensitive optical force

    CERN Document Server

    Cui, Xiaohan; Chan, C T

    2016-01-01

    A slab with relative permittivity $\\epsilon = - 1 + i\\delta$ and permeability $\\mu = - 1 + i\\delta $ has a critical distance away from the slab where a small particle will either be cloaked or imaged depending on whether it is located inside or outside that critical distance. We find that the optical force acting on a small cylinder under plane wave illumination reaches a maximum value at this critical distance. Contrary to the usual observation that superlens systems should be highly loss-sensitive, this maximum optical force remains a constant when loss is changed within a certain range. For a fixed particle-slab distance, increasing loss can even amplify the optical force acting on the small cylinder, contrary to the usual belief that loss compromises the response of supenlens.

  19. Transcription upregulation via force-induced direct stretching of chromatin

    Science.gov (United States)

    Tajik, Arash; Zhang, Yuejin; Wei, Fuxiang; Sun, Jian; Jia, Qiong; Zhou, Wenwen; Singh, Rishi; Khanna, Nimish; Belmont, Andrew S.; Wang, Ning

    2016-12-01

    Mechanical forces play critical roles in the function of living cells. However, the underlying mechanisms of how forces influence nuclear events remain elusive. Here, we show that chromatin deformation as well as force-induced transcription of a green fluorescent protein (GFP)-tagged bacterial-chromosome dihydrofolate reductase (DHFR) transgene can be visualized in a living cell by using three-dimensional magnetic twisting cytometry to apply local stresses on the cell surface via an Arg-Gly-Asp-coated magnetic bead. Chromatin stretching depended on loading direction. DHFR transcription upregulation was sensitive to load direction and proportional to the magnitude of chromatin stretching. Disrupting filamentous actin or inhibiting actomyosin contraction abrogated or attenuated force-induced DHFR transcription, whereas activating endogenous contraction upregulated force-induced DHFR transcription. Our findings suggest that local stresses applied to integrins propagate from the tensed actin cytoskeleton to the LINC complex and then through lamina-chromatin interactions to directly stretch chromatin and upregulate transcription.

  20. Subsurface Airflow Induced by Natural Forcings

    Institute of Scientific and Technical Information of China (English)

    Jiu J. Jiao; LI Hai-long

    2004-01-01

    Subsurface air flow can be induced by natural processes, such as atmospheric or barometric pressure changes, water table fluctuations, topographic effects, and rainfall infiltration. Barometric pressure fluctuations are the most common cause of subsurface air flow, which can be significant under favourable geological conditions. This process has been studied most extensively because of its application to passive soil vapor extraction. Soil air flow induced by water table fluctuations can be significant, particularly where the fluctuations are of high frequency, for example, in tidal-influenced coastal areas. Topographic effects can lead to strong subsoil air flow in areas with great elevation differences. Rainfall infiltration usually produces only weak airflow. Air flow induced by these natural processes has important environmental and engineering implications. Among the different processes, air flow induced by tidal fluctuations has been studied the least, although it has exciting applications to coastal engineering projects and environmental remediation.

  1. Gene duplication as a major force in evolution

    Indian Academy of Sciences (India)

    Santoshkumar Magadum; Urbi Banerjee; Priyadharshini Murugan; Doddabhimappa Gangapur; Rajasekar Ravikesavan

    2013-04-01

    Gene duplication is an important mechanism for acquiring new genes and creating genetic novelty in organisms. Many new gene functions have evolved through gene duplication and it has contributed tremendously to the evolution of developmental programmes in various organisms. Gene duplication can result from unequal crossing over, retroposition or chromosomal (or genome) duplication. Understanding the mechanisms that generate duplicate gene copies and the subsequent dynamics among gene duplicates is vital because these investigations shed light on localized and genomewide aspects of evolutionary forces shaping intra-specific and inter-specific genome contents, evolutionary relationships, and interactions. Based on whole-genome analysis of Arabidopsis thaliana, there is compelling evidence that angiosperms underwent two whole-genome duplication events early during their evolutionary history. Recent studies have shown that these events were crucial for creation of many important developmental and regulatory genes found in extant angiosperm genomes. Recent studies also provide strong indications that even yeast (Saccharomyces cerevisiae), with its compact genome, is in fact an ancient tetraploid. Gene duplication can provide new genetic material for mutation, drift and selection to act upon, the result of which is specialized or new gene functions. Without gene duplication the plasticity of a genome or species in adapting to changing environments would be severely limited. Whether a duplicate is retained depends upon its function, its mode of duplication, (i.e. whether it was duplicated during a whole-genome duplication event), the species in which it occurs, and its expression rate. The exaptation of preexisting secondary functions is an important feature in gene evolution, just as it is in morphological evolution.

  2. Measurement of laterally induced optical forces at the nanoscale

    Science.gov (United States)

    Huang, Fei; Tamma, Venkata Ananth; Rajaei, Mohsen; Almajhadi, Mohammad; Kumar Wickramasinghe, H.

    2017-02-01

    We demonstrate the measurement of laterally induced optical forces using an Atomic Force Microscope (AFM). The lateral electric field distribution between a gold coated AFM probe and a single nano-aperture in a gold film is mapped by measuring the lateral optical force between the apex of the AFM probe and the nano-aperture. The fundamental torsional eigen-mode of an AFM cantilever probe was used to detect the laterally induced optical forces. We engineered the cantilever shape using focused ion beam milling to improve the detected signal to noise ratio. The measured distributions of lateral optical force agree well with electromagnetic simulations of the metal coated AFM probe interacting with the nano-aperture. This technique can be extended to simultaneously detect both lateral and longitudinal optical forces at the nanoscale by using an AFM cantilever as a multi-channel detector. This will enable simultaneous Photon Induced Force Microscopy detection of molecular responses with different incident field polarizations. The technique can be implemented on both cantilever and tuning fork based AFMs.

  3. Excitation-induced force recovery in potassium-inhibited rat soleus muscle

    Science.gov (United States)

    Nielsen, Ole Bækgaard; Hilsted, Linda; Clausen, Torben

    1998-01-01

    Excitation markedly stimulates the Na+-K+ pump in skeletal muscle. The effect of this stimulation on contractility was examined in rat soleus muscles exposed to high extracellular K+ concentration ([K+]o).At a [K+]o of 10 mm, tetanic force declined to 58 % of the force in standard buffer with 5.9 mm K+. Subsequent direct stimulation of the muscle at 1 min intervals with 30 Hz pulse trains of 2 s duration induced a 97 % recovery of force within 14 min. Force recovery could also be elicited by stimulation via the nerve. In muscles exposed to 12.5 mm K+, 30 Hz pulse trains of 2 s duration at 1 min intervals induced a recovery of force from 16 ± 2 to 62 ± 4 % of the initial control force at a [K+]o of 5.9 mm.The recovery of force was associated with a decrease in intracellular Na+ and was blocked by ouabain. This indicates that the force recovery was secondary to activation of the Na+-K+ pump.Excitation stimulates the release of calcitonin gene-related peptide (CGRP) from nerves in the muscle. Since CGRP stimulates the Na+-K+ pump, this may contribute to the excitation-induced force recovery. Indeed, reducing CGRP content by capsaicin pre-treatment or prior denervation prevented both the excitation-induced force recovery and the drop in intracellular Na+.The data suggest that activation of the Na+-K+ pump in contracting muscles counterbalances the depressing effect of reductions in the chemical gradients for Na+ and K+ on excitability. PMID:9769424

  4. Image reconstruction with acoustic radiation force induced shear waves

    Science.gov (United States)

    McAleavey, Stephen A.; Nightingale, Kathryn R.; Stutz, Deborah L.; Hsu, Stephen J.; Trahey, Gregg E.

    2003-05-01

    Acoustic radiation force may be used to induce localized displacements within tissue. This phenomenon is used in Acoustic Radiation Force Impulse Imaging (ARFI), where short bursts of ultrasound deliver an impulsive force to a small region. The application of this transient force launches shear waves which propagate normally to the ultrasound beam axis. Measurements of the displacements induced by the propagating shear wave allow reconstruction of the local shear modulus, by wave tracking and inversion techniques. Here we present in vitro, ex vivo and in vivo measurements and images of shear modulus. Data were obtained with a single transducer, a conventional ultrasound scanner and specialized pulse sequences. Young's modulus values of 4 kPa, 13 kPa and 14 kPa were observed for fat, breast fibroadenoma, and skin. Shear modulus anisotropy in beef muscle was observed.

  5. Vortex ring breakdown induced by topographic forcing

    Energy Technology Data Exchange (ETDEWEB)

    Geiser, J; Kiger, K T, E-mail: kkiger@umd.edu [Department of Mechanical Engineering, University of Maryland, College Park, MD 20910 (United States)

    2011-12-22

    Detailed measurements of the vortex breakdown within a strongly forced impinging jet are presented, with the goal of studying the effects of a small topographic disturbance on the breakdown and turbulence structure. This work is related to an ongoing effort to understand the dynamics of sediment suspension within a landing rotorcraft where a mobile boundary is subject to rapid erosion and deposition. The current work compares the results of a uniform surface to that of a small radial fence placed upstream of the vortex impingement location. The result is a dramatic increase in the coherence of the three-dimensional looping exhibited by the secondary vortex, leading to a more organized and strongly perturbed mean flow. Specifically, a triple decomposition of the velocity fluctuations indicates a very intense periodic stress in the vicinity of the impingement site, followed by a significant decay. Conversely, the random component of the fluctuating stresses gradually increases to modest levels as the coherent contributions decrease, eventually becoming greater than the coherent stress. The fence produces a bifurcation in the flow through the perturbation of the secondary vortex, which in turn creates a high-and low-speed streak on either side of the fence. The subsequent dynamics leads to increased fluctuating stress in the high-speed region, and a dramatically lower stress in the low-speed region, favoring preferential erosion on either side of the topographic disturbance.

  6. Fatigue-induced dissociation between rate of force development and maximal force across repeated rapid contractions.

    Science.gov (United States)

    Boccia, Gennaro; Dardanello, Davide; Tarperi, Cantor; Festa, Luca; La Torre, Antonio; Pellegrini, Barbara; Schena, Federico; Rainoldi, Alberto

    2017-08-01

    We examined whether the presence of fatigue induced by prolonged running influenced the time courses of force generating capacities throughout a series of intermittent rapid contractions. Thirteen male amateur runners performed a set of 15 intermittent isometric rapid contractions of the knee extensor muscles, (3s/5s on/off) the day before (PRE) and immediately after (POST) a half marathon. The maximal voluntary contraction force, rate of force development (RFDpeak), and their ratio (relative RFDpeak) were calculated. At POST, considering the first (out of 15) repetition, the maximal force and RFDpeak decreased (p<0.0001) at the same extent (by 22±6% and 24±22%, respectively), resulting in unchanged relative RFDpeak (p=0.6). Conversely, the decline of RFDpeak throughout the repetitions was more pronounced at POST (p=0.02), thus the decline of relative RFDpeak was more pronounced (p=0.007) at POST (-25±13%) than at PRE (-3±13%). The main finding of this study was that the fatigue induced by a half-marathon caused a more pronounced impairment of rapid compared to maximal force in the subsequent intermittent protocol. Thus, the fatigue-induced impairment in rapid muscle contractions may have a greater effect on repeated, rather than on single, attempts of maximal force production. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Centrifugal force induces human ligamentum flavum fibroblasts inflammation through activation of JNK and p38 pathways.

    Science.gov (United States)

    Chao, Yuan-Hung; Tsuang, Yang-Hwei; Sun, Jui-Sheng; Sun, Man-Ger; Chen, Ming-Hong

    2012-01-01

    Inflammation has been proposed to be an important causative factor in ligamentum flavum hypertrophy. However, the mechanisms of mechanical load on inflammation of ligamentum flavum remain unclear. In this study, we used an in vitro model of human ligamentum flavum fibroblasts subjected to centrifugal force to elucidate the effects of mechanical load on cultured human ligamentum flavum fibroblasts; we further studied its molecular and biochemical mechanisms. Human ligamentum flavum fibroblasts were obtained from six patients undergoing lumbar spine surgery. Monolayer cultures of human ligamentum flavum fibroblasts were subjected to different magnitudes of centrifugal forces. Cell viability, cell death, biochemical response, and molecular response to centrifugal forces were analyzed. It was found that centrifugal stress significantly suppressed cell viability without inducing cell death. Centrifugal force at 67.1 g/cm(2) for 60 min significantly increases the production of prostaglandin E2 and nitric oxide as well as gene expression of proinflammatory cytokines, including interleukin (IL)-1α, IL-1β and IL-6, showed that centrifugal force-dependent induction of cyclooxygense-2 and inducible NO synthase required JNK and p38 mitogen-activated protein kinase, but not ERK 1/2 activities. This study suggested that centrifugal force does induce inflammatory responses in human ligamentum flavum fibroblasts. The activation of both JNK and p38 mitogen-activated protein kinase mechanotransduction cascades is a crucial intracellular mechanism that mediates cyclooxygense-2/prostaglandin E2 and inducible NO synthase/nitric oxide production.

  8. Microtubule depolymerization induces traction force increase through two distinct pathways.

    Science.gov (United States)

    Rape, Andrew; Guo, Wei-hui; Wang, Yu-li

    2011-12-15

    Traction forces increase after microtubule depolymerization; however, the signaling mechanisms underlying this, in particular the dependence upon myosin II, remain unclear. We investigated the mechanism of traction force increase after nocodazole-induced microtubule depolymerization by applying traction force microscopy to cells cultured on micropatterned polyacrylamide hydrogels to obtain samples of homogeneous shape and size. Control cells and cells treated with a focal adhesion kinase (FAK) inhibitor showed similar increases in traction forces, indicating that the response is independent of FAK. Surprisingly, pharmacological inhibition of myosin II did not prevent the increase of residual traction forces upon nocodazole treatment. This increase was abolished upon pharmacological inhibition of FAK. These results suggest two distinct pathways for the regulation of traction forces. First, microtubule depolymerization activates a myosin-II-dependent mechanism through a FAK-independent pathway. Second, microtubule depolymerization also enhances traction forces through a myosin-II-independent, FAK-regulated pathway. Traction forces are therefore regulated by a complex network of complementary signals and force-generating mechanisms.

  9. Nonconservative current-induced forces: A physical interpretation

    Directory of Open Access Journals (Sweden)

    Tchavdar N. Todorov

    2011-10-01

    Full Text Available We give a physical interpretation of the recently demonstrated nonconservative nature of interatomic forces in current-carrying nanostructures. We start from the analytical expression for the curl of these forces, and evaluate it for a point defect in a current-carrying system. We obtain a general definition of the capacity of electrical current flow to exert a nonconservative force, and thus do net work around closed paths, by a formal noninvasive test procedure. Second, we show that the gain in atomic kinetic energy over time, generated by nonconservative current-induced forces, is equivalent to the uncompensated stimulated emission of directional phonons. This connection with electron–phonon interactions quantifies explicitly the intuitive notion that nonconservative forces work by angular momentum transfer.

  10. Nonconservative current-induced forces: A physical interpretation.

    Science.gov (United States)

    Todorov, Tchavdar N; Dundas, Daniel; Paxton, Anthony T; Horsfield, Andrew P

    2011-01-01

    We give a physical interpretation of the recently demonstrated nonconservative nature of interatomic forces in current-carrying nanostructures. We start from the analytical expression for the curl of these forces, and evaluate it for a point defect in a current-carrying system. We obtain a general definition of the capacity of electrical current flow to exert a nonconservative force, and thus do net work around closed paths, by a formal noninvasive test procedure. Second, we show that the gain in atomic kinetic energy over time, generated by nonconservative current-induced forces, is equivalent to the uncompensated stimulated emission of directional phonons. This connection with electron-phonon interactions quantifies explicitly the intuitive notion that nonconservative forces work by angular momentum transfer.

  11. Radiation-induced gene responses

    Energy Technology Data Exchange (ETDEWEB)

    Woloschak, G.E.; Paunesku, T.; Shearin-Jones, P.; Oryhon, J.

    1996-12-31

    In the process of identifying genes that are differentially regulated in cells exposed to ultraviolet radiation (UV), we identified a transcript that was repressed following the exposure of cells to a combination of UV and salicylate, a known inhibitor of NF-kappaB. Sequencing this band determined that it has identify to lactate dehydrogenase, and Northern blots confirmed the initial expression pattern. Analysis of the sequence of the LDH 5` region established the presence of NF-kappaB, Sp1, and two Ap-2 elements; two partial AP- 1; one partial RE, and two halves of E-UV elements were also found. Electromobility shift assays were then performed for the AP-1, NF- kappaB, and E-UV elements. These experiments revealed that binding to NF-kappaB was induced by UV but repressed with salicylic acid; UV did not affect AP-1 binding, but salicylic acid inhibited it alone or following UV exposure; and E-UV binding was repressed by UV, and salicylic acid had little effect. Since the binding of no single element correlated with the expression pattern of LDH, it is likely that multiple elements govern UV/salicylate-mediated expression.

  12. Force-gradient-induced mechanical dissipation of quartz tuning fork force sensors used in atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Castellanos-Gomez, A. [Departamento de Fisica de la Materia Condensada (C-III), Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid (Spain); Agrait, N. [Departamento de Fisica de la Materia Condensada (C-III), Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid (Spain); Instituto Universitario de Ciencia de Materiales ' Nicolas Cabrera' , Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid (Spain); Instituto Madrileno de Estudios Avanzados en Nanociencia, IMDEA-Nanociencia, 28049 Madrid (Spain); Rubio-Bollinger, G., E-mail: gabino.rubio@uam.es [Departamento de Fisica de la Materia Condensada (C-III), Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid (Spain); Instituto Universitario de Ciencia de Materiales ' Nicolas Cabrera' , Universidad Autonoma de Madrid, Campus de Cantoblanco, 28049 Madrid (Spain)

    2011-02-15

    We have studied the dynamics of quartz tuning fork resonators used in atomic force microscopy taking into account the mechanical energy dissipation through the attachment of the tuning fork base. We find that the tuning fork resonator quality factor changes even in the case of a purely elastic sensor-sample interaction. This is due to the effective mechanical imbalance of the tuning fork prongs induced by the sensor-sample force gradient, which in turn has an impact on dissipation through the attachment of the resonator base. This effect may yield a measured dissipation signal that can be different from the one exclusively related to the dissipation between the sensor and the sample. We also find that there is a second-order term in addition to the linear relationship between the sensor-sample force gradient and the resonance frequency shift of the tuning fork that is significant even for force gradients usually present in atomic force microscopy, which are in the range of tens of N/m. -- Research Highlights: {yields} Dynamics of miniature tuning fork force sensors: a mechanical model. {yields} Non-linear relationship between resonance frequency shift and applied force gradient. {yields} An apparent mechanical dissipation channel opens even for purely conservative tip-sample interactions. {yields} qPlus tuning forks configuration has lower Q factor but straightforward dynamics.

  13. Mechanochemistry Induced Using Force Exerted by a Functionalized Microscope Tip

    DEFF Research Database (Denmark)

    Zhang, Yajie; Wang, Yongfeng; Lü, Jing-Tao

    2017-01-01

    Atomic-scale mechanochemistry is realized from force exerted by a C60 -functionalized scanning tunneling microscope tip. Two conformers of tin phthalocyanine can be prepared on coinage-metal surfaces. A transition between these conformers is induced on Cu(111) and Ag(100). Density-functional calc......Atomic-scale mechanochemistry is realized from force exerted by a C60 -functionalized scanning tunneling microscope tip. Two conformers of tin phthalocyanine can be prepared on coinage-metal surfaces. A transition between these conformers is induced on Cu(111) and Ag(100). Density...

  14. Force-Induced Rupture of a DNA Duplex: From Fundamentals to Force Sensors.

    Science.gov (United States)

    Mosayebi, Majid; Louis, Ard A; Doye, Jonathan P K; Ouldridge, Thomas E

    2015-12-22

    The rupture of double-stranded DNA under stress is a key process in biophysics and nanotechnology. In this article, we consider the shear-induced rupture of short DNA duplexes, a system that has been given new importance by recently designed force sensors and nanotechnological devices. We argue that rupture must be understood as an activated process, where the duplex state is metastable and the strands will separate in a finite time that depends on the duplex length and the force applied. Thus, the critical shearing force required to rupture a duplex depends strongly on the time scale of observation. We use simple models of DNA to show that this approach naturally captures the observed dependence of the force required to rupture a duplex within a given time on duplex length. In particular, this critical force is zero for the shortest duplexes, before rising sharply and then plateauing in the long length limit. The prevailing approach, based on identifying when the presence of each additional base pair within the duplex is thermodynamically unfavorable rather than allowing for metastability, does not predict a time-scale-dependent critical force and does not naturally incorporate a critical force of zero for the shortest duplexes. We demonstrate that our findings have important consequences for the behavior of a new force-sensing nanodevice, which operates in a mixed mode that interpolates between shearing and unzipping. At a fixed time scale and duplex length, the critical force exhibits a sigmoidal dependence on the fraction of the duplex that is subject to shearing.

  15. Applications of Lorentz force in medical acoustics: Lorentz force hydrophone, Lorentz Force Electrical Impedance Tomography, Imaging of shear waves induced by Lorentz force

    CERN Document Server

    Grasland-Mongrain, Pol

    2014-01-01

    The ability of the Lorentz force to link a mechanical displacement to an electrical current presents a strong interest for medical acoustics, and three applications were studied in this thesis. In the first part of this work, a hydrophone was developed for mapping the particle velocity of an acoustic field. This hydrophone was constructed using a thin copper wire and an external magnetic field. A model was elaborated to determine the relationship between the acoustic pressure and the measured electrical current, which is induced by Lorentz force when the wire vibrates in the acoustic field of an ultrasound transducer. The built prototype was characterized and its spatial resolution, frequency response, sensitivity, robustness and directivity response were investigated. An imaging method called Lorentz Force Electrical Impedance Tomography was also studied. In this method, a biological tissue is vibrated by ultrasound in a magnetic field, which induces an electrical current by Lorentz force. The electrical imp...

  16. Earth's Inner Core dynamics induced by the Lorentz force

    CERN Document Server

    Lasbleis, M; Cardin, P; Labrosse, S

    2015-01-01

    Seismic studies indicate that the Earth's inner core has a complex structure and exhibits a strong elastic anisotropy with a cylindrical symmetry. Among the various models which have been proposed to explain this anisotropy, one class of models considers the effect of the Lorentz force associated with the magnetic field diffused within the inner core. In this paper we extend previous studies and use analytical calculations and numerical simulations to predict the geometry and strength of the flow induced by the poloidal component of the Lorentz force in a neutrally or stably stratified growing inner core, exploring also the effect of different types of boundary conditions at the inner core boundary (ICB). Unlike previous studies, we show that the boundary condition that is most likely to produce a significant deformation and seismic anisotropy is impermeable, with negligible radial flow through the boundary. Exact analytical solutions are found in the case of a negligible effect of buoyancy forces in the inne...

  17. Force-Driven Polymerization and Turgor-Induced Wall Expansion.

    Science.gov (United States)

    Ali, Olivier; Traas, Jan

    2016-05-01

    While many molecular players involved in growth control have been identified in the past decades, it is often unknown how they mechanistically act to induce specific shape changes during development. Plant morphogenesis results from the turgor-induced yielding of the extracellular and load-bearing cell wall. Its mechanochemical equilibrium appears as a fundamental link between molecular growth regulation and the effective shape evolution of the tissue. We focus here on force-driven polymerization of the cell wall as a central process in growth control. We propose that mechanical forces facilitate the insertion of wall components, in particular pectins, a process that can be modulated through genetic regulation. We formalize this idea in a mathematical model, which we subsequently test with published experimental results.

  18. Characterization of an induced pressure pumping force for microfluidics

    Science.gov (United States)

    Jiang, Hai; Fan, Na; Peng, Bei; Weng, Xuan

    2017-05-01

    The electro-osmotic pumping and pressure-driven manipulation of fluids are considered as the most common strategies in microfluidic devices. However, both of them exhibit major disadvantages such as hard integration and high reagent consumption, and they are destructive methods for detection and photo bleaching. In this paper, an electric field-effect flow control approach, combining the electro-osmotic pumping force and the pressure-driven pumping force, was developed to generate the induced pressure-driven flow in a T-shaped microfluidic chip. Electro-osmotic flow between the T-intersection and two reservoirs was demonstrated, and it provided a stable, continuous, and electric field-free flow in the section of the microchannel without the electrodes. The velocity of the induced pressure-driven flow was linearly proportional to the applied voltages. Both numerical and experimental investigations were conducted to prove the concept, and the experimental results showed good agreement with the numerical simulations. In comparison to other induced pressure pumping methods, this approach can induce a high and controllable pressure drop in the electric field-free segment, subsequently causing an induced pressure-driven flow for transporting particles or biological cells. In addition, the generation of bubbles and the blocking of the microchannel are avoided.

  19. Heat induces gene amplification in cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Bin, E-mail: yanbin@mercyhealth.com [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Mercy Cancer Center, Mercy Medical Center-North Iowa, Mason City, IA 50401 (United States); Ouyang, Ruoyun [Department of Respiratory Medicine, The Second Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410011 (China); Huang, Chenghui [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Department of Oncology, The Third Xiangya Hospital, Xinagya School of Medicine, Central South University, Changsha 410013 (China); Liu, Franklin [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States); Neill, Daniel [Department of Radiation Oncology, University of Mississippi Medical Center, Jackson, MS 39213 (United States); Li, Chuanyuan [Dermatology, Duke University Medical Center, Durham, NC 27710 (United States); Dewhirst, Mark [Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710 (United States)

    2012-10-26

    Highlights: Black-Right-Pointing-Pointer This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. Black-Right-Pointing-Pointer Hyperthermia induces DNA double strand breaks. Black-Right-Pointing-Pointer DNA double strand breaks are considered to be required for the initiation of gene amplification. Black-Right-Pointing-Pointer The underlying mechanism of heat-induced gene amplification is generation of DNA double strand breaks. -- Abstract: Background: Hyperthermia plays an important role in cancer therapy. However, as with radiation, it can cause DNA damage and therefore genetic instability. We studied whether hyperthermia can induce gene amplification in cancer cells and explored potential underlying molecular mechanisms. Materials and methods: (1) Hyperthermia: HCT116 colon cancer cells received water-submerged heating treatment at 42 or 44 Degree-Sign C for 30 min; (2) gene amplification assay using N-(phosphoacetyl)-L-aspartate (PALA) selection of cabamyl-P-synthetase, aspartate transcarbarmylase, dihydro-orotase (cad) gene amplified cells; (3) southern blotting for confirmation of increased cad gene copies in PALA-resistant cells; (4) {gamma}H2AX immunostaining to detect {gamma}H2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 Degree-Sign C for 30 min induces gene amplification. The frequency of cad gene amplification increased by 2.8 and 6.5 folds respectively; (2) heat exposure at both 42 and 44 Degree-Sign C for 30 min induces DNA double strand breaks in HCT116 cells as shown by {gamma}H2AX immunostaining. Conclusion: This study shows that heat exposure can induce gene amplification in cancer cells, likely through the generation of DNA double strand breaks, which are believed to be required for the initiation of gene amplification. This process may be promoted by heat when cellular proteins that are responsible for checkpoints, DNA replication, DNA repair and

  20. Stratospheric Annular Modes Induced By Stationary Wave Forcing

    Science.gov (United States)

    Körnich, H.; Schmitz, G.

    The variability of the winter stratosphere shows distinguishable features in the north- ern and southern hemisphere. Since these differences are based on the different plan- etary waves of the underlying atmosphere, we explore the mechanism how stationary wave forcing in the troposphere can induce a stratospheric Annular Mode using a simple GCM. The model KMCM (Kühlungsborn Mechanistic Circulation Model) extends from the ground up to 60 km height and produces a reasonable winter climate. It takes into account the different large-scale wave forcings in the troposphere as prescribed pro- cesses. This allows us to examine the stratospheric Annular-Mode generation depend- ing on different wave forcings under perpetual January conditions. Principal com- ponent analysis is applied to identify the variability patterns of the geopotential and of the zonally averaged zonal wind. By this way, it is shown that the amplitude and composition of the orographic and thermal eddy forcing determines the stratospheric Annular Mode and the related downward propagation in the temperature field. Further model simplifications are introduced in order to understand the mechanism of the stratospheric AM-generation. Using a linear model version we illuminate the influence of the different wave forcing processes on the Annular Modes. Addition- ally, a constant-troposphere model is used to clarify the importance of transient and stationary waves. Finally, the Annular Mode is interpreted in terms of the dynamical coupling of the troposphere and stratosphere.

  1. Hypergravity-induced changes in gene expression in Arabidopsis hypocotyls.

    Science.gov (United States)

    Yoshioka, R; Soga, K; Wakabayashi, K; Takeba, G; Hoson, T

    2003-01-01

    Under hypergravity conditions, the cell wall of stem organs becomes mechanically rigid and elongation growth is suppressed, which can be recognized as the mechanism for plants to resist gravitational force. The changes in gene expression by hypergravity treatment were analyzed in Arabidopsis hypocotyls by the differential display method, for identifying genes involved in hypergravity-induced growth suppression. Sixty-two cDNA clones were expressed differentially between the control and 300 g conditions: the expression levels of 39 clones increased, whereas those of 23 clones decreased under hypergravity conditions. Sequence analysis and database searching revealed that 12 clones, 9 up-regulated and 3 down-regulated, have homology to known proteins. The expression of these genes was further analyzed using RT-PCR. Finally, six genes were confirmed to be up-regulated by hypergravity. One of such genes encoded 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR), which catalyzes a reaction producing mevalonic acid, a key precursor of terpenoids such as membrane sterols and several types of hormones. The expression of HMGR gene increased within several hours after hypergravity treatment. Also, compactin, an inhibitor of HMGR, prevented hypergravity-induced growth suppression, suggesting that HMGR is involved in suppression of Arabidopsis hypocotyl growth by hypergravity. In addition, hypergravity increased the expression levels of genes encoding CCR1 and ERD15, which were shown to take part in the signaling pathway of environmental stimuli such as temperature and water, and those of the alpha-tubulin gene. These genes may be involved in a series of cellular events leading to growth suppression of stem organs under hypergravity conditions.

  2. Current-induced forces and hot spots in biased nanojunctions.

    Science.gov (United States)

    Lü, Jing-Tao; Christensen, Rasmus B; Wang, Jian-Sheng; Hedegård, Per; Brandbyge, Mads

    2015-03-06

    We investigate theoretically the interplay of current-induced forces (CIFs), Joule heating, and heat transport inside a current-carrying nanoconductor. We find that the CIFs, due to the electron-phonon coherence, can control the spatial heat dissipation in the conductor. This yields a significant asymmetric concentration of excess heating (hot spot) even for a symmetric conductor. When coupled to the electrode phonons, CIFs drive different phonon heat flux into the two electrodes. First-principles calculations on realistic biased nanojunctions illustrate the importance of the effect.

  3. Current-Induced Forces and Hot Spots in Biased Nanojunctions

    DEFF Research Database (Denmark)

    Lu, Jing Tao; Christensen, Rasmus Bjerregaard; Wang, Jian-Sheng;

    2015-01-01

    We investigate theoretically the interplay of current-induced forces (CIFs), Joule heating, and heat transport inside a current-carrying nanoconductor. We find that the CIFs, due to the electron-phonon coherence, can control the spatial heat dissipation in the conductor. This yields a significant...... asymmetric concentration of excess heating (hot spot) even for a symmetric conductor. When coupled to the electrode phonons, CIFs drive different phonon heat flux into the two electrodes. First-principles calculations on realistic biased nanojunctions illustrate the importance of the effect....

  4. Modeling of movement-induced and flow-induced fluid forces in fast switching valves

    DEFF Research Database (Denmark)

    Roemer, Daniel Beck; Johansen, Per; Schmidt, Lasse

    2015-01-01

    Fast switching fluid power valves set strict requirements on performance, size and energy efficiency and simulation models are therefore needed to obtain good designs of such components. The valve moving member is subject to fluid forces depending on the valve flow rate and movement of the valve...... valve design. Simulated results of the total fluid force are presented showing the movement-induced fluid force to be significant for a reference application. The model form established is useful for valve designers during development and for accurate operation simulation....... member itself. These fluid forces may be accurately simulated using Computational Fluid Dynamics (CFD) analysis, but such models suffer from being computationally expensive and is not suited for optimization routines. In this paper, a computationally inexpensive method for modeling the fluid forces...

  5. Growth increase of Arabidopsis by forced expression of rice 45S rRNA gene.

    Science.gov (United States)

    Makabe, So; Motohashi, Reiko; Nakamura, Ikuo

    2017-02-01

    Forced expression of rice 45S rRNA gene conferred ca. 2-fold increase of above-ground growth in transgenic Arabidopsis . This growth increase was probably brought by cell proliferation, not by cell enlargement. Recent increase in carbon dioxide emissions is causing global climate change. The use of plant biomass as alternative energy source is one way to reduce these emissions. Therefore, reinforcement of plant biomass production is an urgent key issue to overcome both depletion of fossil energies and emission of carbon dioxide. Here, we created transgenic Arabidopsis with a 2-fold increase in above-ground growth by forced expression of the rice 45S rRNA gene using the maize ubiquitin promoter. Although the size of guard cells and ploidy of leaf-cells were similar between transgenic and control plants, numbers of stomata and pavement cells were much increased in the transgenic leaf. This data suggested that cell number, not cell expansion, was responsible for the growth increase, which might be brought by the forced expression of exogenous and full-length 45S rRNA gene. The expression level of rice 45S rRNA transcripts was very low, possibly triggering unknown machinery to enhance cell proliferation. Although microarray analysis showed enhanced expression of ethylene-responsive transcription factors, these factors might respond to ethylene induced by abiotic/biotic stresses or genomic incompatibility, which might be involved in the expression of species-specific internal transcribed spacer (ITS) sequences within rice 45S rRNA transcripts. Further analysis of the mechanism underlying the growth increase will contribute to understanding the regulation of the cell proliferation and the mechanism of hybrid vigor.

  6. Orthodontic Forces Induce the Cytoprotective Enzyme Heme Oxygenase-1 in Rats.

    Science.gov (United States)

    Suttorp, Christiaan M; Xie, Rui; Lundvig, Ditte M S; Kuijpers-Jagtman, Anne Marie; Uijttenboogaart, Jasper Tom; Van Rheden, René; Maltha, Jaap C; Wagener, Frank A D T G

    2016-01-01

    Orthodontic forces disturb the microenvironment of the periodontal ligament (PDL), and induce craniofacial bone remodeling which is necessary for tooth movement. Unfortunately, orthodontic tooth movement is often hampered by ischemic injury and cell death within the PDL (hyalinization) and root resorption. Large inter-individual differences in hyalinization and root resorption have been observed, and may be explained by differential protection against hyalinization. Heme oxygenase-1 (HO-1) forms an important protective mechanism by breaking down heme into the strong anti-oxidants biliverdin/bilirubin and the signaling molecule carbon monoxide. These versatile HO-1 products protect against ischemic and inflammatory injury. We postulate that orthodontic forces induce HO-1 expression in the PDL during experimental tooth movement. Twenty-five 6-week-old male Wistar rats were used in this study. The upper three molars at one side were moved mesially using a Nickel-Titanium coil spring, providing a continuous orthodontic force of 10 cN. The contralateral side served as control. After 6, 12, 72, 96, and 120 h groups of rats were killed. On parasagittal sections immunohistochemical staining was performed for analysis of HO-1 expression and quantification of osteoclasts. Orthodontic force induced a significant time-dependent HO-1 expression in mononuclear cells within the PDL at both the apposition- and resorption side. Shortly after placement of the orthodontic appliance HO-1 expression was highly induced in PDL cells but dropped to control levels within 72 h. Some osteoclasts were also HO-1 positive but this induction was shown to be independent of time- and mechanical stress. It is tempting to speculate that differential induction of tissue protecting- and osteoclast activating genes in the PDL determine the level of bone resorption and hyalinization and, subsequently, "fast" and "slow" tooth movers during orthodontic treatment.

  7. Optofluidic lens actuated by laser-induced solutocapillary forces

    Science.gov (United States)

    Malyuk, A. Yu.; Ivanova, N. A.

    2017-06-01

    We demonstrate an adaptive liquid lens controlled by laser-induced solutocapillary forces. The liquid droplet serving as a lens is formed in a thin layer of binary liquid mixture by surface tension driven flows caused by the thermal action of laser irradiation. The shape of droplet, its aperture and the focal length are reversibly changed without hysteresis by varying the intensity of the laser beam. The focal length variation range of the droplet-lens lies in between infinity (a flat layer) to 15 mm (a curved interface). The droplet-lens is capable to adjust the in-plane lateral position in response to a displacement of the laser beam. The proposed laser controlled droplet-lens will enable to develop smart liquid optical devices, which can imitate the accommodation reflex and pupillary light reflex of the eye.

  8. Tetracycline inducible gene manipulation in serotonergic neurons.

    Directory of Open Access Journals (Sweden)

    Tillmann Weber

    Full Text Available The serotonergic (5-HT neuronal system has important and diverse physiological functions throughout development and adulthood. Its dysregulation during development or later in adulthood has been implicated in many neuropsychiatric disorders. Transgenic animal models designed to study the contribution of serotonergic susceptibility genes to a pathological phenotype should ideally allow to study candidate gene overexpression or gene knockout selectively in serotonergic neurons at any desired time during life. For this purpose, conditional expression systems such as the tet-system are preferable. Here, we generated a transactivator (tTA mouse line (TPH2-tTA that allows temporal and spatial control of tetracycline (Ptet controlled transgene expression as well as gene deletion in 5-HT neurons. The tTA cDNA was inserted into a 196 kb PAC containing a genomic mouse Tph2 fragment (177 kb by homologous recombination in E. coli. For functional analysis of Ptet-controlled transgene expression, TPH2-tTA mice were crossed to a Ptet-regulated lacZ reporter line (Ptet-nLacZ. In adult double-transgenic TPH2-tTA/Ptet-nLacZ mice, TPH2-tTA founder line L62-20 showed strong serotonergic β-galactosidase expression which could be completely suppressed with doxycycline (Dox. Furthermore, Ptet-regulated gene expression could be reversibly activated or inactivated when Dox was either withdrawn or added to the system. For functional analysis of Ptet-controlled, Cre-mediated gene deletion, TPH2-tTA mice (L62-20 were crossed to double transgenic Ptet-Cre/R26R reporter mice to generate TPH2-tTA/Ptet-Cre/R26R mice. Without Dox, 5-HT specific recombination started at E12.5. With permanent Dox administration, Ptet-controlled Cre-mediated recombination was absent. Dox withdrawal either postnatally or during adulthood induced efficient recombination in serotonergic neurons of all raphe nuclei, respectively. In the enteric nervous system, recombination could not be detected. We

  9. Gravity-Induced Gene Expression in Plants.

    Science.gov (United States)

    Sederoff, Heike; Heber, Steffen; Howard, Brian; Myburg-Nichols, Henrietta; Hammond, Rebecca; Salinas-Mondragon, Raul; Brown, Christopher S.

    Plants sense changes in their orientation towards the vector of gravity and respond with directional growth. Several metabolites in the signal transduction cascade have been identified. However, very little is known about the interaction between these sensing and signal transduction events and even less is known about their role in the differential growth response. Gravity induced changes in transcript abundance have been identified in Arabidopsis whole seedlings and root apices (Moseyko et al. 2002; Kimbrough et al. 2004). Gravity induced transcript abundance changes can be observed within less than 1 min after stimulation (Salinas-Mondragon et al. 2005). Gene expression however requires not only transcription but also translation of the mRNA. Translation can only occur when mRNA is associated with ribosomes, even though not all mRNA associated with ribosomes is actively translated. To approximate translational capacity we quantified whole genome transcript abundances in corn stem pulvini during the first hour after gravity stimulation in total and poly-ribosomal fractions. As in Arabidopsis root apices, transcript abundances of several clusters of genes responded to gravity stimulation. The vast majority of these transcripts were also found to associate with polyribosomes in the same temporal and quantitative pattern. These genes are transcriptionally regulated by gravity stimulation, but do not exhibit translational regulation. However, a small group of genes showed increased transcriptional regulation after gravity stimulation, but no association with polysomes. These transcripts likely are translationally repressed. The mechanism of translational repression for these transcripts is unknown. Based on the hypothesis that the genes essential for gravitropic responses should be expressed in most or all species, we compared the temporal gravity induced expression pattern of all orthologs identified between maize and Arabidopsis. A small group of genes showed high

  10. Photorhabdus luminescens genes induced upon insect infection

    Directory of Open Access Journals (Sweden)

    Jung Kirsten

    2008-05-01

    Full Text Available Abstract Background Photorhabdus luminescens is a Gram-negative luminescent enterobacterium and a symbiote to soil nematodes belonging to the species Heterorhabditis bacteriophora. P.luminescens is simultaneously highly pathogenic to insects. This bacterium exhibits a complex life cycle, including one symbiotic stage characterized by colonization of the upper nematode gut, and a pathogenic stage, characterized by release from the nematode into the hemocoel of insect larvae, resulting in rapid insect death caused by bacterial toxins. P. luminescens appears to sense and adapt to the novel host environment upon changing hosts, which facilitates the production of factors involved in survival within the host, host-killing, and -exploitation. Results A differential fluorescence induction (DFI approach was applied to identify genes that are up-regulated in the bacterium after infection of the insect host Galleria mellonella. For this purpose, a P. luminescens promoter-trap library utilizing the mCherry fluorophore as a reporter was constructed, and approximately 13,000 clones were screened for fluorescence induction in the presence of a G. mellonella larvae homogenate. Since P. luminescens has a variety of regulators that potentially sense chemical molecules, like hormones, the screen for up-regulated genes or operons was performed in vitro, excluding physicochemical signals like oxygen, temperature or osmolarity as variables. Clones (18 were obtained exhibiting at least 2.5-fold induced fluorescence and regarded as specific responders to insect homogenate. In combination with a bioinformatics approach, sequence motifs were identified in these DNA-fragments that are similar to 29 different promoters within the P. luminescens genome. By cloning each of the predicted promoters upstream of the reporter gene, induction was verified for 27 promoters in vitro, and for 24 promoters in viable G. mellonella larvae. Among the validated promoters are some known

  11. Force

    CERN Document Server

    Graybill, George

    2007-01-01

    Forces are at work all around us. Discover what a force is, and different kinds of forces that work on contact and at a distance. We use simple language and vocabulary to make this invisible world easy for students to ""see"" and understand. Examine how forces ""add up"" to create the total force on an object, and reinforce concepts and extend learning with sample problems.

  12. A numerical study of microparticle acoustophoresis driven by acoustic radiation forces and streaming-induced drag forces

    DEFF Research Database (Denmark)

    Muller, Peter Barkholt; Barnkob, Rune; Jensen, Mads Jakob Herring;

    2012-01-01

    We present a numerical study of the transient acoustophoretic motion of microparticles suspended in a liquid-filled microchannel and driven by the acoustic forces arising from an imposed standing ultrasound wave: the acoustic radiation force from the scattering of sound waves on the particles...... parameters using a numerical particle-tracking scheme. The model shows the transition in the acoustophoretic particle motion from being dominated by streaming-induced drag to being dominated by radiation forces as a function of particle size, channel geometry, and material properties....

  13. Induction by fructose force-feeding of histone H3 and H4 acetylation at their lysine residues around the Slc2a5 gene and its expression in mice.

    Science.gov (United States)

    Honma, Kazue; Mochizuki, Kazuki; Goda, Toshinao

    2013-01-01

    It has been reported that fructose force-feeding rapidly induced jejunal Slc2a5 gene expression in rodents. We demonstrate in this study that acetylation at lysine (K) 9 of histone H3 and acetylation at K5 and K16 of histone H4 were more enhanced in the promoter/enhancer to transcribed regions of the Slc2a5 gene in fructose force-fed mice than in glucose force-fed mice. However, fructose force-feeding did not induce acetylation at K14 of histone H3, or at K8 and K12 of histone H4 around the Slc2a5 gene. These results suggest that fructose force-feeding induced selective histone acetylation, particularly of H3 and H4, around the jejunal Slc2a5 gene in mice.

  14. Harmonic tracking of acoustic radiation force-induced displacements.

    Science.gov (United States)

    Doherty, Joshua R; Dahl, Jeremy J; Trahey, Gregg E

    2013-11-01

    Ultrasound-based elasticity imaging methods rely upon accurate estimates of tissue deformation to characterize the mechanical properties of soft tissues. These methods are corrupted by clutter, which can bias and/or increase variance in displacement estimates. Harmonic imaging methods are routinely used for clutter suppression and improved image quality in conventional B-mode ultrasound, but have not been utilized in ultrasound-based elasticity imaging methods. We introduce a novel, fully-sampled pulse-inversion harmonic method for tracking tissue displacements that corrects the loss in temporal sampling frequency associated with conventional pulse-inversion techniques. The method is implemented with acoustic radiation force impulse (ARFI) imaging to monitor the displacements induced by an impulsive acoustic radiation force excitation. Custom pulse sequences were implemented on a diagnostic ultrasound scanner to collect spatially-matched fundamental and harmonic information within a single acquisition. B-mode and ARFI images created from fundamental data collected at 4 MHz and 8 MHz are compared with 8-MHz harmonic images created using a band-pass filter approach and the fully sampled pulse-inversion method. In homogeneous, tissue-mimicking phantoms, where no visible clutter was observed, there was little difference in the axial displacements, estimated jitter, and normalized cross-correlation among the fundamental and harmonic tracking methods. The similarity of the lower- and higher-frequency methods suggests that any improvement resulting from the increased frequency of the harmonic components is negligible. The harmonic tracking methods demonstrated a marked improvement in B-mode and ARFI image quality of in vivo carotid arteries. Improved feature detection and decreased variance in estimated displacements were observed in the arterial walls of harmonic ARFI images, especially in the pulse-inversion harmonic ARFI images. Within the lumen, the harmonic tracking

  15. Harmonic Tracking of Acoustic Radiation Force Induced Displacements

    Science.gov (United States)

    Doherty, Joshua R.; Dahl, Jeremy J.; Trahey, Gregg E.

    2014-01-01

    Ultrasound-based elasticity imaging methods rely upon accurate estimates of tissue deformation to characterize the mechanical properties of soft tissues. These methods are corrupted by clutter, which can bias and/or increase variance in displacement estimates. Harmonic imaging methods are routinely used for clutter suppression and improved image quality in conventional B-mode ultrasound, but have not been utilized in ultrasound-based elasticity imaging methods. We introduce a novel, fully-sampled pulse inversion harmonic method for tracking tissue displacements that corrects the loss in temporal sampling frequency associated with conventional pulse inversion techniques. The method is implemented with Acoustic Radiation Force Impulse (ARFI) imaging to monitor the displacements induced by an impulsive acoustic radiation force excitation. Custom pulse sequences were implemented on a diagnostic ultrasound scanner to collect spatially-matched fundamental and harmonic information within a single acquisition. B-mode and ARFI images created from fundamental data collected at 4 MHz and 8 MHz are compared with 8 MHz harmonic images created using a bandpass filter approach and the fully sampled pulse inversion method. In homogeneous, tissue-mimicking phantoms, where no visible clutter was observed, there was little difference in the axial displacements, estimated jitter, and normalized cross-correlation among the fundamental and harmonic tracking methods. The similarity of the lower and higher frequency methods suggests that any improvement due to the increased frequency of the harmonic components is negligible. The harmonic tracking methods demonstrated a marked improvement in B-mode and ARFI image quality of in vivo carotid arteries. Improved feature detection and decreased variance in estimated displacements were observed in the arterial walls of harmonic ARFI images, especially in the pulse inversion harmonic ARFI images. Within the lumen, the harmonic tracking methods

  16. The Risk of Noise-Induced Hearing Loss During Simulated Dives in Canadian Forces Hyperbaric Facilities

    Science.gov (United States)

    2012-10-01

    The risk of noise-induced hearing loss during simulated dives in Canadian Forces hyperbaric facilities Sharon M...2012-084 October 2012 The risk of noise-induced hearing loss during simulated dives in Canadian Forces hyperbaric ...transferred into the dive chamber of a hyperbaric facility. The mechanism is audible and sufficiently high in level in adjacent areas to warrant the

  17. Origin of current-induced forces in an atomic gold wire: A first-principles study

    DEFF Research Database (Denmark)

    Brandbyge, Mads; Stokbro, Kurt; Taylor, Jeremy Philip;

    2003-01-01

    We address the microscopic origin of the current-induced forces by analyzing results of first principles density functional calculations of atomic gold wires connected to two gold electrodes with different electrochemical potentials. We find that current induced forces are closely related...

  18. Force-Induced H2S by PDLSCs Modifies Osteoclastic Activity during Tooth Movement.

    Science.gov (United States)

    Liu, F; Wen, F; He, D; Liu, D; Yang, R; Wang, X; Yan, Y; Liu, Y; Kou, X; Zhou, Y

    2017-06-01

    Hydrogen sulfide (H2S), a gasotransmitter, has been recently linked to mesenchymal stem cell (MSC) function and bone homeostasis. Periodontal ligament stem cells (PDLSCs) are the main MSCs in PDL, which respond to mechanical force to induce physiological activities during orthodontic tooth movement (OTM). However, it is unknown whether mechanical force might induce endogenous H2S production by PDLSCs to regulate alveolar bone homeostasis. Here, we used a mouse OTM model to demonstrate that orthodontic force-induced endogenous H2S production in PDL tissue was associated with macrophage accumulation and osteoclastic activity in alveolar bone. Then, we showed that mechanical force application induced cystathionine β-synthase (CBS) expression and endogenous H2S production by PDLSCs. Moreover, blocking endogenous H2S or systemically increasing H2S levels could decrease or enhance force-induced osteoclastic activities to control tooth movement. We further revealed how force-induced H2S production by PDLSCs contributed to the secretion of monocyte chemoattractant protein-1 (MCP-1) and the expression of receptor activator of nuclear factor-κB ligand/osteoprotegerin (RANKL/OPG) system by PDLSCs. The secretion and expression of these factors controlled macrophage migration and osteoclast differentiation. This study demonstrated that PDLSCs produced H2S to respond to and transduce force signals. Force-induced gasotransmitter H2S production in PDLSCs therefore regulated osteoclastic activities in alveolar bone and controlled the OTM process through the MCP-1 secretion and RANKL/OPG system.

  19. Heat-inducible RNAi for gene functional analysis in plants.

    Science.gov (United States)

    Masclaux, Frédéric; Galaud, Jean-Philippe

    2011-01-01

    Controlling gene expression during plant development is an efficient method to explore gene function and RNA interference (RNAi) is now considered as a powerful technology for gene functional analysis. However, constitutive gene silencing cannot be used with genes involved in fundamental processes such as embryo viability or plant growth and alternative silencing strategies avoiding these limitations should be preferred. Tissue-specific and inducible promoters, able to control gene expression at spatial and/or temporal level, can be used to circumvent viability problems. In this chapter, after a rapid overview of the inducible promoters currently used for transgenic approaches in plants, we describe a method we have developed to study gene function by heat-inducible RNAi. This system is easy to use and complementary to those based on chemical gene inducer treatments and might be useful for both research and biotechnological applications.

  20. Repeat-induced gene silencing in mammals.

    Science.gov (United States)

    Garrick, D; Fiering, S; Martin, D I; Whitelaw, E

    1998-01-01

    In both plants and Drosophila melanogaster, expression from a transgenic locus may be silenced when repeated transgene copies are arranged as a concatameric array. This repeat-induced gene silencing is frequently manifested as a decrease in the proportion of cells that express the transgene, resulting in a variegated pattern of expression. There is also some indication that, in transgenic mammals, the number of transgene copies within an array can exert a repressive influence on expression, with several mouse studies reporting a decrease in the level of expression per copy as copy number increases. However, because these studies compare different sites of transgene integration as well as arrays with different numbers of copies, the expression levels observed may be subject to varying position effects as well as the influence of the multicopy array. Here we describe use of the lox/Cre system of site-specific recombination to generate transgenic mouse lines in which different numbers of a transgene are present at the same chromosomal location, thereby eliminating the contribution of position effects and allowing analysis of the effect of copy number alone on transgene silencing. Reduction in copy number results in a marked increase in expression of the transgene and is accompanied by decreased chromatin compaction and decreased methylation at the transgene locus. These findings establish that the presence of multiple homologous copies of a transgene within a concatameric array can have a repressive effect upon gene expression in mammalian systems.

  1. Residual force enhancement following eccentric induced muscle damage.

    Science.gov (United States)

    Power, Geoffrey A; Rice, Charles L; Vandervoort, Anthony A

    2012-06-26

    During lengthening of an activated skeletal muscle, the force maintained following the stretch is greater than the isometric force at the same muscle length. This is termed residual force enhancement (RFE), but it is unknown how muscle damage following repeated eccentric contractions affects RFE. Using the dorsiflexors, we hypothesised muscle damage will impair the force generating sarcomeric structures leading to a reduction in RFE. Following reference maximal voluntary isometric contractions (MVC) in 8 young men (26.5±2.8y) a stretch was performed at 30°/s over a 30° ankle excursion ending at the same muscle length as the reference MVCs (30° plantar flexion). Surface electromyography (EMG) of the tibialis anterior and soleus muscles was recorded during all tasks. The damage protocol involved 4 sets of 25 isokinetic (30°/s) lengthening contractions. The same measures were collected at baseline and immediately post lengthening contractions, and for up to 10min recovery. Following the lengthening contraction task, there was a 30.3±6.4% decrease in eccentric torque (Pmuscle damage (Pmuscle function compared to isometric actions succeeding damage. Thus, active force of cross-bridges is decreased because of impaired excitation-contraction coupling but force generated during stretch remains intact because force contribution from stretched sarcomeric structures is less impaired.

  2. Polarization force-induced changes in the dust sheath formation

    Energy Technology Data Exchange (ETDEWEB)

    Mayout, Saliha; Bentabet, Karima; Tribeche, Mouloud [Plasma Physics Group (PPG), Theoretical Physics Laboratory (TPL), Faculty of Physics, University of Bab-Ezzouar, USTHB, BP 32, El Alia, Algiers 16111 (Algeria)

    2015-09-15

    The modifications arising in the dusty plasma sheath structure due to the presence of polarization forces acting on the dust grains are investigated. The corresponding appropriate Bohm criterion for sheath formation is obtained. It is found that the critical Mach number, beyond which the dusty plasma electrostatic sheath sets in, decreases whenever the polarization effects become important. In addition, when the polarization force dominates over the electrical one, the dust plasma sheath cannot set in. This happens whenever the dust grain size exceeds a critical threshold. Moreover, the sheath electrostatic potential-gradient becomes abruptly steep, and the sheath thickness becomes broader as the polarization force effects strengthen.

  3. EMG and peak force responses to PNF stretching and the relationship between stretching-induced force deficits and bilateral deficits.

    Science.gov (United States)

    Cengiz, Asim

    2015-03-01

    [Purpose] The aim of the present study was to investigate the possibility of an interaction between stretching induced deficit (SFD) and bilateral deficits (BLD) during maximal voluntary isometric hand flexion under PNF stretch and no-stretch conditions through measurement of EMG and force production. [Subjects and Methods] Ten physically active male Caucasian students (age, 24.1±2.38 years; body mass, 79.48±11.40 kg; height, 174.15±0.8 cm) volunteered to participate in this study. EMG and force measurements of the subjects were recorded during either unilateral or bilateral 3-second maximal voluntary isometric hand flexion (MVC) against a force transducer. The paired sample t-test was used to examine the significance of differences among several conditions. Pearson product-moment correlation was used to evaluate the associations between different parameters. [Results] Stretching-induced deficits correlated with bilateral deficits in both force (r=0.85) and iEMG (r=0.89). PNF stretching caused significant decrements in the bilateral and unilateral conditions for both the right and left sides. [Conclusion] Since both force and iEMG decreases were observed in most measurements; it suggests there is a neural mechanism behinnd both the BLD and the SFD.

  4. EMG and peak force responses to PNF stretching and the relationship between stretching-induced force deficits and bilateral deficits

    Science.gov (United States)

    Cengiz, Asim

    2015-01-01

    [Purpose] The aim of the present study was to investigate the possibility of an interaction between stretching induced deficit (SFD) and bilateral deficits (BLD) during maximal voluntary isometric hand flexion under PNF stretch and no-stretch conditions through measurement of EMG and force production. [Subjects and Methods] Ten physically active male Caucasian students (age, 24.1±2.38 years; body mass, 79.48±11.40 kg; height, 174.15±0.8 cm) volunteered to participate in this study. EMG and force measurements of the subjects were recorded during either unilateral or bilateral 3-second maximal voluntary isometric hand flexion (MVC) against a force transducer. The paired sample t-test was used to examine the significance of differences among several conditions. Pearson product-moment correlation was used to evaluate the associations between different parameters. [Results] Stretching-induced deficits correlated with bilateral deficits in both force (r=0.85) and iEMG (r=0.89). PNF stretching caused significant decrements in the bilateral and unilateral conditions for both the right and left sides. [Conclusion] Since both force and iEMG decreases were observed in most measurements; it suggests there is a neural mechanism behinnd both the BLD and the SFD. PMID:25931696

  5. Investigations on the Effects of Vortex-Induced Vibration with Different Distributions of Lorentz Forces

    Directory of Open Access Journals (Sweden)

    Hui Zhang

    2017-01-01

    Full Text Available The control of vortex-induced vibration (VIV in shear flow with different distributions of Lorentz force is numerically investigated based on the stream function–vorticity equations in the exponential-polar coordinates exerted on moving cylinder for Re = 150. The cylinder motion equation coupled with the fluid, including the mathematical expressions of the lift force coefficient C l , is derived. The initial and boundary conditions as well as the hydrodynamic forces on the surface of cylinder are also formulated. The Lorentz force applied to suppress the VIV has no relationship with the flow field, and involves two categories, i.e., the field Lorentz force and the wall Lorentz force. With the application of symmetrical Lorentz forces, the symmetric field Lorentz force can amplify the drag, suppress the flow separation, decrease the lift fluctuation, and then suppress the VIV while the wall Lorentz force decreases the drag only. With the application of asymmetrical Lorentz forces, besides the above-mentioned effects, the field Lorentz force can increase additional lift induced by shear flow, whereas the wall Lorentz force can counteract the additional lift, which is dominated on the total effect.

  6. Water’s role in the force-induced unfolding of ubiquitin

    OpenAIRE

    2010-01-01

    In atomic force spectroscopic studies of the elastomeric protein ubiquitin, the β-strands 1-5 serve as the force clamp. Simulations show how the rupture force in the force-induced unfolding depends on the kinetics of water molecule insertion into positions where they can eventually form hydrogen bonding bridges with the backbone hydrogen bonds in the force-clamp region. The intrusion of water into this region is slowed down by the hydrophobic shielding effect of carbonaceous groups on the sur...

  7. Bi-Force: large-scale bicluster editing and its application to gene expression data biclustering.

    Science.gov (United States)

    Sun, Peng; Speicher, Nora K; Röttger, Richard; Guo, Jiong; Baumbach, Jan

    2014-05-01

    The explosion of the biological data has dramatically reformed today's biological research. The need to integrate and analyze high-dimensional biological data on a large scale is driving the development of novel bioinformatics approaches. Biclustering, also known as 'simultaneous clustering' or 'co-clustering', has been successfully utilized to discover local patterns in gene expression data and similar biomedical data types. Here, we contribute a new heuristic: 'Bi-Force'. It is based on the weighted bicluster editing model, to perform biclustering on arbitrary sets of biological entities, given any kind of pairwise similarities. We first evaluated the power of Bi-Force to solve dedicated bicluster editing problems by comparing Bi-Force with two existing algorithms in the BiCluE software package. We then followed a biclustering evaluation protocol in a recent review paper from Eren et al. (2013) (A comparative analysis of biclustering algorithms for gene expressiondata. Brief. Bioinform., 14:279-292.) and compared Bi-Force against eight existing tools: FABIA, QUBIC, Cheng and Church, Plaid, BiMax, Spectral, xMOTIFs and ISA. To this end, a suite of synthetic datasets as well as nine large gene expression datasets from Gene Expression Omnibus were analyzed. All resulting biclusters were subsequently investigated by Gene Ontology enrichment analysis to evaluate their biological relevance. The distinct theoretical foundation of Bi-Force (bicluster editing) is more powerful than strict biclustering. We thus outperformed existing tools with Bi-Force at least when following the evaluation protocols from Eren et al. Bi-Force is implemented in Java and integrated into the open source software package of BiCluE. The software as well as all used datasets are publicly available at http://biclue.mpi-inf.mpg.de. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  8. [Effect of Pinch-3 gene interference of glomerular podocytes on cell morphology and cell traction force].

    Science.gov (United States)

    Yang, Yu; Niu, Qingyuan; Ji, Zhenling; Zhang, Jingjing; Li, Jianting; Ma, Deshun

    2013-06-01

    Pinch-3 protein is an important constituent of cell membranes, which directly affects the cell morphology and mechanical properties. We observed and compared the change of morphology and cell traction force of glomerular podocytes before and after Pinch-3 gene inhibition by gene interference technology in this experiment. We found that a number of pores appeared on the cell surface, and the cell projected area were increased at the same time, with an approximate average about an increase of 40% after Pinch-3 gene inhibition. The results showed that the cell traction force of glomerular podocytes was significantly reduced, with an approximate average decrease of 40%, the maximum value of the cell traction force was reduced and the distribution of cell traction force became dispersive. All this suggested that after Pinch-3 gene inhibition, some pores created on the cell surface influenced the physical properties of glomerular podocytes and then affected the cell projected area and influenced the formation and distribution of cell traction force of the glomerular podocytes as well.

  9. Seizure-induced muscle force can caused lumbar spine fracture

    DEFF Research Database (Denmark)

    Mehlhorn, A T; Strohm, P C; Hausschildt, O;

    2007-01-01

    Patients suffering form epilepsy have an increased risk for fractures. Beside fractures caused by fall or accident muscles forces alone generated during tonic-clonic seizure can result in severe musculoskeletal injury. Contractions of strong paraspinal muscles can lead to compression fracture of ...

  10. Experimental identification of pedestrian-induced lateral forces on footbridges

    DEFF Research Database (Denmark)

    Ingólfsson, Einar Thór; Georgakis, Christos; Ricciardelli, Francesco

    2011-01-01

    This paper presents a comprehensive experimental analysis of lateral forces generated by single pedestrians during continuous walking on a treadmill. Two different conditions are investigated; initially the treadmill is fixed and then it is laterally driven in a sinusoidal motion at varying combi...

  11. Attosecond Electro-Magnetic Forces Acting on Metal Nanospheres Induced By Relativistic Electrons

    Science.gov (United States)

    Lagos, M. J.; Batson, P. E.; Reyes-Coronado, A.; Echenique, P. M.; Aizpurua, J.

    2014-03-01

    Swift electron scattering near nanoscale materials provides information about light-matter behavior, including induced forces. We calculate time-dependent electromagnetic forces acting on 1-1.5 nm metal nanospheres induced by passing swift electrons, finding both impulse-like and oscillatory response forces. Initially, impulse-like forces are generated by a competition between attractive electric forces and repulsive magnetic forces, lasting a few attoseconds (5-10 as). Oscillatory, plasmonic response forces take place later in time, last a few femtoseconds (1- 5 fs), and apparently rely on photon emission by decay of the electron-induced surface plasmons. A comparison of the strength of these two forces suggests that the impulse-like behavior dominates the process, and can transfer significant linear momentum to the sphere. Our results advance understanding of the physics behind the observation of both attractive and repulsive behavior of gold nano-particles induced by electron beams in aberration-corrected electron microscopy. Work supported under DOE, Award # DE-SC0005132, Basque Gov. project ETORTEK inano, Spanish Ministerio de Ciencia e Innovacion, No. FIS2010-19609-C02-01.

  12. Analysis of Disturbance Source Inducing by The Variable Speed Wind Turbine System Forced Power Oscillations

    DEFF Research Database (Denmark)

    Tan, Jin; Hu, Weihao; Wang, Xiaoru

    2015-01-01

    The main focus of forced low frequency oscillations is to analyze the disturbance source and the origin of forced oscillations. In this paper, the origin of low-frequency periodical oscillations induced by wind turbines’ mechanical power is investigated and the mechanism is studied of fluctuating...

  13. Two-phase flow-induced forces on bends in small scale tubes

    NARCIS (Netherlands)

    Cargnelutti, M.F.; Belfroid, S.P.C.; Schiferli, W.

    2010-01-01

    Two-phase flow occurs in many situations in industry. Under certain circumstances, it can be a source of flow-induced vibrations. The forces generated can be sufficiently large to affect the performance or efficiency of an industrial device. In the worst-case scenario, the mechanical forces that ari

  14. Is transcription the dominant force during dynamic changes in gene expression?

    Science.gov (United States)

    Turner, Martin

    2011-01-01

    Dynamic changes in gene expression punctuate lymphocyte development and are a characteristic of lymphocyte activation. A prevailing view has been that these changes are driven by DNA transcription factors, which are the dominant force in gene expression. Accumulating evidence is challenging this DNA centric view and has highlighted the prevalence and dynamic nature of RNA handling mechanisms. Alternative splicing and differential polyadenylation appear to be more widespread than first thought. Changes in mRNA decay rates also affect the abundance of transcripts and this mechanism may contribute significantly to gene expression. Additional RNA handling mechanisms that control the intracellular localization of mRNA and association with translating ribosomes are also important. Thus, gene expression is regulated through the coordination of transcriptional and post-transcriptional mechanisms. Developing a more "RNA centric" view of gene expression will allow a more systematic understanding of how gene expression and cell function are integrated.

  15. Force Responses and Sarcomere Dynamics of Cardiac Myofibrils Induced by Rapid Changes in [Pi].

    Science.gov (United States)

    Stehle, Robert

    2017-01-24

    The second phase of the biphasic force decay upon release of phosphate from caged phosphate was previously interpreted as a signature of kinetics of the force-generating step in the cross-bridge cycle. To test this hypothesis without using caged compounds, force responses and individual sarcomere dynamics upon rapid increases or decreases in concentration of inorganic phosphate [Pi] were investigated in calcium-activated cardiac myofibrils. Rapid increases in [Pi] induced a biphasic force decay with an initial slow decline (phase 1) and a subsequent 3-5-fold faster major decay (phase 2). Phase 2 started with the distinct elongation of a single sarcomere, the so-called sarcomere "give". "Give" then propagated from sarcomere to sarcomere along the myofibril. Propagation speed and rate constant of phase 2 (k+Pi(2)) had a similar [Pi]-dependence, indicating that the kinetics of the major force decay (phase 2) upon rapid increase in [Pi] is determined by sarcomere dynamics. In contrast, no "give" was observed during phase 1 after rapid [Pi]-increase (rate constant k+Pi(1)) and during the single-exponential force rise (rate constant k-Pi) after rapid [Pi]-decrease. The values of k+Pi(1) and k-Pi were similar to the rate constant of mechanically induced force redevelopment (kTR) and Ca(2+)-induced force development (kACT) measured at same [Pi]. These results indicate that the major phase 2 of force decay upon a Pi-jump does not reflect kinetics of the force-generating step but results from sarcomere "give". The other phases of Pi-induced force kinetics that occur in the absence of "give" yield the same information as mechanically and Ca(2+)-induced force kinetics (k+Pi(1) ∼ k-Pi ∼ kTR ∼ kACT). Model simulations indicate that Pi-induced force kinetics neither enable the separation of Pi-release from the rate-limiting transition f into force states nor differentiate whether the "force-generating step" occurs before, along, or after the Pi-release.

  16. The percentage of bacterial genes on leading versus lagging strands is influenced by multiple balancing forces

    Science.gov (United States)

    Mao, Xizeng; Zhang, Han; Yin, Yanbin; Xu, Ying

    2012-01-01

    The majority of bacterial genes are located on the leading strand, and the percentage of such genes has a large variation across different bacteria. Although some explanations have been proposed, these are at most partial explanations as they cover only small percentages of the genes and do not even consider the ones biased toward the lagging strand. We have carried out a computational study on 725 bacterial genomes, aiming to elucidate other factors that may have influenced the strand location of genes in a bacterium. Our analyses suggest that (i) genes of some functional categories such as ribosome have higher preferences to be on the leading strands; (ii) genes of some functional categories such as transcription factor have higher preferences on the lagging strands; (iii) there is a balancing force that tends to keep genes from all moving to the leading and more efficient strand and (iv) the percentage of leading-strand genes in an bacterium can be accurately explained based on the numbers of genes in the functional categories outlined in (i) and (ii), genome size and gene density, indicating that these numbers implicitly contain the information about the percentage of genes on the leading versus lagging strand in a genome. PMID:22735706

  17. Association of Forced Vital Capacity with the Developmental Gene NCOR2

    NARCIS (Netherlands)

    C. Minelli (Cosetta); C.H. Dean (Charlotte H.); M. Hind (Matthew); A.C. Alves (Alexessander Couto); A.F.S. Amaral (André F.S.); V. Siroux (V.); V. Huikari (Ville); M.S. Artigas; D.M. Evans (David M.); D.W. Loth (Daan); Y. Bossé (Yohan); D.S. Postma (Dirkje); D.D. Sin; J.R. Thompson (John); F. Demenais (Florence); J. Henderson (John); E. Bouzigon (Emmanuelle); D.L. Jarvis (Deborah); M.-R. Jarvelin (Marjo-Riitta); P.G. Burney; S.A. Gharib (Sina); L.V. Wain (Louise); N. Franceschini (Nora); B. Koch (Beate); T.D. Pottinger (Tess); A.V. Smith; Q. Duan (Qing); C. Oldmeadow (Christopher); M.K. Lee (Mi Kyeong); D.P. Strachan (David P.); A.L. James (Alan L.); J.E. Huffman (Jennifer); V. Vitart (Veronique); A. Ramasamy (Adaikalavan); N.J. Wareham (Nicholas J.); J. Kaprio (Jaakko); X.-Q. Wang (Xin-Qun); H. Trochet (Holly); M. Kähönen (Mika); C. Flexeder (Claudia); E. Albrecht (Eva); L.M. Lopez (Lorna M.); K. de Jong (Kim); B. Thyagarajan (Bharat); S. Enroth (Stefan); E. Omenaas (Ernst); P.K. Joshi (Peter); M. Fall (Magnus); A. Viñuela (Ana); L.J. Launer (Lenore); L.R. Loehr (Laura); M. Fornage (Myriam); G. Li (Guo); J.B. Wilk (Jemma); W. Tang (Wenbo); A. Manichaikul (Ani); L. Lahousse (Lies); T.B. Harris (Tamara); K.E. North (Kari E.); A.R. Rudnicka (Alicja); J. Hui (Jennie); X. Gu (Xiangjun); T. Lumley (Thomas); A.F. Wright (Alan F.); N. Hastie (Nick); S. Campbell (Susan); R. Kumar (Rajesh); I. Pin (Isabelle); R.A. Scott (Robert); K.H. Pietilainen (Kirsi Hannele); I. Surakka (Ida); Y. Liu (Yongmei); E.G. Holliday (Elizabeth); H. Schulz (Holger); J. Heinrich (Joachim); G. Davies (Gail); J. MVonk (Judith); M.K. Wojczynski (Mary ); A. Pouta (Anneli); A. Johansson (Åsa); S. Wild (Sarah); E. Ingelsson (Erik); F. Rivadeneira Ramirez (Fernando); H. Völzke (Henry); P.G. Hysi (Pirro); G. Eiriksdottir (Gudny); A.C. Morrison (Alanna); J.I. Rotter (Jerome I.); W. Gao (Wei); W.B. White (Wendy); S.S. Rich (Stephen S.); A. Hofman (Albert); T. Aspelund (Thor); D.J. Couper (David); L.J. Smith (Lewis J.); B.M. Psaty (Bruce); K. Lohman (Kurt); E.G. Burchard (Esteban); A.G. Uitterlinden (André); M. Garcia (Melissa); B.R. Joubert (Bonnie); W.L. McArdle (Wendy); A.B. Musk (A Bill); C.R.W. Hansel (Christian); S.R. Heckbert (Susan); L. Zgaga (Lina); J.B.J. van Meurs (Joyce); P. Navarro (Pau); I. Rudan (Igor); Y.-M. Oh (Yeon-Mok); S. Redline (Susan); D.L. Jarvis (Deborah L.); J.H. Zhao (Jing Hua); T. Rantanen (Taina); G.T. O Connor (George T.); S. Ripatti (Samuli); R.J. Scott (Rodney); S. Karrasch (Stefan); H. Grallert (Harald); N.C. Gaddis (Nathan C.); J. MStarr (John); C. Wijmenga (Cisca); R.L. Minster (Ryan); C.W. Lederer (Carsten); J. Pekkanen (Juha); U. Gyllensten (Ulf); H. Campbell (Harry); A.P. Morris (Andrew); S. Gläser (Sven); C.J. Hammond (Christopher J.); K. MBurkart (Kristin); J. Beilby (John); S.B. Kritchevsky (Stephen B.); V. Gudnason (Vilmundur); D.B. Hancock (Dana); O. Williams (O'Dale); O. Polasek (Ozren); T. Zemunik (Tatijana); I. Kolcic (Ivana); M.F. Petrini (Marcy); M. Wjst (Matthias); W.J. Kim (Woo Jin); D.J. Porteous (David J.); G. Scotland (Generation); B.H. Smith (Blair); A. Viljanen (Anne); M. Heliovaara (Markku); J. Attia (John); I. Sayers (Ian); R. Hampel (Regina); C. Gieger (Christian); I.J. Deary (Ian J.); H.M. Boezen (Marike); A.B. Newman (Anne B.); J.F. Wilson (James F); L. Lind (Lars); B.H.Ch. Stricker (Bruno); A. Teumer (Alexander); T.D. Spector (Timothy); E. Melén (Erik); M.J. Peters (Marjolein); L.A. Lange (Leslie); R.G. Barr (R. Graham); K.R. Bracke (Ken); F. MVerhamme (Fien); J. Sung (Joohon); P.S. Hiemstra (Pieter); P.A. Cassano (Patricia); A. Sood (Akshay); C. Hayward (Caroline); J. Dupuis (Josée); I.P. Hall (Ian); G.G. Brusselle (Guy); M.D. Tobin (Martin); S.J. London (Stephanie J)

    2016-01-01

    textabstractBackground Forced Vital Capacity (FVC) is an important predictor of all-cause mortality in the absence of chronic respiratory conditions. Epidemiological evidence highlights the role of early life factors on adult FVC, pointing to environmental exposures and genes affecting lung developm

  18. Association of Forced Vital Capacity with the Developmental Gene NCOR2

    NARCIS (Netherlands)

    C. Minelli (Cosetta); C.H. Dean (Charlotte H.); M. Hind (Matthew); A.C. Alves (Alexessander Couto); A.F.S. Amaral (André F.S.); V. Siroux (V.); V. Huikari (Ville); M.S. Artigas; D.M. Evans (David M.); D.W. Loth (Daan); Y. Bossé (Yohan); D.S. Postma (Dirkje); D.D. Sin; J.R. Thompson (John); F. Demenais (Florence); J. Henderson (John); E. Bouzigon (Emmanuelle); D.L. Jarvis (Deborah); M.-R. Jarvelin (Marjo-Riitta); P.G. Burney; S.A. Gharib (Sina); L.V. Wain (Louise); N. Franceschini (Nora); B. Koch (Beate); T.D. Pottinger (Tess); A.V. Smith; Q. Duan (Qing); C. Oldmeadow (Christopher); M.K. Lee (Mi Kyeong); D.P. Strachan (David P.); A.L. James (Alan L.); J.E. Huffman (Jennifer); V. Vitart (Veronique); A. Ramasamy (Adaikalavan); N.J. Wareham (Nicholas J.); J. Kaprio (Jaakko); X.-Q. Wang (Xin-Qun); H. Trochet (Holly); M. Kähönen (Mika); C. Flexeder (Claudia); E. Albrecht (Eva); L.M. Lopez (Lorna M.); K. de Jong (Kim); B. Thyagarajan (Bharat); S. Enroth (Stefan); E. Omenaas (Ernst); P.K. Joshi (Peter); M. Fall (Magnus); A. Viñuela (Ana); L.J. Launer (Lenore); L.R. Loehr (Laura); M. Fornage (Myriam); G. Li (Guo); J.B. Wilk (Jemma); W. Tang (Wenbo); A. Manichaikul (Ani); L. Lahousse (Lies); T.B. Harris (Tamara); K.E. North (Kari E.); A.R. Rudnicka (Alicja); J. Hui (Jennie); X. Gu (Xiangjun); T. Lumley (Thomas); A.F. Wright (Alan F.); N. Hastie (Nick); S. Campbell (Susan); R. Kumar (Rajesh); I. Pin (Isabelle); R.A. Scott (Robert); K.H. Pietilainen (Kirsi Hannele); I. Surakka (Ida); Y. Liu (Yongmei); E.G. Holliday (Elizabeth); H. Schulz (Holger); J. Heinrich (Joachim); G. Davies (Gail); J. MVonk (Judith); M.K. Wojczynski (Mary ); A. Pouta (Anneli); A. Johansson (Åsa); S. Wild (Sarah); E. Ingelsson (Erik); F. Rivadeneira Ramirez (Fernando); H. Völzke (Henry); P.G. Hysi (Pirro); G. Eiriksdottir (Gudny); A.C. Morrison (Alanna); J.I. Rotter (Jerome I.); W. Gao (Wei); W.B. White (Wendy); S.S. Rich (Stephen S.); A. Hofman (Albert); T. Aspelund (Thor); D.J. Couper (David); L.J. Smith (Lewis J.); B.M. Psaty (Bruce); K. Lohman (Kurt); E.G. Burchard (Esteban); A.G. Uitterlinden (André); M. Garcia (Melissa); B.R. Joubert (Bonnie); W.L. McArdle (Wendy); A.B. Musk (A Bill); C.R.W. Hansel (Christian); S.R. Heckbert (Susan); L. Zgaga (Lina); J.B.J. van Meurs (Joyce); P. Navarro (Pau); I. Rudan (Igor); Y.-M. Oh (Yeon-Mok); S. Redline (Susan); D.L. Jarvis (Deborah L.); J.H. Zhao (Jing Hua); T. Rantanen (Taina); G.T. O Connor (George T.); S. Ripatti (Samuli); R.J. Scott (Rodney); S. Karrasch (Stefan); H. Grallert (Harald); N.C. Gaddis (Nathan C.); J. MStarr (John); C. Wijmenga (Cisca); R.L. Minster (Ryan); C.W. Lederer (Carsten); J. Pekkanen (Juha); U. Gyllensten (Ulf); H. Campbell (Harry); A.P. Morris (Andrew); S. Gläser (Sven); C.J. Hammond (Christopher J.); K. MBurkart (Kristin); J. Beilby (John); S.B. Kritchevsky (Stephen B.); V. Gudnason (Vilmundur); D.B. Hancock (Dana); O. Williams (O'Dale); O. Polasek (Ozren); T. Zemunik (Tatijana); I. Kolcic (Ivana); M.F. Petrini (Marcy); M. Wjst (Matthias); W.J. Kim (Woo Jin); D.J. Porteous (David J.); G. Scotland (Generation); B.H. Smith (Blair); A. Viljanen (Anne); M. Heliovaara (Markku); J. Attia (John); I. Sayers (Ian); R. Hampel (Regina); C. Gieger (Christian); I.J. Deary (Ian J.); H.M. Boezen (Marike); A.B. Newman (Anne B.); J.F. Wilson (James F); L. Lind (Lars); B.H.Ch. Stricker (Bruno); A. Teumer (Alexander); T.D. Spector (Timothy); E. Melén (Erik); M.J. Peters (Marjolein); L.A. Lange (Leslie); R.G. Barr (R. Graham); K.R. Bracke (Ken); F. MVerhamme (Fien); J. Sung (Joohon); P.S. Hiemstra (Pieter); P.A. Cassano (Patricia); A. Sood (Akshay); C. Hayward (Caroline); J. Dupuis (Josée); I.P. Hall (Ian); G.G. Brusselle (Guy); M.D. Tobin (Martin); S.J. London (Stephanie J)

    2016-01-01

    textabstractBackground Forced Vital Capacity (FVC) is an important predictor of all-cause mortality in the absence of chronic respiratory conditions. Epidemiological evidence highlights the role of early life factors on adult FVC, pointing to environmental exposures and genes affecting lung

  19. Lateral-drag Casimir forces induced by anisotropy

    CERN Document Server

    Nefedov, Igor S

    2016-01-01

    We predict the existence of lateral drag forces near the flat surface of an absorbing slab of an anisotropic material. The forces originate from the fluctuations of the electromagnetic field, when the anisotropy axis of the material forms a certain angle with the surface. In this situation, the spatial spectra of the fluctuating electromagnetic fields becomes asymmetric, different for positive and negative transverse wave vectors components. Differently from the case of van der Waals interactions in which the forward-backward symmetry is broken due to the particle movement or in quantum noncontact friction where it is caused by the mutual motion of the bodies, in our case the lateral motion results merely from the anisotropy of the slab. This new effect, of particular significance in hyperbolic materials, could be used for the manipulation of nanoparticles.

  20. Lateral-drag propulsion forces induced by anisotropy.

    Science.gov (United States)

    Nefedov, Igor S; Rubi, J Miguel

    2017-07-21

    We predict the existence of lateral drag forces near the flat surface of an absorbing slab made of an anisotropic material. The forces originate from the fluctuations of the electromagnetic field, when the anisotropy axis of the material forms a certain angle with the surface. In this situation, the spatial spectra of the fluctuating electromagnetic fields becomes asymmetric, different for positive and negative transverse wave vectors components. Differently from the case of van der Waals interactions in which the forward-backward symmetry is broken due to the particle movement, in our case the lateral motion results merely from the anisotropy of the slab. This new effect, of particular significance in hyperbolic materials, could be used for the manipulation of nanoparticles.

  1. A complex social-ecological disaster: Environmentally induced forced migration.

    Science.gov (United States)

    Rechkemmer, Andreas; O'Connor, Ashley; Rai, Abha; Decker Sparks, Jessica L; Mudliar, Pranietha; Shultz, James M

    2016-01-01

    In the 21(st) century, global issues are increasingly characterized by inter-connectedness and complexity. Global environmental change, and climate change in particular, has become a powerful driver and catalyst of forced migration and internal displacement of people. Environmental migrants may far outnumber any other group of displaced people and refugees in the years to come. Deeper scientific integration, especially across the social sciences, is a prerequisite to tackle this issue.

  2. A complex social-ecological disaster: Environmentally induced forced migration

    Science.gov (United States)

    Rechkemmer, Andreas; O'Connor, Ashley; Rai, Abha; Decker Sparks, Jessica L.; Mudliar, Pranietha; Shultz, James M.

    2016-01-01

    ABSTRACT In the 21st century, global issues are increasingly characterized by inter-connectedness and complexity. Global environmental change, and climate change in particular, has become a powerful driver and catalyst of forced migration and internal displacement of people. Environmental migrants may far outnumber any other group of displaced people and refugees in the years to come. Deeper scientific integration, especially across the social sciences, is a prerequisite to tackle this issue.

  3. Virus-induced gene silencing in soybean and common bean.

    Science.gov (United States)

    Zhang, Chunquan; Whitham, Steven A; Hill, John H

    2013-01-01

    Plant viral vectors are useful for transient gene expression as well as for downregulation of gene expression via virus-induced gene silencing (VIGS). When used in reverse genetics approaches, VIGS offers a convenient way of transforming genomic information into knowledge of gene function. Efforts to develop and improve plant viral vectors have expanded their applications and have led to substantial advances needed to facilitate gene function studies in major row crops. Here, we describe a DNA-based Bean pod mottle virus (BPMV) vector system for both gene expression and VIGS in soybean and common bean.

  4. Electron fluctuation induced resonance broadening in nano electromechanical systems: the origin of shear force in vacuum.

    Science.gov (United States)

    Siria, A; Barois, T; Vilella, K; Perisanu, S; Ayari, A; Guillot, D; Purcell, S T; Poncharal, P

    2012-07-11

    This article presents a study of the poorly understood "shear-force" used in an important class of near-field instruments that use mechanical resonance feedback detection. In the case of a metallic probe near a metallic surface in vacuum, we show that in the 10-60 nm range there is no such a thing as a shear-force in the sense of the nonconservative friction force. Fluctuations of the oscillator resonance frequency, likely induced by local charge variations, could account for the reported effects in the literature without introducing a dissipative force.

  5. Current-induced forces in mesoscopic systems: A scattering-matrix approach

    Directory of Open Access Journals (Sweden)

    Niels Bode

    2012-02-01

    Full Text Available Nanoelectromechanical systems are characterized by an intimate connection between electronic and mechanical degrees of freedom. Due to the nanoscopic scale, current flowing through the system noticeably impacts upons the vibrational dynamics of the device, complementing the effect of the vibrational modes on the electronic dynamics. We employ the scattering-matrix approach to quantum transport in order to develop a unified theory of nanoelectromechanical systems out of equilibrium. For a slow mechanical mode the current can be obtained from the Landauer–Büttiker formula in the strictly adiabatic limit. The leading correction to the adiabatic limit reduces to Brouwer’s formula for the current of a quantum pump in the absence of a bias voltage. The principal results of the present paper are the scattering-matrix expressions for the current-induced forces acting on the mechanical degrees of freedom. These forces control the Langevin dynamics of the mechanical modes. Specifically, we derive expressions for the (typically nonconservative mean force, for the (possibly negative damping force, an effective “Lorentz” force that exists even for time-reversal-invariant systems, and the fluctuating Langevin force originating from Nyquist and shot noise of the current flow. We apply our general formalism to several simple models that illustrate the peculiar nature of the current-induced forces. Specifically, we find that in out-of-equilibrium situations the current-induced forces can destabilize the mechanical vibrations and cause limit-cycle dynamics.

  6. Flexural waves induced by electro-impulse deicing forces

    Science.gov (United States)

    Gien, P. H.

    1990-01-01

    The generation, reflection and propagation of flexural waves created by electroimpulsive deicing forces are demonstrated both experimentally and analytically in a thin circular plate and a thin semicylindrical shell. Analytical prediction of these waves with finite element models shows good correlation with acceleration and displacement measurements at discrete points on the structures studied. However, sensitivity to spurious flexural waves resulting from the spatial discretization of the structures is shown to be significant. Consideration is also given to composite structures as an extension of these studies.

  7. Depletion forces on circular and elliptical obstacles induced by active matter.

    Science.gov (United States)

    Leite, L R; Lucena, D; Potiguar, F Q; Ferreira, W P

    2016-12-01

    Depletion forces exerted by self-propelled particles on circular and elliptical passive objects are studied using numerical simulations. We show that a bath of active particles can induce repulsive and attractive forces which are sensitive to the shape and orientation of the passive objects (either horizontal or vertical ellipses). The resultant force on the passive objects due to the active particles is studied as a function of the shape and orientation of the passive objects, magnitude of the angular noise, and distance between the passive objects. By increasing the distance between obstacles the magnitude of the repulsive depletion force increases, as long as such a distance is less than one active particle diameter. For longer distances, the magnitude of the force always decreases with increasing distance. We also found that attractive forces may arise for vertical ellipses at high enough area fraction.

  8. Virus-induced gene silencing in detached tomatoes and biochemical effects of phytoene desaturase gene silencing

    NARCIS (Netherlands)

    Romero, I.; Tikunov, Y.M.; Bovy, A.G.

    2011-01-01

    Virus-induced gene silencing (VIGS) is a technology that has rapidly emerged for gene function studies in plants. Many advances have been made in applying this technique in an increasing number of crops. Recently, VIGS has been successfully used to silence genes in tomato fruit through agroinfiltrat

  9. Insect and wound induced GUS gene expression from a Beta vulgaris proteinase inhibitor gene promoter

    Science.gov (United States)

    Inducible gene promoters that are specifically activated by pathogen invasion or insect pest attack are needed for effective expression of resistance genes to control plant diseases. In the present study, a promoter from a serine proteinase inhibitor gene (BvSTI) shown to be up-regulated in resist...

  10. Correlative atomic force and confocal fluorescence microscopy: single molecule imaging and force induced spectral shifts (Conference Presentation)

    Science.gov (United States)

    Basché, Thomas; Hinze, Gerald; Stöttinger, Sven

    2016-09-01

    A grand challenge in nanoscience is to correlate structure or morphology of individual nano-sized objects with their photo-physical properties. An early example have been measurements of the emission spectra and polarization of single semiconductor quantum dots as well as their crystallographic structure by a combination of confocal fluorescence microscopy and transmission electron microscopy.[1] Recently, the simultaneous use of confocal fluorescence and atomic force microscopy (AFM) has allowed for correlating the morphology/conformation of individual nanoparticle oligomers or molecules with their photo-physics.[2, 3] In particular, we have employed the tip of an AFM cantilever to apply compressive stress to single molecules adsorbed on a surface and follow the effect of the impact on the electronic states of the molecule by fluorescence spectroscopy.[3] Quantum mechanical calculations corroborate that the spectral changes induced by the localized force can be associated to transitions among the different possible conformers of the adsorbed molecule.

  11. Optimization of single-cell electroporation protocol for forced gene expression in primary neuronal cultures.

    Science.gov (United States)

    Nishikawa, Shin; Hirashima, Naohide; Tanaka, Masahiko

    2014-09-01

    The development and function of the central nervous system (CNS) are realized through interactions between many neurons. To investigate cellular and molecular mechanisms of the development and function of the CNS, it is thus crucial to be able to manipulate the gene expression of single neurons in a complex cell population. We recently developed a technique for gene silencing by introducing small interfering RNA into single neurons in primary CNS cultures using single-cell electroporation. However, we had not succeeded in forced gene expression by introducing expression plasmids using single-cell electroporation. In the present study, we optimized the experimental conditions to enable the forced expression of green fluorescent protein (GFP) in cultured cerebellar Purkinje neurons using single-cell electroporation. We succeeded in strong GFP expression in Purkinje neurons by increasing the inside diameter of micropipettes or by making the size of the original plasmid smaller by digestion and cyclizing it by ligation. Strong GFP expression in Purkinje neurons electroporated under the optimal conditions continued to be observed for more than 25 days after electroporation. Thus, this technique could be used for forced gene expression in single neurons to investigate cellular and molecular mechanisms of the development, function, and disease of the CNS.

  12. EMG and peak force responses to PNF stretching and the relationship between stretching-induced force deficits and bilateral deficits

    OpenAIRE

    2015-01-01

    [Purpose] The aim of the present study was to investigate the possibility of an interaction between stretching induced deficit (SFD) and bilateral deficits (BLD) during maximal voluntary isometric hand flexion under PNF stretch and no-stretch conditions through measurement of EMG and force production. [Subjects and Methods] Ten physically active male Caucasian students (age, 24.1±2.38 years; body mass, 79.48±11.40 kg; height, 174.15±0.8 cm) volunteered to participate in this study. EMG and fo...

  13. Resonance in Interacting Induced-Dipole Polarizing Force Fields: Application to Force-Field Derivatives

    Directory of Open Access Journals (Sweden)

    Francisco Torrens

    2009-03-01

    Full Text Available The Silberstein model of the molecular polarizability of diatomic molecules, generalized by Applequist et al. for polyatomic molecules, is analyzed. The atoms are regarded as isotropically polarizable points located at their nuclei, interacting via the fields of their induced dipoles. The use of additive values for atom polarizabilities gives poor results, in some cases leading to artificial predictions of absorption bands. The molecular polarizability of methane and its derivative are computed. The agreement with experimental mean molecular polarizabilities is within 1–5%. A hypothesis is indispensable for a suitable representation of polarizability derivative.

  14. A riboswitch-based inducible gene expression system for mycobacteria.

    Directory of Open Access Journals (Sweden)

    Jessica C Seeliger

    Full Text Available Research on the human pathogen Mycobacterium tuberculosis (Mtb would benefit from novel tools for regulated gene expression. Here we describe the characterization and application of a synthetic riboswitch-based system, which comprises a mycobacterial promoter for transcriptional control and a riboswitch for translational control. The system was used to induce and repress heterologous protein overexpression reversibly, to create a conditional gene knockdown, and to control gene expression in a macrophage infection model. Unlike existing systems for controlling gene expression in Mtb, the riboswitch does not require the co-expression of any accessory proteins: all of the regulatory machinery is encoded by a short DNA segment directly upstream of the target gene. The inducible riboswitch platform has the potential to be a powerful general strategy for creating customized gene regulation systems in Mtb.

  15. Duplication and diversification of the hypoxia-inducible IGFBP-1 gene in zebrafish.

    Directory of Open Access Journals (Sweden)

    Hiroyasu Kamei

    Full Text Available BACKGROUND: Gene duplication is the primary force of new gene evolution. Deciphering whether a pair of duplicated genes has evolved divergent functions is often challenging. The zebrafish is uniquely positioned to provide insight into the process of functional gene evolution due to its amenability to genetic and experimental manipulation and because it possess a large number of duplicated genes. METHODOLOGY/PRINCIPAL FINDINGS: We report the identification and characterization of two hypoxia-inducible genes in zebrafish that are co-ortholgs of human IGF binding protein-1 (IGFBP-1. IGFBP-1 is a secreted protein that binds to IGF and modulates IGF actions in somatic growth, development, and aging. Like their human and mouse counterparts, in adult zebrafish igfbp-1a and igfbp-1b are exclusively expressed in the liver. During embryogenesis, the two genes are expressed in overlapping spatial domains but with distinct temporal patterns. While zebrafish IGFBP-1a mRNA was easily detected throughout embryogenesis, IGFBP-1b mRNA was detectable only in advanced stages. Hypoxia induces igfbp-1a expression in early embryogenesis, but induces the igfbp-1b expression later in embryogenesis. Both IGFBP-1a and -b are capable of IGF binding, but IGFBP-1b has much lower affinities for IGF-I and -II because of greater dissociation rates. Overexpression of IGFBP-1a and -1b in zebrafish embryos caused significant decreases in growth and developmental rates. When tested in cultured zebrafish embryonic cells, IGFBP-1a and -1b both inhibited IGF-1-induced cell proliferation but the activity of IGFBP-1b was significantly weaker. CONCLUSIONS/SIGNIFICANCE: These results indicate subfunction partitioning of the duplicated IGFBP-1 genes at the levels of gene expression, physiological regulation, protein structure, and biological actions. The duplicated IGFBP-1 may provide additional flexibility in fine-tuning IGF signaling activities under hypoxia and other catabolic

  16. Repulsive forces between looping chromosomes induce entropy-driven segregation.

    Directory of Open Access Journals (Sweden)

    Manfred Bohn

    Full Text Available One striking feature of chromatin organization is that chromosomes are compartmentalized into distinct territories during interphase, the degree of intermingling being much smaller than expected for linear chains. A growing body of evidence indicates that the formation of loops plays a dominant role in transcriptional regulation as well as the entropic organization of interphase chromosomes. Using a recently proposed model, we quantitatively determine the entropic forces between chromosomes. This Dynamic Loop Model assumes that loops form solely on the basis of diffusional motion without invoking other long-range interactions. We find that introducing loops into the structure of chromatin results in a multi-fold higher repulsion between chromosomes compared to linear chains. Strong effects are observed for the tendency of a non-random alignment; the overlap volume between chromosomes decays fast with increasing loop number. Our results suggest that the formation of chromatin loops imposes both compartmentalization as well as order on the system without requiring additional energy-consuming processes.

  17. Opposing Shear-Induced Forces Dominate Inertial Focusing in Curved Channels and High Reynolds Numbers

    CERN Document Server

    Keinan, Eliezer; Nahmias, Yaakov

    2015-01-01

    Inertial focusing is the migration of particles in fluid toward equilibrium, where current theory predicts that shear-induced and wall-induced lift forces are balanced. First reported in 1961, this Segre-Silberberg effect is particularly useful for microfluidic isolation of cells and particles. Interestingly, recent work demonstrated particle focusing at high Reynolds numbers that cannot be explained by current theory. In this work, we show that non-monotonous velocity profiles, such as those developed in curved channels, create peripheral velocity maxima around which opposing shear-induced forces dominate over wall effects. Similarly, entry effects amplified in high Reynolds flow produce an equivalent trapping mechanism in short, straight channels. This new focusing mechanism in the developing flow regime enables a 10-fold miniaturization of inertial focusing devices, while our model corrects long-standing misconceptions about the nature of mechanical forces governing inertial focusing in curved channels.

  18. Imaging of shear waves induced by Lorentz force in soft tissues.

    Science.gov (United States)

    Grasland-Mongrain, P; Souchon, R; Cartellier, F; Zorgani, A; Chapelon, J Y; Lafon, C; Catheline, S

    2014-07-18

    This study presents the first observation of elastic shear waves generated in soft solids using a dynamic electromagnetic field. The first and second experiments of this study showed that Lorentz force can induce a displacement in a soft phantom and that this displacement was detectable by an ultrasound scanner using speckle-tracking algorithms. For a 100 mT magnetic field and a 10 ms, 100 mA peak-to-peak electrical burst, the displacement reached a magnitude of 1 μm. In the third experiment, we showed that Lorentz force can induce shear waves in a phantom. A physical model using electromagnetic and elasticity equations was proposed. Computer simulations were in good agreement with experimental results. The shear waves induced by Lorentz force were used in the last experiment to estimate the elasticity of a swine liver sample.

  19. Imaging of Shear Waves Induced by Lorentz Force in Soft Tissues

    CERN Document Server

    Grasland-Mongrain, Pol; Cartellier, Florian; Zorgani, Ali; Chapelon, Jean-Yves; Lafon, Cyril; Catheline, Stefan

    2014-01-01

    This study presents the first observation of elastic shear waves generated in soft solids using a dynamic electromagnetic field. The first and second experiments of this 5 study showed that Lorentz force can induce a displacement in a soft phantom and that this displacement was detectable by an ultrasound scanner using speckle-tracking algorithms. For a 100 mT magnetic field and a 10 ms, 100 mA peak-to-peak electrical burst, the displacement reached a magnitude of 1 um. In the third experiment, we showed that Lorentz force can induce shear waves in a phantom. A physical model 10 using electromagnetic and elasticity equations was proposed. Computer simulations were in good agreement with experimental results. The shear waves induced by Lorentz force were used in the last experiment to estimate the elasticity of a swine liver sample.

  20. Divergent global precipitation changes induced by natural versus anthropogenic forcing.

    Science.gov (United States)

    Liu, Jian; Wang, Bin; Cane, Mark A; Yim, So-Young; Lee, June-Yi

    2013-01-31

    As a result of global warming, precipitation is likely to increase in high latitudes and the tropics and to decrease in already dry subtropical regions. The absolute magnitude and regional details of such changes, however, remain intensely debated. As is well known from El Niño studies, sea-surface-temperature gradients across the tropical Pacific Ocean can strongly influence global rainfall. Palaeoproxy evidence indicates that the difference between the warm west Pacific and the colder east Pacific increased in past periods when the Earth warmed as a result of increased solar radiation. In contrast, in most model projections of future greenhouse warming this gradient weakens. It has not been clear how to reconcile these two findings. Here we show in climate model simulations that the tropical Pacific sea-surface-temperature gradient increases when the warming is due to increased solar radiation and decreases when it is due to increased greenhouse-gas forcing. For the same global surface temperature increase the latter pattern produces less rainfall, notably over tropical land, which explains why in the model the late twentieth century is warmer than in the Medieval Warm Period (around AD 1000-1250) but precipitation is less. This difference is consistent with the global tropospheric energy budget, which requires a balance between the latent heat released in precipitation and radiative cooling. The tropospheric cooling is less for increased greenhouse gases, which add radiative absorbers to the troposphere, than for increased solar heating, which is concentrated at the Earth's surface. Thus warming due to increased greenhouse gases produces a climate signature different from that of warming due to solar radiation changes.

  1. Excited and enhanced twinborn acoustic-induced mutual forces in oblique grating structures

    Science.gov (United States)

    Lu, Shuifang; Zhang, Xin; Wu, Fugen; Yao, Yuanwei; Chen, Zongwang

    2016-07-01

    We propose a water-immersed geometrically oblique grating structure patterned with a 1D periodic array of oblique rhombuses. Twin acoustic-induced mutual forces (both repulsive and attractive) between coupled steel plates were realized in this system when the external plane wave normally impacted the plates. Calculations showed that the emerging forces are more than an order of magnitude larger than the corresponding induced force of a conventional grating structure. We also found that the strong acoustic-induced mutual forces stem from the resonant excitation of nonleaky flexural Lamb modes in the coupled plates, and that these forces couple more strongly with the external incident acoustic waves. Furthermore, the amplitudes and resonant wavelengths of these forces can be coarsely controlled by changing the symmetry of the system and finely adjusted by varying the slant angle and the edge-length of the oblique rhombus. The proposed acoustic system could potentially be applied in sensors and in the ultrasonic detection of weak signals in water.

  2. Does walking in a virtual environment induce unstable gait? An examination of vertical ground reaction forces.

    Science.gov (United States)

    Hollman, John H; Brey, Robert H; Bang, Tami J; Kaufman, Kenton R

    2007-07-01

    Virtual reality (VR) can induce postural instability in standing and walking, as quantified with kinematic parameters. This study examines the effect of a VR environment on kinetic gait parameters. Ten healthy volunteers walked on an instrumented treadmill in a VR environment and a non-VR environment. In the VR environment, a corridor with colored vertical stripes comprising the walls was projected onto a concave screen placed in front of the treadmill. The speed of the moving image was perceptually equivalent to the speed of the treadmill, creating an illusion that subjects walked through the corridor. Vertical ground reaction forces were sampled. Kinetic parameters that reflect gait stability (weight acceptance peak force, weight acceptance rate, push-off peak force and push-off rate) were compared between the VR and non-VR environments. Subjects walked in the VR environment with increased magnitudes and rates of weight acceptance force and with increased rates of push-off force. Variability in weight acceptance rates and peak forces, and variability in push-off peak forces, were also increased in the VR environment. The gait deviations reflect a compensatory response to visual stimulation that occurs in the VR environment, suggesting that walking in a VR environment may induce gait instability in healthy subjects.

  3. Regulatory systems for hypoxia-inducible gene expression in ischemic heart disease gene therapy.

    Science.gov (United States)

    Kim, Hyun Ah; Rhim, Taiyoun; Lee, Minhyung

    2011-07-18

    Ischemic heart diseases are caused by narrowed coronary arteries that decrease the blood supply to the myocardium. In the ischemic myocardium, hypoxia-responsive genes are up-regulated by hypoxia-inducible factor-1 (HIF-1). Gene therapy for ischemic heart diseases uses genes encoding angiogenic growth factors and anti-apoptotic proteins as therapeutic genes. These genes increase blood supply into the myocardium by angiogenesis and protect cardiomyocytes from cell death. However, non-specific expression of these genes in normal tissues may be harmful, since growth factors and anti-apoptotic proteins may induce tumor growth. Therefore, tight gene regulation is required to limit gene expression to ischemic tissues, to avoid unwanted side effects. For this purpose, various gene expression strategies have been developed for ischemic-specific gene expression. Transcriptional, post-transcriptional, and post-translational regulatory strategies have been developed and evaluated in ischemic heart disease animal models. The regulatory systems can limit therapeutic gene expression to ischemic tissues and increase the efficiency of gene therapy. In this review, recent progresses in ischemic-specific gene expression systems are presented, and their applications to ischemic heart diseases are discussed.

  4. Screening of hypoxia-inducible genes in sporadic ALS.

    LENUS (Irish Health Repository)

    Cronin, Simon

    2008-10-01

    Genetic variations in two hypoxia-inducible angiogenic genes, VEGF and ANG, have been linked with sporadic amyotrophic lateral sclerosis (SALS). Common variations in these genes may reduce the levels or functioning of their products. VEGF and ANG belong to a larger group of angiogenic genes that are up-regulated under hypoxic conditions. We hypothesized that common genetic variation across other members of this group may also predispose to sporadic ALS. To screen other hypoxia-inducible angiogenic genes for association with SALS, we selected 112 tagging single nucleotide polymorphisms (tgSNPs) that captured the common genetic variation across 16 VEGF-like and eight ANG-like hypoxia-inducible genes. Screening for association was performed in 270 Irish individuals with typical SALS and 272 ethnically matched unrelated controls. SNPs showing association in the Irish phase were genotyped in a replication sample of 281 Swedish sporadic ALS patients and 286 Swedish controls. Seven markers showed association in the Irish. The one modest replication signal observed in the Swedish replication sample, at rs3801158 in the gene inhibin beta A, was for the opposite allele vs. the Irish cohort. We failed to detect association of common variation across 24 candidate hypoxia-inducible angiogenic genes with SALS.

  5. Highly asymmetric interaction forces induced by acoustic waves in coupled plate structures

    CERN Document Server

    Fan, Xiying; Zhang, Shenwei; Ke, Manzhu; Liu, Zhengyou

    2015-01-01

    Mutual forces can be induced between coupled structures when illuminated by external acoustic waves. In this Letter, we propose a concept of asymmetric interaction between two coupled plate-like structures, which is generated by oppositely incident plane waves. Besides the striking contrast in magnitude, the mutual force induced by one of the incidences can be tuned extremely strong due to the resonant excitation of the flexural plate modes. The highly asymmetric interaction with enhanced strength in single side should be potentially useful, such as in designing ultrasound instruments and sensors.

  6. Grounding-Induced Sectional Forces and Residual Strength of Grounded Ship Hulls

    DEFF Research Database (Denmark)

    Paik, Jeom Kee; Pedersen, Preben Terndrup

    1996-01-01

    The aim of the present study is to determine the sectional forces induced by ship grounding and also to assess the residual strength of groundedship hulls. An analytical approach is used to estimate the grounding-induced sectional forces of ships. The extent and location of structural damage due...... to grounding is defined based on the ABS Safe Hull guide. The residual strength of damaged hulls is calculated by using a simple analytical formula. The method is applied to residual strength assessment of a damaged double hull tanker of 38,400 dwt due to grounding....

  7. Zscan4 transiently reactivates early embryonic genes during the generation of induced pluripotent stem cells.

    Science.gov (United States)

    Hirata, Tetsuya; Amano, Tomokazu; Nakatake, Yuhki; Amano, Misa; Piao, Yulan; Hoang, Hien G; Ko, Minoru S H

    2012-01-01

    The generation of induced pluripotent stem cells (iPSCs) by the forced expression of defined transcription factors in somatic cells holds great promise for the future of regenerative medicine. However, the initial reprogramming mechanism is still poorly understood. Here we show that Zscan4, expressed transiently in2-cell embryos and embryonic stem cells (ESCs), efficiently produces iPSCs from mouse embryo fibroblasts when coexpressed with Klf4, Oct4, and Sox2. Interestingly, the forced expression of Zscan4 is required onlyfor the first few days of iPSC formation. Microarray analysis revealed transient and early induction of preimplantation-specific genes in a Zscan4-dependent manner. Our work indicates that Zscan4 is a previously unidentified potent natural factor that facilitates the reprogramming process and reactivates early embryonic genes.

  8. Mechanisms of radiation-induced gene responses

    Energy Technology Data Exchange (ETDEWEB)

    Woloschak, G.E.; Paunesku, T.

    1996-10-01

    In the process of identifying genes differentially expressed in cells exposed ultraviolet radiation, we have identified a transcript having a 26-bp region that is highly conserved in a variety of species including Bacillus circulans, yeast, pumpkin, Drosophila, mouse, and man. When the 5` region (flanking region or UTR) of a gene, the sequence is predominantly in +/+ orientation with respect to the coding DNA strand; while in the coding region and the 3` region (UTR), the sequence is most frequently in the +/-orientation with respect to the coding DNA strand. In two genes, the element is split into two parts; however, in most cases, it is found only once but with a minimum of 11 consecutive nucleotides precisely depicting the original sequence. The element is found in a large number of different genes with diverse functions (from human ras p21 to B. circulans chitonase). Gel shift assays demonstrated the presence of a protein in HeLa cell extracts that binds to the sense and antisense single-stranded consensus oligomers, as well as to the double- stranded oligonucleotide. When double-stranded oligomer was used, the size shift demonstrated as additional protein-oligomer complex larger than the one bound to either sense or antisense single-stranded consensus oligomers alone. It is speculated either that this element binds to protein(s) important in maintaining DNA is a single-stranded orientation for transcription or, alternatively that this element is important in the transcription-coupled DNA repair process.

  9. Active shortening protects against stretch-induced force deficits in human skeletal muscle.

    Science.gov (United States)

    Saripalli, Anjali L; Sugg, Kristoffer B; Mendias, Christopher L; Brooks, Susan V; Claflin, Dennis R

    2017-02-23

    Skeletal muscle contraction results from molecular interactions of myosin "crossbridges" with adjacent actin filament binding sites. The binding of myosin to actin can be "weak" or "strong", and only strong binding states contribute to force production. During active shortening, the number of strongly-bound crossbridges declines with increasing shortening velocity. Forcibly stretching a muscle that is actively shortening at high velocity results in no apparent negative consequences whereas stretch of an isometrically (fixed-length) contracting muscle causes ultrastructural damage and a decline in force-generating capability. Our working hypothesis is that stretch-induced damage is uniquely attributable to the population of crossbridges that are strongly-bound. We tested the hypothesis that stretch-induced force deficits decline as the prevailing shortening velocity is increased. Experiments were performed on permeabilized segments of individual skeletal muscle fibers obtained from human subjects. Fibers were maximally activated and either allowed to generate maximum isometric force (Fo), or to shorten at velocities that resulted in force maintenance of ≈50% Fo or ≈2% Fo. For each test condition, a rapid stretch equivalent to 0.1 x optimal fiber length was applied. Relative to pre-stretch Fo, force deficits resulting from stretches applied during force maintenance of 100%, ≈50%, and ≈2% Fo were 23.2 ± 8.6%, 7.8 ± 4.2% and 0.3 ± 3.3%, respectively (mean ± SD, n=20). We conclude that stretch-induced damage declines with increasing shortening velocity, consistent with the working hypothesis that the fraction of strongly-bound crossbridges is a causative factor in the susceptibility of skeletal muscle to stretch-induced damage.

  10. Forced expression of OCT4 influences the expression of pluripotent genes in human mesenchymal stem cells and fibroblasts.

    Science.gov (United States)

    Palma, C S; Tannous, M A; Malta, T M; Russo, E M S; Covas, D T; Picanço-Castro, V

    2013-04-02

    Genetic reprogramming of adult cells to generate induced pluripotent stem (iPS) cells is a new and important step in sidestepping some of the ethical issues and risks involved in the use of embryonic stem cells. iPS cells can be generated by introduction of transcription factors, such as OCT4, SOX2, KLF4, and CMYC. iPS cells resemble embryonic stem cells in their properties and differentiation potential. The mechanisms that lead to induced pluripotency and the effect of each transcription factor are not completely understood. We performed a critical evaluation of the effect of overexpressing OCT4 in mesenchymal stem cells and fibroblasts and found that OCT4 can activate the expression of other stemness genes, such as SOX2, NANOG, CMYC, FOXD3, KLF4, and βCATENIN, which are not normally or are very weakly expressed in mesenchymal stem cells. Transient expression of OCT4 was also performed to evaluate whether these genes are affected by its overexpression in the first 48 h. Transfected fibroblast cells expressed around 275-fold more OCT4 than non-transfected cells. In transient expression, in which cells were analyzed after 48 h, we detected only the up-regulation of FOXD3, SOX2, and KLF4 genes, suggesting that these genes are the earlier targets of OCT4 in this cellular type. We conclude that forced expression of OCT4 can alter cell status and activate the pluripotent network. Knowledge gained through study of these systems may help us to understand the kinetics and mechanism of cell reprogramming.

  11. Reprogramming A375 cells to induced-resembled neuronal cells by structured overexpression of specific transcription genes

    OpenAIRE

    ZHANG, HENGZHU; Wei, Min; Jiang, Yangyang; Wang, Xiaodong; SHE, LEI; Yan, Zhengcun; Dong, Lun; Pang, Lujun; Wang, Xingdong

    2016-01-01

    Induced-resembled neuronal cells (irNCs) are generated by reprogramming human melanoma cells through the introduction of key transcription factors, providing novel concepts in the treatment of malignant tumor cells and making it possible to supply neural cells for laboratory use. In the present study, irNCs were derived from A375 cells by inducing the 'forced' overexpression of specific genes, including achaete-scute homolog 1 (Ascl1), neuronal differentiation factor 1 (Neurod1), myelin trans...

  12. Irrigation Induced Surface Cooling in the Context of Modern and Increased Greenhouse Gas Forcing

    Science.gov (United States)

    Cook, Benjamin I.; Puma, Michael J.; Krakauer, Nir Y.

    2010-01-01

    There is evidence that expected warming trends from increased greenhouse gas (GHG) forcing have been locally masked by irrigation induced cooling, and it is uncertain how the magnitude of this irrigation masking effect will change in the future. Using an irrigation dataset integrated into a global general circulation model, we investigate the equilibrium magnitude of irrigation induced cooling under modern (Year 2000) and increased (A1B Scenario, Year 2050) GHG forcing, using modern irrigation rates in both scenarios. For the modern scenario, the cooling is largest over North America, India, the Middle East, and East Asia. Under increased GHG forcing, this cooling effect largely disappears over North America, remains relatively unchanged over India, and intensifies over parts of China and the Middle East. For North America, irrigation significantly increases precipitation under modern GHG forcing; this precipitation enhancement largely disappears under A1B forcing, reducing total latent heat fluxes and the overall irrigation cooling effect. Over India, irrigation rates are high enough to keep pace with increased evaporative demand from the increased GHG forcing and the magnitude of the cooling is maintained. Over China, GHG forcing reduces precipitation and shifts the region to a drier evaporative regime, leading to a relatively increased impact of additional water from irrigation on the surface energy balance. Irrigation enhances precipitation in the Middle East under increased GHG forcing, increasing total latent heat fluxes and enhancing the irrigation cooling effect. Ultimately, the extent to which irrigation will continue to compensate for the warming from increased GHG forcing will primarily depend on changes in the background evaporative regime, secondary irrigation effects (e.g. clouds, precipitation), and the ability of societies to maintain (or increase) current irrigation rates.

  13. Salmonella induces prominent gene expression in the rat colon

    Directory of Open Access Journals (Sweden)

    Roosing Susanne

    2007-09-01

    Full Text Available Abstract Background Salmonella enteritidis is suggested to translocate in the small intestine. In vivo it induces gene expression changes in the ileal mucosa and Peyer's patches. Stimulation of Salmonella translocation by dietary prebiotics fermented in colon suggests involvement of the colon as well. However, effects of Salmonella on colonic gene expression in vivo are largely unknown. We aimed to characterize time dependent Salmonella-induced changes of colonic mucosal gene expression in rats using whole genome microarrays. For this, rats were orally infected with Salmonella enteritidis to mimic a foodborne infection and colonic gene expression was determined at days 1, 3 and 6 post-infection (n = 8 rats per time-point. As fructo-oligosaccharides (FOS affect colonic physiology, we analyzed colonic mucosal gene expression of FOS-fed versus cellulose-fed rats infected with Salmonella in a separate experiment. Colonic mucosal samples were isolated at day 2 post-infection. Results Salmonella affected transport (e.g. Chloride channel calcium activated 6, H+/K+ transporting Atp-ase, antimicrobial defense (e.g. Lipopolysaccharide binding protein, Defensin 5 and phospholipase A2, inflammation (e.g. calprotectin, oxidative stress related genes (e.g. Dual oxidase 2 and Glutathione peroxidase 2 and Proteolysis (e.g. Ubiquitin D and Proteosome subunit beta type 9. Furthermore, Salmonella translocation increased serum IFNγ and many interferon-related genes in colonic mucosa. The gene most strongly induced by Salmonella infection was Pancreatitis Associated Protein (Pap, showing >100-fold induction at day 6 after oral infection. Results were confirmed by Q-PCR in individual rats. Stimulation of Salmonella translocation by dietary FOS was accompanied by enhancement of the Salmonella-induced mucosal processes, not by induction of other processes. Conclusion We conclude that the colon is a target tissue for Salmonella, considering the abundant changes in

  14. Mechanical force alters morphogenetic movements and segmental gene expression patterns during Drosophila embryogenesis.

    Directory of Open Access Journals (Sweden)

    Abhishek Kumar

    Full Text Available The development of an organism is accompanied by various cellular morphogenetic movements, changes in cellular as well as nuclear morphology and transcription programs. Recent evidence suggests that intra and inter-cellular connections mediated by various adhesion proteins contribute to defining nuclear morphology. In addition, three dimensional organization of the cell nucleus regulate the transcription programs. However the link between cellular morphogenetic movements and its coupling to nuclear function in a developmental context is poorly understood. In this paper we use a point perturbation by tissue level laser ablation and sheet perturbation by application of force using magnetic tweezers to alter cellular morphogenetic movements and probe its impact on nuclear morphology and segmental gene expression patterns. Mechanical perturbations during blastoderm stage in a developing Drosophila embryo resulted in localized alterations in nuclear morphology and cellular movement. In addition, global defects in germ-band (GB extension and retraction are observed when external force is applied during morphogenetic movements, suggesting a long-range physical coupling within the GB layer of cells. Further local application of force resulted in redistribution of non muscle myosin-II in the GB layer. Finally these perturbations lead to altered segmental gene (engrailed expression patterns later during the development. Our observations suggest that there exists a tight regulation between nuclear morphology and cellular adhesive connections during morphogenetic movement of cells in the embryo. The observed spatial changes in patterning genes, with perturbation, highlight the importance of nuclear integrity to cellular movement in establishing gene expression program in a developmental system.

  15. Inducible gene expression and environmentally regulated genes in lactic acid bacteria

    NARCIS (Netherlands)

    Kok, Jan

    1996-01-01

    Relatively recently, a number of genes and operons have been identified in lactic acid bacteria that are inducible and respond to environmental factors. Some of these genes/operons had been isolated and analysed because of their importance in the fermentation industry and, consequently, their transc

  16. Lithium ions induce prestalk-associated gene expression and inhibit prespore gene expression in Dictyostelium discoideum

    NARCIS (Netherlands)

    Peters, Dorien J.M.; Lookeren Campagne, Michiel M. van; Haastert, Peter J.M. van; Spek, Wouter; Schaap, Pauline

    1989-01-01

    We investigated the effect of Li+ on two types of cyclic AMP-regulated gene expression and on basal and cyclic AMP-stimulated inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) levels. Li+ effectively inhibits cyclic AMP-induced prespore gene expression, half-maximal inhibition occurring at about 2mM-LiCl.

  17. Regulation of Masticatory Force During Cortically Induced Rhythmic Jaw Movements in the Anesthetized Rabbit

    National Research Council Canada - National Science Library

    Hidaka, O; Morimoto, T; Masuda, Y; Kato, T; Matsuo, R; Inoue, T; Kobayashi, M; Takada, K

    1997-01-01

    ... Japan Hidaka, O., T. Morimoto, Y. Masuda, T. Kato, R. Matsuo, T. Inoue, M. Kobayashi, and K. Takada. Regulation of masticatory force during cortically induced rhythmic jaw movements in the anesthetized rabbit...

  18. Near-field light detection of a photo induced force by atomic force microscopy with frequency modulation

    Science.gov (United States)

    Satoh, Nobuo; Kobayashi, Kei; Matsushige, Kazumi; Yamada, Hirofumi

    2017-08-01

    We demonstrated near-field light detection using a non contact-mode atomic force microscope (nc-AFM). This system obtains molecular-level resolution by reducing noise in the displacement detection of a Si cantilever. The Si cantilever probe tip was brought close to a glass with a patterned chromium film on a dove prism. The backside of the prism was irradiated by an intensity-modulated laser light to create an evanescent field at the glass surface. We obtained a near-field optical image of the chromium-patterned glass by detecting the amplitude modulation induced by the near-field light while the tip-sample distance was regulated by the frequency modulation method under atmospheric conditions.

  19. Water’s role in the force-induced unfolding of ubiquitin

    Science.gov (United States)

    Li, Jingyuan; Fernandez, Julio M.; Berne, B. J.

    2010-01-01

    In atomic force spectroscopic studies of the elastomeric protein ubiquitin, the β-strands 1-5 serve as the force clamp. Simulations show how the rupture force in the force-induced unfolding depends on the kinetics of water molecule insertion into positions where they can eventually form hydrogen bonding bridges with the backbone hydrogen bonds in the force-clamp region. The intrusion of water into this region is slowed down by the hydrophobic shielding effect of carbonaceous groups on the surface residues of β-strands 1-5, which thereby regulates water insertion prior to hydrogen bond breakage. The experiments show that the unfolding of the mechanically stressed protein is nonexponential due to static disorder. Our simulations show that different numbers and/or locations of bridging water molecules give rise to a long-lived distribution of transition states and static disorder. We find that slowing down the translational (not rotational) motions of the water molecules by increasing the mass of their oxygen atoms, which leaves the force field and thereby the equilibrium structure of the solvent unchanged, increases the average rupture force; however, the early stages of the force versus time behavior are very similar for our “normal” and fictitious “heavy” water models. Finally, we construct six mutant systems to regulate the hydrophobic shielding effect of the surface residues in the force-clamp region. The mutations in the two termini of β-sheets 1-5 are found to determine a preference for different unfolding pathways and change mutant’s average rupture force. PMID:20974922

  20. A programmable DNA origami nanospring that reveals force-induced adjacent binding of myosin VI heads

    Science.gov (United States)

    Iwaki, M.; Wickham, S. F.; Ikezaki, K.; Yanagida, T.; Shih, W. M.

    2016-01-01

    Mechanosensitive biological nanomachines such as motor proteins and ion channels regulate diverse cellular behaviour. Combined optical trapping with single-molecule fluorescence imaging provides a powerful methodology to clearly characterize the mechanoresponse, structural dynamics and stability of such nanomachines. However, this system requires complicated experimental geometry, preparation and optics, and is limited by low data-acquisition efficiency. Here we develop a programmable DNA origami nanospring that overcomes these issues. We apply our nanospring to human myosin VI, a mechanosensory motor protein, and demonstrate nanometre-precision single-molecule fluorescence imaging of the individual motor domains (heads) under force. We observe force-induced transitions of myosin VI heads from non-adjacent to adjacent binding, which correspond to adapted roles for low-load and high-load transport, respectively. Our technique extends single-molecule studies under force and clarifies the effect of force on biological processes. PMID:27941751

  1. A programmable DNA origami nanospring that reveals force-induced adjacent binding of myosin VI heads.

    Science.gov (United States)

    Iwaki, M; Wickham, S F; Ikezaki, K; Yanagida, T; Shih, W M

    2016-12-12

    Mechanosensitive biological nanomachines such as motor proteins and ion channels regulate diverse cellular behaviour. Combined optical trapping with single-molecule fluorescence imaging provides a powerful methodology to clearly characterize the mechanoresponse, structural dynamics and stability of such nanomachines. However, this system requires complicated experimental geometry, preparation and optics, and is limited by low data-acquisition efficiency. Here we develop a programmable DNA origami nanospring that overcomes these issues. We apply our nanospring to human myosin VI, a mechanosensory motor protein, and demonstrate nanometre-precision single-molecule fluorescence imaging of the individual motor domains (heads) under force. We observe force-induced transitions of myosin VI heads from non-adjacent to adjacent binding, which correspond to adapted roles for low-load and high-load transport, respectively. Our technique extends single-molecule studies under force and clarifies the effect of force on biological processes.

  2. Fundamental study on gene transfer utilizing magnetic force and jet injector

    Energy Technology Data Exchange (ETDEWEB)

    Hasegawa, T.; Nakagami, H.; Akiyama, Y.; Nishjima, S. [Osaka University, Osaka (Japan)

    2017-03-15

    Recently, DNA vaccination is attracting attentions as a new therapeutic method for lifestyle diseases and autoimmune diseases. However, its clinical applications are limited because a safe and efficient gene transfer method has not been established yet. In this study, a new method of gene transfer was proposed which utilizes the jet injection and the magnetic transfection. The jet injection is a method to inject medical liquid by momentary high pressure without needle. The injected liquid diffuses in the bio tissue and the endocytosis is considered to be improved by the diffusion. The magnetic transfection is a method to deliver the conjugates of plasmid DNA and magnetic particles to the desired site by external magnetic field. It is expected that jet injection of the conjugates causes slight membrane disruptions and the traction of the conjugates by magnetic field induces the efficient gene transfer. In conclusion, the possibility of improvement of the gene expression by the combination of jet injection and magnetic transfection was confirmed.

  3. Lift Enhancement and Oscillatory Suppression of Vortex-induced Vibration in Shear Flow by Loentz Force

    Institute of Scientific and Technical Information of China (English)

    张辉; 范宝春; 李鸿志

    2012-01-01

    The flow of the weak electrolyte solution can be controlled by Lorentz force achieved with the suitable magnetic and electric fields, and it has the advantages of vortex street suppression, drag reduction, lift enhancement and oscillatory suppression for the flow over a bluff body. The electro-magnetic control of vortex-induced vibration (VIV) of a circular cyl- inder in the shear flow was investigated numerically in the exponential-polar coordinates attached on the moving cylinder for Re = 150. With the effect of background vorticity, the vortex street of VIV cylinder was composed of two parallel rows with an opposite sign of the vortices which inclines toward the lower side and the strength of upper vortex is larger than that of lower vortex. The lift force vibrated periodically with the effect of vortex shedding and the mean value was negative due to the background vorticity. The Lorentz force for controlling the VIV cylinder was classified into the field Lorentz force and the wall Lorentz force. The field Lorentz force suppresses the lift oscillation, and in turn, suppresses the VIV, whereas the wall Loreutz force increases the lift.

  4. Tip-force induced surface deformation in the layered commensurate tellurides NbA xTe 2 (A = Si, Ge) during atomic force microscopy measurements

    Science.gov (United States)

    Bengel, H.; Cantow, H.-J.; Magonov, S. N.; Monconduit, L.; Evain, M.; Whangbo, M.-H.

    1994-12-01

    The Te-atom surfaces of commensurate layered tellurides NbA xTe 2 ( A = Si, x = {1}/{2}; A = Ge, x = {1}/{3}, {2}/{5}, {3}/{7}) were examined by atomic force microscopy (AFM) at different applied forces. Although the bulk crystal structures show a negligible height corrugation in the surface Te-atom sheets, the AFM images exhibit dark linear patterns that become strongly pronounced at high applied forces (several hundreds nN). This feature comes about because the tip-sample force interactions induce a surface corrugation according to the local hardness variation of the surface.

  5. INDUCIBLE RNAi-MEDIATED GENE SILENCING USING NANOSTRUCTURED GENE DELIVERY ARRAYS

    Energy Technology Data Exchange (ETDEWEB)

    Mann, David George James [ORNL; McKnight, Timothy E [ORNL; Mcpherson, Jackson [University of Tennessee, Knoxville (UTK); Hoyt, Peter R [ORNL; Melechko, Anatoli Vasilievich [ORNL; Simpson, Michael L [ORNL; Sayler, Gary Steven [ORNL

    2008-01-01

    RNA interference has become a powerful biological tool over the last decade. In this study, a tetracycline-inducible shRNA vector system was designed for silencing CFP expression and introduced alongside the yfp marker gene into Chinese hamster ovary cells using spatially indexed vertically aligned carbon nanofiber arrays (VACNFs) in a gene delivery process termed impalefection. The VACNF architecture provided simultaneous delivery of multiple genes, subsequent adherence and proliferation of interfaced cells, and repeated monitoring of single cells over time. 24 hours after nanofiber-mediated delivery, 53.1% 10.4% of the cells that expressed the yfp marker gene were also fully silenced by the inducible CFP-silencing shRNA vector. Additionally, efficient CFP-silencing was observed in single cells among a population of cells that remained CFP-expressing. This effective transient expression system enables rapid analysis of gene silencing effects using RNAi in single cells and cell populations.

  6. Temporary threshold shift of vibratory sensation induced by a vibrating handle and its gripping force.

    Science.gov (United States)

    Nishiyama, K; Taoda, K; Yamashita, H; Watanabe, S

    1996-01-01

    This study examines the effect of the force with which a vibrating handle is gripped on the temporary threshold shift of vibratory sensation (TTSv) induced by hand-arm vibration. Six healthy subjects gripped a handle vibrating with a 1.3 octave-band vibration, with a central frequency of 200 Hz and an intensity of 39.2 m/s2. Exposure was for 1 min and 10 min, respectively. Gripping forces for the 1-min exposure were 5 N, 10 N, 40 N and 80 N, respectively, with 0 N push-pull force. Gripping forces for the 10-min exposure were the same as for the 1-min exposure but omitting 80 N. The vibratory sensation threshold at 125 Hz was measured before and after exposure of an exposed fingertip to vibration. The differences measured determine TTSv.t at time t. TTSv.t determines TTSv.0, that is, the temporary threshold shift of vibratory sensation immediately after exposure to vibration according to the estimate made on the basis of the preceding study. The same experimental conditions were repeated 3 times on different days in a soundproof and thermoregulated room. Our findings show that TTSv increases significantly with increasing gripping force. We also determined the quantitative relationships between TTSv.0 and gripping force as described by the equation TTSv.0 = exp(kf x F + Cf). where kt and Cf are constants and F is gripping force. This study revealed the importance of ergonomic design in reducing the force with which a vibrating handle is gripped to prevent an adverse effect of local vibration. The equation devised may help in the quantitative assessment of the effect of reduced gripping force.

  7. Remanent cell traction force in renal vascular smooth muscle cells induced by integrin-mediated mechanotransduction.

    Science.gov (United States)

    Balasubramanian, Lavanya; Lo, Chun-Min; Sham, James S K; Yip, Kay-Pong

    2013-02-15

    It was previously demonstrated in isolated renal vascular smooth muscle cells (VSMCs) that integrin-mediated mechanotransduction triggers intracellular Ca(2+) mobilization, which is the hallmark of myogenic response in VSMCs. To test directly whether integrin-mediated mechanotransduction results in the myogenic response-like behavior in renal VSMCs, cell traction force microscopy was used to monitor cell traction force when the cells were pulled with fibronectin-coated or low density lipoprotein (LDL)-coated paramagnetic beads. LDL-coated beads were used as a control for nonintegrin-mediated mechanotransduction. Pulling with LDL-coated beads increased the cell traction force by 61 ± 12% (9 cells), which returned to the prepull level after the pulling process was terminated. Pulling with noncoated beads had a minimal increase in the cell traction force (12 ± 9%, 8 cells). Pulling with fibronectin-coated beads increased the cell traction force by 56 ± 20% (7 cells). However, the cell traction force was still elevated by 23 ± 14% after the pulling process was terminated. This behavior is analogous to the changes of vascular resistance in pressure-induced myogenic response, in which vascular resistance remains elevated after myogenic constriction. Fibronectin is a native ligand for α(5)β(1)-integrins in VSMCs. Similar remanent cell traction force was found when cells were pulled with beads coated with β(1)-integrin antibody (Ha2/5). Activation of β(1)-integrin with soluble antibody also triggered variations of cell traction force and Ca(2+) mobilization, which were abolished by the Src inhibitor. In conclusion, mechanical force transduced by α(5)β(1)-integrins triggered a myogenic response-like behavior in isolated renal VSMCs.

  8. Transcription dynamics of inducible genes modulated by negative regulations.

    Science.gov (United States)

    Li, Yanyan; Tang, Moxun; Yu, Jianshe

    2015-06-01

    Gene transcription is a stochastic process in single cells, in which genes transit randomly between active and inactive states. Transcription of many inducible genes is also tightly regulated: It is often stimulated by extracellular signals, activated through signal transduction pathways and later repressed by negative regulations. In this work, we study the nonlinear dynamics of the mean transcription level of inducible genes modulated by the interplay of the intrinsic transcriptional randomness and the repression by negative regulations. In our model, we integrate negative regulations into gene activation process, and make the conventional assumption on the production and degradation of transcripts. We show that, whether or not the basal transcription is temporarily terminated when cells are stimulated, the mean transcription level grows in the typical up and down pattern commonly observed in immune response genes. With the help of numerical simulations, we clarify the delicate impact of the system parameters on the transcription dynamics, and demonstrate how our model generates the distinct temporal gene-induction patterns in mouse fibroblasts discerned in recent experiments.

  9. Roles of factorial noise in inducing bimodal gene expression

    Science.gov (United States)

    Liu, Peijiang; Yuan, Zhanjiang; Huang, Lifang; Zhou, Tianshou

    2015-06-01

    Some gene regulatory systems can exhibit bimodal distributions of mRNA or protein although the deterministic counterparts are monostable. This noise-induced bimodality is an interesting phenomenon and has important biological implications, but it is unclear how different sources of expression noise (each source creates so-called factorial noise that is defined as a component of the total noise) contribute separately to this stochastic bimodality. Here we consider a minimal model of gene regulation, which is monostable in the deterministic case. Although simple, this system contains factorial noise of two main kinds: promoter noise due to switching between gene states and transcriptional (or translational) noise due to synthesis and degradation of mRNA (or protein). To better trace the roles of factorial noise in inducing bimodality, we also analyze two limit models, continuous and adiabatic approximations, apart from the exact model. We show that in the case of slow gene switching, the continuous model where only promoter noise is considered can exhibit bimodality; in the case of fast switching, the adiabatic model where only transcriptional or translational noise is considered can also exhibit bimodality but the exact model cannot; and in other cases, both promoter noise and transcriptional or translational noise can cooperatively induce bimodality. Since slow gene switching and large protein copy numbers are characteristics of eukaryotic cells, whereas fast gene switching and small protein copy numbers are characteristics of prokaryotic cells, we infer that eukaryotic stochastic bimodality is induced mainly by promoter noise, whereas prokaryotic stochastic bimodality is induced primarily by transcriptional or translational noise.

  10. A silicon-nanowire memory driven by optical gradient force induced bistability

    Energy Technology Data Exchange (ETDEWEB)

    Dong, B. [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), Singapore 117685 (Singapore); Cai, H., E-mail: caih@ime.a-star.edu.sg; Gu, Y. D.; Kwong, D. L. [Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), Singapore 117685 (Singapore); Chin, L. K.; Ng, G. I.; Ser, W. [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Huang, J. G. [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), Singapore 117685 (Singapore); School of Mechanical Engineering, Xi' an Jiaotong University, Xi' an 710049 (China); Yang, Z. C. [School of Electronics Engineering and Computer Science, Peking University, Beijing 100871 (China); Liu, A. Q., E-mail: eaqliu@ntu.edu.sg [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); School of Electronics Engineering and Computer Science, Peking University, Beijing 100871 (China)

    2015-12-28

    In this paper, a bistable optical-driven silicon-nanowire memory is demonstrated, which employs ring resonator to generate optical gradient force over a doubly clamped silicon-nanowire. Two stable deformation positions of a doubly clamped silicon-nanowire represent two memory states (“0” and “1”) and can be set/reset by modulating the light intensity (<3 mW) based on the optical force induced bistability. The time response of the optical-driven memory is less than 250 ns. It has applications in the fields of all optical communication, quantum computing, and optomechanical circuits.

  11. Plant nodulation inducers enhance horizontal gene transfer of Azorhizobium caulinodans symbiosis island.

    Science.gov (United States)

    Ling, Jun; Wang, Hui; Wu, Ping; Li, Tao; Tang, Yu; Naseer, Nawar; Zheng, Huiming; Masson-Boivin, Catherine; Zhong, Zengtao; Zhu, Jun

    2016-11-29

    Horizontal gene transfer (HGT) of genomic islands is a driving force of bacterial evolution. Many pathogens and symbionts use this mechanism to spread mobile genetic elements that carry genes important for interaction with their eukaryotic hosts. However, the role of the host in this process remains unclear. Here, we show that plant compounds inducing the nodulation process in the rhizobium-legume mutualistic symbiosis also enhance the transfer of symbiosis islands. We demonstrate that the symbiosis island of the Sesbania rostrata symbiont, Azorhizobium caulinodans, is an 87.6-kb integrative and conjugative element (ICE(Ac)) that is able to excise, form a circular DNA, and conjugatively transfer to a specific site of gly-tRNA gene of other rhizobial genera, expanding their host range. The HGT frequency was significantly increased in the rhizosphere. An ICE(Ac)-located LysR-family transcriptional regulatory protein AhaR triggered the HGT process in response to plant flavonoids that induce the expression of nodulation genes through another LysR-type protein, NodD. Our study suggests that rhizobia may sense rhizosphere environments and transfer their symbiosis gene contents to other genera of rhizobia, thereby broadening rhizobial host-range specificity.

  12. Gamma globulins-induced interaction between two red blood cells: forces measurement with optical tweezers

    Science.gov (United States)

    Lee, Kisung; Muravyov, Alexei; Semenov, Alexei; Wagner, Christian; Priezzhev, Alexander; Lyubin, Eugeny; Fedyanin, Andrey

    2017-03-01

    The protein contribution to the red blood cell (RBC) aggregation is studied using the in-house made two-channeled optical tweezers. The cells interaction was characterized using two forces: the force required for separating two cells (FD - disaggregating force) and the force required for holding them from their spontaneous aggregation (FA - aggregating force). The gamma globulin solutions with/without albumin were used to induce the RBC aggregation. The strong interaction (3-10 pN) between the cells was measured within the contact formed using optical tweezers. We found that FD becomes stronger as the gamma globulin concentration increases, while the addition of albumin to the solution led to the significant (few fold) enhancement of the cells interaction forces. However, despite of the strong interaction between the cells their spontaneous overlapping was not observed, unlike the case in plasma, where the cells did increase their overlapping surface, when attached with small interacting surface and released from optical traps. This work in addition to our previous work with model solutions of fibrinogen allows us to conclude that the synergy of blood components is one of the most important features that contribute to the reversible RBC aggregation.

  13. Comparison of rotordynamic fluid forces in axial inducers and centrifugal turbopump impellers

    Science.gov (United States)

    d'Agostino, Luca

    2016-05-01

    The paper illustrates and compares the results of the experimental campaigns carried out in the Cavitating Pump Rotordynamic Test Facility (CPRTF) at Alta, Italy, under ESA funding for the characterization of the lateral rotordynamic fluid forces acting on high-head axial inducers and centrifugal turbopump impellers for space propulsion applications. The configurations presented here refer to a three-bladed tapered-hub, variable-pitch, inducer (DAPROT3) and a single-stage centrifugal pump (VAMPIRE) with vaneless diffuser and single spiral volute. Both the centrifugal pump and the inducer have been designed by means of reduced order models specifically developed by the author and his collaborators for the geometric definition and performance prediction of this kind of hydraulic turbomachinery. Continuous spectra of the rotordynamic forces acting on the impellers as functions of the whirl frequency have been obtained by means of the novel technique recently developed and demonstrated at Alta. The influence of the rotor whirl motion, flow rate, cavitating conditions, and liquid temperature (thermal cavitation effects) on the rotordynamic fluid forces is illustrated and the observed differences in their behavior in axial inducers and centrifugal turbpumps are discussed and interpreted in the light of the outcome of recent cavitation visualization experiments carried out by the Chemical Propulsion Team at Alta.

  14. Maxwell stress analysis of displacement induced electric force and its application to natural self-oscillation of a free electron

    CERN Document Server

    Yu, Zhixian

    2013-01-01

    The Maxwell stress analysis is used for the calculation of displacement induced temporary electric force of a single charge. This force comes from the variation of the charge's electric intensities that follow Coulomb's inverse square law, and is a kind of displacement dependent temporary restore force. The possibility of natural self-oscillation of a free electron which is driven by this self-force is also suggested.

  15. A gene-trap strategy identifies quiescence-induced genes in synchronized myoblasts

    Indian Academy of Sciences (India)

    Ramkumar Sambasivan; Grace K Pavlath; Jyotsna Dhawan

    2008-03-01

    Cellular quiescence is characterized not only by reduced mitotic and metabolic activity but also by altered gene expression. Growing evidence suggests that quiescence is not merely a basal state but is regulated by active mechanisms. To understand the molecular programme that governs reversible cell cycle exit, we focused on quiescence-related gene expression in a culture model of myogenic cell arrest and activation. Here we report the identification of quiescence-induced genes using a gene-trap strategy. Using a retroviral vector, we generated a library of gene traps in C2C12 myoblasts that were screened for arrest-induced insertions by live cell sorting (FACS-gal). Several independent genetrap lines revealed arrest-dependent induction of gal activity, confirming the efficacy of the FACS screen. The locus of integration was identified in 15 lines. In three lines, insertion occurred in genes previously implicated in the control of quiescence, i.e. EMSY – a BRCA2-interacting protein, p8/com1– a p300HAT-binding protein and MLL5 – a SET domain protein. Our results demonstrate that expression of chromatin modulatory genes is induced in G0, providing support to the notion that this reversibly arrested state is actively regulated.

  16. The force analysis for superparamagnetic nanoparticles-based gene delivery in an oscillating magnetic field

    Science.gov (United States)

    Sun, Jiajia; Shi, Zongqian; Jia, Shenli; Zhang, Pengbo

    2017-04-01

    Due to the peculiar magnetic properties and the ability to function in cell-level biological interaction, superparamagnetic nanoparticles (SMNP) have been being the attractive carrier for gene delivery. The superparamagnetic nanoparticles with surface-bound gene vector can be attracted to the surface of cells by the Kelvin force provided by external magnetic field. In this article, the influence of the oscillating magnetic field on the characteristics of magnetofection is studied in terms of the magnetophoretic velocity. The magnetic field of a cylindrical permanent magnet is calculated by equivalent current source (ECS) method, and the Kelvin force is derived by using the effective moment method. The results show that the static magnetic field accelerates the sedimentation of the particles, and drives the particles inward towards the axis of the magnet. Based on the investigation of the magnetophoretic velocity of the particle under horizontally oscillating magnetic field, an oscillating velocity within the amplitude of the magnet oscillation is observed. Furthermore, simulation results indicate that the oscillating amplitude plays an important role in regulating the active region, where the particles may present oscillating motion. The analysis of the magnetophoretic velocity gives us an insight into the physical mechanism of the magnetofection. It's also helpful to the optimal design of the magnetofection system.

  17. Association of Forced Vital Capacity with the Developmental Gene NCOR2.

    Directory of Open Access Journals (Sweden)

    Cosetta Minelli

    Full Text Available Forced Vital Capacity (FVC is an important predictor of all-cause mortality in the absence of chronic respiratory conditions. Epidemiological evidence highlights the role of early life factors on adult FVC, pointing to environmental exposures and genes affecting lung development as risk factors for low FVC later in life. Although highly heritable, a small number of genes have been found associated with FVC, and we aimed at identifying further genetic variants by focusing on lung development genes.Per-allele effects of 24,728 SNPs in 403 genes involved in lung development were tested in 7,749 adults from three studies (NFBC1966, ECRHS, EGEA. The most significant SNP for the top 25 genes was followed-up in 46,103 adults (CHARGE and SpiroMeta consortia and 5,062 children (ALSPAC. Associations were considered replicated if the replication p-value survived Bonferroni correction (p<0.002; 0.05/25, with a nominal p-value considered as suggestive evidence. For SNPs with evidence of replication, effects on the expression levels of nearby genes in lung tissue were tested in 1,111 lung samples (Lung eQTL consortium, with further functional investigation performed using public epigenomic profiling data (ENCODE.NCOR2-rs12708369 showed strong replication in children (p = 0.0002, with replication unavailable in adults due to low imputation quality. This intronic variant is in a strong transcriptional enhancer element in lung fibroblasts, but its eQTL effects could not be tested due to low imputation quality in the eQTL dataset. SERPINE2-rs6754561 replicated at nominal level in both adults (p = 0.036 and children (p = 0.045, while WNT16-rs2707469 replicated at nominal level only in adults (p = 0.026. The eQTL analyses showed association of WNT16-rs2707469 with expression levels of the nearby gene CPED1. We found no statistically significant eQTL effects for SERPINE2-rs6754561.We have identified a new gene, NCOR2, in the retinoic acid signalling pathway pointing

  18. Association of Forced Vital Capacity with the Developmental Gene NCOR2

    Science.gov (United States)

    Minelli, Cosetta; Dean, Charlotte H.; Hind, Matthew; Alves, Alexessander Couto; Amaral, André F. S.; Siroux, Valerie; Huikari, Ville; Soler Artigas, María; Evans, David M.; Loth, Daan W.; Bossé, Yohan; Postma, Dirkje S.; Sin, Don; Thompson, John; Demenais, Florence; Henderson, John; Bouzigon, Emmanuelle; Jarvis, Deborah; Järvelin, Marjo-Riitta; Burney, Peter

    2016-01-01

    Background Forced Vital Capacity (FVC) is an important predictor of all-cause mortality in the absence of chronic respiratory conditions. Epidemiological evidence highlights the role of early life factors on adult FVC, pointing to environmental exposures and genes affecting lung development as risk factors for low FVC later in life. Although highly heritable, a small number of genes have been found associated with FVC, and we aimed at identifying further genetic variants by focusing on lung development genes. Methods Per-allele effects of 24,728 SNPs in 403 genes involved in lung development were tested in 7,749 adults from three studies (NFBC1966, ECRHS, EGEA). The most significant SNP for the top 25 genes was followed-up in 46,103 adults (CHARGE and SpiroMeta consortia) and 5,062 children (ALSPAC). Associations were considered replicated if the replication p-value survived Bonferroni correction (p<0.002; 0.05/25), with a nominal p-value considered as suggestive evidence. For SNPs with evidence of replication, effects on the expression levels of nearby genes in lung tissue were tested in 1,111 lung samples (Lung eQTL consortium), with further functional investigation performed using public epigenomic profiling data (ENCODE). Results NCOR2-rs12708369 showed strong replication in children (p = 0.0002), with replication unavailable in adults due to low imputation quality. This intronic variant is in a strong transcriptional enhancer element in lung fibroblasts, but its eQTL effects could not be tested due to low imputation quality in the eQTL dataset. SERPINE2-rs6754561 replicated at nominal level in both adults (p = 0.036) and children (p = 0.045), while WNT16-rs2707469 replicated at nominal level only in adults (p = 0.026). The eQTL analyses showed association of WNT16-rs2707469 with expression levels of the nearby gene CPED1. We found no statistically significant eQTL effects for SERPINE2-rs6754561. Conclusions We have identified a new gene, NCOR2, in the retinoic

  19. Universal laws in the force-induced unraveling of biological bonds

    Science.gov (United States)

    Pereverzev, Yuriy V.; Prezhdo, Oleg V.

    2007-01-01

    Universal laws in the force-induced unbinding of receptor-ligand complexes are established for a general functional dependence of the dissociation rate constant on the applied force and are detailed with the two-pathway model that describes the recently discovered biological catch bond. The relationships link the data obtained with constant and time-dependent forces in different regimes, provide common representation for the previously unrelated data sets, and, thereby, greatly facilitate analysis and interpretation of experiments. The universal laws are demonstrated with the monomeric and dimeric catch-slip bonds between P-selectins and P-selectin glycoprotein ligands-1, and the slip bond between E-selectin and sialyl Lewisx antigen.

  20. Effects of wave-induced forcing on a circulation model of the North Sea

    Science.gov (United States)

    Staneva, Joanna; Alari, Victor; Breivik, Øyvind; Bidlot, Jean-Raymond; Mogensen, Kristian

    2017-04-01

    The effect of wind waves on water level and currents during two storms in the North Sea is investigated using a high-resolution NEMO model forced with fluxes and fields from a high-resolution wave model. The additional terms accounting for wave-current interaction that are considered in this study are the Stokes-Coriolis force and the sea-state dependent energy and momentum fluxes. The individual and collective role of these processes is quantified and the results are compared with a control run without wave effects as well as against current and water level measurements from coastal stations. We find a better agreement with observations when the circulation model is forced by sea-state dependent fluxes, especially in extreme events. The two extreme events, the storm Christian (25-27 October 2013), and about a month later, the storm Xaver (5-7 December 2013), induce different wave and surge conditions over the North Sea. Including the wave effects in the circulation model for the storm Xaver raises the modelled surge by more than 40 cm compared with the control run in the German Bight area. For the storm Christian, a difference of 20-30 cm in the surge level between the wave-forced and the stand-alone ocean model is found over the whole southern part of the North Sea. Moreover, the modelled vertical velocity profile fits the observations very well when the wave forcing is accounted for. The contribution of wave-induced forcing has been quantified indicating that this represents an important mechanism for improving water level and current predictions.

  1. Effects of wave-induced forcing on a circulation model of the North Sea

    Science.gov (United States)

    Staneva, Joanna; Alari, Victor; Breivik, Øyvind; Bidlot, Jean-Raymond; Mogensen, Kristian

    2017-01-01

    The effect of wind waves on water level and currents during two storms in the North Sea is investigated using a high-resolution Nucleus for European Modelling of the Ocean (NEMO) model forced with fluxes and fields from a high-resolution wave model. The additional terms accounting for wave-current interaction that are considered in this study are the Stokes-Coriolis force, the sea-state-dependent energy and momentum fluxes. The individual and collective role of these processes is quantified and the results are compared with a control run without wave effects as well as against current and water-level measurements from coastal stations. We find a better agreement with observations when the circulation model is forced by sea-state-dependent fluxes, especially in extreme events. The two extreme events, the storm Christian (25-27 October 2013), and about a month later, the storm Xaver (5-7 December 2013), induce different wave and surge conditions over the North Sea. Including the wave effects in the circulation model for the storm Xaver raises the modelled surge by more than 40 cm compared with the control run in the German Bight area. For the storm Christian, a difference of 20-30 cm in the surge level between the wave-forced and the stand-alone ocean model is found over the whole southern part of the North Sea. Moreover, the modelled vertical velocity profile fits the observations very well when the wave forcing is accounted for. The contribution of wave-induced forcing has been quantified indicating that this represents an important mechanism for improving water-level and current predictions.

  2. Schlieren High Speed Imaging on Fluid Flow in Liquid Induced by Plasma-driven Interfacial Forces

    Science.gov (United States)

    Lai, Janis; Foster, John

    2016-10-01

    Effective plasma-based water purification depends heavily on the transport of plasma-derived reactive species from the plasma into the liquid. Plasma interactions at the liquid-gas boundary are known to drive circulation in the bulk liquid. This forced circulation is not well understood. A 2-D plasma- in-liquid water apparatus is currently being investigated as a means to study the plasma-liquid interface to understand not only reactive species flows but to also understand plasma- driven fluid dynamic effects in the bulk fluid. Using Schlieren high speed imaging, plasma-induced density gradients near the interfacial region and into the bulk solution are measured to investigate the nature of these interfacial forces. Plasma-induced flow was also measured using particle imaging velocimetry. NSF CBET 1336375 and DOE DE-SC0001939.

  3. Gene expression profiling reveals multiple toxicity endpoints induced by hepatotoxicants

    Energy Technology Data Exchange (ETDEWEB)

    Huang Qihong; Jin Xidong; Gaillard, Elias T.; Knight, Brian L.; Pack, Franklin D.; Stoltz, James H.; Jayadev, Supriya; Blanchard, Kerry T

    2004-05-18

    Microarray technology continues to gain increased acceptance in the drug development process, particularly at the stage of toxicology and safety assessment. In the current study, microarrays were used to investigate gene expression changes associated with hepatotoxicity, the most commonly reported clinical liability with pharmaceutical agents. Acetaminophen, methotrexate, methapyrilene, furan and phenytoin were used as benchmark compounds capable of inducing specific but different types of hepatotoxicity. The goal of the work was to define gene expression profiles capable of distinguishing the different subtypes of hepatotoxicity. Sprague-Dawley rats were orally dosed with acetaminophen (single dose, 4500 mg/kg for 6, 24 and 72 h), methotrexate (1 mg/kg per day for 1, 7 and 14 days), methapyrilene (100 mg/kg per day for 3 and 7 days), furan (40 mg/kg per day for 1, 3, 7 and 14 days) or phenytoin (300 mg/kg per day for 14 days). Hepatic gene expression was assessed using toxicology-specific gene arrays containing 684 target genes or expressed sequence tags (ESTs). Principal component analysis (PCA) of gene expression data was able to provide a clear distinction of each compound, suggesting that gene expression data can be used to discern different hepatotoxic agents and toxicity endpoints. Gene expression data were applied to the multiplicity-adjusted permutation test and significantly changed genes were categorized and correlated to hepatotoxic endpoints. Repression of enzymes involved in lipid oxidation (acyl-CoA dehydrogenase, medium chain, enoyl CoA hydratase, very long-chain acyl-CoA synthetase) were associated with microvesicular lipidosis. Likewise, subsets of genes associated with hepatotocellular necrosis, inflammation, hepatitis, bile duct hyperplasia and fibrosis have been identified. The current study illustrates that expression profiling can be used to: (1) distinguish different hepatotoxic endpoints; (2) predict the development of toxic endpoints; and

  4. Intronic L1 retrotransposons and nested genes cause transcriptional interference by inducing intron retention, exonization and cryptic polyadenylation.

    Directory of Open Access Journals (Sweden)

    Kristel Kaer

    Full Text Available BACKGROUND: Transcriptional interference has been recently recognized as an unexpectedly complex and mostly negative regulation of genes. Despite a relatively few studies that emerged in recent years, it has been demonstrated that a readthrough transcription derived from one gene can influence the transcription of another overlapping or nested gene. However, the molecular effects resulting from this interaction are largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: Using in silico chromosome walking, we searched for prematurely terminated transcripts bearing signatures of intron retention or exonization of intronic sequence at their 3' ends upstream to human L1 retrotransposons, protein-coding and noncoding nested genes. We demonstrate that transcriptional interference induced by intronic L1s (or other repeated DNAs and nested genes could be characterized by intron retention, forced exonization and cryptic polyadenylation. These molecular effects were revealed from the analysis of endogenous transcripts derived from different cell lines and tissues and confirmed by the expression of three minigenes in cell culture. While intron retention and exonization were comparably observed in introns upstream to L1s, forced exonization was preferentially detected in nested genes. Transcriptional interference induced by L1 or nested genes was dependent on the presence or absence of cryptic splice sites, affected the inclusion or exclusion of the upstream exon and the use of cryptic polyadenylation signals. CONCLUSIONS/SIGNIFICANCE: Our results suggest that transcriptional interference induced by intronic L1s and nested genes could influence the transcription of the large number of genes in normal as well as in tumor tissues. Therefore, this type of interference could have a major impact on the regulation of the host gene expression.

  5. Fluctuation-induced forces between rings threaded around a polymer chain under tension

    CERN Document Server

    Gilles, F M; Pastorino, C

    2016-01-01

    We characterize the fluctuation properties of a polymer chain under external tension and the fluctuation-induced forces between two ring molecules threaded around the chain. The problem is relevant in the context of fluctuation-induced forces in soft matter systems, features of liquid interfaces and to describe properties of polyrotaxanes and slide-ring materials. We perform molecular dynamics simulations of the Kremer-Grest bead-spring model for the polymer and a simple ring-molecule model, in the canonical ensemble. We study transverse fluctuations of the stretched chain, as a function of chain stretching and in the presence of ring-shaped threaded molecules. The fluctuation spectra of the chains are analyzed in equilibrium at constant temperature and the differences in presence of two ring molecules are compared. For the rings located at fixed distances, we find an attractive fluctuation-induced force between the rings, proportional to the temperature and decaying with the ring distance. We characterize th...

  6. Substrates with engineered step changes in rigidity induce traction force polarity and durotaxis.

    Science.gov (United States)

    Breckenridge, Mark T; Desai, Ravi A; Yang, Michael T; Fu, Jianping; Chen, Christopher S

    2014-03-01

    Rigidity sensing plays a fundamental role in multiple cell functions ranging from migration, to proliferation and differentiation(1-5). During migration, single cells have been reported to preferentially move toward more rigid regions of a substrate in a process termed durotaxis. Durotaxis could contribute to cell migration in wound healing and gastrulation, where local gradients in tissue rigidity have been described. Despite the potential importance of this phenomenon to physiology and disease, it remains unclear how rigidity guides these behaviors and the underlying cellular and molecular mechanisms. To investigate the functional role of subcellular distribution and dynamics of cellular traction forces during durotaxis, we developed a unique microfabrication strategy to generate elastomeric micropost arrays patterned with regions exhibiting two different rigidities juxtaposed next to each other. After initial cell attachment on the rigidity boundary of the micropost array, NIH 3T3 fibroblasts were observed to preferentially migrate toward the rigid region of the micropost array, indicative of durotaxis. Additionally, cells bridging two rigidities across the rigidity boundary on the micropost array developed stronger traction forces on the more rigid side of the substrate indistinguishable from forces generated by cells exclusively seeded on rigid regions of the micropost array. Together, our results highlighted the utility of step-rigidity micropost arrays to investigate the functional role of traction forces in rigidity sensing and durotaxis, suggesting that cells could sense substrate rigidity locally to induce an asymmetrical intracellular traction force distribution to contribute to durotaxis.

  7. Prediction Model for Vortex-Induced Vibration of Circular Cylinder with Data of Forced Vibration

    Institute of Scientific and Technical Information of China (English)

    PAN Zhi-yuan; CUI Wei-cheng; LIU Ying-zhong

    2007-01-01

    A model based on the data from forced vibration experiments is developed for predicting the vortex-induced vibrations (VIV) of elastically mounted circular cylinders in flow. The assumptions for free and forced vibration tests are explored briefly. Energy equilibrium is taken into account to set up the relationship between the dynamic response of self-excited oscillations and the force coefficients from forced vibration experiments. The gap between these two cases is bridged straightforwardly with careful treatment of key parameters. Given reduced mass m* and material damping ratio ζ of an elastically mounted circular cylinder in flow, the response characteristics such as amplitude, frequency, lock-in range, added mass coefficient, cross-flow fluid force and the corresponding phase angle can be predicted all at once. Instances with different combination of reduced mass and material damping ratio are compared to investigate their effects on VIV. The hysteresis phenomenon can be interpreted reasonably. The predictions and the results from recent experiments carried out by Williamson's group are in rather good agreement.

  8. Effects of amplitude and phase-duration modification on electrically induced contraction force and discomfort.

    Science.gov (United States)

    Springer, Shmuel; Shapiro, Maxim

    2017-08-09

    Neuromuscular electrical stimulation (NMES) is commonly used in rehabilitation. However, the optimal combination of phase-duration and amplitude for enhancing motor output is not yet resolved. To test the effects of increasing phase-duration and amplitude on isometric knee extension force and discomfort, while controlling the effects of electrode-skin resistance and body mass index (BMI). Twenty-one healthy volunteers participated in the study. Stimulation was set at 250 μsec phase-duration and 45 Hz to evoke 10% of maximal voluntary isometric contraction of the quadriceps. Electrode-skin resistance was measured. Then, electrically induced contraction (EIC) forces and discomfort level were measured under four conditions: Moderate (25%) or substantial increase (50%) from baseline amplitude with constant phase-duration and moderate (25%) or substantial increase (50%) in phase-duration with amplitude constant. Compared with baseline, EIC force was significantly higher in all intensification conditions, while discomfort was significantly greater in all conditions except for moderate increase in phase-duration (p= 0.44). Amplitude intensification produced significantly higher force and greater discomfort than phase-duration. Electrode-skin resistance and BMI were not significant covariates. Greater force is elicited by increasing amplitude than by similar increase in phase-duration; however, the associated discomfort is also higher. Clinicians may use phase-duration while conditioning for NMES.

  9. Interparticle Capillary Forces at a Fluid-Fluid Interface with Strong Polymer-Induced Aging.

    Science.gov (United States)

    Cappelli, Stefano; de Jong, Arthur M; Baudry, Jean; Prins, Menno W J

    2017-01-24

    We report on a measurement of forces between particles adsorbed at a water-oil interface in the presence of an oil-soluble polymer. The cationic polymer interacts electrostatically with the negatively charged particles, thereby modulating the particle contact angle and the magnitude of capillary attraction between the particles. However, polymer adsorption to the interface also generates an increase in the apparent interfacial viscosity over several orders of magnitude in a time span of a few hours. We have designed an experiment in which repeated motion trajectories are measured on pairs of particles. The experiment gives an independent quantification of the interfacial drag coefficient (10(-7)-10(-4) Ns/m) and of the interparticle capillary forces (0.1-10 pN). We observed that the attractive capillary force depends on the amount of polymer in the oil phase and on the particle pair. However, the attraction appears to be independent of the surface rheology, with changes over a wide range of apparent viscosity values due to aging. Given the direction (attraction), the range (∼μm), and the distance dependence (∼1/S(5)) of the observed interparticle force, we interpret the force as being caused by quadrupolar deformations of the fluid-fluid interface induced by particle surface roughness. The results suggest that capillary forces are equilibrated in the early stages of interface aging and thereafter do not change anymore, even though strong changes in surface rheology still occur. The described experimental approach is powerful for studying dissipative as well as conservative forces of micro- and nanoparticles at fluid-fluid interfaces for systems out of equilibrium.

  10. Differential Gene Expression in Chemically Induced Mouse Lung Adenomas

    Directory of Open Access Journals (Sweden)

    Ruisheng Yao

    2003-01-01

    Full Text Available Because of similarities in histopathology and tumor progression stages between mouse and human lung adenocarcinomas, the mouse lung tumor model with lung adenomas as the endpoint has been used extensively to evaluate the efficacy of putative lung cancer chemopreventive agents. In this study, a competitive cDNA library screening (CCLS was employed to determine changes in the expression of mRNA in chemically induced lung adenomas compared with paired normal lung tissues. A total of 2555 clones having altered expression in tumors were observed following competitive hybridization between normal lung and lung adenomas after primary screening of over 160,000 clones from a mouse lung cDNA library. Among the 755 clones confirmed by dot blot hybridization, 240 clones were underexpressed, whereas 515 clones were overexpressed in tumors. Sixty-five clones with the most frequently altered expression in six individual tumors were confirmed by semiquantitative RT-PCR. When examining the 58 known genes, 39 clones had increased expression and 19 had decreased expression, whereas the 7 novel genes showed overexpression. A high percentage (>60% of overexpressed or underexpressed genes was observed in at least two or three of the lesions. Reproducibly overexpressed genes included ERK-1, JAK-1, surfactant proteins A, B, and C, NFAT1, α-1 protease inhibitor, helix-loop-helix ubiquitous kinase (CHUK, α-adaptin, α-1 PI2, thioether S-methyltransferase, and CYP2C40. Reproducibly underexpressed genes included paroxanase, ALDH II, CC10, von Ebner salivary gland protein, and α- and β-globin. In addition, CCLS identified several novel genes or genes not previously associated with lung carcinogenesis, including a hypothetical protein (FLJ11240 and a guanine nucleotide exchange factor homologue. This study shows the efficacy of this methodology for identifying genes with altered expression. These genes may prove to be helpful in our understanding of the genetic basis of

  11. Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing.

    Science.gov (United States)

    Nagamatsu, Atsushi; Masuta, Chikara; Senda, Mineo; Matsuura, Hideyuki; Kasai, Atsushi; Hong, Jin-Sung; Kitamura, Keisuke; Abe, Jun; Kanazawa, Akira

    2007-11-01

    Virus-induced gene silencing (VIGS) is a powerful tool for functional analysis of genes in plants. A wide-host-range VIGS vector, which was developed based on the Cucumber mosaic virus (CMV), was tested for its ability to silence endogenous genes involved in flavonoid biosynthesis in soybean. Symptomless infection was established using a pseudorecombinant virus, which enabled detection of specific changes in metabolite content by VIGS. It has been demonstrated that the yellow seed coat phenotype of various cultivated soybean lines that lack anthocyanin pigmentation is induced by natural degradation of chalcone synthase (CHS) mRNA. When soybean plants with brown seed coats were infected with a virus that contains the CHS gene sequence, the colour of the seed coats changed to yellow, which indicates that the naturally occurring RNA silencing is reproduced by VIGS. In addition, CHS VIGS consequently led to a decrease in isoflavone content in seeds. VIGS was also tested on the putative flavonoid 3'-hydroxylase (F3'H) gene in the pathway. This experiment resulted in a decrease in the content of quercetin relative to kaempferol in the upper leaves after viral infection, which suggests that the putative gene actually encodes the F3'H protein. In both experiments, a marked decrease in the target mRNA and accumulation of short interfering RNAs were detected, indicating that sequence-specific mRNA degradation was induced. The present report is a successful demonstration of the application of VIGS for genes involved in flavonoid biosynthesis in plants; the CMV-based VIGS system provides an efficient tool for functional analysis of soybean genes.

  12. Topographic analysis of the skull vibration-induced nystagmus test with piezoelectric accelerometers and force sensors.

    Science.gov (United States)

    Dumas, Georges; Lion, Alexis; Perrin, Philippe; Ouedraogo, Evariste; Schmerber, Sébastien

    2016-03-23

    Vibration-induced nystagmus is elicited by skull or posterior cervical muscle stimulations in patients with vestibular diseases. Skull vibrations delivered by the skull vibration-induced nystagmus test are known to stimulate the inner ear structures directly. This study aimed to measure the vibration transfer at different cranium locations and posterior cervical regions to contribute toward stimulus topographic optimization (experiment 1) and to determine the force applied on the skull with a hand-held vibrator to study the test reproducibility and provide recommendations for good clinical practices (experiment 2). In experiment 1, a 100 Hz hand-held vibrator was applied on the skull (vertex, mastoids) and posterior cervical muscles in 11 healthy participants. Vibration transfer was measured by piezoelectric sensors. In experiment 2, the vibrator was applied 30 times by two experimenters with dominant and nondominant hands on a mannequin equipped to measure the force. Experiment 1 showed that after unilateral mastoid vibratory stimulation, the signal transfer was higher when recorded on the contralateral mastoid than on the vertex or posterior cervical muscles (P<0.001). No difference was observed between the different vibratory locations when vibration transfer was measured on vertex and posterior cervical muscles. Experiment 2 showed that the force applied to the mannequin varied according to the experimenters and the handedness, higher forces being observed with the most experienced experimenter and with the dominant hand (10.3 ± 1.0 and 7.8 ± 2.9 N, respectively). The variation ranged from 9.8 to 29.4% within the same experimenter. Bone transcranial vibration transfer is more efficient from one mastoid to the other mastoid than other anatomical sites. The mastoid is therefore the optimal site for skull vibration-induced nystagmus test in patients with unilateral vestibular lesions and enables a stronger stimulation of the healthy side. In clinical practice

  13. Evaluating the ability of the barley stripe mosaic virus-induced gene silencing system to simultaneously silence two wheat genes

    Science.gov (United States)

    Virus-induced gene silencing (VIGS) is an important tool for rapid assessment of gene function in plants. The ability of the Barley Stripe Mosaic Virus (BSMV) VIGS system to simultaneously silence two genes was assessed by comparing the extent of down-regulation of the wheat PDS and SGT1 genes afte...

  14. INDUCIBLE RNAi-MEDIATED GENE SILENCING USING NANOSTRUCTURED GENE DELIVERY ARRAYS

    Energy Technology Data Exchange (ETDEWEB)

    Mann, David George James [ORNL; McKnight, Timothy E [ORNL; Mcpherson, Jackson [University of Tennessee, Knoxville (UTK); Hoyt, Peter R [ORNL; Melechko, Anatoli Vasilievich [ORNL; Simpson, Michael L [ORNL; Sayler, Gary Steven [ORNL

    2008-01-01

    RNA interference has become a powerful biological tool over the last decade. In this study, a tetracycline-inducible shRNA vector system was designed for silencing CFP expression and delivered alongside the yfp marker gene into Chinese hamster ovary cells using impalefection on spatially indexed vertically aligned carbon nanofiber arrays (VACNFs). The VACNF architecture provided simultaneous delivery of multiple genes, subsequent adherence and proliferation of interfaced cells, and repeated monitoring of single cells over time. Following impalefection and tetracycline induction, 53.1% 10.4% of impalefected cells were fully silenced by the inducible CFP-silencing shRNA vector. Additionally, efficient CFP-silencing was observed in single cells among a population of cells that remained CFP-expressing. This effective transient expression system enables rapid analysis of gene silencing effects using RNAi in single cells and cell populations.

  15. The dilp2/5 genes control diapause inducibility

    OpenAIRE

    Schiesari, Luca

    2015-01-01

    Many holometabolous insects hibernate by triggering diapause, an “actively-induced” dormancy that blocks developmental functions. Yet, the nature of signals enhancing the plasticity of developmental system and underlying diapause inducibility is still elusive. We show that the “Insulin/IGF” dilp2/5 genes, encoding for developmental hormones, antagonize diapause switch in D. melanogaster and their modulation is pivotal in sensitizing the developmental system to environmental perturbations. Fun...

  16. Screening Helicobacter pylori genes induced during infection of mouse stomachs

    Institute of Scientific and Technical Information of China (English)

    Aparna Singh; Nathaniel Hodgson; Ming Yan; Jungsoo Joo; Lei Gu; Hong Sang; Emmalena Gregory-Bryson

    2012-01-01

    AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori (H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology (IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction (PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced (ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of all ivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only

  17. Cold acclimation induced genes of trifoliate orange (Poncirus trifoliata).

    Science.gov (United States)

    Zhang, Can-kui; Lang, Ping; Dane, Fenny; Ebel, Robert C; Singh, Narendra K; Locy, Robert D; Dozier, William A

    2005-03-01

    Commercial citrus varieties are sensitive to low temperature. Poncirus trifoliata is a close relative of Citrus species and has been widely used as a cold-hardy rootstock for citrus production in low-temperature environments. mRNA differential display-reverse transcription (DDRT)-PCR and quantitative relative-RT-PCR were used to study gene expression of P. trifoliata under a gradual cold-acclimation temperature regime. Eight up-regulated cDNA fragments were isolated and sequenced. These fragments showed high similarities at the amino acid level to the following genes with known functions: betaine/proline transporter, water channel protein, aldo-keto reductase, early light-induced protein, nitrate transporter, tetratricopeptide-repeat protein, F-box protein, and ribosomal protein L15. These cold-acclimation up-regulated genes in P. trifoliata are also regulated by osmotic and photo-oxidative signals in other plants.

  18. Duplication and Diversification of the Hypoxia-Inducible IGFBP-1 Gene in Zebrafish

    DEFF Research Database (Denmark)

    Kamei, Hiroyasu; Lu, Ling; Jiao, Shuang

    2008-01-01

    Background: Gene duplication is the primary force of new gene evolution. Deciphering whether a pair of duplicated genes has evolved divergent functions is often challenging. The zebrafish is uniquely positioned to provide insight into the process of functional gene evolution due to its amenabilit...

  19. Mapping interfacial chemistry induced variations in protein adsorption with scanning force microscopy.

    Science.gov (United States)

    Ta, T C; McDermott, M T

    2000-06-01

    In this work, we demonstrate the sensitivity of scanning force microscopy (SFM), operated in friction force mode, to adsorbed protein conformation or orientation. We employ patterned films of methyl- and carboxylate-terminated alkanethiolate monolayers on gold as substrates for protein adsorption to observe the effect of each functional group in the same image. Infrared spectroscopic and SFM studies of bovine fibrinogen (BFG) adsorption to single-component monolayers indicate that complete films of BFG that are stable to imaging are formed at each functional group. After adsorption of BFG to a patterned monolayer, we observe a contrast in friction images due to differences in adsorbed BFG conformation or orientation induced by each functional group. We also observe frictional contrast in films of other proteins adsorbed on patterned monolayers. These observations lead to the conclusion that SFM-measured friction is sensitive to adsorbed protein state.

  20. On combining Thole's induced point dipole model with fixed charge distributions in molecular mechanics force fields.

    Science.gov (United States)

    Antila, Hanne S; Salonen, Emppu

    2015-04-15

    The Thole induced point dipole model is combined with three different point charge fitting methods, Merz-Kollman (MK), charges from electrostatic potentials using a grid (CHELPG), and restrained electrostatic potential (RESP), and two multipole algorithms, distributed multipole analysis (DMA) and Gaussian multipole model (GMM), which can be used to describe the electrostatic potential (ESP) around molecules in molecular mechanics force fields. This is done to study how the different methods perform when intramolecular polarizability contributions are self-consistently removed from the fitting done in the force field parametrization. It is demonstrated that the polarizable versions of the partial charge models provide a good compromise between accuracy and computational efficiency in describing the ESP of small organic molecules undergoing conformational changes. For the point charge models, the inclusion of polarizability reduced the the average root mean square error of ESP over the test set by 4-10%.

  1. a Simple Model to Estimate the Impact Force Induced by Piston Slap

    Science.gov (United States)

    CHO, S.-H.; AHN, S.-T.; KIM, Y.-H.

    2002-08-01

    The dynamics of piston's secondary motion (lateral and rotational motion) across the clearance between piston and cylinder inner wall of reciprocating machines are analyzed. This paper presents an analytical model, which can predict the impact forces and vibratory response of engine block surface induced by the piston slap of an internal combustion engine. A piston is modelled on a three-degree-of-freedom system to represent its planar motion. When slap occurs, the impact point between piston skirt and cylinder inner wall is modelled on a two-degree-of-freedom vibratory system. The equivalent parameters such as mass, spring constant, and damping constant of piston and cylinder inner wall are estimated by using measured (driving) point mobility. Those parameters are used to calculate the impact force and for estimating the vibration level of engine block surfaces. The predicted results are compared with experimental results to verify the model.

  2. Force-induced unfolding of fibronectin in the extracellular matrix of living cells.

    Directory of Open Access Journals (Sweden)

    Michael L Smith

    2007-10-01

    Full Text Available Whether mechanically unfolded fibronectin (Fn is present within native extracellular matrix fibrils is controversial. Fn extensibility under the influence of cell traction forces has been proposed to originate either from the force-induced lengthening of an initially compact, folded quaternary structure as is found in solution (quaternary structure model, where the dimeric arms of Fn cross each other, or from the force-induced unfolding of type III modules (unfolding model. Clarification of this issue is central to our understanding of the structural arrangement of Fn within fibrils, the mechanism of fibrillogenesis, and whether cryptic sites, which are exposed by partial protein unfolding, can be exposed by cell-derived force. In order to differentiate between these two models, two fluorescence resonance energy transfer schemes to label plasma Fn were applied, with sensitivity to either compact-to-extended conformation (arm separation without loss of secondary structure or compact-to-unfolded conformation. Fluorescence resonance energy transfer studies revealed that a significant fraction of fibrillar Fn within a three-dimensional human fibroblast matrix is partially unfolded. Complete relaxation of Fn fibrils led to a refolding of Fn. The compactly folded quaternary structure with crossed Fn arms, however, was never detected within extracellular matrix fibrils. We conclude that the resting state of Fn fibrils does not contain Fn molecules with crossed-over arms, and that the several-fold extensibility of Fn fibrils involves the unfolding of type III modules. This could imply that Fn might play a significant role in mechanotransduction processes.

  3. Virus-induced Gene Silencing in Eggplant (Solanum melongena)

    Institute of Scientific and Technical Information of China (English)

    HaipingLiu; Daqi Fu; Benzhong Zhu; Huaxue Yan; Xiaoying Shen; Jinhua Zuo; Yi Zhu; Yunbo Luo

    2012-01-01

    Eggplant (Solanum melongena) is an economically important vegetable requiring investigation into its various genomic functions.The current limitation in the investigation of genomic function in eggplant is the lack of effective tools available for conducting functional assays.Virus-induced gene silencing (VIGS) has played a critical role in the functional genetic analyses.In this paper,TRV-mediated VIGS was successfully elicited in eggplant.We first cloned the CDS sequence of PDS (PHYTOENE DESATURASE) in eggplant and then silenced the PDS gene.Photo-bleaching was shown on the newly-developed leaves four weeks after agroinoculation,indicating that VIGS can be used to silence genes in eggplant.To further illustrate the reliability of VIGS in eggplant,we selected Chl H,Su and CLA1 as reporters to elicit VIGS using the high-pressure spray method.Suppression of Chl H and Su led to yellow leaves,while the depletion of CLA1 resulted in albino.In conclusion,four genes,PDS,Chl H,Su (Sulfur),CLA1,were down-regulated significantly by VIGS,indicating that the VIGS system can be successfully applied in eggplant and is a reliable tool for the study of gene function.

  4. Octylphenol induced gene expression in testes of Frog, Rana chensinensis.

    Science.gov (United States)

    Li, Xinyi; Liu, Jia; Zhang, Yuhui

    2016-06-01

    Octylphenol (OP) is an endocrine-disrupting chemical (EDC), which can disrupt the reproductive system. To understand the effect of OP, a subtractive cDNA library was constructed using suppression subtractive hybridization (SSH) to identify alterations of gene transcription in the testes of the frog Rana chensinensis after OP exposure. Two hundred positive clones were selected and 134 sequences of gene fragments were produced from the subtractive library randomly. These genes were identified to be involved in metabolic process, cellular process, biological regulation, stimulus, immune system and female pregnancy process. In order to verify the efficiency of the subtractive cDNA library, PSG9 and PAPP-A were analyzed further as two representatives of differentially expressed transcription genes using semi-quantitative RT-PCR. Our result was the first successful construction of the subtractive cDNA library in frog testes after OP treatment. Based on this cDNA library, OP was shown to affect multiple physiological processes including inducing immune response, disrupting the steroid hormone synthesis and influencing spermatogenesis in the testis by up-regulation of specific genes.

  5. Efficient Virus-Induced Gene Silencing in Solanum rostratum.

    Directory of Open Access Journals (Sweden)

    Lan-Huan Meng

    Full Text Available Solanum rostratum is a "super weed" that grows fast, is widespread, and produces the toxin solanine, which is harmful to both humans and other animals. To our knowledge, no study has focused on its molecular biology owing to the lack of available transgenic methods and sequence information for S. rostratum. Virus-induced gene silencing (VIGS is a powerful tool for the study of gene function in plants; therefore, in the present study, we aimed to establish tobacco rattle virus (TRV-derived VIGS in S. rostratum. The genes for phytoene desaturase (PDS and Chlorophyll H subunit (ChlH of magnesium protoporphyrin chelatase were cloned from S. rostratum and used as reporters of gene silencing. It was shown that high-efficiency VIGS can be achieved in the leaves, flowers, and fruit of S. rostratum. Moreover, based on our comparison of three different types of infection methods, true leaf infection was found to be more efficient than cotyledon and sprout infiltration in long-term VIGS in multiple plant organs. In conclusion, the VIGS technology and tomato genomic sequences can be used in the future to study gene function in S. rostratum.

  6. Efficient Virus-Induced Gene Silencing in Solanum rostratum

    Science.gov (United States)

    Meng, Lan-Huan; Wang, Rui-Heng; Zhu, Ben-Zhong; Zhu, Hong-Liang; Luo, Yun-Bo; Fu, Da-Qi

    2016-01-01

    Solanum rostratum is a “super weed” that grows fast, is widespread, and produces the toxin solanine, which is harmful to both humans and other animals. To our knowledge, no study has focused on its molecular biology owing to the lack of available transgenic methods and sequence information for S. rostratum. Virus-induced gene silencing (VIGS) is a powerful tool for the study of gene function in plants; therefore, in the present study, we aimed to establish tobacco rattle virus (TRV)-derived VIGS in S. rostratum. The genes for phytoene desaturase (PDS) and Chlorophyll H subunit (ChlH) of magnesium protoporphyrin chelatase were cloned from S. rostratum and used as reporters of gene silencing. It was shown that high-efficiency VIGS can be achieved in the leaves, flowers, and fruit of S. rostratum. Moreover, based on our comparison of three different types of infection methods, true leaf infection was found to be more efficient than cotyledon and sprout infiltration in long-term VIGS in multiple plant organs. In conclusion, the VIGS technology and tomato genomic sequences can be used in the future to study gene function in S. rostratum. PMID:27258320

  7. Efficient Virus-Induced Gene Silencing in Solanum rostratum.

    Science.gov (United States)

    Meng, Lan-Huan; Wang, Rui-Heng; Zhu, Ben-Zhong; Zhu, Hong-Liang; Luo, Yun-Bo; Fu, Da-Qi

    2016-01-01

    Solanum rostratum is a "super weed" that grows fast, is widespread, and produces the toxin solanine, which is harmful to both humans and other animals. To our knowledge, no study has focused on its molecular biology owing to the lack of available transgenic methods and sequence information for S. rostratum. Virus-induced gene silencing (VIGS) is a powerful tool for the study of gene function in plants; therefore, in the present study, we aimed to establish tobacco rattle virus (TRV)-derived VIGS in S. rostratum. The genes for phytoene desaturase (PDS) and Chlorophyll H subunit (ChlH) of magnesium protoporphyrin chelatase were cloned from S. rostratum and used as reporters of gene silencing. It was shown that high-efficiency VIGS can be achieved in the leaves, flowers, and fruit of S. rostratum. Moreover, based on our comparison of three different types of infection methods, true leaf infection was found to be more efficient than cotyledon and sprout infiltration in long-term VIGS in multiple plant organs. In conclusion, the VIGS technology and tomato genomic sequences can be used in the future to study gene function in S. rostratum.

  8. Bitumen fume-induced gene expression profile in rat lung.

    Science.gov (United States)

    Gate, Laurent; Langlais, Cristina; Micillino, Jean-Claude; Nunge, Hervé; Bottin, Marie-Claire; Wrobel, Richard; Binet, Stéphane

    2006-08-15

    Exposure to bitumen fumes during paving and roofing activities may represent an occupational health risk. To date, most of the studies performed on the biological effect of asphalt fumes have been done with regard to their content in carcinogenic polycyclic aromatic hydrocarbons (PAH). In order to gain an additional insight into the mechanisms of action of bitumen fumes, we studied their pulmonary effects in rodents following inhalation using the microarray technology. Fisher 344 rats were exposed for 5 days, 6 h/day to bitumen fumes generated at road paving temperature (170 degrees C) using a nose-only exposition device. With the intention of studying the early transcriptional events induced by asphalt fumes, lung tissues were collected immediately following exposure and gene expression profiles in control and exposed rats were determined by using oligonucleotide microarrays. Data analysis revealed that genes involved in lung inflammatory response as well as genes associated with PAH metabolization and detoxification were highly expressed in bitumen-exposed animals. In addition, the expression of genes related to elastase activity and its inhibition which are associated with emphysema was also modulated. More interestingly genes coding for monoamine oxidases A and B involved in the metabolism of neurotransmitters and xenobiotics were downregulated in exposed rats. Altogether, these data give additional information concerning the bitumen fumes biological effects and would allow to better review the health effects of occupational asphalt fumes exposure.

  9. Single molecule studies of force-induced S2 site exposure in the mammalian Notch negative regulatory domain.

    Science.gov (United States)

    Ploscariu, Nicoleta; Kuczera, Krzysztof; Malek, Katarzyna E; Wawrzyniuk, Magdalena; Dey, Ashim; Szoszkiewicz, Robert

    2014-05-08

    Notch signaling in metazoans is responsible for key cellular processes related to embryonic development and tissue homeostasis. Proteolitic cleavage of the S2 site within an extracellular NRR domain of Notch is a key early event in Notch signaling. We use single molecule force-extension (FX) atomic force microscopy (AFM) to study force-induced exposure of the S2 site in the NRR domain from mouse Notch 1. Our FX AFM measurements yield a histogram of N-to-C termini lengths, which we relate to conformational transitions within the NRR domain. We detect four classes of such conformational transitions. From our steered molecular dynamics (SMD) results, we associate first three classes of such events with the S2 site exposure. AFM experiments yield their mean unfolding forces as 69 ± 42, 79 ± 45, and 90 ± 50 pN, respectively, at 400 nm/s AFM pulling speeds. These forces are matched by the SMD results recalibrated to the AFM force loading rates. Next, we provide a conditional probability analysis of the AFM data to support the hypothesis that a whole sequence of conformational transitions within those three clases is the most probable pathway for the force-induced S2 site exposure. Our results support the hypothesis that force-induced Notch activation requires ligand binding to exert mechanical force not in random but in several strokes and over a substantial period of time.

  10. The relationship of host-mediated induced resistance to polymorphism in gene-for-gene relationships.

    Science.gov (United States)

    Tellier, Aurélien; Brown, James K M

    2008-01-01

    Gene-for-gene relationships are a common feature of plant-parasite interactions. Polymorphism at host resistance and parasite avirulence loci is maintained if there is negative, direct frequency-dependent selection on alleles of either gene. More specifically, selection of this kind is generated when the disease is polycyclic with frequent auto-infection. When an incompatible interaction occurs between a resistant host and an avirulent parasite, systemic defenses are triggered, rendering the plant more resistant to a later attack by another parasite. However, induced resistance (IR) incurs a fitness cost to the plant. Here, the effect of IR on polymorphism in gene-for-gene interactions is investigated. First, in an infinite population model in which parasites have two generations per host generation, increasing the fitness cost of IR increases selection for susceptible plants at low disease severity, while increasing the effectiveness of IR against further parasite attacks enhances selection for resistant plants at high disease severity. This reduces the possibility of polymorphism being maintained in host and parasite populations. In finite population models, the number of plants varies over time as a function of the disease burden of the population. Polymorphism in gene-for-gene relationships is then more stable at high disease prevalence and severity if IR reactions are more costly when there is competition for resources between plants.

  11. Metabolic costs of force generation for constant-frequency and catchlike-inducing electrical stimulation in human tibialis anterior muscle

    DEFF Research Database (Denmark)

    Ratkevicius, Aivaras; Quistorff, Bjørn

    2002-01-01

    -frequency trains, catchlike-inducing trains produced a faster force generation and were more effective in maintaining the force--time integral as well as peak force. However, ATP costs of force generation were similar for the catchlike-inducing and constant-frequency stimulation (6.7 plus/minus 1.1 and 6.6 plus......Metabolic costs of force generation were compared for constant-frequency and catchlike-inducing electrical stimulation. Repetitive catchlike-inducing trains consisted of 2 interpulse intervals (IPIs) at 12.5 ms, 1 IPI at 25 ms, and 5 IPIs at 50 ms. Constant-frequency trains consisted of 8 IPIs....../minus 1.0 micromol ATP/kg wet weight/Ncenter dots, respectively, P = 0.601). This suggests that the positive effects of catchlike-inducing stimulation on force maintenance are mediated by potentiated Ca(2+) release from the sarcoplasmic reticulum rather than by lower metabolic costs of muscle force...

  12. Inducible gene manipulations in brain serotonergic neurons of transgenic rats.

    Directory of Open Access Journals (Sweden)

    Tillmann Weber

    Full Text Available The serotonergic (5-HT system has been implicated in various physiological processes and neuropsychiatric disorders, but in many aspects its role in normal and pathologic brain function is still unclear. One reason for this might be the lack of appropriate animal models which can address the complexity of physiological and pathophysiological 5-HT functioning. In this respect, rats offer many advantages over mice as they have been the animal of choice for sophisticated neurophysiological and behavioral studies. However, only recently technologies for the targeted and tissue specific modification of rat genes - a prerequisite for a detailed study of the 5-HT system - have been successfully developed. Here, we describe a rat transgenic system for inducible gene manipulations in 5-HT neurons. We generated a Cre driver line consisting of a tamoxifen-inducible CreERT2 recombinase under the control of mouse Tph2 regulatory sequences. Tissue-specific serotonergic Cre recombinase expression was detected in four transgenic TPH2-CreERT2 rat founder lines. For functional analysis of Cre-mediated recombination, we used a rat Cre reporter line (CAG-loxP.EGFP, in which EGFP is expressed after Cre-mediated removal of a loxP-flanked lacZ STOP cassette. We show an in-depth characterisation of this rat Cre reporter line and demonstrate its applicability for monitoring Cre-mediated recombination in all major neuronal subpopulations of the rat brain. Upon tamoxifen induction, double transgenic TPH2-CreERT2/CAG-loxP.EGFP rats show selective and efficient EGFP expression in 5-HT neurons. Without tamoxifen administration, EGFP is only expressed in few 5-HT neurons which confirms minimal background recombination. This 5-HT neuron specific CreERT2 line allows Cre-mediated, inducible gene deletion or gene overexpression in transgenic rats which provides new opportunities to decipher the complex functions of the mammalian serotonergic system.

  13. Putting the diet back into diet-induced obesity: diet-induced hypothalamic gene expression.

    Science.gov (United States)

    Mercer, Julian G; Archer, Zoë A

    2008-05-06

    A wealth of detailed mechanistic information relating to obesity and body weight regulation has emerged from study of single gene mutation models, and continues to be generated by engineered rodent models targeting specific genes. However, as an early step in translational research, many researchers are turning to models of diet-induced obesity. Interpretation of data generated from such models is not aided by the variety of diets and rodent strains employed in these studies and a strong case could be made for rationalisation. Differences in experimental protocol, which may deploy a single obligatory solid diet, a choice of solid diets, or liquid/solid combinations, and which may or may not allow a preferred macronutrient composition to be selected, mean that different models of diet-induced obesity achieve that obesity by different routes. The priority should be to mimic the palatability- and choice-driven over-consumption that probably underlies the majority of human obesity. Some of the hypothalamic energy balance genes apparently 'recognise' developing diet-induced obesity as indicated by counter-regulatory changes in expression levels. However, substantial changes in gene expression on long-term exposure to obesogenic diets are not able to prevent weight gain. Forebrain reward systems are widely assumed to be overriding hypothalamic homeostatic energy balance systems under these circumstances. More mechanism-based research at the homeostatic/reward/diet interface may enable diets to be manipulated with therapeutic benefit, or define the contribution of these interactions to susceptibility to diet-induced obesity.

  14. The Torsional Vibration of Turbo Axis Induced by Unsteady Aerodynamic Force on Rotor blade

    Institute of Scientific and Technical Information of China (English)

    ChenZuoyi; WuXiaofeng

    1998-01-01

    An algorithm for computing the 3-D oscillating flow field of the balde passage under the torsional vibration of the rotor is applied to analyze the stability in turbomachines.The induced flow field responding to blade vibration is computed by Oscillating Fluid Mechanics Method and parametric Polynomial Method.After getting the solution of the unsteady flow field,the work done by the unsteay aerodynamic force acting on the blade can be obtained.The negative or positive work is the criterion of the aeroelastic stability.Numerical results indicate that there are instabilities of the torsional vibration in some frequency bands.

  15. Enhancing thermally induced effects on atomic force microscope cantilevers using optical microcavities

    Science.gov (United States)

    Duy Vy, Nguyen; Iida, Takuya

    2016-12-01

    A theory of enhancing thermally induced effects on atomic force microscope cantilevers with respect to the input power is proposed. An optical microcavity is used to increase the absorbed power and radiation pressure on thin films. We show that the response to the input power is enhanced up to an order of magnitude for cantilevers of ∼200 µm in length and ∼0.5 µm in thickness. A decrease in the absorbed power in the presence of cantilever deflection increases system endurability with respect to the input power. The study gives methods for amplifying and tuning vibration amplitudes in amplitude modulation modes with high sensitivity and low controlling input power.

  16. [Study of long-lasting effects of acute prenatal stress induced forced swimming].

    Science.gov (United States)

    Volodina, M A; Sebentsova, E A; Levitskaia, N G; Kamenskiĭ, A A

    2010-01-01

    The aim of the present work was to assess long-lasting effects of acute prenatal stress in white rats. Forced swimming in cold water on the 7th or the 14th gestational day was used as a prenatal stressor. The prenatal stress led to low birthweight of offspring and their delayed growth rate during the second month of life. Prenatally stressed animals showed abnormalities in exploratory behavior and anxiety, increased emotionality and impaired learning capabilities at the age of 1-2 month. Consequently, acute stress on the 7th and at the 14th day of pregnancy induced long-lasting negative behavioral changes in offspring of stressed white rats.

  17. Cold-atom physics using ultrathin optical fibers: light-induced dipole forces and surface interactions.

    Science.gov (United States)

    Sagué, G; Vetsch, E; Alt, W; Meschede, D; Rauschenbeutel, A

    2007-10-19

    The strong evanescent field around ultrathin unclad optical fibers bears a high potential for detecting, trapping, and manipulating cold atoms. Introducing such a fiber into a cold-atom cloud, we investigate the interaction of a small number of cold cesium atoms with the guided fiber mode and with the fiber surface. Using high resolution spectroscopy, we observe and analyze light-induced dipole forces, van der Waals interaction, and a significant enhancement of the spontaneous emission rate of the atoms. The latter can be assigned to the modification of the vacuum modes by the fiber.

  18. Rotator cuff tear reduces muscle fiber specific force production and induces macrophage accumulation and autophagy.

    Science.gov (United States)

    Gumucio, Jonathan P; Davis, Max E; Bradley, Joshua R; Stafford, Patrick L; Schiffman, Corey J; Lynch, Evan B; Claflin, Dennis R; Bedi, Asheesh; Mendias, Christopher L

    2012-12-01

    Full-thickness tears to the rotator cuff can cause severe pain and disability. Untreated tears progress in size and are associated with muscle atrophy and an infiltration of fat to the area, a condition known as "fatty degeneration." To improve the treatment of rotator cuff tears, a greater understanding of the changes in the contractile properties of muscle fibers and the molecular regulation of fatty degeneration is essential. Using a rat model of rotator cuff injury, we measured the force generating capacity of individual muscle fibers and determined changes in muscle fiber type distribution that develop after a full thickness rotator cuff tear. We also measured the expression of mRNA and miRNA transcripts involved in muscle atrophy, lipid accumulation, and matrix synthesis. We hypothesized that a decrease in specific force of rotator cuff muscle fibers, an accumulation of type IIb fibers, and an upregulation in fibrogenic, adipogenic, and inflammatory gene expression occur in torn rotator cuff muscles. Thirty days following rotator cuff tear, we observed a reduction in muscle fiber force production, an induction of fibrogenic, adipogenic, and autophagocytic mRNA and miRNA molecules, and a dramatic accumulation of macrophages in areas of fat accumulation. Copyright © 2012 Orthopaedic Research Society.

  19. Forced FOG1 expression in erythroleukemia cells: Induction of erythroid genes and repression of myelo-lymphoid transcription factor PU.1.

    Science.gov (United States)

    Fujiwara, Tohru; Sasaki, Katsuyuki; Saito, Kei; Hatta, Shunsuke; Ichikawa, Satoshi; Kobayashi, Masahiro; Okitsu, Yoko; Fukuhara, Noriko; Onishi, Yasushi; Harigae, Hideo

    2017-02-16

    The transcription factor GATA-1-interacting protein Friend of GATA-1 (FOG1) is essential for proper transcriptional activation and repression of GATA-1 target genes; yet, the mechanisms by which FOG1 exerts its activating and repressing functions remain unknown. Forced FOG1 expression in human K562 erythroleukemia cells induced the expression of erythroid genes (SLC4A1, globins) but repressed that of GATA-2 and PU.1. A quantitative chromatin immunoprecipitation (ChIP) analysis demonstrated increased GATA-1 chromatin occupancy at both FOG1-activated as well as FOG1-repressed gene loci. However, while TAL1 chromatin occupancy was significantly increased at FOG1-activated gene loci, it was significantly decreased at FOG1-repressed gene loci. When FOG1 was overexpressed in TAL1-knocked down K562 cells, FOG1-mediated activation of HBA, HBG, and SLC4A1 was significantly compromised by TAL1 knockdown, suggesting that FOG1 may require TAL1 to activate GATA-1 target genes. Promoter analysis and quantitative ChIP analysis demonstrated that FOG1-mediated transcriptional repression of PU.1 would be mediated through a GATA-binding element located at its promoter, accompanied by significantly decreased H3 acetylation at lysine 4 and 9 (K4 and K9) as well as H3K4 trimethylation. Our results provide important mechanistic insight into the role of FOG1 in the regulation of GATA-1-regulated genes and suggest that FOG1 has an important role in inducing cells to differentiate toward the erythroid lineage rather than the myelo-lymphoid one by repressing the expression of PU.1.

  20. Experimental study of radiation induced electromotive force effects on mineral insulated cables

    Science.gov (United States)

    Van Nieuwenhove, R.; Vermeeren, L.

    2003-11-01

    Measurements of radiation induced electromotive force (RIEMF) effects on mineral insulated cables in a pure gamma field and in a combined neutron and gamma field are presented and compared to model calculations. The effect of materials in the immediate surroundings of the cable, as predicted by the model calculations, is clearly demonstrated. In a fission reactor environment, delayed current contributions due to the neutron activation and subsequent beta emission in base materials as well as in impurities such as Mn are clearly observed and are well reproduced by model calculations. The prediction of the gamma induced current component was severely complicated by its strong sensitivity to the detailed geometry and the spectrum and the directivity of the gamma field. Although the RIEMF effect on MI cables can therefore in general not be completely eliminated, some guidelines are provided to minimize them.

  1. Reducing friction-induced vibration using intelligent active force control (AFC) with piezoelectric actuators

    Indian Academy of Sciences (India)

    S M Hashemi-Dehkordi; A R Abu-Bakar; M Mailah

    2012-12-01

    In this paper, a novel approach to reduce the effect of mode coupling that causes friction induced vibration (FIV) is proposed by applying an intelligent active force control (AFC)-based strategy employing piezoelectric actuators with hysteresis effect to a simplified two degree-of-freedom mathematical model of a friction-induced vibration system. At first, the model is simulated and analysed using a closed loop pure Proportional-Integral-Derivative (PID) controller. Later, it is integrated with the intelligent AFC with fuzzy logic (FL) estimator and simulated under similar operating condition. After running several tests with different sets of operating and loading conditions, the results both in time and frequency domains show that the PID controller with the intelligent AFC is much more effective in reducing the vibration, compared to the pure PID controller alone.

  2. Tip-bias-induced domain evolution in PMN-PT transparent ceramics via piezoresponse force microscopy

    Science.gov (United States)

    Zhao, K. Y.; Zhao, W.; Zeng, H. R.; Yu, H. Z.; Ruan, W.; Xu, K. Q.; Li, G. R.

    2015-05-01

    Piezoresponse force microscopy (PFM) was employed to investigate ferroelectric domain structures and their dynamic behavior of lead magnesium niobate-lead titanate [Pb(Mg1/3Nb2/3)O3-xPbTiO3 (PMN-PT)] transparent ceramics under an tip-bias-induced electric field. A remarkable effect of fluctuation of PT content on the domain configurations and domain dynamic response in PMN-PT transparent ferroelectric ceramics were found by PFM. Comparing with PMN-10%PT and PMN-20%PT, the reversed polarization of macrodomain area in PMN-35%PT and PMN-25%PT exhibits a relatively higher response behavior and better polarization retention performance under the PFM tip-bias-induced electric field, which correspond to their unique macroscopic electro-optic properties.

  3. The effect of temperature dependent tissue parameters on acoustic radiation force induced displacements

    CERN Document Server

    Suomi, Visa; Konofagou, Elisa; Cleveland, Robin

    2016-01-01

    Multiple ultrasound elastography techniques rely on acoustic radiation force (ARF) in monitoring high-intensity focused ultrasound (HIFU) therapy. However, ARF is dependent on tissue attenuation and sound speed, both of which are also known to change with temperature making the therapy monitoring more challenging. Furthermore, the viscoelastic properties of tissue are also temperature dependent, which affects the displacements induced by ARF. The aim of this study is to quantify the temperature dependent changes in the acoustic and viscoelastic properties of liver and investigate their effect on ARF induced displacements by using both experimental methods and simulations. Furthermore, the temperature dependent viscoelastic properties of liver are experimentally measured over a frequency range of 0.1-200 Hz at temperatures reaching 80 C, and both conventional and fractional Zener models are used to fit the data. The fractional Zener model was found to fit better with the experimental viscoelasticity data with ...

  4. Multivariate nonlinear regression analysis of trajectory tracking performance using force reflecting joystick in chronic stroke-induced hemiparesis.

    Science.gov (United States)

    Feng, Xin; Winters, Jack M

    2011-01-01

    Individualizing a neurorehabilitation training protocol requires understanding the performance of subjects with various capabilities under different task settings. We use multivariate regression to evaluate the performance of subjects with stroke-induced hemiparesis in trajectory tracking tasks using a force-reflecting joystick. A nonlinear effect was consistently shown in both dimensions of force field strength and impairment level for selected kinematic performance measures, with greatest sensitivity at lower force fields. This suggests that the form of a force field may play a different "role" for subjects with various impairment levels, and confirms that to achieve optimized therapeutic benefit, it is necessary to personalize interfaces.

  5. Tension and drag forces of flexible risers undergoing vortex-induced vibration

    Science.gov (United States)

    Song, Lei-jian; Fu, Shi-xiao; Li, Man; Gao, Yun; Ma, Lei-xin

    2017-03-01

    This paper presents the results of an experimental investigation on the variation in the tension and the distribution of drag force coefficients along flexible risers under vortex-induced vibration (VIV) in a uniform flow for Reynolds numbers ( Re) up to 2.2×105. The results show that the mean tension is proportional to the square of the incoming current speed, and the tension coefficient of a flexible riser undergoing VIV can be up to 12. The mean drag force is uniformly and symmetrically distributed along the axes of the risers undergoing VIV. The corresponding drag coefficient can vary between 1.6 and 2.4 but is not a constant value of 1.2, as it is for a fixed cylinder in the absence of VIV. These experimental results are used to develop a new empirical prediction model to estimate the drag force coefficient for flexible risers undergoing VIV for Reynolds number on the order of 105, which accounts for the effects of the incoming current speed, the VIV dominant modal number and the frequency.

  6. Propagation of the state change induced by external forces in local interactions

    Science.gov (United States)

    Lu, Jianjun; Tokinaga, Shozo

    2016-10-01

    This paper analyses the propagation of the state changes of agents that are induced by external forces applied to a plane. In addition, we propose two models for the behavior of the agents placed on a lattice plane, both of which are affected by local interactions. We first assume that agents are allowed to move to another site to maximise their satisfaction. Second, we utilise a model in which the agents choose activities on each site. The results show that the migration (activity) patterns of agents in both models achieve stability without any external forces. However, when we apply an impulsive external force to the state of the agents, we then observe the propagation of the changes in the agents' states. Using simulation studies, we show the conditions for the propagation of the state changes of the agents. We also show the propagation of the state changes of the agents allocated in scale-free networks and discuss the estimation of the agents' decisions in real state changes. Finally, we discuss the estimation of the agents' decisions in real state temporal changes using economic and social data from Japan and the United States.

  7. The effect of temperature on eccentric contraction-induced isometric force loss in isolated perfused rat medial gastrocnemius muscle

    Directory of Open Access Journals (Sweden)

    Vasaghi Gharamaleki B

    2008-09-01

    Full Text Available "nBackground: The typical features of eccentric exercise-induced muscle damage are delayed-onset muscle soreness (DOMS and prolonged loss of muscle strength. It has been shown that passive warmth is effective in reducing muscle injury. Due to the interaction of different systems in vivo, we used isolated perfused medial gastrocnemius skeletal muscle to study the direct effect of temperature on the eccentric contraction-induced force loss. "nMethods: After femoral artery cannulation of a rat, the left medial gastrocnemius muscle was separated and then the entire lower limb was transferred into a prewarmed (35oC chamber. With the chamber temperature at 31, 35 and 39oC before and during eccentric contraction. Isometric force loss was measured after 15 eccentric contractions (N=7-9. "nResults: Maximum contraction force reduction has been used as an index for eccentric contraction-induced force loss. In this study eccentric contraction caused a significant reduction in maximum isometric tension (p<0.01, but no significant difference was seen in isometric force loss at 31oC and 39oC compared with that at 35oC. "nConclusions: Our results suggest that temperature changes before or during eccentric contractions have no effect on eccentric contraction-induced force loss. "nKeywords: Isolated perfused muscle, skeletal muscle, eccentric contractions, isometric force, gastrocnemius muscle, temperature.

  8. Tomato leaf spatial expression of stress-induced Asr genes.

    Science.gov (United States)

    Maskin, Laura; Maldonado, Sara; Iusem, Norberto D

    2008-12-01

    Asr1 and Asr2 are water stress-inducible genes belonging to the Asr gene family, which transcriptionally regulate a sugar transporter gene, at least in grape. Using an in situ RNA hybridization methodology, we determined that, in basal conditions, expression of Asr2 in tomato leaves is detected in the phloem tissue, particularly in companion phloem cells. When plants are exposed to water stress, Asr2 expression is contained in companion cells but expands occasionally to mesophyll cells. In contrast, Asr1 transcript localization seems to be sparse in leaf vascular tissue under both non-stress and stress conditions. The occurrence of Asr transcripts precisely in companion cells is in accordance with the cell type specificity reported for hexose-transporter protein molecules in grape encoded by the only Asr-target gene known to date. The results are discussed in light of the reported scarcity of plasmodesmata between companion cells and the rest of leaf tissue in the family Solanaceae.

  9. Sensing nanometric displacement of a micro-/nano-fiber induced by optical forces by use of white light interferometry

    Science.gov (United States)

    Qiu, Weiqia; Huang, Hankai; Yu, Jianhui; Dong, Huazhuo; Chen, Zhe; Lu, Huihui

    2015-07-01

    Sensing the nanometric displacement of a micro-/nano-fiber induced by optical forces is a key technology to study optical forces and optical momentum. When the gap between a micro-/nano-fiber and glass substrate becomes down to micrometer scale or less, a white light interference was observed. The gap changes when optical force arising from the propagating pump light along the micro-/nano-fiber causes a transversal nanometric displacement of a micro-/nanofiber, resulting in movement of the interferometric fringes. Therefore this movement of the interferometric fringes can be used to sense the nanometric displacement of the micro-/nano-fiber induced by optical forces. Experimental results show that the resolutions of this method can reach 7.27nm/pixel for tilted angle 0.8o between the micro-/nano-fiber and substrate. It is concluded that the white light interferometry method is suitable for measuring the weak optical force.

  10. Stimulated Raman spectroscopy and nanoscopy of molecules using near field photon induced forces without resonant electronic enhancement gain

    Science.gov (United States)

    Tamma, Venkata Ananth; Huang, Fei; Nowak, Derek; Kumar Wickramasinghe, H.

    2016-06-01

    We report on stimulated Raman spectroscopy and nanoscopy of molecules, excited without resonant electronic enhancement gain, and recorded using near field photon induced forces. Photon-induced interaction forces between the sharp metal coated silicon tip of an Atomic Force Microscope (AFM) and a sample resulting from stimulated Raman excitation were detected. We controlled the tip to sample spacing using the higher order flexural eigenmodes of the AFM cantilever, enabling the tip to come very close to the sample. As a result, the detection sensitivity was increased compared with previous work on Raman force microscopy. Raman vibrational spectra of azobenzene thiol and l-phenylalanine were measured and found to agree well with published results. Near-field force detection eliminates the need for far-field optical spectrometer detection. Recorded images show spatial resolution far below the optical diffraction limit. Further optimization and use of ultrafast pulsed lasers could push the detection sensitivity towards the single molecule limit.

  11. Charge-Induced Force Noise on Free-Falling Test Masses: Results from LISA Pathfinder

    Science.gov (United States)

    Armano, M.; Audley, H.; Auger, G.; Baird, J. T.; Binetruy, P.; Born, M.; Bortoluzzi, D.; Brandt, N.; Bursi, A.; Caleno, M.; Cavalleri, A.; Cesarini, A.; Cruise, M.; Danzmann, K.; de Deus Silva, M.; Diepholz, I.; Dolesi, R.; Dunbar, N.; Ferraioli, L.; Ferroni, V.; Fitzsimons, E. D.; Flatscher, R.; Freschi, M.; Gallegos, J.; García Marirrodriga, C.; Gerndt, R.; Gesa, L.; Gibert, F.; Giardini, D.; Giusteri, R.; Grimani, C.; Grzymisch, J.; Harrison, I.; Heinzel, G.; Hewitson, M.; Hollington, D.; Hueller, M.; Huesler, J.; Inchauspé, H.; Jennrich, O.; Jetzer, P.; Johlander, B.; Karnesis, N.; Kaune, B.; Killow, C. J.; Korsakova, N.; Lloro, I.; Liu, L.; López-Zaragoza, J. P.; Maarschalkerweerd, R.; Madden, S.; Mance, D.; Martín, V.; Martin-Polo, L.; Martino, J.; Martin-Porqueras, F.; Mateos, I.; McNamara, P. W.; Mendes, J.; Mendes, L.; Moroni, A.; Nofrarias, M.; Paczkowski, S.; Perreur-Lloyd, M.; Petiteau, A.; Pivato, P.; Plagnol, E.; Prat, P.; Ragnit, U.; Ramos-Castro, J.; Reiche, J.; Romera Perez, J. A.; Robertson, D. I.; Rozemeijer, H.; Rivas, F.; Russano, G.; Sarra, P.; Schleicher, A.; Slutsky, J.; Sopuerta, C.; Sumner, T. J.; Texier, D.; Thorpe, J. I.; Trenkel, C.; Vetrugno, D.; Vitale, S.; Wanner, G.; Ward, H.; Wass, P. J.; Wealthy, D.; Weber, W. J.; Wittchen, A.; Zanoni, C.; Ziegler, T.; Zweifel, P.; LISA Pathfinder Collaboration

    2017-04-01

    We report on electrostatic measurements made on board the European Space Agency mission LISA Pathfinder. Detailed measurements of the charge-induced electrostatic forces exerted on free-falling test masses (TMs) inside the capacitive gravitational reference sensor are the first made in a relevant environment for a space-based gravitational wave detector. Employing a combination of charge control and electric-field compensation, we show that the level of charge-induced acceleration noise on a single TM can be maintained at a level close to 1.0 fm s-2 Hz-1 /2 across the 0.1-100 mHz frequency band that is crucial to an observatory such as the Laser Interferometer Space Antenna (LISA). Using dedicated measurements that detect these effects in the differential acceleration between the two test masses, we resolve the stochastic nature of the TM charge buildup due to interplanetary cosmic rays and the TM charge-to-force coupling through stray electric fields in the sensor. All our measurements are in good agreement with predictions based on a relatively simple electrostatic model of the LISA Pathfinder instrument.

  12. Factors of force potentiation induced by stretch-shortening cycle in plantarflexors.

    Directory of Open Access Journals (Sweden)

    Atsuki Fukutani

    Full Text Available Muscle force is potentiated by countermovement; this phenomenon is called stretch-shortening cycle (SSC effect. In this study, we examined the factors strongly related to SSC effect in vivo, focusing on tendon elongation, preactivation, and residual force enhancement. Twelve healthy men participated in this study. Ankle joint angle was passively moved by a dynamometer, with a range of motion from 15° dorsiflexion (DF to 15° plantarflexion (PF. Muscle contraction was evoked by electrical stimulation, with stimulation timing adjusted to elicit three types of contraction: (1 concentric contraction without preliminary contraction (CON, (2 concentric contraction after preliminary eccentric contraction (ECC, and (3 concentric contraction after preliminary isometric contraction (ISO. Joint torque was recorded at DF5°, PF0°, and PF5°, respectively. SSC effect was calculated as the ratio of joint torque obtained in ECC or ISO with respect to that obtained in CON at the aforementioned three joint angles. SSC effect was prominent in the first half of movement in both ECC (DF5°, 329.3 ± 101.2%; PF0°, 159.2 ± 29.4%; PF5°, 125.5 ± 20.8% and ISO (DF5°, 276.4 ± 87.0%; PF0°, 134.5 ± 24.5%; PF5°, 106.8 ± 18.0% conditions. SSC effect was significantly larger in ECC than in ISO at all joint angles (P < 0.001. Even without preliminary eccentric contraction (i.e., ISO condition, SSC effect was clearly large, indicating that a significant part of SSC effect is derived from preactivation. However, the active lengthening-induced force potentiation mechanism (residual force enhancement also contributes to SSC effect.

  13. A Critical Review of Constraint-Induced Movement Therapy and Forced Use in Children With Hemiplegia

    Science.gov (United States)

    Charles, Jeanne; Gordon, Andrew M.

    2005-01-01

    Hemiplegia is a physical impairment that can occur in childhood following head trauma, cerebral vascular accident or transient ischemic attack (stroke), brain tumor, or congenital or perinatal injury. One of the most disabling symptoms of hemiplegia is unilaterally impaired hand and arm function. Sensory and motor impairments in children with hemiplegia compromise movement efficiency. Such children often tend not to use the affected extremity, which may further exacerbate the impairments, resulting in a developmentally learned non-use of the involved upper extremity, termed ‘developmental disuse’. Recent studies suggest that children with hemiplegia benefit from intensive practice. Forced use and Constraint-lnduced Movement Therapy (CI therapy) are recent therapeutic interventions involving the restraint of the non-involved upper extremity and intensive practice with the involved upper extremity. These approaches were designed for adults with hemiplegia, and increasing evidence suggests that they are efficacious in this population. Recently, forced use and constraint-induced therapy have been applied to children with hemiplegia. In this review, we provide a brief description of forced use and CI therapy and their historical basis, provide a summary of studies of these interventions in children, and discuss a number of important theoretical considerations, as well as implications for postural control. We will show that whereas the studies to date suggest that both forced use and CI therapy appear to be promising for improving hand function in children with hemiplegia, the data are limited. Substantially more work must be performed before this approach can be advocated for general clinical use. PMID:16097492

  14. A Critical Review of Constraint-Induced Movement Therapy and Forced Use in Children With Hemiplegia

    Directory of Open Access Journals (Sweden)

    Jeanne Charles

    2005-01-01

    Full Text Available Hemiplegia is a physical impairment that can occur in childhood following head trauma, cerebral vascular accident or transient ischemic attack (stroke, brain tumor, or congenital or perinatal injury. One of the most disabling symptoms of hemiplegia is unilaterally impaired hand and arm function. Sensory and motor impairments in children with hemiplegia compromise movement efficiency. Such children often tend not to use the affected extremity, which may further exacerbate the impairments, resulting in a developmentally learned non-use of the involved upper extremity, termed ‘developmental disuse’. Recent studies suggest that children with hemiplegia benefit from intensive practice. Forced use and Constraint-lnduced Movement Therapy (CI therapy are recent therapeutic interventions involving the restraint of the non-involved upper extremity and intensive practice with the involved upper extremity. These approaches were designed for adults with hemiplegia, and increasing evidence suggests that they are efficacious in this population. Recently, forced use and constraint-induced therapy have been applied to children with hemiplegia. In this review, we provide a brief description of forced use and CI therapy and their historical basis, provide a summary of studies of these interventions in children, and discuss a number of important theoretical considerations, as well as implications for postural control. We will show that whereas the studies to date suggest that both forced use and CI therapy appear to be promising for improving hand function in children with hemiplegia, the data are limited. Substantially more work must be performed before this approach can be advocated for general clinical use.

  15. Effect of thioperamide on modified forced swimming test-induced oxidative stress in mice.

    Science.gov (United States)

    Akhtar, Mohd; Pillai, K K; Vohora, Divya

    2005-10-01

    This study was designed i) to investigate the role of histamine H3-receptor ligands on mouse modified forced swimming test, a method that distinguishes the catecholaminergic behaviour with that of serotonergic compounds and ii) to evaluate the role of free radicals in mediation of such effects. Swiss strain albino mice were treated with different doses of histamine H3-receptor antagonist thioperamide (3.75, 7.5 and 15 mg/kg intraperitoneally) and agonist (R)-alpha-methylhistamine (5 microg intracerebroventricularly). The climbing, swimming and immobility times were recorded for 6 min. Immediately after modified forced swimming test, the animals were sacrificed and parameters of oxidative stress were assessed in the brain by measuring the thiobarbituric acid reactive substance (TBARS), glutathione (GSH) and catalase levels. Thioperamide (7.5 and 15 mg/kg intraperitoneally) dose-dependently decreased immobility time and increased swimming time but not climbing time. The behaviour of mice treated with (R)-alpha-methylhistamine was similar to that of control mice. A significant reduction in GSH and an increase in catalase levels were observed in brains of mice exposed to modified forced swimming test. Thioperamide pretreatment dose-dependently reversed such an alteration in oxidative stress parameters. (R)-alpha-methylhistamine caused a reversal of altered catalase but not GSH levels. Thioperamide shows antidepressant effects in the modified forced swimming test and causes a reversal of the test-induced oxidative stress indicating its antioxidant potential. The antidepressant effect of thioperamide appears to be mediated via serotonergic and/or antioxidant mechanisms.

  16. Virus-induced gene silencing and transient gene expression in soybean using Bean pod mottle virus infectious clones

    Science.gov (United States)

    Virus-induced gene silencing (VIGS) is a powerful and rapid approach for determining the functions of plant genes. The basis of VIGS is that a viral genome is engineered so that it can carry fragments of plant genes, typically in the 200-300 base pair size range. The recombinant viruses are used to ...

  17. Creatine supplementation enhances muscle force recovery after eccentrically-induced muscle damage in healthy individuals

    Directory of Open Access Journals (Sweden)

    Cribb Paul J

    2009-06-01

    Full Text Available Abstract Background Eccentric exercise-induced damage leads to reductions in muscle force, increased soreness, and impaired muscle function. Creatine monohydrate's (Cr ergogenic potential is well established; however few studies have directly examined the effects of Cr supplementation on recovery after damage. We examined the effects of Cr supplementation on muscle proteins and force recovery after eccentrically-induced muscle damage in healthy individuals. Methods Fourteen untrained male participants (22.1 ± 2.3 yrs, 173 ± 7.7 cm, 76.2 ± 9.3 kg were randomly separated into 2 supplement groups: i Cr and carbohydrate (Cr-CHO; n = 7; or ii carbohydrate (CHO; n = 7. Participants consumed their supplement for a period of 5 days prior to, and 14 days following a resistance exercise session. Participants performed 4 sets of 10 eccentric-only repetitions at 120% of their maximum concentric 1-RM on the leg press, leg extension and leg flexion exercise machine. Plasma creatine kinase (CK and lactate dehydrogenase (LDH activity were assessed as relevant blood markers of muscle damage. Muscle strength was examined by voluntary isokinetic knee extension using a Cybex dynamometer. Data were analyzed using repeated measures ANOVA with an alpha of 0.05. Results The Cr-supplemented group had significantly greater isokinetic (10% higher and isometric (21% higher knee extension strength during recovery from exercise-induced muscle damage. Furthermore, plasma CK activity was significantly lower (by an average of 84% after 48 hrs (P Conclusion The major finding of this investigation was a significant improvement in the rate of recovery of knee extensor muscle function after Cr supplementation following injury.

  18. Inducible gene expression and environmentally regulated genes in lactic acid bacteria.

    Science.gov (United States)

    Kok, J

    1996-10-01

    Relatively recently, a number of genes and operons have been identified in lactic acid bacteria that are inducible and respond to environmental factors. Some of these genes/operons had been isolated and analysed because of their importance in the fermentation industry and, consequently, their transcription was studied and found to be regulatable. Examples are the lactose operon, the operon for nisin production, and genes in the proteolytic pathway of Lactococcus lactis, as well as xylose metabolism in Lactobacillus pentosus. Some other operons were specifically targetted with the aim to compare their mode of regulation with known regulatory mechanisms in other well-studied bacteria. These studies, dealing with the biosynthesis of histidine, tryptophan, and of the branched chain amino acids in L. lactis, have given new insights in gene regulation and in the occurrence of auxotrophy in these bacteria. Also, nucleotide sequence analyses of a number of lactococcal bacteriophages was recently initiated to, among other things, specifically learn more about regulation of the phage life cycle. Yet another approach in the analysis of regulated genes is the 'random' selection of genetic elements that respond to environmental stimuli and the first of such sequences from lactic acid bacteria have been identified and characterized. The potential of these regulatory elements in fundamental research and practical (industrial) applications will be discussed.

  19. Forced chromatin looping raises fetal hemoglobin in adult sickle cells to higher levels than pharmacologic inducers.

    Science.gov (United States)

    Breda, Laura; Motta, Irene; Lourenco, Silvia; Gemmo, Chiara; Deng, Wulan; Rupon, Jeremy W; Abdulmalik, Osheiza Y; Manwani, Deepa; Blobel, Gerd A; Rivella, Stefano

    2016-08-25

    Overcoming the silencing of the fetal γ-globin gene has been a long-standing goal in the treatment of sickle cell disease (SCD). The major transcriptional enhancer of the β-globin locus, called the locus control region (LCR), dynamically interacts with the developmental stage-appropriate β-type globin genes via chromatin looping, a process requiring the protein Ldb1. In adult erythroid cells, the LCR can be redirected from the adult β- to the fetal γ-globin promoter by tethering Ldb1 to the human γ-globin promoter with custom-designed zinc finger (ZF) proteins (ZF-Ldb1), leading to reactivation of γ-globin gene expression. To compare this approach to pharmacologic reactivation of fetal hemoglobin (HbF), hematopoietic cells from patients with SCD were treated with a lentivirus expressing the ZF-Ldb1 or with chemical HbF inducers. The HbF increase in cells treated with ZF-Ldb1 was more than double that observed with decitabine and pomalidomide; butyrate had an intermediate effect whereas tranylcypromine and hydroxyurea showed relatively low HbF reactivation. ZF-Ldb1 showed comparatively little toxicity, and reduced sickle hemoglobin (HbS) synthesis as well as sickling of SCD erythroid cells under hypoxic conditions. The efficacy and low cytotoxicity of lentiviral-mediated ZF-Ldb1 gene transfer compared with the drug regimens support its therapeutic potential for the treatment of SCD.

  20. Virus-induced gene silencing in detached tomatoes and biochemical effects of phytoene desaturase gene silencing.

    Science.gov (United States)

    Romero, Irene; Tikunov, Yury; Bovy, Arnaud

    2011-07-01

    Virus-induced gene silencing (VIGS) is a technology that has rapidly emerged for gene function studies in plants. Many advances have been made in applying this technique in an increasing number of crops. Recently, VIGS has been successfully used to silence genes in tomato fruit through agroinfiltration of fruit attached to the plant. The phytoene desaturase (Pds) gene has been widely used as a reporter gene in VIGS experiments, although little is known about the changes that occur due to its silencing in plants. In this paper, we describe the efficient silencing of the Pds gene through the VIGS approach in detached tomato fruits, which makes the VIGS procedure even more versatile and applicable. After 16 days of agroinfiltration, approximately 75% of the tomatoes showed Pds silencing symptoms, although the distribution of silenced areas was variable among fruits. To study the potential effects caused by Pds silencing in detached tomatoes, carotenoids and other semi-polar secondary metabolites were analyzed using Liquid Chromatography-Mass Spectrometry. In addition, potential differences in primary metabolites were analyzed using Gas Chromatography-Mass Spectrometry. The results indicated that the yellow phenotype observed in Pds-silenced fruit was mainly due to the lack of the red-colored lycopene and therefore to a more pronounced contribution of the yellow-orange carotenoids (lutein, violaxanthin, and zeaxanthin) to the final color of the fruits. Furthermore, the biochemical changes observed in Pds-silenced detached tomatoes suggested that carotenoid and other pathways, e.g. leading to alkaloids and flavonoids, might be affected by the silencing of this reporter gene, and this should be taken into consideration for future experimental designs.

  1. Virus-induced silencing of a tobacco deoxyhypusine synthase gene

    Institute of Scientific and Technical Information of China (English)

    WANG Hongzhi; MA Rongcai; LI Ruifen; WANG Guoying; WEI Jianhua

    2005-01-01

    A cDNA fragment corresponding to deoxyhypusine synthase gene NbDHS was isolated and cloned into potato virus X (PVX) vector for functional analysis in Nicotiana benthamiana by using virus-induced gene silencing (VIGS). Plants agroinfected with recombinant virus vector PVX-NbDHS exhibited an increase in leaf biomass, delay in natural leaf senescence and flowering time, and decrease in leaf chlorophyll content. Semi-quantitative RT-PCR and Northern analysis showed that the transcript level of DHS was significantly lower in PVX-NbDHS infected plants. At the same time, the expression for eIF-5A, the target proteins of DHS in N. benthamiana, was concomitantly suppressed by semi-quantitative RT-PCR and Western analysis. From the phenotypic feature of the infected plants and the reduced expression abundance of DHS and eIF-5A, we concluded that NbDHS plays important roles in plant growth, development and senescence. The possible application of DHS gene in genetic modification of crops and horticultural plants was discussed.

  2. Intracerebroventricular administration of neuronostatin induces depression-like effect in forced swim test of mice.

    Science.gov (United States)

    Yang, Ai-min; Ji, Yue-ke; Su, Shu-fang; Yang, Shao-bin; Lu, Song-song; Mi, Ze-yun; Yang, Qing-zhen; Chen, Qiang

    2011-09-01

    Neuronostatin is a recently discovered endogenous bioactive peptide that is encoded by pro-mRNA of somatostatin. In the present study, we investigated the effect of neuronostatin on mood regulation in the forced swim test of mice. Our results showed intracerebroventricular (i.c.v.) administration of neuronostatin produced an increase in the immobility time, suggesting that neuronostatin induced depression-like effect. In order to rule out the possibility that neuronostatin had increased immobility time by a non-specific reduction in general activity, the effect of neuronostatin on locomotor activity was examined. Neuronostatin had no influence on locomotor activity in mice. In addition, the depression-like effect of neuronostatin was completely reversed by melanocortin 3/4 receptor antagonist SHU9119 or GABAA receptor antagonist bicuculline, but not by opioid receptor antagonist naloxone. These data suggested that the depression-like effect induced by i.c.v. administered neuronostatin was dependent upon the central melanocortin system and GABAA receptor. In conclusion, the results of this study report that neuronostatin induces depression-like effect. These findings reveal that neuronostatin is a new neuropeptide with an important role in regulating depressive behavior.

  3. Forced IFIT-2 expression represses LPS induced TNF-alpha expression at posttranscriptional levels

    Directory of Open Access Journals (Sweden)

    Autenrieth Ingo B

    2008-12-01

    Full Text Available Abstract Background Interferon induced tetratricopeptide repeat protein 2 (IFIT-2, P54 belongs to the type I interferon response genes and is highly induced after stimulation with LPS. The biological function of this protein is so far unclear. Previous studies indicated that IFIT-2 binds to the initiation factor subunit eIF-3c, affects translation initiation and inhibits protein synthesis. The aim of the study was to further characterize the function of IFIT-2. Results Stimulation of RAW264.7 macrophages with LPS or IFN-γ leads to the expression of IFIT-2 in a type I interferon dependent manner. By using stably transfected RAW264.7 macrophages overexpressing IFIT-2 we found that IFIT-2 inhibits selectively LPS induced expression of TNF-α, IL-6, and MIP-2 but not of IFIT-1 or EGR-1. In IFIT-2 overexpressing cells TNF-α mRNA expression was lower after LPS stimulation due to reduced mRNA stability. Further experiments suggest that characteristics of the 3'UTR of transcripts discriminate whether IFIT-2 has a strong impact on protein expression or not. Conclusion Our data suggest that IFIT-2 may affect selectively LPS induced protein expression probably by regulation at different posttranscriptional levels.

  4. Myosin Light Chain Kinase (MLCK) Gene Influences Exercise Induced Muscle Damage during a Competitive Marathon.

    Science.gov (United States)

    Del Coso, Juan; Valero, Marjorie; Lara, Beatriz; Salinero, Juan José; Gallo-Salazar, César; Areces, Francisco

    2016-01-01

    Myosin light chain kinase (MLCK) phosphorylates the regulatory light chain (RLC) of myosin producing increases in force development during skeletal muscle contraction. It has been suggested that MLCK gene polymorphisms might alter RLC phosphorylation thereby decreasing the ability to produce force and to resist strain during voluntary muscle contractions. Thus, the genetic variations in the MLCK gene might predispose some individuals to higher values of muscle damage during exercise, especially during endurance competitions. The aim of this investigation was to determine the influence of MLCK genetic variants on exercise-induced muscle damage produced during a marathon. Sixty-seven experienced runners competed in a marathon race. The MLCK genotype (C37885A) of these marathoners was determined. Before and after the race, a sample of venous blood was obtained to assess changes in serum myoglobin concentrations and leg muscle power changes were measured during a countermovement jump. Self-reported leg muscle pain and fatigue were determined by questionnaires. A total of 59 marathoners (88.1%) were CC homozygotes and 8 marathoners (11.9%) were CA heterozygotes. The two groups of participants completed the race with a similar time (228 ± 33 vs 234 ± 39 min; P = 0.30) and similar self-reported values for fatigue (15 ± 2 vs 16 ± 2 A.U.; P = 0.21) and lower-limb muscle pain (6.2 ± 1.7 vs 6.6 ± 1.8 cm; P = 0.29). However, CC marathoners presented higher serum myoglobin concentrations (739 ± 792 vs 348 ± 144 μg·mL-1; P = 0.03) and greater pre-to-post- race leg muscle power reduction (-32.7 ± 15.7 vs -21.2 ± 21.6%; P = 0.05) than CA marathoners. CA heterozygotes for MLCK C37885A might present higher exercise-induced muscle damage after a marathon competition than CC counterparts.

  5. L-glutamine Induces Expression of Listeria monocytogenes Virulence Genes

    Science.gov (United States)

    Lobel, Lior; Burg-Golani, Tamar; Sigal, Nadejda; Rose, Jessica; Livnat-Levanon, Nurit; Lewinson, Oded; Herskovits, Anat A.

    2017-01-01

    The high environmental adaptability of bacteria is contingent upon their ability to sense changes in their surroundings. Bacterial pathogen entry into host poses an abrupt and dramatic environmental change, during which successful pathogens gauge multiple parameters that signal host localization. The facultative human pathogen Listeria monocytogenes flourishes in soil, water and food, and in ~50 different animals, and serves as a model for intracellular infection. L. monocytogenes identifies host entry by sensing both physical (e.g., temperature) and chemical (e.g., metabolite concentrations) factors. We report here that L-glutamine, an abundant nitrogen source in host serum and cells, serves as an environmental indicator and inducer of virulence gene expression. In contrast, ammonia, which is the most abundant nitrogen source in soil and water, fully supports growth, but fails to activate virulence gene transcription. We demonstrate that induction of virulence genes only occurs when the Listerial intracellular concentration of L-glutamine crosses a certain threshold, acting as an on/off switch: off when L-glutamine concentrations are below the threshold, and fully on when the threshold is crossed. To turn on the switch, L-glutamine must be present, and the L-glutamine high affinity ABC transporter, GlnPQ, must be active. Inactivation of GlnPQ led to complete arrest of L-glutamine uptake, reduced type I interferon response in infected macrophages, dramatic reduction in expression of virulence genes, and attenuated virulence in a mouse infection model. These results may explain observations made with other pathogens correlating nitrogen metabolism and virulence, and suggest that gauging of L-glutamine as a means of ascertaining host localization may be a general mechanism. PMID:28114430

  6. High g-Force Rollercoaster Rides Induce Sinus Tachycardia but No Cardiac Arrhythmias in Healthy Children.

    Science.gov (United States)

    Pieles, Guido E; Husk, Victoria; Blackwell, Teresa; Wilson, Deirdre; Collin, Simon M; Williams, Craig A; Stuart, A Graham

    2017-01-01

    Theme park operators and medical professionals advise children with heart conditions against using rollercoaster rides, but these recommendations are not evidence-based. The underlying assumption is that the combination of adrenergic stimulation through stress and acceleration might trigger arrhythmias in susceptible individuals. We conducted a cross-sectional observational study to assess heart rate and rhythm in healthy children during commercial rollercoaster rides. Twenty healthy children (9 male) aged 11-15 (mean 13.3 ± 1.4) years underwent continuous heart rate and rhythm monitoring (2-lead ECG) from 5 min before until 10 min after each of 4 high speed (>50 km h(-1)), high g-force (>4) commercial rollercoaster rides. Total recording time was 13 h 20 min. No arrhythmic events were detected. Resting heart rate was 81 ± 10 b min(-1) and increased to 158 ± 20 b·min(-1) during rides. The highest mean HR (165 ± 23 b min(-1)) was observed on the ride with the lowest g-force (4.5 g), but one of the highest speeds (100 km h(-1)). Anticipatory tachycardia (126 ± 15 b min(-1)) within 5 min was frequently observed. A 10 min recovery HR (124 ± 17 b min(-1)) was 56 % greater than resting HR. The speed and g-force experienced on roller coasters induce sinus tachycardia but do not elicit pathological arrhythmias in healthy children.

  7. Force-induced chemical reactions on the metal centre in a single metalloprotein molecule.

    Science.gov (United States)

    Zheng, Peng; Arantes, Guilherme M; Field, Martin J; Li, Hongbin

    2015-06-25

    Metalloproteins play indispensable roles in biology owing to the versatile chemical reactivity of metal centres. However, studying their reactivity in many metalloproteins is challenging, as protein three-dimensional structure encloses labile metal centres, thus limiting their access to reactants and impeding direct measurements. Here we demonstrate the use of single-molecule atomic force microscopy to induce partial unfolding to expose metal centres in metalloproteins to aqueous solution, thus allowing for studying their chemical reactivity in aqueous solution for the first time. As a proof-of-principle, we demonstrate two chemical reactions for the FeS4 centre in rubredoxin: electrophilic protonation and nucleophilic ligand substitution. Our results show that protonation and ligand substitution result in mechanical destabilization of the FeS4 centre. Quantum chemical calculations corroborated experimental results and revealed detailed reaction mechanisms. We anticipate that this novel approach will provide insights into chemical reactivity of metal centres in metalloproteins under biologically more relevant conditions.

  8. Force-induced chemical reactions on the metal centre in a single metalloprotein molecule

    Science.gov (United States)

    Zheng, Peng; Arantes, Guilherme M.; Field, Martin J.; Li, Hongbin

    2015-06-01

    Metalloproteins play indispensable roles in biology owing to the versatile chemical reactivity of metal centres. However, studying their reactivity in many metalloproteins is challenging, as protein three-dimensional structure encloses labile metal centres, thus limiting their access to reactants and impeding direct measurements. Here we demonstrate the use of single-molecule atomic force microscopy to induce partial unfolding to expose metal centres in metalloproteins to aqueous solution, thus allowing for studying their chemical reactivity in aqueous solution for the first time. As a proof-of-principle, we demonstrate two chemical reactions for the FeS4 centre in rubredoxin: electrophilic protonation and nucleophilic ligand substitution. Our results show that protonation and ligand substitution result in mechanical destabilization of the FeS4 centre. Quantum chemical calculations corroborated experimental results and revealed detailed reaction mechanisms. We anticipate that this novel approach will provide insights into chemical reactivity of metal centres in metalloproteins under biologically more relevant conditions.

  9. Artificial Force Induced Reaction (AFIR) Method for Exploring Quantum Chemical Potential Energy Surfaces.

    Science.gov (United States)

    Maeda, Satoshi; Harabuchi, Yu; Takagi, Makito; Taketsugu, Tetsuya; Morokuma, Keiji

    2016-10-01

    In this account, a technical overview of the artificial force induced reaction (AFIR) method is presented. The AFIR method is one of the automated reaction-path search methods developed by the authors, and has been applied extensively to a variety of chemical reactions, such as organocatalysis, organometallic catalysis, and photoreactions. There are two modes in the AFIR method, i.e., a multicomponent mode and a single-component mode. The former has been applied to bimolecular and multicomponent reactions and the latter to unimolecular isomerization and dissociation reactions. Five numerical examples are presented for an Aldol reaction, a Claisen rearrangement, a Co-catalyzed hydroformylation, a fullerene structure search, and a nonradiative decay path search in an electronically excited naphthalene molecule. Finally, possible applications of the AFIR method are discussed.

  10. Probing weak force induced parity violation by high resolution mid-infrared molecular spectroscopy

    CERN Document Server

    Tokunaga, Sean K; Auguste, Frédéric; Shelkovnikov, Alexander; Daussy, Christophe; Amy-Klein, Anne; Chardonnet, Christian; Darquié, Benoît

    2013-01-01

    To date no experiment has reached the level of sensitivity required to observe weak nuclear force induced parity violation (PV) energy differences in chiral molecules. In this paper, we present the approach, adopted at Laboratoire de Physique des Lasers (LPL), to measure frequency differences in the vibrational spectrum of enantiomers. We review different spectroscopic methods developed at LPL leading to the highest resolutions, as well as 20 years of CO2 laser stabilization work enabling such precise measurements. After a first attempt to observe PV vibrational frequency shifts using sub-Doppler saturated absorption spectroscopy in a cell, we are currently aiming at an experiment based on Doppler-free two-photon Ramsey interferometry on a supersonic beam. We report on our latest progress towards observing PV with chiral organo-metallic complexes containing a heavy rhenium atom.

  11. A stochastic load model for pedestrian-induced lateral forces on footbridges

    DEFF Research Database (Denmark)

    Ingólfsson, Einar Thór; Georgakis, Christos

    2011-01-01

    between limited and excessive lateral vibrations occurs for a small increase in the number of people occupying the bridge. This disproportionate increase in the lateral vibration amplitude is caused by a dynamic interaction between the pedestrian and the laterally moving structure, although the governing......). It was shown that large amplitude vibrations are the result of correlated pedestrian forces in the form of “negative damping”, with magnitudes that depend on the relationship between the pacing frequency and the frequency of the lateral movement.Herewith, a novel stochastic load model for the frequency......In the past decade, several researchers have studied the phenomenon of excessive pedestrian-induced lateral vibrations and full-scale measurements of various bridges under crowd loading have been carried out. These tests have verified the existence of a form of instability for which a transition...

  12. Effect of Polysaccharide from Cordyceps militaris (Ascomycetes) on Physical Fatigue Induced by Forced Swimming.

    Science.gov (United States)

    Xu, Yan-Feng

    2016-01-01

    Cordyceps militaris is the one of the most important medicinal mushrooms, widely used in East Asian countries. Polysaccharide is considered to be the principal active component in C. militaris and has a wide range of biological and pharmacological properties. This study was undertaken to investigate the effect of polysaccharide from C. militaris (PCM) on physical fatigue induced in animals through a forced swimming test. The mice were divided into 4 groups receiving 28 days' treatment with drinking water (exercise control) or low-, medium-, and high-dose PCM (40, 80, and 160 mg/kg/day, respectively). After 28 days, the mice were subjected to the forced swimming test; the exhaustive swimming time was measured and fatigue-related biochemical parameters, including serum lactic acid, urea nitrogen, creatine kinase, alanine aminotransferase, aspartate aminotransferase, superoxide dismutase, glutathi- one peroxidase, catalase, malondialdehyde, liver glycogen, and muscle glycogen, were analyzed. The results showed that PCM could significantly prolong the exhaustive swimming time of mice; decrease concentrations of serum lactic acid, urea nitrogen, creatine kinase, aspartate aminotransferase, alanine aminotransferase, and malondialdehyde; and increase liver and muscle glycogen contents and the concentrations of serum superoxide dismutase, glutathione per- oxidase, and catalase. The data suggest that PCM has an antifatigue effect, and it might become a new functional food or medicine for fatigue resistance.

  13. Simultaneous Knee Extensor Muscle Action Induces an Increase in Voluntary Force Generation of Plantar Flexor Muscles.

    Science.gov (United States)

    Suzuki, Takahito; Shioda, Kohei; Kinugasa, Ryuta; Fukashiro, Senshi

    2017-02-01

    Suzuki, T, Shioda, K, Kinugasa, R, and Fukashiro, S. Simultaneous knee extensor muscle action induces an increase in voluntary force generation of plantar flexor muscles. J Strength Cond Res 31(2): 365-371, 2017-Maximum activation of the plantar flexor muscles is required for various sporting activities that involve simultaneous plantar flexion and knee extension. During a multi-joint movement, activation of the plantar flexor muscles is affected by the activity of the knee extensor muscles. We hypothesized that coactivation of the plantar flexor muscles and knee extensor muscles would result in a higher plantar flexion torque. To test this hypothesis, 8 male volunteers performed maximum voluntary isometric action of the plantar flexor muscles with and without isometric action of the knee extensor muscles. Surface electromyographic data were collected from 8 muscles of the right lower limb. Voluntary activation of the triceps surae muscles, evaluated using the interpolated twitch technique, significantly increased by 6.4 percentage points with intentional knee extensor action (p = 0.0491). This finding is in line with a significant increase in the average rectified value of the electromyographic activity of the vastus lateralis, fibularis longus, and soleus muscles (p = 0.013, 0.010, and 0.045, respectively). The resultant plantar flexion torque also significantly increased by 11.5% of the predetermined maximum (p = 0.031). These results suggest that higher plantar flexor activation coupled with knee extensor activation facilitates force generation during a multi-joint task.

  14. Limitations to photosynthesis by proton motive force-induced photosystem II photodamage

    Science.gov (United States)

    Davis, Geoffry A; Kanazawa, Atsuko; Schöttler, Mark Aurel; Kohzuma, Kaori; Froehlich, John E; Rutherford, A William; Satoh-Cruz, Mio; Minhas, Deepika; Tietz, Stefanie; Dhingra, Amit; Kramer, David M

    2016-01-01

    The thylakoid proton motive force (pmf) generated during photosynthesis is the essential driving force for ATP production; it is also a central regulator of light capture and electron transfer. We investigated the effects of elevated pmf on photosynthesis in a library of Arabidopsis thaliana mutants with altered rates of thylakoid lumen proton efflux, leading to a range of steady-state pmf extents. We observed the expected pmf-dependent alterations in photosynthetic regulation, but also strong effects on the rate of photosystem II (PSII) photodamage. Detailed analyses indicate this effect is related to an elevated electric field (Δψ) component of the pmf, rather than lumen acidification, which in vivo increased PSII charge recombination rates, producing singlet oxygen and subsequent photodamage. The effects are seen even in wild type plants, especially under fluctuating illumination, suggesting that Δψ-induced photodamage represents a previously unrecognized limiting factor for plant productivity under dynamic environmental conditions seen in the field. DOI: http://dx.doi.org/10.7554/eLife.16921.001 PMID:27697149

  15. Kinetic Defects Induced by Melittin in Model Lipid Membranes: A Solution Atomic Force Microscopy Study.

    Science.gov (United States)

    Pan, Jianjun; Khadka, Nawal K

    2016-05-26

    Quantitative characterization of membrane defects (pores) is important for elucidating the molecular basis of many membrane-active peptides. We study kinetic defects induced by melittin in vesicular and planar lipid bilayers. Fluorescence spectroscopy measurements indicate that melittin induces time-dependent calcein leakage. Solution atomic force microscopy (AFM) is used to visualize melittin-induced membrane defects. After initial equilibration, the most probable defect radius is ∼3.8 nm in 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC) bilayers. Unexpectedly, defects become larger with longer incubation, accompanied by substantial shape transformation. The initial defect radius is ∼4.7 nm in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayers. Addition of 30 mol % cholesterol to DOPC bilayers suppresses defect kinetics, although the inhibitory impact is negated by longer incubation. Overall, the kinetic rate of defect development follows DLPC > DOPC > DOPC/cholesterol. Kinetic defects are also observed when anionic lipids are present. Based on the observation that defects can occupy as large as 40% of the bilayer surface, we propose a kinetic defect growth model. We also study the effect of melittin on the phase behavior of DOPC/egg-sphingomyelin/cholesterol bilayers. We find that melittin initially suppresses or eliminates liquid-ordered (Lo) domains; Lo domains gradually emerge and become the dominant species with longer incubation; and defects in phase-coexisting bilayers have a most probable radius of ∼5 nm and are exclusively localized in the liquid-disordered (Ld) phase. Our experimental data highlight that melittin-induced membrane defects are not static; conversely, spontaneous defect growth is intrinsically associated with membrane permeabilization exerted by melittin.

  16. Transcription-induced DNA double strand breaks: both oncogenic force and potential therapeutic target?

    Science.gov (United States)

    Haffner, Michael C; De Marzo, Angelo M; Meeker, Alan K; Nelson, William G; Yegnasubramanian, Srinivasan

    2011-06-15

    An emerging model of transcriptional activation suggests that induction of transcriptional programs, for instance by stimulating prostate or breast cells with androgens or estrogens, respectively, involves the formation of DNA damage, including DNA double strand breaks (DSB), recruitment of DSB repair proteins, and movement of newly activated genes to transcription hubs. The DSB can be mediated by the class II topoisomerase TOP2B, which is recruited with the androgen receptor and estrogen receptor to regulatory sites on target genes and is apparently required for efficient transcriptional activation of these genes. These DSBs are recognized by the DNA repair machinery triggering the recruitment of repair proteins such as poly(ADP-ribose) polymerase 1 (PARP1), ATM, and DNA-dependent protein kinase (DNA-PK). If illegitimately repaired, such DSBs can seed the formation of genomic rearrangements like the TMPRSS2-ERG fusion oncogene in prostate cancer. Here, we hypothesize that these transcription-induced, TOP2B-mediated DSBs can also be exploited therapeutically and propose that, in hormone-dependent tumors like breast and prostate cancers, a hormone-cycling therapy, in combination with topoisomerase II poisons or inhibitors of the DNA repair components PARP1 and DNA-PK, could overwhelm cancer cells with transcription-associated DSBs. Such strategies may find particular utility in cancers, like prostate cancer, which show low proliferation rates, in which other chemotherapeutic strategies that target rapidly proliferating cells have had limited success.

  17. MicroRNA-138 enhances TRAIL-induced apoptosis through interferon-stimulated gene 15 downregulation in hepatocellular carcinoma cells.

    Science.gov (United States)

    Zuo, Chaohui; Sheng, Xinyi; Liu, Zhuo; Ma, Min; Xiong, Shuhan; Deng, Hongyu; Li, Sha; Yang, Darong; Wang, Xiaohong; Xiao, Hua; Quan, Hu; Xia, Man

    2017-06-01

    Hepatocellular carcinoma is a leading cause of cancer-related mortality worldwide. TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a potential target for cancer therapy. However, many cancer cells are resistant to TRAIL-induced apoptosis and its mechanism is not well understood. In this study, to identify potential therapeutic targets for TRAIL-resistant cancer cells, we compared the expression levels of interferon-stimulated gene 15 in TRAIL-sensitive and TRAIL-resistant hepatocellular carcinoma cell lines. Western blot analysis showed that interferon-stimulated gene 15 expression levels were significantly higher in resistant HLCZ01and Huh7 cells than in sensitive LH86 and SMMC-7721 cells. Interferon-stimulated gene 15 knockdown in resistance cells led to TRAIL sensitivity. Conversely, interferon-stimulated gene 15 overexpression in sensitive cells resulted in TRAIL resistance. Our bioinformatics search detected a putative target sequence for microRNA miR-138 in the 3' untranslated region of the interferon-stimulated gene 15. Real-time quantitative polymerase chain reaction analysis demonstrated that miR-138 was significantly downregulated in TRAIL-resistant cells compared to TRAIL-sensitive cells. Forced expression of miR-138 in resistant cells decreased both messenger RNA and protein levels of interferon-stimulated gene 15, and when exposed to TRAIL, activated poly(adenosine diphosphate-ribose) polymerase, indicating sensitization to TRAIL. The results suggested that miR-138 regulates the interferon-stimulated gene 15 expression by directly targeting the 3' untranslated region of interferon-stimulated gene 15 and modulates the sensitivity to TRAIL-induced apoptosis. MiR-138 may be a target for therapeutic intervention in TRAIL-based drug treatments of resistant hepatocellular carcinoma or could be a biomarker to select patients who may benefit from the treatment.

  18. Murine bone cell lines as models for spaceflight induced effects on differentiation and gene expression

    Science.gov (United States)

    Lau, P.; Hellweg, C. E.; Baumstark-Khan, C.; Reitz, G.

    Critical health factors for space crews especially on long-term missions are radiation exposure and the absence of gravity DNA double strand breaks DSB are presumed to be the most deleterious DNA lesions after radiation as they disrupt both DNA strands in close proximity Besides radiation risk the absence of gravity influences the complex skeletal apparatus concerning muscle and especially bone remodelling which results from mechanical forces exerting on the body Bone is a dynamic tissue which is life-long remodelled by cells from the osteoblast and osteoclast lineage Any imbalance of this system leads to pathological conditions such as osteoporosis or osteopetrosis Osteoblastic cells play a crucial role in bone matrix synthesis and differentiate either into bone-lining cells or into osteocytes Premature terminal differentiation has been reported to be induced by a number of DNA damaging or cell stress inducing agents including ionising and ultraviolet radiation as well as treatment with mitomycin C In the present study we compare the effects of sequential differentiation by adding osteoinductive substances ss -glycerophosphate and ascorbic acid Radiation-induced premature differentiation was investigated regarding the biosynthesis of specific osteogenic marker molecules and the differentiation dependent expression of marker genes The bone cell model established in our laboratory consists of the osteocyte cell line MLO-Y4 the osteoblast cell line OCT-1 and the subclones 4 and 24 of the osteoblast cell line MC3T3-E1 expressing several

  19. Changes in Gene Expression of Arabidopsis Thaliana Cell Cultures Upon Exposure to Real and Simulated Partial- g Forces

    Science.gov (United States)

    Fengler, Svenja; Spirer, Ina; Neef, Maren; Ecke, Margret; Hauslage, Jens; Hampp, Rüdiger

    2016-06-01

    Cell cultures of the plant model organism Arabidopsis thaliana were exposed to partial- g forces during parabolic flight and clinostat experiments (0.16 g, 0.38 g and 0.5 g were tested). In order to investigate gravity-dependent alterations in gene expression, samples were metabolically quenched by the fixative RNA later Ⓡ to stabilize nucleic acids and used for whole-genome microarray analysis. An attempt to identify the potential threshold acceleration for the gravity-dependent response showed that the smaller the experienced g-force, the greater was the susceptibility of the cell cultures. Compared to short-term μ g during a parabolic flight, the number of differentially expressed genes under partial- g was lower. In addition, the effect on the alteration of amounts of transcripts decreased during partial- g parabolic flight due to the sequence of the different parabolas (0.38 g, 0.16 g and μ g). A time-dependent analysis under simulated 0.5 g indicates that adaptation occurs within minutes. Differentially expressed genes (at least 2-fold up- or down-regulated in expression) under real flight conditions were to some extent identical with those affected by clinorotation. The highest number of homologuous genes was detected within seconds of exposure to 0.38 g (both flight and clinorotation). To a considerable part, these genes deal with cell wall properties. Additionally, responses specific for clinorotation were observed.

  20. Structural basis and kinetics of force-induced conformational changes of an αA domain-containing integrin.

    Directory of Open Access Journals (Sweden)

    Xue Xiang

    Full Text Available BACKGROUND: Integrin α(Lβ₂ (lymphocyte function-associated antigen, LFA-1 bears force upon binding to its ligand intercellular adhesion molecule 1 (ICAM-1 when a leukocyte adheres to vascular endothelium or an antigen presenting cell (APC during immune responses. The ligand binding propensity of LFA-1 is related to its conformations, which can be regulated by force. Three conformations of the LFA-1 αA domain, determined by the position of its α₇-helix, have been suggested to correspond to three different affinity states for ligand binding. METHODOLOGY/PRINCIPAL FINDINGS: The kinetics of the force-driven transitions between these conformations has not been defined and dynamically coupled to the force-dependent dissociation from ligand. Here we show, by steered molecular dynamics (SMD simulations, that the αA domain was successively transitioned through three distinct conformations upon pulling the C-terminus of its α₇-helix. Based on these sequential transitions, we have constructed a mathematical model to describe the coupling between the αA domain conformational changes of LFA-1 and its dissociation from ICAM-1 under force. Using this model to analyze the published data on the force-induced dissociation of single LFA-1/ICAM-1 bonds, we estimated the force-dependent kinetic rates of interstate transition from the short-lived to intermediate-lived and from intermediate-lived to long-lived states. Interestingly, force increased these transition rates; hence activation of LFA-1 was accelerated by pulling it via an engaged ICAM-1. CONCLUSIONS/SIGNIFICANCE: Our study defines the structural basis for mechanical regulation of the kinetics of LFA-1 αA domain conformational changes and relates these simulation results to experimental data of force-induced dissociation of single LFA-1/ICAM-1 bonds by a new mathematical model, thus provided detailed structural and kinetic characterizations for force-stabilization of LFA-1/ICAM-1 interaction.

  1. BiForce Toolbox: powerful high-throughput computational analysis of gene-gene interactions in genome-wide association studies.

    Science.gov (United States)

    Gyenesei, Attila; Moody, Jonathan; Laiho, Asta; Semple, Colin A M; Haley, Chris S; Wei, Wen-Hua

    2012-07-01

    Genome-wide association studies (GWAS) have discovered many loci associated with common disease and quantitative traits. However, most GWAS have not studied the gene-gene interactions (epistasis) that could be important in complex trait genetics. A major challenge in analysing epistasis in GWAS is the enormous computational demands of analysing billions of SNP combinations. Several methods have been developed recently to address this, some using computers equipped with particular graphical processing units, most restricted to binary disease traits and all poorly suited to general usage on the most widely used operating systems. We have developed the BiForce Toolbox to address the demand for high-throughput analysis of pairwise epistasis in GWAS of quantitative and disease traits across all commonly used computer systems. BiForce Toolbox is a stand-alone Java program that integrates bitwise computing with multithreaded parallelization and thus allows rapid full pairwise genome scans via a graphical user interface or the command line. Furthermore, BiForce Toolbox incorporates additional tests of interactions involving SNPs with significant marginal effects, potentially increasing the power of detection of epistasis. BiForce Toolbox is easy to use and has been applied in multiple studies of epistasis in large GWAS data sets, identifying interesting interaction signals and pathways.

  2. Atomic Force Microscopy of spermidine-induced DNA condensates on silicon surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Carrasco, I.S.S.; Bastos, F.M.; Munford, M.L.; Ramos, E.B.; Rocha, M.S., E-mail: marcios.rocha@ufv.br

    2012-01-01

    In the present work, we show that oxidized silicon may be successfully used to image multivalent cation-induced DNA condensates under the Atomic Force Microscope (AFM). The images thus obtained are good enough, allowing us to distinguish between different condensate forms and to perform nanometer-sized measurements. Qualitative results previously obtained using mica as a substrate are recovered here. We additionally show that the interactions between the cation spermidine (the condensing agent) and the DNA molecules are not significantly disturbed by the silicon surface, since the phase behavior of an ensemble of DNA molecules deposited on the silicon substrate as a function of the cation concentration is very similar to that found in solution. - Highlights: Black-Right-Pointing-Pointer We developed a protocol do deposit condensed DNA on oxidized silicon substrates. Black-Right-Pointing-Pointer We measured the sizes of the spermidine-induced DNA condensates under the AFM. Black-Right-Pointing-Pointer We sketch the phase diagram of an ensemble of DNA molecules in a spermidine solution.

  3. Chemical memory reactions induced bursting dynamics in gene expression.

    Science.gov (United States)

    Tian, Tianhai

    2013-01-01

    Memory is a ubiquitous phenomenon in biological systems in which the present system state is not entirely determined by the current conditions but also depends on the time evolutionary path of the system. Specifically, many memorial phenomena are characterized by chemical memory reactions that may fire under particular system conditions. These conditional chemical reactions contradict to the extant stochastic approaches for modeling chemical kinetics and have increasingly posed significant challenges to mathematical modeling and computer simulation. To tackle the challenge, I proposed a novel theory consisting of the memory chemical master equations and memory stochastic simulation algorithm. A stochastic model for single-gene expression was proposed to illustrate the key function of memory reactions in inducing bursting dynamics of gene expression that has been observed in experiments recently. The importance of memory reactions has been further validated by the stochastic model of the p53-MDM2 core module. Simulations showed that memory reactions is a major mechanism for realizing both sustained oscillations of p53 protein numbers in single cells and damped oscillations over a population of cells. These successful applications of the memory modeling framework suggested that this innovative theory is an effective and powerful tool to study memory process and conditional chemical reactions in a wide range of complex biological systems.

  4. Fetal Globin Gene Inducers: Novel Agents & New Potential

    Science.gov (United States)

    Perrine, Susan P.; Castaneda, Serguei A.; Chui, David H.; Faller, Douglas V.; Berenson, Ronald J.; Fucharoen, Suthat

    2013-01-01

    Inducing expression of endogenous fetal globin (γ-globin) gene expression to 60-70% of alpha globin synthesis produces β-thalassemia trait globin synthetic ratios and can reduce anemia to a mild level. Several classes of therapeutics have induced γ-globin expression in beta thalassemia patients and subsequently raised total hemoglobin levels, demonstrating proof-of-concept of the approach. Butyrate treatment eliminated transfusion requirements in formerly transfusion-dependent patients with treatment for as long as 7 years. However, prior generations were not readily applicable for widespread use. Currently, a novel oral dual-action therapeutic sodium 2,2-dimethylbutyrate is in clinical trials, an oral decitabine formulation is under development, and agents with complementary mechanisms of action can be applied in combined regimens. Identification of 3 major genetic trait loci which modulate clinical severity provides avenues for developing tailored regimens. These refinements offer renewed potential to apply fetal globin induction as a treatment approach in patient-friendly regimens that can be used world-wide. PMID:20712788

  5. Deoxynivalenol-Induced Proinflammatory Gene Expression: Mechanisms and Pathological Sequelae

    Directory of Open Access Journals (Sweden)

    James J. Pestka

    2010-06-01

    Full Text Available The trichothecene mycotoxin deoxynivalenol (DON is commonly encountered in human cereal foods throughout the world as a result of infestation of grains in the field and in storage by the fungus Fusarium. Significant questions remain regarding the risks posed to humans from acute and chronic DON ingestion, and how to manage these risks without imperiling access to nutritionally important food commodities. Modulation of the innate immune system appears particularly critical to DON’s toxic effects. Specifically, DON induces activation of mitogen-activated protein kinases (MAPKs in macrophages and monocytes, which mediate robust induction of proinflammatory gene expression—effects that can be recapitulated in intact animals. The initiating mechanisms for DON-induced ribotoxic stress response appear to involve the (1 activation of constitutive protein kinases on the damaged ribosome and (2 autophagy of the chaperone GRP78 with consequent activation of the ER stress response. Pathological sequelae resulting from chronic low dose exposure include anorexia, impaired weight gain, growth hormone dysregulation and aberrant IgA production whereas acute high dose exposure evokes gastroenteritis, emesis and a shock-like syndrome. Taken together, the capacity of DON to evoke ribotoxic stress in mononuclear phagocytes contributes significantly to its acute and chronic toxic effects in vivo. It is anticipated that these investigations will enable the identification of robust biomarkers of effect that will be applicable to epidemiological studies of the human health effects of this common mycotoxin.

  6. Steered molecular dynamics simulations of a type IV pilus probe initial stages of a force-induced conformational transition.

    Directory of Open Access Journals (Sweden)

    Joseph L Baker

    2013-04-01

    Full Text Available Type IV pili are long, protein filaments built from a repeating subunit that protrudes from the surface of a wide variety of infectious bacteria. They are implicated in a vast array of functions, ranging from bacterial motility to microcolony formation to infection. One of the most well-studied type IV filaments is the gonococcal type IV pilus (GC-T4P from Neisseria gonorrhoeae, the causative agent of gonorrhea. Cryo-electron microscopy has been used to construct a model of this filament, offering insights into the structure of type IV pili. In addition, experiments have demonstrated that GC-T4P can withstand very large tension forces, and transition to a force-induced conformation. However, the details of force-generation, and the atomic-level characteristics of the force-induced conformation, are unknown. Here, steered molecular dynamics (SMD simulation was used to exert a force in silico on an 18 subunit segment of GC-T4P to address questions regarding the nature of the interactions that lead to the extraordinary strength of bacterial pili. SMD simulations revealed that the buried pilin α1 domains maintain hydrophobic contacts with one another within the core of the filament, leading to GC-T4P's structural stability. At the filament surface, gaps between pilin globular head domains in both the native and pulled states provide water accessible routes between the external environment and the interior of the filament, allowing water to access the pilin α1 domains as reported for VC-T4P in deuterium exchange experiments. Results were also compared to the experimentally observed force-induced conformation. In particular, an exposed amino acid sequence in the experimentally stretched filament was also found to become exposed during the SMD simulations, suggesting that initial stages of the force induced transition are well captured. Furthermore, a second sequence was shown to be initially hidden in the native filament and became exposed upon

  7. Host-Induced Gene Silencing of Rice Blast Fungus Magnaporthe oryzae Pathogenicity Genes Mediated by the Brome Mosaic Virus.

    Science.gov (United States)

    Zhu, Lin; Zhu, Jian; Liu, Zhixue; Wang, Zhengyi; Zhou, Cheng; Wang, Hong

    2017-09-26

    Magnaportheoryzae is a devastating plant pathogen, which has a detrimental impact on rice production worldwide. Despite its agronomical importance, some newly-emerging pathotypes often overcome race-specific disease resistance rapidly. It is thus desirable to develop a novel strategy for the long-lasting resistance of rice plants to ever-changing fungal pathogens. Brome mosaic virus (BMV)-induced RNA interference (RNAi) has emerged as a useful tool to study host-resistance genes for rice blast protection. Planta-generated silencing of targeted genes inside biotrophic pathogens can be achieved by expression of M.oryzae-derived gene fragments in the BMV-mediated gene silencing system, a technique termed host-induced gene silencing (HIGS). In this study, the effectiveness of BMV-mediated HIGS in M.oryzae was examined by targeting three predicted pathogenicity genes, MoABC1,MoMAC1 and MoPMK1. Systemic generation of fungal gene-specific small interfering RNA (siRNA) molecules induced by inoculation of BMV viral vectors inhibited disease development and reduced the transcription of targeted fungal genes after subsequent M.oryzae inoculation. Combined introduction of fungal gene sequences in sense and antisense orientation mediated by the BMV silencing vectors significantly enhanced the efficiency of this host-generated trans-specific RNAi, implying that these fungal genes played crucial roles in pathogenicity. Collectively, our results indicated that BMV-HIGS system was a great strategy for protecting host plants against the invasion of pathogenic fungi.

  8. Subchronic treatment with aldosterone induces depression-like behaviours and gene expression changes relevant to major depressive disorder.

    Science.gov (United States)

    Hlavacova, Natasa; Wes, Paul D; Ondrejcakova, Maria; Flynn, Marianne E; Poundstone, Patricia K; Babic, Stanislav; Murck, Harald; Jezova, Daniela

    2012-03-01

    The potential role of aldosterone in the pathophysiology of depression is unclear. The aim of this study was to test the hypothesis that prolonged elevation of circulating aldosterone induces depression-like behaviour accompanied by disease-relevant changes in gene expression in the hippocampus. Subchronic (2-wk) treatment with aldosterone (2 μg/100 g body weight per day) or vehicle via subcutaneous osmotic minipumps was used to induce hyperaldosteronism in male rats. All rats (n = 20/treatment group) underwent a modified sucrose preference test. Half of the animals from each treatment group were exposed to the forced swim test (FST), which served both as a tool to assess depression-like behaviour and as a stress stimulus. Affymetrix microarray analysis was used to screen the entire rat genome for gene expression changes in the hippocampus. Aldosterone treatment induced an anhedonic state manifested by decreased sucrose preference. In the FST, depressogenic action of aldosterone was manifested by decreased latency to immobility and increased time spent immobile. Aldosterone treatment resulted in transcriptional changes of genes in the hippocampus involved in inflammation, glutamatergic activity, and synaptic and neuritic remodelling. Furthermore, aldosterone-regulated genes substantially overlapped with genes affected by stress in the FST. This study demonstrates the existence of a causal relationship between the hyperaldosteronism and depressive behaviour. In addition, aldosterone treatment induced changes in gene expression that may be relevant to the aetiology of major depressive disorder. Subchronic treatment with aldosterone represents a new animal model of depression, which may contribute to the development of novel targets for the treatment of depression.

  9. Genome-based identification of cancer genes by proviral tagging in mouse retrovirus-induced T-cell lymphomas.

    Science.gov (United States)

    Kim, Rachel; Trubetskoy, Alla; Suzuki, Takeshi; Jenkins, Nancy A; Copeland, Neal G; Lenz, Jack

    2003-02-01

    The identification of tumor-inducing genes is a driving force for elucidating the molecular mechanisms underlying cancer. Many retroviruses induce tumors by insertion of viral DNA adjacent to cellular oncogenes, resulting in altered expression and/or structure of the encoded proteins. The availability of the mouse genome sequence now allows analysis of retroviral common integration sites in murine tumors to be used as a genetic screen for identification of large numbers of candidate cancer genes. By positioning the sequences of inverse PCR-amplified, virus-host junction fragments within the mouse genome, 19 target genes were identified in T-cell lymphomas induced by the retrovirus SL3-3. The candidate cancer genes included transcription factors (Fos, Gfi1, Lef1, Myb, Myc, Runx3, and Sox3), all three D cyclins, Ras signaling pathway components (Rras2/TC21 and Rasgrp1), and Cmkbr7/CCR7. The most frequent target was Rras2. Insertions as far as 57 kb away from the transcribed portion were associated with substantially increased transcription of Rras2, and no coding sequence mutations, including those typically involved in Ras activation, were detected. These studies demonstrate the power of genome-based analysis of retroviral insertion sites for cancer gene discovery, identify several new genes worth examining for a role in human cancer, and implicate the pathways in which those genes act in lymphomagenesis. They also provide strong genetic evidence that overexpression of unmutated Rras2 contributes to tumorigenesis, thus suggesting that it may also do so if it is inappropriately expressed in human tumors.

  10. An Agent-Based Discrete Collagen Fiber Network Model of Dynamic Traction Force-Induced Remodeling.

    Science.gov (United States)

    Reinhardt, James W; Gooch, Keith

    2017-09-21

    We developed an agent-based model that incorporates repetitively applied traction force within a discrete fiber network to understand how microstructural properties of the network influence mechanical properties and traction force-induced remodeling. An important difference between our model and similar finite-element models is that by implementing more biologically-realistic dynamic traction, we can explore a greater range of matrix remodeling. Here, we validated our model by reproducing qualitative trends observed in three sets of experimental data reported by others: tensile and shear testing of cell-free collagen gels, collagen remodeling around a single isolated cell, and collagen remodeling between pairs of cells. In response to tensile and shear strain, simulated acellular networks exhibited biphasic stress-strain curves indicative of strain-stiffening. Our data support the notion that strain-stiffening might occur as individual fibrils successively align along the axis of strain and become engaged in tension. In simulations with a single, contractile cell, peak collagen displacement occurred closest to the cell and decreased with increasing distance. In simulations with two cells, compaction of collagen between cells appeared inversely related to the initial distance between cells. Further analysis revealed strain energy was relatively uniform around the outer surface of cells separated by 250 microns, but became increasingly non-uniform as the distance between cells decreased. This pattern was partly attributable to the pattern of collagen compaction. These findings are of interest because fibril alignment, density, and strain energy may each contribute to contact guidance during tissue morphogenesis.

  11. Expression of myosin heavy-chain mRNA in cultured myoblasts induced by centrifugal force.

    Science.gov (United States)

    Kurokawa, Katsuhide; Sakiyama, Koji; Abe, Shinichi; Hiroki, Emi; Naito, Kaoru; Nakajima, Kazunori; Takeda, Tomotaka; Inoue, Takashi; Ide, Yoshinobu; Ishigami, Keiichi

    2008-11-01

    Ballistic muscle training leads to hypertrophy of fast type fibers and training for endurance induces that of slow type fibers. Numerous studies have been conducted on electrical, extending and magnetic stimulation of cells, but the effect of centrifugal force on cells remains to be investigated. In this study, we investigated the effect of stimulating cultured myoblasts with centrifugal force at different speeds on cell proliferation and myosin heavy-chain (MyHC) mRNA expression in muscle fiber. Stimulation of myoblasts was carried out at 2 different speeds for 20 min using the Himac CT6D, a desk centrifuge, and cells were observed at 1, 3 and 5 days later. Number of cells 1 and 5 days after centrifugal stimulation was significantly larger in the 62.5 x g and 4,170 x g stimulation groups than in the control group. Expression of MyHC-2b mRNA 1 day after centrifugal stimulation was significantly higher in the 2 stimulation groups than in the control group. Almost no expression of MyHC-2a was observed in any group at 1 and 3 days after centrifugal stimulation. However, 5 days after stimulation, MyHC-2a was strongly expressed in the 2 stimulation groups in comparison to the control group. Three days after centrifugal stimulation, expression of MyHC-1 was significantly higher in the 2 stimulation groups than in the control group. The results of this study clarified the effect of different centrifugal stimulation speeds on muscle fiber characteristics, and suggest that centrifugal stimulation of myoblasts enhances cell proliferation.

  12. Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene

    NARCIS (Netherlands)

    Kop, D.A.M. van der; Schuyer, M.; Pinas, J.E.; Zaal, B.J. van der; Hooykaas, P.J.J.

    1999-01-01

    Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the β-glucuronidase (gusA) reporter gene. Subsequently, seeds were tr

  13. The forced walking test: a novel test for pinpointing the anesthetic-induced transition in consciousness in mouse.

    Science.gov (United States)

    Hwang, Eunjin; Kim, Seunghwan; Shin, Hee-Sup; Choi, Jee Hyun

    2010-04-30

    In consciousness or anesthesia studies, pinpointing the precise moment of consciousness or anesthetic transition has been challenging because of the variable lag time between a treatment and its induced response. Here, we describe a novel behavioral method, a forced walking test, which pinpoints the moment of the anesthetic-induced loss of motion (LOM) without handling the animals manually. The mouse is forced to walk on a treadmill, and an anesthetic drug is administered into the peritoneum via a previously secured injection route. The physical activity and the angle of head posture are tracked using a motion sensor preinstalled on the animal's head. The moments of LOM and recovery of motion (ROM) are identified from the physical activity parameters obtained by the sensor. Comparison of our method with the conventional loss-of-righting-reflex assay showed that the time point of LOM was not significantly different between the two methods when examined with two different types of anesthetic agents, propofol and ketamine/xylazine cocktail. In addition, the electrophysiological signals simultaneously acquired in the cortex and the thalamus of the mouse during the forced walking test showed that the brain rhythms induced by ketamine/xylazine anesthesia were generated and terminated in a time-locked manner with respect to LOM and ROM, respectively. In conclusion, the forced walking test allows an objective and precise detection of anesthetic-induced LOM, as well as ROM during awakening from anesthesia, in test animals.

  14. Identification of differentially expressed radiation-induced genes in cervix carcinoma cells using suppression subtractive hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jun Sang; Lee, Young Sook; Lee, Jeung Hoon; Lee, Woong Hee; Seo, Eun Young; Cho, Moon June [Chungnam National University, Daejeon (Korea, Republic of)

    2005-03-15

    A number of genes and their products are induced early or late following exposure of cells to ionizing radiation. These radiation-induced genes have various effects of irradiated cells and tissues. Suppression subtractive hybridization (SSH) based on PCR was used to identify the differentially expressed genes by radiation in cervix carcinoma cells. Total RNA and poly (A){sup +} mRNA were isolated from irradiated and non-irradiated HeLa cells. Forward-and reverse-subtracted cDNA libraries were constructed using SSH. Eighty-eight clones of each were used to randomly select differentially expressed genes using reverse Northern blotting (dot blot analysis). Northern blotting was used to verify the screened genes. Of the 176 clones, 10 genes in the forward-subtracted library and 9 genes in the reverse-subtracted library were identified as differentially expressed radiation-induced genes by PCR-select differential screening. Three clones from the forward-subtracted library were confirmed by Northern blotting, and showed increased expression in a dose-dependent manner, including a telomerase catalytic subunit and sodium channel-like protein gene, and an ESTs (expressed sequence tags) gene. We identified differentially expressed radiation-induced genes with low-abundance genes with SSH, but further characterization of theses genes are necessary to clarify the biological functions of them.

  15. A linear description of shortening induced changes in isometric length-force characteristics of rat muscle

    OpenAIRE

    Meijer, K; Grootenboer, H.J.; Koopman, H.F.J.M.; Huijing, P.A.

    1996-01-01

    Active muscle shortening reduces the isometric force potential of muscle. This observation indicates that the isometric length-force characteristics are altered during muscle shortening. Post-shortening decrease in isometric force depends on starting length, shortening amplitude and shortening velocity. In the present study, post-shortening decrease in isometric force was determined after isokinetic contractions with various shortening amplitudes initiated from different lengths of rat medial...

  16. A linear description of shortening induced changes in isometric length-force characteristics of rat muscle

    NARCIS (Netherlands)

    Meijer, K.; Grootenboer, H.J.; Koopman, H.F.J.M.; Huijing, P.A.

    1996-01-01

    Active muscle shortening reduces the isometric force potential of muscle. This observation indicates that the isometric length-force characteristics are altered during muscle shortening. Post-shortening decrease in isometric force depends on starting length, shortening amplitude and shortening veloc

  17. Shortening induced effects on force (re)development in pig urinary smooth muscle

    NARCIS (Netherlands)

    E. van Asselt (Els); J.J.M. Pel (Johan); R. van Mastrigt (Ron)

    2007-01-01

    textabstractIntroduction: When muscle is allowed to shorten during an active contraction, the maximum force that redevelops after shortening is smaller than the isometric force at the same muscle length without prior shortening. We studied the course of force redevelopment after shortening in smooth

  18. The effect of temperature dependent tissue parameters on acoustic radiation force induced displacements

    Science.gov (United States)

    Suomi, Visa; Han, Yang; Konofagou, Elisa; Cleveland, Robin O.

    2016-10-01

    Multiple ultrasound elastography techniques rely on acoustic radiation force (ARF) in monitoring high-intensity focused ultrasound (HIFU) therapy. However, ARF is dependent on tissue attenuation and sound speed, both of which are also known to change with temperature making the therapy monitoring more challenging. Furthermore, the viscoelastic properties of tissue are also temperature dependent, which affects the displacements induced by ARF. The aim of this study is to quantify the temperature dependent changes in the acoustic and viscoelastic properties of liver and investigate their effect on ARF induced displacements by using both experimental methods and simulations. Furthermore, the temperature dependent viscoelastic properties of liver are experimentally measured over a frequency range of 0.1-200 Hz at temperatures reaching 80 °C, and both conventional and fractional Zener models are used to fit the data. The fractional Zener model was found to fit better with the experimental viscoelasticity data with respect to the conventional model with up to two orders of magnitude lower sum of squared errors (SSE). The characteristics of experimental displacement data were also seen in the simulations due to the changes in attenuation coefficient and lesion development. At low temperatures before thermal ablation, attenuation was found to affect the displacement amplitude. At higher temperature, the decrease in displacement amplitude occurs approximately at 60-70 °C due to the combined effect of viscoelasticity changes and lesion growth overpowering the effect of attenuation. The results suggest that it is necessary to monitor displacement continuously during HIFU therapy in order to ascertain when ablation occurs.

  19. Protocol: using virus-induced gene silencing to study the arbuscular mycorrhizal symbiosis in Pisum sativum

    DEFF Research Database (Denmark)

    Grønlund, Mette; Olsen, Anne; Johansen, Elisabeth

    2010-01-01

    Virus-induced gene silencing (VIGS) is an alternative reverse genetics tool for silencing of genes in some plants, which are difficult to transform. The pea early-browning virus (PEBV) has been developed as a VIGS vector and used in pea for functional analysis of several genes. However, the avail......Virus-induced gene silencing (VIGS) is an alternative reverse genetics tool for silencing of genes in some plants, which are difficult to transform. The pea early-browning virus (PEBV) has been developed as a VIGS vector and used in pea for functional analysis of several genes. However......, the available PEBV-VIGS protocols are inadequate for studying genes involved in the symbiosis with arbuscular mycorrhizal fungi (AMF). Here we describe a PEBV-VIGS protocol suitable for reverse genetics studies in pea of genes involved in the symbiosis with AMF and show its effectiveness in silencing genes...... involved in the early and late stages of AMF symbiosis....

  20. Heavy-ion induced modification of lithium fluoride observed by scanning force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, A.; Neumann, R.; Schwartz, K.; Steckenreiter, T.; Trautmann, C.

    1997-12-01

    To study ion-induced damages in single crystals of lithium fluoride with scanning force microscopy (SFM), samples were irradiated with several heavy-ion species of a kinetic energy of 11.4 MeV per nucleon at the linear accelerator UNILAC of GSI. As concluded from a previous analysis of ion tracks in LiF by optical absorption spectroscopy and small-angle X-ray scattering, single point defects occur in a track halo with a radius of about 15-30 nm, whereas defect aggregates are formed in a track core region possessing a radius of only about 1-2 nm. These aggregates can be attacked by chemical etching if the energy loss along the ion trajectory surpasses a critical value of about 1 keV/Aa. SFM images of etched as well as unetched sample surfaces revealed new damage characteristics: Etched ion track profiles directed parallel to the ion trajectories exhibit a sequence of single etch pits with an average distance of about 140 nm. After exposure to heavy-ion irradiation at normal incidence, the unetched LiF surface is covered with round hillocks with a mean diameter of 55(8) nm and heights in the order of 3 nm. (orig.)

  1. Turbulence-induced melting of a nonequilibrium vortex crystal in a forced thin fluid film

    CERN Document Server

    Perlekar, Prasad

    2009-01-01

    To develop an understanding of recent experiments on the turbulence-induced melting of a periodic array of vortices in a thin fluid film, we perform a direct numerical simulation of the two-dimensional Navier-Stokes equations forced such that, at low Reynolds numbers, the steady state of the film is a square lattice of vortices. We find that, as we increase the Reynolds number, this lattice undergoes a series of nonequilibrium phase transitions, first to a crystal with a different reciprocal lattice and then to a sequence of crystals that oscillate in time. Initially the temporal oscillations are periodic; this periodic behaviour becomes more and more complicated, with increasing Reynolds number, until the film enters a spatially disordered nonequilibrium statistical steady that is turbulent. We study this sequence of transitions by using fluid-dynamics measures, such as the Okubo-Weiss parameter that distinguishes between vortical and extensional regions in the flow, ideas from nonlinear dynamics, e.g., \\Poi...

  2. Depletion forces induce visco-elasto-capillary thinning of non-Brownian suspensions

    Science.gov (United States)

    Harich, R.; Deblais, A.; Colin, A.; Kellay, H.

    2016-06-01

    Droplet pinch-off, which occurs when a drop of liquid detaches from a capillary, can be strongly modified in the presence of complex fluids such as polymer solutions and suspensions giving rise to long and slender filaments that thin slowly in time. While for polymers, the molecular conformations of the molecules in the filament are responsible for such a behavior, in suspensions the mechanisms at play remain to be deciphered. Here we show, experimentally, that while liquid bridges of non-Brownian suspensions of moderate concentrations have a thinning behavior very close to that of the solvent, the addition of short-chain polymers inducing depletion attractions between the particles in the suspension changes the thinning dynamics and gives rise to exponential thinning in time. The characteristic time of this dynamics increases with polymer concentration and therefore the intensity of the depletion forces at play. The tunability of this dynamics may be important for injket and 3D printing applications where short rupture times are sought for or in other situations where drop formation has to be minimized or inhibited.

  3. +GZ-induced neck injuries in Royal Australian Air Force fighter pilots.

    Science.gov (United States)

    Newman, D G

    1997-06-01

    +GZ-induced neck injuries are a relatively common occurrence in pilots of high performance fighter aircraft. We surveyed 52 fighter pilots from the Royal Australian Air Force Base at Williamtown via an anonymous questionnaire in order to determine the prevalence and operational significance of these injuries. The pilots flew either the F/A-18 Hornet or the MB326H Macchi. Of the respondents, 44 reported having had a neck injury under +GZ. A higher rate was reported in pilots of the F/A-18. Most of these injuries were simple muscle sprains. There were 20 pilots who reported their neck injury as having interfered with mission completion. Only 12 pilots reported doing any regular neck strengthening exercises, while 33 pilots reported doing preflight neck stretches immediately prior to high +GZ exposure. There were 14 pilots who sought medical attention for their injury, with 9 being taken off flight status for an average of 2 weeks. Air combat maneuvering sorties and the "check six" head position were identified as causal factors by most pilots. This study demonstrates the operational significance of these injuries, and highlights the need for more research into this important aerospace medicine issue.

  4. Activation-Induced Cytidine Deaminase Contributes to Pancreatic Tumorigenesis by Inducing Tumor-Related Gene Mutations.

    Science.gov (United States)

    Sawai, Yugo; Kodama, Yuzo; Shimizu, Takahiro; Ota, Yuji; Maruno, Takahisa; Eso, Yuji; Kurita, Akira; Shiokawa, Masahiro; Tsuji, Yoshihisa; Uza, Norimitsu; Matsumoto, Yuko; Masui, Toshihiko; Uemoto, Shinji; Marusawa, Hiroyuki; Chiba, Tsutomu

    2015-08-15

    Pancreatic ductal adenocarcinoma (PDAC) develops via an accumulation of various gene mutations. The mechanism underlying the mutations in PDAC development, however, is not fully understood. Recent insight into the close association between the mutation pattern of various cancers and specific mutagens led us to investigate the possible involvement of activation-induced cytidine deaminase (AID), a DNA editing enzyme, in pancreatic tumorigenesis. Our immunohistochemical findings revealed AID protein expression in human acinar ductal metaplasia, pancreatic intraepithelial neoplasia, and PDAC. Both the amount and intensity of the AID protein expression increased with the progression from precancerous to cancerous lesions in human PDAC tissues. To further assess the significance of ectopic epithelial AID expression in pancreatic tumorigenesis, we analyzed the phenotype of AID transgenic (AID Tg) mice. Consistent with our hypothesis that AID is involved in the mechanism of the mutations underlying pancreatic tumorigenesis, we found precancerous lesions developing in the pancreas of AID Tg mice. Using deep sequencing, we also detected Kras and c-Myc mutations in our analysis of the whole pancreas of AID Tg mice. In addition, Sanger sequencing confirmed the presence of Kras, c-Myc, and Smad4 mutations, with the typical mutational footprint of AID in precancerous lesions in AID Tg mice separated by laser capture microdissection. Taken together, our findings suggest that AID contributes to the development of pancreatic precancerous lesions by inducing tumor-related gene mutations. Our new mouse model without intentional manipulation of specific tumor-related genes provides a powerful system for analyzing the mutations involved in PDAC.

  5. Cyclic AMP-inducible genes respond uniformly to seasonal lighting conditions in the rat pineal gland.

    Science.gov (United States)

    Spessert, R; Gupta, B B P; Rohleder, N; Gerhold, S; Engel, L

    2006-12-01

    The encoding of photoperiodic information ensues in terms of the daily profile in the expression of cyclic AMP (cAMP)-inducible genes such as the arylalkylamine N-acetyltransferase (AA-NAT) gene that encodes the rate-limiting enzyme in melatonin formation. In the present study, we compared the influence of the photoperiodic history on the cAMP-inducible genes AA-NAT, inducible cyclic AMP early repressor (ICER), fos-related antigen-2 (FRA-2), mitogen-activated protein kinase phosphatase-1 (MKP-1), nerve growth factor inducible gene-A (NGFI-A) and nerve growth factor inducible gene-B (NGFI-B) in the pineal gland of rats. For this purpose, we monitored the daily profiles of each gene in the same pineal gland under a long (light/dark 16:8) and a short (light/dark 8:16) photoperiod by measuring the respective mRNA amounts by real-time polymerase chain reaction analysis. We found that, for all genes under investigation, the duration of increased nocturnal expression is lengthened and, in relation to light onset, the nocturnal rise is earlier under the long photoperiod (light/dark 16:8). Furthermore, with the exception of ICER, all other cAMP-inducible genes tend to display higher maximum expression under light/dark 8:16 than under light/dark 16:8. Photoperiod-dependent changes persist for all of the cAMP-inducible genes when the rats are kept for two cycles under constant darkness. Therefore, all cAMP-inducible genes are also influenced by the photoperiod of prior entrained cycles. Our study indicates that, despite differences regarding the expressional control and the temporal phasing of the daily profile, cAMP-inducible genes are uniformly influenced by photoperiodic history in the rat pineal gland.

  6. The effect of heating and cooling on time course of voluntary and electrically induced muscle force variation.

    Science.gov (United States)

    Brazaitis, Marius; Skurvydas, Albertas; Vadopalas, Kazys; Daniusevičiūtė, Laura; Senikienė, Zibuoklė

    2011-01-01

    The aim of this study was to investigate the effect of heating and cooling on time course of voluntary and electrically induced muscle force variation. Ten volunteers performed 50 maximal voluntary and electrically induced contractions of the knee extensors at an angle of 120 degrees under the control conditions and after passive lower body heating and cooling in the control, heating, and cooling experiments. Peak torque, torque variation, and half-relaxation time were assessed during the exercise. Passive lower body heating increased muscle and core temperatures, while cooling lowered muscle temperature, but did not affect core temperature. We observed significantly lower muscle fatigue during voluntary contraction compared with electrically induced contractions. Body heating (opposite to cooling) increased involuntarily induced muscle force, but caused greater electrically induced muscle fatigue. In the middle of the exercise, the coefficient of correlation for electrically induced muscle torque decreased significantly as compared with the beginning of the exercise, while during maximal voluntary contractions, this relation for torque remained significant until the end of the exercise. It was shown that time course of voluntary contraction was more stable than in electrically induced contractions.

  7. Dexamethasone-Inducible Green Fluorescent Protein Gene Expression in Transgenic Plant Cells

    Institute of Scientific and Technical Information of China (English)

    Wei Tang; Hilary Collver; Katherine Kinken

    2004-01-01

    Genomic research has made a large number of sequences of novel genes or expressed sequence tags available. To investigate functions of these genes, a system for conditional control of gene expression would be a useful tool. Inducible transgene expression that uses green fluorescent protein gene (gfp) as a reporter gene has been investigated in transgenic cell lines of cotton (COT; Gossypium hirsutum L.), Fraser fir [FRA; Abies fraseri (Pursh) Poir], Nordmann fir (NOR; Abies nordmanniana Lk.), and rice (RIC; Oryza sativa L. Cv. Radon). Transgenic cell lines were used to test the function of the chemical inducer dexamethasone. Inducible transgene expression was observed with fluorescence and confocal microscopy, and was confirmed by northern blot analyses. Dexamethasone at 5 mg/L induced gfp expression to the nearly highest level 48 h after treatment in COT, FRA, NOR, and RIC. Dexamethasone at 10 mg/L inhibited the growth of transgenic cells in FRA and NOR, but not COT and RIC. These results demonstrated that concentrations of inducer for optimum inducible gene expression system varied among transgenic cell lines. The inducible gene expression system described here was very effective and could be valuable in evaluating the function of novel gene.

  8. Accelerated greenhouse gases versus slow insolation forcing induced climate changes in southern South America since the Mid-Holocene

    Science.gov (United States)

    Berman, Ana Laura; Silvestri, Gabriel E.; Rojas, Maisa; Tonello, Marcela S.

    2017-01-01

    This paper is a pioneering analysis of past climates in southern South America combining multiproxy reconstructions and the state-of-the-art CMIP5/PMIP3 paleoclimatic models to investigate the time evolution of regional climatic conditions from the Mid-Holocene (MH) to the present. This analysis allows a comparison between the impact of the long term climate variations associated with insolation changes and the more recent effects of anthropogenic forcing on the region. The PMIP3 multimodel experiments suggest that changes in precipitation over almost all southern South America between MH and pre-industrial (PI) times due to insolation variations are significantly larger than those between PI and the present, which are due to changes in greenhouse gas concentrations. Anthropogenic forcing has been particularly intense over western Patagonia inducing reduction of precipitation in summer, autumn and winter as a consequence of progressively weaker westerly winds over the region, which have moved further poleward, between ca. 35-55°S and have become stronger south of about 50°S. Orbital variations between the MH to the PI period increased insolation over southern South America during summer and autumn inducing warmer conditions in the PI, accentuated by the effect of anthropogenic forcing during the last century. On the other hand, changes in orbital parameters from the MH to the PI period reduced insolation during winter and spring inducing colder conditions, which have been reversed by the anthropogenic forcing.

  9. Gene Therapy Induces Antigen-Specific Tolerance in Experimental Collagen-Induced Arthritis.

    Directory of Open Access Journals (Sweden)

    Sara Tengvall

    Full Text Available Here, we investigate induction of immunological tolerance by lentiviral based gene therapy in a mouse model of rheumatoid arthritis, collagen II-induced arthritis (CIA. Targeting the expression of the collagen type II (CII to antigen presenting cells (APCs induced antigen-specific tolerance, where only 5% of the mice developed arthritis as compared with 95% of the control mice. In the CII-tolerized mice, the proportion of Tregs as well as mRNA expression of SOCS1 (suppressors of cytokine signaling 1 increased at day 3 after CII immunization. Transfer of B cells or non-B cell APC, as well as T cells, from tolerized to naïve mice all mediated a certain degree of tolerance. Thus, sustainable tolerance is established very early during the course of arthritis and is mediated by both B and non-B cells as APCs. This novel approach for inducing tolerance to disease specific antigens can be used for studying tolerance mechanisms, not only in CIA but also in other autoimmune diseases.

  10. Preventive Effects of Forced Exercise against Alcohol-induced Physical Dependency and Reduction of Pain Perception Threshold

    Directory of Open Access Journals (Sweden)

    Majid Motaghinejad

    2014-01-01

    Full Text Available Background: Treatment of postabstinence syndrome of alcohol is one of the major strategies of alcoholism treatment. Exercise can be modulated major brain pathways such as a reward system and pain perception centers. The aim of this study was to evaluation the effects of forced exercise in the management of alcohol dependence and pain perception alteration which induced by alcoholism. Methods: 72 adult male rats were divided into 2 major groups: (1 40 of them was divided into groups of positive control (alcohol dependent negative control and alcohol dependent groups under treatment by forced exercise, diazepam (0.4 mg/kg and forced exercise in combination with diazepam and alcohol withdrawal signs, and blood cortisols, were measured in this groups. (2 32 rats were divided into control, alcohol dependent (without treatment, and alcohol-dependent groups under treatment by forced exercise or indometacin (5 mg/kg and then pain perception was assessed by using writhing test, tail-flick and hot plate test. Results: Forced exercise, diazepam, and their combinations significantly attenuates withdrawal syndrome to 20 ± 2, 22 ± 1.3 and 16 ± 2 and blood cortisol level to 6.8 ± 1.3,7.9 ± 1.2 and 5.8 ± 1.1, respectively, in comparison with the positive control group (P < 0.05 and P < 0.001. In alcohol dependent animal under treatment by forced exercise, pain response significantly inhibited with 37%, 57% and 38% decreases in writhing test, hot plate, and tail-flick test, respectively, in comparison with alcohol dependent (without treatment group (P < 0.05. Conclusions: This study suggested that forced exercise can be useful as adjunct therapy in alcoholism patient and also can be effective in modulation of pain threshold reduction that was induced by alcohol dependency.

  11. Modelling and experimental verification of tip-induced polarization in Kelvin probe force microscopy measurements on dielectric surfaces

    DEFF Research Database (Denmark)

    Nielsen, Dennis Achton; Popok, Vladimir; Pedersen, Kjeld

    2015-01-01

    Kelvin probe force microscopy is a widely used technique for measuring surface potential distributions on the micro- and nanometer scale. The data are, however, often analyzed qualitatively, especially for dielectrics. In many cases, the phenomenon of polarization and its influence on the measured...... signals is disregarded leading to misinterpretation of the results. In this work, we present a model that allows prediction of the surface potential on a metal/polymer heterostructure as measured by Kelvin probe force microscopy by including the tip-induced polarization of the dielectric that arises...

  12. The jet impact force of laser-induced bubble under the water-film with different thickness

    Science.gov (United States)

    Li, Beibei; Wang, Bingyang; Liu, Xiumei; He, Jie; Lu, Jian

    2015-05-01

    The effect of water-film on the laser-induced bubble was investigated by a piezoelectric ceramic transducer (PZT) sensor. Both of the collapse time and liquid-jet impact force of the bubble under the water-film were obtained, and the experiments were also completed in different laser energy. The collapse time increase with the thickness of the waterfilm, but the liquid-jet impact force decrease. We consider that the collapse time was affected by both of the rigid boundary and surface, and the increasing of the collapse time is the reason the decreasing of the liquid-jet impact force. The velocity of bubble wall is lower with the longer collapse time for the uniform bubbles energy, so the liquid-jet impact force is lower. For the other reasons, more laser energy would be absorbed by the thicker water-film, but the water was also splashed for the thinner water-film. So, for the thinner water-film, the bubble energy is higher, the liquidjet impact force is higher, but the maximal radius is smaller because of the splash process. In the other hand, both of the collapse time and the liquid-jet impact force are increase with the laser energy. These researches are useful for the laser processing under water.

  13. Shortening induced effects on force (re)development in pig urinary smooth muscle

    OpenAIRE

    van Asselt, Els; Pel, Johan; van Mastrigt, Ron

    2007-01-01

    textabstractIntroduction: When muscle is allowed to shorten during an active contraction, the maximum force that redevelops after shortening is smaller than the isometric force at the same muscle length without prior shortening. We studied the course of force redevelopment after shortening in smooth muscle to unravel the mechanism responsible for this deactivation. Method: In a first series of measurements the shortening velocity was varied resulting in different shortening amplitudes. In a s...

  14. Modeling of the intermolecular Force-Induced Adhesion in Freestanding Nanostructures Made of Nano-beams

    Directory of Open Access Journals (Sweden)

    Alireza Yekrangi

    2015-11-01

    Full Text Available Among the intermolecular interactions, the Casimir and van der Waals forces are the most important forces that highly affect the behavior of nanostructures. This paper studies the effect of such forces on the adhesion of cantilever freestanding nanostructures. The nanostructures are made of a freestanding nano-beam which is suspended between two upper and lower conductive surfaces. The linear spring model is applied to derive the elastic force. The Lumped Parameter Model (LPM is used to obtain constitutive equations of the systems. The maximum length of the nano-beam which prevents the adhesion is computed. Results of this study are useful for design and development of miniature devices.

  15. Asian summer monsoon anomalies induced by aerosol direct forcing: the role of the Tibetan Plateau

    Energy Technology Data Exchange (ETDEWEB)

    Lau, K.M. [NASA Goddard Space Flight Center, Laboratory for Atmospheres, Greenbelt, MD (United States); Kim, M.K. [Kongju National University, Department of Atmospheric Science, Gongju (Korea); Kim, K.M. [Science Systems and Applications, Inc, Lanham, MD (United States)

    2006-06-15

    In this paper we present results of a numerical study using the NASA finite-volume GCM to elucidate a plausible mechanism for aerosol impact on the Asian summer monsoon involving interaction with physical processes over the Tibetan Plateau (TP). During the pre-monsoon season of March-April, dusts from the deserts of western China, Afghanistan/Pakistan, and the Middle East are transported into and stacked up against the northern and southern slopes of the TP. The absorption of solar radiation by dust heats up the elevated surface air over the slopes. On the southern slopes, the atmospheric heating is reinforced by black carbon from local emission. The heated air rises via dry convection, creating a positive temperature anomaly in the mid-to-upper troposphere over the TP relative to the region to the south. In May through early June in a manner akin to an ''elevated heat pump'', the rising hot air forced by the increasing heating in the upper troposphere, draws in warm and moist air over the Indian subcontinent, setting the stage for the onset of the South Asia summer monsoon. Our results suggest that increased dust loading coupled with black carbon emission from local sources in northern India during late spring may lead to an advance of the rainy periods and subsequently an intensification of the Indian summer monsoon. The enhanced rainfall over India is associated with the development of an aerosol-induced large-scale sea level pressure anomaly pattern, which causes the East Asia (Mei-yu) rain belt to shift northwestward, suppressing rainfall over East Asia and the adjacent oceanic regions. (orig.)

  16. Coordinate gene regulation by fimbriae-induced signal transduction

    DEFF Research Database (Denmark)

    Schembri, Mark; Klemm, Per

    2001-01-01

    whether fimbriae expression can affect expression of other genes, Analysis of gene expression in two E.coli strains, differing in the fim locus, indicated the flu gene to be affected. The flu gene encodes the antigen 43 (Ag43) surface protein, specifically involved in bacterial aggregation...... of Ag43 production. No effect was observed in an oxyR mutant. We conclude that fimbriae expression per se constitutes a signal transduction mechanism that affects a number of unrelated genes via the thiol-disulfide status of OxyR. Thus, phase variation in fimbrial expression is coordinated...

  17. Glomerulonephritis-induced changes in kidney gene expression in rats

    Directory of Open Access Journals (Sweden)

    Mira Pavkovic

    2015-12-01

    Full Text Available We investigated a glomerulonephritis (GN model in rats induced by nephrotoxic serum (NTS which contains antibodies against the glomerular basement membrane (GBM. The anti-GBM GN model in rats is widely used since its biochemical and histopathological characteristics are similar to crescentic nephritis and Goodpasture's disease in humans (Pusey, 2003 [2]. Male Wistar Kyoto (WKY and Sprague–Dawley (SD rats were dosed once with 1, 2.5 and 5 ml/kg nephrotoxic serum (NTS or 1.5 and 5 ml/kg NTS, respectively. GN and tubular damage were observed histopathologically in all treated rats after 14 days. To obtain insight into molecular processes during GN pathogenesis, mRNA expression was investigated in WKY and SD kidneys using Affymetrix's GeneChip Rat genome 230_2.0 arrays (GSE64265. The immunopathological processes during GN are still not fully understood and likely involve both innate and adaptive immunity. In the present study, several hundred mRNAs were found deregulated, which functionally were mostly associated with inflammation and regeneration. The β-chain of the major histocompatibility complex class II RT1.B (Rt1-Bb and complement component 6 (C6 were identified as two mRNAs differentially expressed between WKY and SD rat strains which could be related to known different susceptibilities to NTS of different rat strains; both were increased in WKY and decreased in SD rats (Pavkovic et al., 2015 [1]. Increased Rt1-Bb expression in WKY rats could indicate a stronger and more persistent cellular reaction of the adaptive immune system in this strain, in line with findings indicating adaptive immune reactions during GN. The complement cascade is also known to be essential for GN development, especially terminal cascade products like C6.

  18. Veratrine blocks the lamotrigine-induced swimming increase and immobility decrease in the modified forced swimming test.

    Science.gov (United States)

    Codagnone, F T; Consoni, F T; Rodrigues, A L S; Vital, M A B F; Andreatini, R

    2007-08-15

    Lamotrigine exhibits an anti-immobility effect in the modified forced swimming test, increasing swimming and climbing, behaviors that are related to serotonergic and noradrenergic effects, respectively. However, these effects could be secondary to lamotrigine blockade of Na(+) sensitive channel. Thus, this study investigated the influence of veratrine (0.1 mg/kg, ip, 10 min before each lamotrigine administration), an Na(+) channel activator, in the effect of lamotrigine (20 mg/kg, ip, 24, 5, 1 h before the test session) in the modified forced swimming test. Veratrine pre-treatment blocked lamotrigine-induced immobility decrease and swimming increase but it did not change the effect of lamotrigine on climbing. These results suggest that the serotonergic effect of lamotrigine in the modified forced swimming test is dependent on Na(+) voltage sensitive channel blockade, whereas its noradrenergic effect is not.

  19. Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene.

    Science.gov (United States)

    van der Kop, D A; Schuyer, M; Pinas, J E; van der Zaal, B J; Hooykaas, P J

    1999-03-01

    Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the beta-glucuronidase (gusA) reporter gene. Subsequently, seeds were treated with EMS to obtain mutants in which both reporter gene fusions were up-regulated. Northern analysis showed that the mRNA level of a related, endogenous auxin-inducible GST gene of Arabidopsis was increased in some of these mutants as well. Two of the gup (GST up-regulated) mutants were characterized in more detail and roughly mapped. Both had epinastic cotyledons and leaves, a phenotype that turned out to be linked to the gup mutation.

  20. Mechanics of the IL2RA Gene Activation Revealed by Modeling and Atomic Force Microscopy

    OpenAIRE

    Pascale Milani; Monique Marilley; Albert Sanchez-Sevilla; Jean Imbert; Cédric Vaillant; Françoise Argoul; Jean-Marc Egly; José Rocca-Serra; Alain Arneodo

    2011-01-01

    Transcription implies recruitment of RNA polymerase II and transcription factors (TFs) by DNA melting near transcription start site (TSS). Combining atomic force microscopy and computer modeling, we investigate the structural and dynamical properties of the IL2RA promoter and identify an intrinsically negative supercoil in the PRRII region (containing Elf-1 and HMGA1 binding sites), located upstream of a curved DNA region encompassing TSS. Conformational changes, evidenced by time-lapse studi...

  1. Hybrid electro-optical stimulation of the rat sciatic nerve induces force generation in the plantarflexor muscles

    Science.gov (United States)

    Duke, Austin R.; Peterson, Erik; Mackanos, Mark A.; Atkinson, James; Tyler, Dustin; Jansen, E. Duco

    2012-12-01

    Objective. Optical methods of neural activation are becoming important tools for the study and treatment of neurological disorders. Infrared nerve stimulation (INS) is an optical technique exhibiting spatially precise activation in the native neural system. While this technique shows great promise, the risk of thermal damage may limit some applications. Combining INS with traditional electrical stimulation, a method known as hybrid electro-optical stimulation, reduces the laser power requirements and mitigates the risk of thermal damage while maintaining spatial selectivity. Here we investigate the capability of inducing force generation in the rat hind limb through hybrid stimulation of the sciatic nerve. Approach. Hybrid stimulation was achieved by combining an optically transparent nerve cuff for electrical stimulation and a diode laser coupled to an optical fiber for infrared stimulation. Force generation in the rat plantarflexor muscles was measured in response to hybrid stimulation with 1 s bursts of pulses at 15 and 20 Hz and with a burst frequency of 0.5 Hz. Main results. Forces were found to increase with successive stimulus trains, ultimately reaching a plateau by the 20th train. Hybrid evoked forces decayed at a rate similar to the rate of thermal diffusion in tissue. Preconditioning the nerve with an optical stimulus resulted in an increase in the force response to both electrical and hybrid stimulation. Histological evaluation showed no signs of thermally induced morphological changes following hybrid stimulation. Our results indicate that an increase in baseline temperature is a likely contributor to hybrid force generation. Significance. Extraneural INS of peripheral nerves at physiologically relevant repetition rates is possible using hybrid electro-optical stimulation.

  2. Evidence for non-conservative current-induced forces in the breaking of Au and Pt atomic chains.

    Science.gov (United States)

    Sabater, Carlos; Untiedt, Carlos; van Ruitenbeek, Jan M

    2015-01-01

    This experimental work aims at probing current-induced forces at the atomic scale. Specifically it addresses predictions in recent work regarding the appearance of run-away modes as a result of a combined effect of the non-conservative wind force and a 'Berry force'. The systems we consider here are atomic chains of Au and Pt atoms, for which we investigate the distribution of break down voltage values. We observe two distinct modes of breaking for Au atomic chains. The breaking at high voltage appears to behave as expected for regular break down by thermal excitation due to Joule heating. However, there is a low-voltage breaking mode that has characteristics expected for the mechanism of current-induced forces. Although a full comparison would require more detailed information on the individual atomic configurations, the systems we consider are very similar to those considered in recent model calculations and the comparison between experiment and theory is very encouraging for the interpretation we propose.

  3. RNA splicing regulates the temporal order of TNF-induced gene expression.

    Science.gov (United States)

    Hao, Shengli; Baltimore, David

    2013-07-16

    When cells are induced to express inflammatory genes by treatment with TNF, the mRNAs for the induced genes appear in three distinct waves, defining gene groups I, II, and III, or early, intermediate, and late genes. To examine the basis for these different kinetic classes, we have developed a PCR-based procedure to distinguish pre-mRNAs from mRNAs. It shows that the three groups initiate transcription virtually simultaneously but that delays in splicing characterize groups II and III. We also examined the elongation times, concluding that pre-mRNA synthesis is coordinate but splicing differences directly regulate the timing of mRNA production.

  4. Relationship between gene responses and symptoms induced by Rice grassy stunt virus

    Directory of Open Access Journals (Sweden)

    Kouji eSatoh

    2013-10-01

    Full Text Available Rice grassy stunt virus (RGSV is a serious threat to rice production in Southeast Asia. RGSV is a member of the genus Tenuivirus, and it induces leaf yellowing, stunting, and excess tillering on rice plants. Here we examined gene responses of rice to RGSV infection to gain insight into the gene responses which might be associated with the disease symptoms. The results indicated that 1 many genes related to cell wall synthesis and chlorophyll synthesis were predominantly suppressed by RGSV infection; 2 RGSV infection induced genes associated with tillering process; 3 RGSV activated genes involved in inactivation of gibberellic acid and indole-3-acetic acid ; and 4 the genes for strigolactone signaling were suppressed by RGSV. These results suggest that these gene responses to RGSV infection account for the excess tillering specific to RGSV infection as well as other symptoms by RGSV, such as stunting and leaf chlorosis.

  5. Rationale for developing new virus vectors to analyze gene function in grasses through virus-induced gene silencing.

    Science.gov (United States)

    Ramanna, Hema; Ding, Xin Shun; Nelson, Richard S

    2013-01-01

    The exploding availability of genome and EST-based sequences from grasses requires a technology that allows rapid functional analysis of the multitude of genes that these resources provide. There are several techniques available to determine a gene's function. For gene knockdown studies, silencing through RNAi is a powerful tool. Gene silencing can be accomplished through stable transformation or transient expression of a fragment of a target gene sequence. Stable transformation in rice, maize, and a few other species, although routine, remains a relatively low-throughput process. Transformation in other grass species is difficult and labor-intensive. Therefore, transient gene silencing methods including Agrobacterium-mediated and virus-induced gene silencing (VIGS) have great potential for researchers studying gene function in grasses. VIGS in grasses already has been used to determine the function of genes during pathogen challenge and plant development. It also can be used in moderate-throughput reverse genetics screens to determine gene function. However, the number of viruses modified to serve as silencing vectors in grasses is limited, and the silencing phenotype induced by these vectors is not optimal: the phenotype being transient and with moderate penetration throughout the tissue. Here, we review the most recent information available for VIGS in grasses and summarize the strengths and weaknesses in current virus-grass host systems. We describe ways to improve current virus vectors and the potential of other grass-infecting viruses for VIGS studies. This work is necessary because VIGS for the foreseeable future remains a higher throughput and more rapid system to evaluate gene function than stable transformation.

  6. Cooperativity in Thermal and Force-Induced Protein Unfolding: Integration of Crack Propagation and Network Elasticity Models

    Science.gov (United States)

    Srivastava, Amit; Granek, Rony

    2013-03-01

    We investigate force-induced and temperature-induced unfolding of proteins using the combination of a Gaussian network model and a crack propagation model based on “bond”-breaking independent events. We assume the existence of threshold values for the mean strain and strain fluctuations that dictate bond rupture. Surprisingly, we find that this stepwise process usually leads to a few cooperative, first-order-like, transitions in which several bonds break simultaneously, reminiscent of the “avalanches” seen in disordered networks.

  7. Ultra-short laser pulse ablation using shear-force feedback: Femtosecond laser induced breakdown spectroscopy feasibility study

    Energy Technology Data Exchange (ETDEWEB)

    Samek, Ota [ISAS-Institute for Analytical Sciences, Bunsen-Kirchhoff Str.11, 44139 Dortmund (Germany)]. E-mail: samek@ansci.de; Kurowski, Andre [ISAS-Institute for Analytical Sciences, Bunsen-Kirchhoff Str.11, 44139 Dortmund (Germany); Kittel, Silke [ISAS-Institute for Analytical Sciences, Bunsen-Kirchhoff Str.11, 44139 Dortmund (Germany); Kukhlevsky, Sergei [Institute of Physics, University of Pecs, Ifjusag u. 6, Pecs 7624 (Hungary); Hergenroeder, Roland [ISAS-Institute for Analytical Sciences, Bunsen-Kirchhoff Str.11, 44139 Dortmund (Germany)

    2005-08-31

    This work reports on a feasibility study of proximity ablation using femtosecond pulses. Ultra-short pulses were launched to a bare tapered optical fiber and delivered to the sample. The tip-sample distance was controlled by means of shear-force feedback. Consequently, ablation craters with submicrometer dimensions were obtained. Potential analytical applications for Laser Induced Breakdown Spectroscopy (LIBS) technique, such as e.g. inclusions in steel or bio cells, are suggested.

  8. Regulatory gene mutation: a driving force behind group a Streptococcus strain- and serotype-specific variation.

    Science.gov (United States)

    Sarkar, Poulomee; Sumby, Paul

    2017-02-01

    Data from multiple bacterial pathogens are consistent with regulator-encoding genes having higher mutation frequencies than the genome average. Such mutations drive both strain- and type- (e.g., serotype, haplotype) specific phenotypic heterogeneity, and may challenge public health due to the potential of variants to circumvent established treatment and/or preventative regimes. Here, using the human bacterial pathogen the group A Streptococcus (GAS; S. pyogenes) as a model organism, we review the types and regulatory-, phenotypic-, and disease-specific consequences of naturally occurring regulatory gene mutations. Strain-specific regulator mutations that will be discussed include examples that transform isolates into hyper-invasive forms by enhancing expression of immunomodulatory virulence factors, and examples that promote asymptomatic carriage of the organism. The discussion of serotype-specific regulator mutations focuses on serotype M3 GAS isolates, and how the identified rewiring of regulatory networks in this serotype may be contributing to a decades old epidemiological association of M3 isolates with particularly severe invasive infections. We conclude that mutation plays an outsized role in GAS pathogenesis and has clinical relevance. Given the phenotypic variability associated with regulatory gene mutations, the rapid examination of these genes in infecting isolates may inform with respect to potential patient complications and treatment options.

  9. Effect of oblique force source induced by laser ablation on ultrasonic generation.

    Science.gov (United States)

    Guo, Yuning; Yang, Dexing; Chang, Ying; Gao, Wei

    2014-01-13

    The effect of asymmetry caused by oblique line-shaped laser ablation on the generation of ultrasonic waves in metal, especially the effect of transverse component of the ablation force source on the ultrasonic waves is analyzed. Due to the oblique force source, the displacements of shear wave increase obviously by the enhanced shear force, the energy concentration area of longitudinal wave deflects to the small range centered on the incident direction while that of shear wave is approximately perpendicular to incident direction. In addition, surface wave enhances in the direction of transverse power flow. Furthermore, some ultrasonic characteristics under vortex laser ablation condition are inferred.

  10. Estradiol-induced gene expression in largemouth bass (Micropterus salmoides)

    Science.gov (United States)

    Bowman, C.J.; Kroll, K.J.; Gross, T.G.; Denslow, N.D.

    2002-01-01

    Vitellogenin (Vtg) and estrogen receptor (ER) gene expression levels were measured in largemouth bass to evaluate the activation of the ER-mediated pathway by estradiol (E2). Single injections of E2 ranging from 0.0005 to 5 mg/kg up-regulated plasma Vtg in a dose-dependent manner. Vtg and ER mRNAs were measured using partial cDNA sequences corresponding to the C-terminal domain for Vtg and the ligand-binding domain of ER?? sequences. After acute E2-exposures (2 mg/kg), Vtg and ER mRNAs and plasma Vtg levels peaked after 2 days. The rate of ER mRNA accumulation peaked 36-42 h earlier than Vtg mRNA. The expression window for ER defines the primary response to E2 in largemouth bass and that for Vtg a delayed primary response. The specific effect of E2 on other estrogen-regulated genes was tested during these same time windows using differential display RT-PCR. Specific up-regulated genes that are expressed in the same time window as Vtg were ERp72 (a membrane-bound disulfide isomerase) and a gene with homology to an expressed gene identified in zebrafish. Genes that were expressed in a pattern that mimics the ER include the gene for zona radiata protein ZP2, and a gene with homology to an expressed gene found in winter flounder. One gene for fibrinogen ?? was down-regulated and an unidentified gene was transiently up-regulated after 12 h of exposure and returned to basal levels by 48 h. Taken together these studies indicate that the acute molecular response to E2 involves a complex network of responses over time. ?? 2002 Elsevier Science Ireland Ltd. All rights reserved.

  11. CTCF-mediated chromatin loops enclose inducible gene regulatory domains

    NARCIS (Netherlands)

    Oti, M.O.; Falck, J.; Huynen, M.A.; Zhou, Huiqing

    2016-01-01

    BACKGROUND: The CCTC-binding factor (CTCF) protein is involved in genome organization, including mediating three-dimensional chromatin interactions. Human patient lymphocytes with mutations in a single copy of the CTCF gene have reduced expression of enhancer-associated genes involved in response to

  12. Fluctuation-induced forces in confined ideal and imperfect Bose gases

    Science.gov (United States)

    Diehl, H. W.; Rutkevich, Sergei B.

    2017-06-01

    Fluctuation-induced ("Casimir") forces caused by thermal and quantum fluctuations are investigated for ideal and imperfect Bose gases confined to d -dimensional films of size ∞d -1×D under periodic (P), antiperiodic (A), Dirichlet-Dirichlet (DD), Neumann-Neumann (NN), and Robin (R) boundary conditions (BCs). The full scaling functions ΥdBC(xλ=D /λth ,xξ=D /ξ ) of the residual reduced grand potential per area φres,dBC(T ,μ ,D ) =D-(d -1 )ΥdBC(xλ,xξ) are determined for the ideal gas case with these BCs, where λth and ξ are the thermal de Broglie wavelength and the bulk correlation length, respectively. The associated limiting scaling functions ΘdBC(xξ) ≡ΥdBC(∞ ,xξ) describing the critical behavior at the bulk condensation transition are shown to agree with those previously determined from a massive free O (2 ) theory for BC=P,A,DD,DN,NN . For d =3 , they are expressed in closed analytical form in terms of polylogarithms. The analogous scaling functions ΥdBC(xλ,xξ,c1D ,c2D ) and ΘdR(xξ,c1D ,c2D ) under the RBCs (∂z-c1) ϕ |z=0=(∂z+c2) ϕ | z =D=0 with c1≥0 and c2≥0 are also determined. The corresponding scaling functions Υ∞,d P(xλ,xξ) and Θ∞,d P(xξ) for the imperfect Bose gas are shown to agree with those of the interacting Bose gas with n internal degrees of freedom in the limit n →∞ . Hence, for d =3 , Θ∞,d P(xξ) is known exactly in closed analytic form. To account for the breakdown of translation invariance in the direction perpendicular to the boundary planes implied by free BCs such as DDBCs, a modified imperfect Bose gas model is introduced that corresponds to the limit n →∞ of this interacting Bose gas. Numerically and analytically exact results for the scaling function Θ∞,3 DD(xξ) therefore follow from those of the O (2 n ) ϕ4 model for n →∞ .

  13. Precise integration of inducible transcriptional elements (PrIITE) enables absolute control of gene expression

    DEFF Research Database (Denmark)

    Pinto, Rita; Hansen, Lars; Hintze, John Birger Hjalmar

    2017-01-01

    Tetracycline-based inducible systems provide powerful methods for functional studies where gene expression can be controlled. However, the lack of tight control of the inducible system, leading to leakiness and adverse effects caused by undesirable tetracycline dosage requirements, has proven...... to be a limitation. Here, we report that the combined use of genome editing tools and last generation Tet-On systems can resolve these issues. Our principle is based on precise integration of inducible transcriptional elements (coined PrIITE) targeted to: (i) exons of an endogenous gene of interest (GOI) and (ii......) a safe harbor locus. Using PrIITE cells harboring a GFP reporter or CDX2 transcription factor, we demonstrate discrete inducibility of gene expression with complete abrogation of leakiness. CDX2 PrIITE cells generated by this approach uncovered novel CDX2 downstream effector genes. Our results provide...

  14. Remanent cell traction force in renal vascular smooth muscle cells induced by integrin-mediated mechanotransduction

    OpenAIRE

    Balasubramanian, Lavanya; Lo, Chun-Min; Sham, James S. K.; Yip, Kay-Pong

    2013-01-01

    It was previously demonstrated in isolated renal vascular smooth muscle cells (VSMCs) that integrin-mediated mechanotransduction triggers intracellular Ca2+ mobilization, which is the hallmark of myogenic response in VSMCs. To test directly whether integrin-mediated mechanotransduction results in the myogenic response-like behavior in renal VSMCs, cell traction force microscopy was used to monitor cell traction force when the cells were pulled with fibronectin-coated or low density lipoprotei...

  15. Investigation on flow and mixing characteristics of supersonic mixing layer induced by forced vibration of cantilever

    Science.gov (United States)

    Zhang, Dongdong; Tan, Jianguo; Lv, Liang

    2015-12-01

    The mixing process has been an important issue for the design of supersonic combustion ramjet engine, and the mixing efficiency plays a crucial role in the improvement of the combustion efficiency. In the present study, nanoparticle-based planar laser scattering (NPLS), particle image velocimetry (PIV) and large eddy simulation (LES) are employed to investigate the flow and mixing characteristics of supersonic mixing layer under different forced vibration conditions. The indexes of fractal dimension, mixing layer thickness, momentum thickness and scalar mixing level are applied to describe the mixing process. Results show that different from the development and evolution of supersonic mixing layer without vibration, the flow under forced vibration is more likely to present the characteristics of three-dimensionality. The laminar flow region of mixing layer under forced vibration is greatly shortened and the scales of rolled up Kelvin-Helmholtz vortices become larger, which promote the mixing process remarkably. The fractal dimension distribution reveals that comparing with the flow without vibration, the turbulent fluctuation of supersonic mixing layer under forced vibration is more intense. Besides, the distribution of mixing layer thickness, momentum thickness and scalar mixing level are strongly influenced by forced vibration. Especially, when the forcing frequency is 4000 Hz, the mixing layer thickness and momentum thickness are 0.0391 m and 0.0222 m at the far field of 0.16 m, 83% and 131% higher than that without vibration at the same position, respectively.

  16. Forced treadmill exercise can induce stress and increase neuronal damage in a mouse model of global cerebral ischemia

    Directory of Open Access Journals (Sweden)

    Martina Svensson

    2016-12-01

    Full Text Available Physical exercise is known to be a beneficial factor by increasing the cellular stress tolerance. In ischemic stroke, physical exercise is suggested to both limit the brain injury and facilitate behavioral recovery. In this study we investigated the effect of physical exercise on brain damage following global cerebral ischemia in mice. We aimed to study the effects of 4.5 weeks of forced treadmill running prior to ischemia on neuronal damage, neuroinflammation and its effect on general stress by measuring corticosterone in feces. We subjected C57bl/6 mice (n = 63 to either treadmill running or a sedentary program prior to induction of global ischemia. Anxious, depressive, and cognitive behaviors were analyzed. Stress levels were analyzed using a corticosterone ELISA. Inflammatory and neurological outcomes were analyzed using immunohistochemistry, multiplex electrochemoluminescence ELISA and Western blot. To our surprise, we found that forced treadmill running induced a stress response, with increased anxiety in the Open Field test and increased levels of corticosterone. In accordance, mice subjected to forced exercise prior to ischemia developed larger neuronal damage in the hippocampus and showed higher cytokine levels in the brain and blood compared to non-exercised mice. The extent of neuronal damage correlated with increased corticosterone levels. To compare forced treadmill with voluntary wheel running, we used a different set of mice that exercised freely on running wheels. These mice did not show any anxiety or increased corticosterone levels. Altogether, our results indicate that exercise pre-conditioning may not be beneficial if the animals are forced to run as it can induce a detrimental stress response.

  17. Temperature jump induced force generation in rabbit muscle fibres gets faster with shortening and shows a biphasic dependence on velocity.

    Science.gov (United States)

    Ranatunga, K W; Roots, H; Offer, G W

    2010-02-01

    We examined the tension responses to ramp shortening and rapid temperature jump (muscle fibres at 8-9 degrees C (the fibre length (L(0)) was approximately 1.5 mm and sarcomere length 2.5 microm). The aim was to investigate the strain sensitivity of crossbridge force generation in muscle. The T-jump induced tension rise was examined during steady shortening over a wide range of velocities (V) approaching the V(max) (V range approximately 0.01 to approximately 1.5 L(0) s(1)). In the isometric state, a T-jump induced a biphasic tension rise consisting of a fast (approximately 50 s(1), phase 2b) and a slow (approximately 10 s(1), phase 3) component, but if treated as monophasic the rate was approximately 20 s(1). During steady shortening the T-jump tension rise was monophasic; the rate of tension rise increased linearly with shortening velocity, and near V(max) it was approximately 200 s(1), approximately 10x faster than in the isometric state. Relative to the tension reached after the T-jump, the amplitude increased with shortening velocity, and near V(max) it was 4x larger than in the isometric state. Thus, the temperature sensitivity of muscle force is markedly increased with velocity during steady shortening, as found in steady state experiments. The rate of tension decline during ramp shortening also increased markedly with increase of velocity. The absolute amplitude of T-jump tension rise was larger than that in the isometric state at the low velocities (shortening velocity is increased, probably enhancement of crossbridge force generation and faster (post-stroke) crossbridge detachment by negative strain. Overall, our results show that T-jump force generation is strain sensitive and becomes considerably faster when exposed to negative strain. Thus the crossbridge force generation step in muscle is both temperature sensitive (endothermic) and strain sensitive.

  18. Identification of novel light-induced genes in the suprachiasmatic nucleus

    Directory of Open Access Journals (Sweden)

    Piontkivska Helen

    2007-11-01

    Full Text Available Abstract Background The transmission of information about the photic environment to the circadian clock involves a complex array of neurotransmitters, receptors, and second messenger systems. Exposure of an animal to light during the subjective night initiates rapid transcription of a number of immediate-early genes in the suprachiasmatic nucleus of the hypothalamus. Some of these genes have known roles in entraining the circadian clock, while others have unknown functions. Using laser capture microscopy, microarray analysis, and quantitative real-time PCR, we performed a comprehensive screen for changes in gene expression immediately following a 30 minute light pulse in suprachiasmatic nucleus of mice. Results The results of the microarray screen successfully identified previously known light-induced genes as well as several novel genes that may be important in the circadian clock. Newly identified light-induced genes include early growth response 2, proviral integration site 3, growth-arrest and DNA-damage-inducible 45 beta, and TCDD-inducible poly(ADP-ribose polymerase. Comparative analysis of promoter sequences revealed the presence of evolutionarily conserved CRE and associated TATA box elements in most of the light-induced genes, while other core clock genes generally lack this combination of promoter elements. Conclusion The photic signalling cascade in the suprachiasmatic nucleus activates an array of immediate-early genes, most of which have unknown functions in the circadian clock. Detected evolutionary conservation of CRE and TATA box elements in promoters of light-induced genes suggest that the functional role of these elements has likely remained the same over evolutionary time across mammalian orders.

  19. Transposon-induced nuclear mutations that alter chloroplast gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Barkan, A.

    1992-01-01

    The goal of this project is to use mutant phenotypes as a guide to nuclear genes that determine the timing and localization of chloroplast development The immediate goals are to identify nuclear mutants with defects in chloroplast gene expression from maize lines harboring active Mu transposons; characterize their phenotypes to determine the precise defect in gene expression; clone several of the most interesting mutations by exploiting the transposon tag; and use the clones to further define the roles of these genes in modulating chloroplast gene expression. Three mutants were described earlier that had global defects in chloroplast gene expression. We have found that two of these mutations are allelic. Both alleles have global defects in chloroplast translation initiation, as revealed by the failure to assemble chloroplast mRNAs into polysomes. We have isolated and characterized three new mutants from Mu lines that have novel defects in chloroplast RNA metabolism. We are now ready to begin the task of cloning several of these genes, by using the Mu transposon tag.

  20. Ebola virus infection induces irregular dendritic cell gene expression.

    Science.gov (United States)

    Melanson, Vanessa R; Kalina, Warren V; Williams, Priscilla

    2015-02-01

    Filoviruses subvert the human immune system in part by infecting and replicating in dendritic cells (DCs). Using gene arrays, a phenotypic profile of filovirus infection in human monocyte-derived DCs was assessed. Monocytes from human donors were cultured in GM-CSF and IL-4 and were infected with Ebola virus Kikwit variant for up to 48 h. Extracted DC RNA was analyzed on SuperArray's Dendritic and Antigen Presenting Cell Oligo GEArray and compared to uninfected controls. Infected DCs exhibited increased expression of cytokine, chemokine, antiviral, and anti-apoptotic genes not seen in uninfected controls. Significant increases of intracellular antiviral and MHC I and II genes were also noted in EBOV-infected DCs. However, infected DCs failed to show any significant difference in co-stimulatory T-cell gene expression from uninfected DCs. Moreover, several chemokine genes were activated, but there was sparse expression of chemokine receptors that enabled activated DCs to home to lymph nodes. Overall, statistically significant expression of several intracellular antiviral genes was noted, which may limit viral load but fails to stop replication. EBOV gene expression profiling is of vital importance in understanding pathogenesis and devising novel therapeutic treatments such as small-molecule inhibitors.

  1. Pregnancy-induced gene expression changes in vivo among women with rheumatoid arthritis

    DEFF Research Database (Denmark)

    Goin, Dana E; Smed, Mette Kiel; Pachter, Lior

    2017-01-01

    of Gene Ontology processes and protein networks. RESULTS: A total of 1296 genes were differentially expressed between T3 and T0 among the 8 pregDASimproved women, with 161 genes showing at least two-fold change (FC) in expression by T3. The majority (108 of 161 genes) were also differentially expressed......BACKGROUND: Little is known about gene expression changes induced by pregnancy in women with rheumatoid arthritis (RA) and healthy women because the few studies previously conducted did not have pre-pregnancy samples available as baseline. We have established a cohort of women with RA and healthy...... women followed prospectively from a pre-pregnancy baseline. In this study, we tested the hypothesis that pregnancy-induced changes in gene expression among women with RA who improve during pregnancy (pregDASimproved) overlap substantially with changes observed among healthy women and differ from changes...

  2. Mechanics of the IL2RA gene activation revealed by modeling and atomic force microscopy.

    Science.gov (United States)

    Milani, Pascale; Marilley, Monique; Sanchez-Sevilla, Albert; Imbert, Jean; Vaillant, Cédric; Argoul, Françoise; Egly, Jean-Marc; Rocca-Serra, José; Arneodo, Alain

    2011-04-13

    Transcription implies recruitment of RNA polymerase II and transcription factors (TFs) by DNA melting near transcription start site (TSS). Combining atomic force microscopy and computer modeling, we investigate the structural and dynamical properties of the IL2RA promoter and identify an intrinsically negative supercoil in the PRRII region (containing Elf-1 and HMGA1 binding sites), located upstream of a curved DNA region encompassing TSS. Conformational changes, evidenced by time-lapse studies, result in the progressive positioning of curvature apex towards the TSS, likely facilitating local DNA melting. In vitro assays confirm specific binding of the General Transcription Factors (GTFs) TBP and TFIIB over TATA-TSS position, where an inhibitory nucleosome prevented preinitiation complex (PIC) formation and uncontrolled DNA melting. These findings represent a substantial advance showing, first, that the structural properties of the IL2RA promoter are encoded in the DNA sequence and second, that during the initiation process DNA conformation is dynamic and not static.

  3. Elicitor and fusarium-induced expression of NPR-1 like genes in banana

    CSIR Research Space (South Africa)

    Endah, R

    2008-11-01

    Full Text Available NPR1 is an essential positive regulator of salicylic acid-induced PR gene expression and systemic acquired resistance. Two novel full-length NPR1-like genes; MNPR1A and MNPR1B, were isolated by application of the PCR and RACE techniques. The two...

  4. Alcohol-induced histone acetylation reveals a gene network involved in alcohol tolerance.

    Directory of Open Access Journals (Sweden)

    Alfredo Ghezzi

    Full Text Available Sustained or repeated exposure to sedating drugs, such as alcohol, triggers homeostatic adaptations in the brain that lead to the development of drug tolerance and dependence. These adaptations involve long-term changes in the transcription of drug-responsive genes as well as an epigenetic restructuring of chromosomal regions that is thought to signal and maintain the altered transcriptional state. Alcohol-induced epigenetic changes have been shown to be important in the long-term adaptation that leads to alcohol tolerance and dependence endophenotypes. A major constraint impeding progress is that alcohol produces a surfeit of changes in gene expression, most of which may not make any meaningful contribution to the ethanol response under study. Here we used a novel genomic epigenetic approach to find genes relevant for functional alcohol tolerance by exploiting the commonalities of two chemically distinct alcohols. In Drosophila melanogaster, ethanol and benzyl alcohol induce mutual cross-tolerance, indicating that they share a common mechanism for producing tolerance. We surveyed the genome-wide changes in histone acetylation that occur in response to these drugs. Each drug induces modifications in a large number of genes. The genes that respond similarly to either treatment, however, represent a subgroup enriched for genes important for the common tolerance response. Genes were functionally tested for behavioral tolerance to the sedative effects of ethanol and benzyl alcohol using mutant and inducible RNAi stocks. We identified a network of genes that are essential for the development of tolerance to sedation by alcohol.

  5. Effect of electro-acupuncture on gene expression in heart of rats with stress-induced pre-hypertension based on gene chip technology

    National Research Council Canada - National Science Library

    Guo Yan Xie Xiaojia Guo Changqing Wang Zhaoyang Liu Qingguo

    2015-01-01

    OBJECTIVE:To explore electro-acupuncture's(EA's)effect on gene expression in heart of rats with stress-induced pre-hypertension and try to reveal its biological mechanism based on gene chip...

  6. Effect of force-induced mechanical stress at the coronary artery bifurcation stenting: Relation to in-stent restenosis

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Cheng-Hung [Division of Cardiology, Department of Internal Medicine, Chang Gung Memorial Hospital, Linkou, Chang Gung University College of Medicine, Tao-Yuan, Taiwan (China); Department of Mechanical Engineering, Chang Gung University, Tao-Yuan, Taiwan (China); Jhong, Guan-Heng [Graduate Institute of Medical Mechatronics, Chang Gung University, Tao-Yuan, Taiwan (China); Hsu, Ming-Yi; Wang, Chao-Jan [Department of Medical Imaging and Intervention, Chang Gung Memorial Hospital, Linkou, Tao-Yuan, Taiwan (China); Liu, Shih-Jung, E-mail: shihjung@mail.cgu.edu.tw [Department of Mechanical Engineering, Chang Gung University, Tao-Yuan, Taiwan (China); Hung, Kuo-Chun [Division of Cardiology, Department of Internal Medicine, Chang Gung Memorial Hospital, Linkou, Chang Gung University College of Medicine, Tao-Yuan, Taiwan (China)

    2014-05-28

    The deployment of metallic stents during percutaneous coronary intervention has become common in the treatment of coronary bifurcation lesions. However, restenosis occurs mostly at the bifurcation area even in present era of drug-eluting stents. To achieve adequate deployment, physicians may unintentionally apply force to the strut of the stents through balloon, guiding catheters, or other devices. This force may deform the struts and impose excessive mechanical stresses on the arterial vessels, resulting in detrimental outcomes. This study investigated the relationship between the distribution of stress in a stent and bifurcation angle using finite element analysis. The unintentionally applied force following stent implantation was measured using a force sensor that was made in the laboratory. Geometrical information on the coronary arteries of 11 subjects was extracted from contrast-enhanced computed tomography scan data. The numerical results reveal that the application of force by physicians generated significantly higher mechanical stresses in the arterial bifurcation than in the proximal and distal parts of the stent (post hoc P < 0.01). The maximal stress on the vessels was significantly higher at bifurcation angle <70° than at angle ≧70° (P < 0.05). The maximal stress on the vessels was negatively correlated with bifurcation angle (P < 0.01). Stresses at the bifurcation ostium may cause arterial wall injury and restenosis, especially at small bifurcation angles. These finding highlight the effect of force-induced mechanical stress at coronary artery bifurcation stenting, and potential mechanisms of in-stent restenosis, along with their relationship with bifurcation angle.

  7. An insulin-induced DNA-binding protein for the human growth hormone gene.

    OpenAIRE

    Prager, D; Gebremedhin, S; Melmed, S

    1990-01-01

    The control of gene transcription is usually mediated by transacting transcriptional factors that bind to upstream regulatory elements. As insulin regulates transcription of the growth hormone (GH) gene, we tested nuclear extracts from unstimulated and insulin-stimulated Chinese hamster ovarian (CHO) cells for binding to four human GH (hGH) gene promoter oligonucleotide fragments identified as target-binding sequences by DNAse I footprinting. Using a mobility shift assay, an insulin-induced D...

  8. A virus-induced gene silencing method to study soybean cyst nematode parasitism in Glycine max

    OpenAIRE

    Kandoth, Pramod K; Heinz, Robert; Yeckel, Greg; Gross, Nathan W; Juvale, Parijat S; Hill, John; Whitham, Steven A.; Baum, Thomas J.; Mitchum, Melissa G.

    2013-01-01

    Background Bean pod mottle virus (BPMV) based virus-induced gene silencing (VIGS) vectors have been developed and used in soybean for the functional analysis of genes involved in disease resistance to foliar pathogens. However, BPMV-VIGS protocols for studying genes involved in disease resistance or symbiotic associations with root microbes have not been developed. Findings Here we describe a BPMV-VIGS protocol suitable for reverse genetic studies in soybean roots. We use this method for anal...

  9. Development of Virus-Induced Gene Expression and Silencing Vector Derived from Grapevine Algerian Latent Virus

    OpenAIRE

    Sang-Ho Park; Hoseong Choi; Semin Kim; Won Kyong Cho; Kook-Hyung Kim

    2016-01-01

    Grapevine Algerian latent virus (GALV) is a member of the genus Tombusvirus in the Tombusviridae and infects not only woody perennial grapevine plant but also herbaceous Nicotiana benthamiana plant. In this study, we developed GALV-based gene expression and virus-induced gene silencing (VIGS) vectors in N. benthamiana. The GALV coat protein deletion vector, pGMG, was applied to express the reporter gene, green fluorescence protein (GFP), but the expression of GFP was not detected due to the n...

  10. Probiotic bacteria change Echherichia coli-induced gene expression in cultured colonocytes: Implications in intestinal pathophysiology

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To investigate the change in eukaryotic gene expression profile in Caco-2 cells after infection with strains of Escherichia coli and commensal probiotic bacteria.METHODS: A 19200 gene/expressed sequence tag gene chip was used to examine expression of genes after infection of Caco-2 cells with strains of normal flora E.coli, Lactobacillus plantarum, and a combination of the two.RESULTS: The cDNA microarray revealed up-regulation of 155 and down-regulation of 177 genes by E. coli. L. plantarum up-regulated 45 and down-regulated 36 genes. During mixed infection, 27 genes were upregulated and 59 were down-regulated, with nullification of stimulatory/inhibitory effects on most of the genes. Expression of several new genes was noted in this group.CONCLUSION: The commensal bacterial strains used in this study induced the expression of a large number of genes in colonocyte-like cultured cells and changed the expression of several genes involved in important cellular processes such as regulation of transcription, protein biosynthesis, metabolism, cell adhesion, ubiquitination,and apoptosis. Such changes induced by the presence of probiotic bacteria may shape the physiologic and pathologic responses they trigger in the host.

  11. Evidence for non-conservative current-induced forces in the breaking of Au and Pt atomic chains

    Directory of Open Access Journals (Sweden)

    Carlos Sabater

    2015-12-01

    Full Text Available This experimental work aims at probing current-induced forces at the atomic scale. Specifically it addresses predictions in recent work regarding the appearance of run-away modes as a result of a combined effect of the non-conservative wind force and a ‘Berry force’. The systems we consider here are atomic chains of Au and Pt atoms, for which we investigate the distribution of break down voltage values. We observe two distinct modes of breaking for Au atomic chains. The breaking at high voltage appears to behave as expected for regular break down by thermal excitation due to Joule heating. However, there is a low-voltage breaking mode that has characteristics expected for the mechanism of current-induced forces. Although a full comparison would require more detailed information on the individual atomic configurations, the systems we consider are very similar to those considered in recent model calculations and the comparison between experiment and theory is very encouraging for the interpretation we propose.

  12. DNA intercalation by ethidium bromide: A quantitative binding study using DNA stretching and force-induced melting

    Science.gov (United States)

    Williams, Mark C.; Rouzina, Ioulia

    2005-03-01

    The interactions between single DNA molecules and the non- covalent binding agent ethidium bromide are investigated using an optical tweezers instrument and the effects of this intercalator on the structure and mechanical stability of DNA molecules are quantitatively analyzed using our model of force- induced melting. The DNA force-extension cycles in the presence and absence of drug are recorded. It is found that the drug binds preferentially to double-stranded DNA and stabilizes the double helix. There is clear evidence of the force induced melting transition at low concentrations of drug, while at higher concentrations the drug is able to prevent the melting transition. The DNA contour length is obtained as a function of ligand concentration directly from the stretching curves. From this data we obtain the complete ethidium bromide dsDNA binding isotherm, which is used to find the binding constant and the binding site size of the intercalator. Out data also allows us to quantify directly the effect of ethidium bromide on the free energy of the helix-coil transition in dsDNA. This single molecule study brings new insights into the molecular mechanisms which drive drug-DNA complex formation.

  13. Quantitative Analyses of Force-Induced Amyloid Formation in Candida albicans Als5p: Activation by Standard Laboratory Procedures.

    Directory of Open Access Journals (Sweden)

    Cho X J Chan

    Full Text Available Candida albicans adhesins have amyloid-forming sequences. In Als5p, these amyloid sequences cluster cell surface adhesins to create high avidity surface adhesion nanodomains. Such nanodomains form after force is applied to the cell surface by atomic force microscopy or laminar flow. Here we report centrifuging and resuspending S. cerevisiae cells expressing Als5p led to 1.7-fold increase in initial rate of adhesion to ligand coated beads. Furthermore, mechanical stress from vortex-mixing of Als5p cells or C. albicans cells also induced additional formation of amyloid nanodomains and consequent activation of adhesion. Vortex-mixing for 60 seconds increased the initial rate of adhesion 1.6-fold. The effects of vortex-mixing were replicated in heat-killed cells as well. Activation was accompanied by increases in thioflavin T cell surface fluorescence measured by flow cytometry or by confocal microscopy. There was no adhesion activation in cells expressing amyloid-impaired Als5pV326N or in cells incubated with inhibitory concentrations of anti-amyloid dyes. Together these results demonstrated the activation of cell surface amyloid nanodomains in yeast expressing Als adhesins, and further delineate the forces that can activate adhesion in vivo. Consequently there is quantitative support for the hypothesis that amyloid forming adhesins act as both force sensors and effectors.

  14. Water spray-induced grooming is negatively correlated with depressive behavior in the forced swimming test in rats.

    Science.gov (United States)

    Shiota, Noboru; Narikiyo, Kimiya; Masuda, Akira; Aou, Shuji

    2016-05-01

    Rodents show grooming, a typical self-care behavior, under stress and non-stress conditions. Previous studies revealed that grooming under stress conditions such as the open-field test (OFT) or the elevated plus-maze test (EPM) is associated with anxiety, but the roles of grooming under non-stress conditions are not well understood. Here, we examined spray-induced grooming as a model of grooming under a non-stress condition to investigate the relationship between this grooming and depression-like behavior in the forced swim test (FST) and tail suspension test, and we compared spray-induced grooming with OFT- and EPM-induced grooming. The main finding was that the duration of spray-induced grooming, but not that of OFT/EPM-induced grooming, was negatively correlated with the duration of immobility in the FST, an index of depression-like behavior. The results suggest that spray-induced grooming is functionally different from the grooming in the OFT and EPM and is related to reduction of depressive behavior.

  15. Classical and fluctuation-induced electromagnetic interactions in micronscale systems: designer bonding, antibonding, and Casimir forces

    CERN Document Server

    Rodriguez, Alejandro W; Woolf, David N; Johnson, Steven G; Loncar, Marko; Capasso, Federico

    2014-01-01

    Whether intentionally introduced to exert control over particles and macroscopic objects, such as for trapping or cooling, or whether arising from the quantum and thermal fluctuations of charges in otherwise neutral bodies, leading to unwanted stiction between nearby mechanical parts, electromagnetic interactions play a fundamental role in many naturally occurring processes and technologies. In this review, we survey recent progress in the understanding and experimental observation of optomechanical and quantum-fluctuation forces. Although both of these effects arise from exchange of electromagnetic momentum, their dramatically different origins, involving either real or virtual photons, lead to different physical manifestations and design principles. Specifically, we describe recent predictions and measurements of attractive and repulsive optomechanical forces, based on the bonding and antibonding interactions of evanescent waves, as well as predictions of modified and even repulsive Casimir forces between n...

  16. Unidirectional Transport of Ferromagnetic Particles in a Viscous LiquidInduced by the Magnus Force

    Directory of Open Access Journals (Sweden)

    S.I. Denisov

    2016-10-01

    Full Text Available We study the unidirectional motion of spherical ferromagnetic particles suspended in a viscous liquid and subjected to the action of an external periodic force and a non-uniformly rotating magnetic field. In the case when the translational and rotational motions of particles are characterized by small Reynolds numbers, we propose a system of equations that describes their dynamics and that accounts for the influence of the Magnus force. Theoretical and numerical analysis of the steady-state solution of this system of equations is carried out, the average velocity of unidirectional (drift motion of particles is calculated, and the dependence of the average velocity on the characteristics of particles, liquid, external force and magnetic field is studied.

  17. Bifidobacterium bifidum Actively Changes the Gene Expression Profile Induced by Lactobacillus acidophilus in Murine Dendritic Cells

    DEFF Research Database (Denmark)

    Weiss, Gudrun Margarethe; Rasmussen, Simon; Fink, Lisbeth Nielsen

    2010-01-01

    cytokine IL-12 in DC, whereas bifidobacteria do not induce IL-12 but inhibit the IL-12 production induced by lactobacilli. In the present study, genome-wide microarrays were used to investigate the gene expression pattern of murine DC stimulated with Lactobacillus acidophilus NCFM and Bifidobacterium...

  18. Global analysis of radiative forcing from fire-induced shortwave albedo change

    Directory of Open Access Journals (Sweden)

    G. López-Saldaña

    2014-05-01

    Full Text Available Land surface albedo, a key parameter to derive Earth's surface energy balance, is used in the parameterization of numerical weather prediction, climate monitoring and climate change impact assessments. Changes in albedo due to fire have not been fully investigated at continental and global scale. The main goal of this study therefore, is to quantify the changes in albedo produced by biomass burning activities and their associated shortwave radiative forcing. The study relies on the Moderate Resolution Imaging Spectroradiometer (MODIS MCD64A1 burned area product to create an annual composite of areas affected by fire and the MCD43C2 BRDF-Albedo snow-free product to compute a bihemispherical reflectance time series. The approximate day of burn is used to calculate the instantaneous change in shortwave Albedo. Using the corresponding National Centers for Environmental Prediction (NCEP monthly mean downward solar radiation flux at the surface, the global radiative forcing associated to fire was computed. The analysis reveals a mean decrease in shortwave albedo of −0.023 (1σ = 0.018 causing a mean positive radiative forcing of 6.31 W m–2 (1σ = 5.04 over the 2002–2012 time period in areas affected by fire. The greatest drop in mean shortwave albedo change occurs in 2002, which corresponds to the highest total area burnt (3.66 Mha observed in the same year and produces the highest mean radiative forcing (6.75 W m–2. Africa is the main contributor in terms of burned area but forests globally are giving the highest radiative forcing per unit area, thus give detectable changes in shortwave albedo. The global mean radiative forcing for the whole studied period ~ 0.04 W m–2 shows that the contribution of fires into the Earth system is not insignificant.

  19. RNA-Seq analysis of high NaCl-induced gene expression.

    Science.gov (United States)

    Izumi, Yuichiro; Yang, Wenjing; Zhu, Jun; Burg, Maurice B; Ferraris, Joan D

    2015-10-01

    High extracellular NaCl is known to change expression of numerous genes, many of which are regulated by the osmoprotective transcription factor nuclear factor of activated T cells-5 (NFAT5). In the present study we employed RNA-Seq to provide a comprehensive, unbiased account of genes regulated by high NaCl in mouse embryonic fibroblast cells (MEFs). To identify genes regulated by NFAT5 we compared wild-type MEFs (WT-MEFs) to MEFs in which mutation of the NFAT5 gene inhibits its transcriptional activity (Null-MEFs). In WT-MEFs adding NaCl to raise osmolality from 300 to 500 mosmol/kg for 24 h increases expression of 167 genes and reduces expression of 412. Raising osmolality through multiple passages (adapted cells) increases expression of 196 genes and reduces expression of 528. In Null-MEFs, after 24 h of high NaCl, expression of 217 genes increase and 428 decrease, while in adapted Null-MEFs 143 increase and 622 decrease. Fewer than 10% of genes are regulated in common between WT- and null-MEFs, indicating a profound difference in regulation of high-NaCl induced genes induced by NFAT5 compared with those induced in the absence of NFAT5. Based on our findings we suggest a mechanism for this phenomenon, which had previously been unexplained. The NFAT5 DNA-binding motif (osmotic response element) is overrepresented in the vicinity of genes that NFAT5 upregulates, but not genes that it downregulates. We used Gene Ontology and manual curation to determine the function of the genes targeted by NFAT5, revealing many novel consequences of NFAT5 transcriptional activity.

  20. Lipopolysaccharide triggers nuclear import of Lpcat1 to regulate inducible gene expression in lung epithelia

    Institute of Scientific and Technical Information of China (English)

    Bryon; Ellis; Leah; Kaercher; Courtney; Snavely

    2012-01-01

    AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus influenza and Escherichia coli infection was analyzed by employing quantitative Reverse Transcription Polymerase Chain Reaction techniques. Nucleofection was used to deliver Lenti-viral system to express or knock down Lpcat1 in MLE cells. Subcellular protein fractionation and Western blotting were utilized to study Lpcat1 nuclear relocation. RESULTS:Lpcat1 translocates into the nucleus from thecytoplasm in murine lung epithelia (MLE) after LPS treatment. Haemophilus influenza and Escherichia coli , two LPS-containing pathogens that cause pneumonia, triggered Lpcat1 nuclear translocation from the cytoplasm. The LPS inducible gene expression profile was determined by quantitative reverse transcription polymerase chain reaction after silencing Lpcat1 or overexpression of the enzyme in MLE cells. We detected that 17 out of a total 38 screened genes were upregulated, 14 genes were suppressed, and 7 genes remained unchanged in LPS treated cells in comparison to controls. Knockdown of Lpcat1 by shRNA dramatically changed the spectrum of the LPS inducible gene transcription, as 18 genes out of 38 genes were upregulated, of which 20 genes were suppressed or unchanged. Notably, in Lpcat1 overex-pressed cells, 25 genes out of 38 genes were reduced in the setting of LPS treatment.CONCLUSION:These observations suggest that Lpcat1 relocates into the nucleus in response to bacterial infection to differentially regulate gene transcriptional repression.

  1. Analysis of target genes induced by IL-13 cytotoxin in human glioblastoma cells.

    Science.gov (United States)

    Han, Jing; Yang, Liming; Puri, Raj K

    2005-03-01

    IL-13 cytotoxin comprised of IL-13 and a mutated form of Pseudomonas exotoxin (fusion protein termed IL-13-PE38QQR) has been shown to inhibit protein synthesis leading to necrotic and apoptotic cell death in glioblastoma cells that express high levels of interleukin-13 receptors (IL-13R). To identify target genes of cell death and other cellular genes with IL-13 receptors in glioblastoma cells, we utilized the cDNA microarrays to analyze global gene expression profiles after IL-13 cytotoxin and IL-13 treatment. IL-13 cytotoxin mediated cytotoxicity to U251 cells in a dose-dependent manner. Hierarchical cluster analysis of differentially expressed genes in U251 glioma cells at different time points after IL-13 cytotoxin treatment showed three major groups, each representing a specific expression pattern. Randomly selected differentially expressed genes from each group were confirmed by RT-PCR analysis. Most down-regulated genes belong to cell adhesion, motility, angiogenesis, DNA repair, and metabolic pathways. While up-regulated genes belong to cell cycle arrest, apoptosis, signaling and various metabolic pathways. Unexpectedly, at early time points, both IL-13 and IL-13 cytotoxin induced several genes belonging to different pathways most notably IL-8, DIO2, END1, and ALDH1A3 indicating that these genes are early response genes and their products may be associated with IL-13R. In addition, IL-13 cytotoxin induced IL-13Ralpha2 mRNA expression during the treatment in glioma cells. Our results indicate that novel cellular genes are involved with IL-13 receptors and that IL-13 cytotoxin induced cell death involves various target genes in human glioblastoma cells. On going studies will determine the role of associated genes and their products in the IL-13R functions in glioma cells.

  2. Estimating Wind and Wave Induced Forces On a Floating Wind Turbine

    DEFF Research Database (Denmark)

    Henriksen, Lars Christian; Natarajan, Anand; Kim, Taeseong

    -principles derived state space model of the floating wind turbine. The ability to estimate aero- and hydrodynamic states could prove crucial for the performance of model-based control methods applied on floating wind turbines. Furthermore, two types of water kinematics have been compared two determine whether......In this work, the basic model for a spar buoy floating wind turbine [1], used by an extended Kalman filter, is presented and results concerning wind speed and wave force estimations are shown. The wind speed and aerodynamic forces are estimated using an extended Kalman filter based on a first...... or not linear and nonlinear water kinematics lead to significantly different loads....

  3. Estimating Wind and Wave Induced Forces On a Floating Wind Turbine

    DEFF Research Database (Denmark)

    Henriksen, Lars Christian; Natarajan, Anand; Kim, Taeseong

    2013-01-01

    -principles derived state space model of the floating wind turbine. The ability to estimate aero- and hydrodynamic states could prove crucial for the performance of model-based control methods applied on floating wind turbines. Furthermore, two types of water kinematics have been compared two determine whether......In this work, the basic model for a spar buoy floating wind turbine [1], used by an extended Kalman filter, is presented and results concerning wind speed and wave force estimations are shown. The wind speed and aerodynamic forces are estimated using an extended Kalman filter based on a first...... or not linear and nonlinear water kinematics lead to significantly different loads....

  4. Characterisation of stochastically-induced climate jumps to other histories paced by the astronomical forcing.

    Science.gov (United States)

    De Saedeleer, B.

    2012-04-01

    The mystery of ice ages induced by a varying incoming solar radiation has drawn ceaseless attention for several decades. A pleiad of paleoclimatic models has been developed in order to have a try at catching the underlying climate dynamics, and their validity is challenged by comparison with typical milestones in paleoclimatic records. In several published works, the astronomical forcing synchronises the climate to a unique climatic attracting trajectory representing the ice volume evolution. Other studies, though, reported multistability, i.e. the fact that several climatic attracting trajectories could coexist for some given set of parameters, in a deterministic framework. More importantly, it has been illustrated that additional disturbances may cause some 'jumps' from one trajectory to other ones in the climatic history over the last millions years of the Pleistocene. These stochastic effects hence indirectly affect the timing of the glacial inceptions and terminations. The jumping mechanism is closely linked to the widely spread hypothesis that the glacial-interglacial cycles could be primarily triggered by random internal climate variability. A conjecture has recently been made that these externally triggered jumps are the most likely when the temporary desynchronisation (positive largest local Lyapunov exponent) due to the loss of local stability coalesces with the weakening of the global stability due to the proximity to the basin boundary. No proof of this conjecture has however been provided sofar; it is precisely the aim of the present research to assess the conditions for such a jump to occur. We uncover the details of the underlying mechanisms by providing a systematic numerical study of the conditions under which these jumps are likely to occur. Extensive Monte Carlo experiments are performed in order to show that the jumps occur preferentially at specific times or locations in the phase space, for a given level of noise. We show how the most critical

  5. Fast and sensitive detection of indels induced by precise gene targeting

    DEFF Research Database (Denmark)

    Yang, Zhang; Steentoft, Catharina; Hauge, Camilla

    2015-01-01

    The nuclease-based gene editing tools are rapidly transforming capabilities for altering the genome of cells and organisms with great precision and in high throughput studies. A major limitation in application of precise gene editing lies in lack of sensitive and fast methods to detect...... and characterize the induced DNA changes. Precise gene editing induces double-stranded DNA breaks that are repaired by error-prone non-homologous end joining leading to introduction of insertions and deletions (indels) at the target site. These indels are often small and difficult and laborious to detect...

  6. Protective effects of L-selenomethionine on space radiation induced changes in gene expression.

    Science.gov (United States)

    Stewart, J; Ko, Y-H; Kennedy, A R

    2007-06-01

    Ionizing radiation can produce adverse biological effects in astronauts during space travel. Of particular concern are the types of radiation from highly energetic, heavy, charged particles known as HZE particles. The aims of our studies are to characterize HZE particle radiation induced biological effects and evaluate the effects of L-selenomethionine (SeM) on these adverse biological effects. In this study, microarray technology was used to measure HZE radiation induced changes in gene expression, as well as to evaluate modulation of these changes by SeM. Human thyroid epithelial cells (HTori-3) were irradiated (1 GeV/n iron ions) in the presence or in the absence of 5 microM SeM. At 6 h post-irradiation, all cells were harvested for RNA isolation. Gene Chip U133Av2 from Affymetrix was used for the analysis of gene expression, and ANOVA and EASE were used for a determination of the genes and biological processes whose differential expression is statistically significant. Results of this microarray study indicate that exposure to small doses of radiation from HZE particles, 10 and 20 cGy from iron ions, induces statistically significant differential expression of 196 and 610 genes, respectively. In the presence of SeM, differential expression of 77 out of 196 genes (exposure to 10 cGy) and 336 out of 610 genes (exposure to 20 cGy) is abolished. In the presence or in the absence of SeM, radiation from HZE particles induces differential expression of genes whose products have roles in the induction of G1/S arrest during the mitotic cell cycle, as well as heat shock proteins. Some of the genes, whose expressions were affected by radiation from HZE particles and were unchanged in irradiated cells treated with SeM, have been shown to have altered expression levels in cancer cells. The conclusions of this report are that radiation from HZE particles can induce differential expression of many genes, some of which are known to play roles in the same processes that have

  7. Copper induces the expression of cholesterogenic genes in human macrophages.

    Science.gov (United States)

    Svensson, Per Arne; Englund, Mikael C O; Markström, Emilia; Ohlsson, Bertil G; Jernås, Margareta; Billig, Håkan; Torgerson, Jarl S; Wiklund, Olov; Carlsson, Lena M S; Carlsson, Björn

    2003-07-01

    Accumulation of lipids and cholesterol by macrophages and subsequent transformation into foam cells are key features in development of atherosclerosis. Serum copper concentrations have been shown to be associated with cardiovascular disease. However, the mechanism behind the proatherogenic effect of copper is not clear. We used DNA microarrays to define the changes in gene expression profile in response to copper exposure of human macrophages. Expression monitoring by DNA microarray revealed 91 genes that were regulated. Copper increased the expression of seven cholesterogenic genes (3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) synthase, IPP isomerase, squalene synthase, squalene epoxidase, methyl sterol oxidase, H105e3 mRNA and sterol-C5-desaturase) and low-density lipoprotein receptor (LDL-R), and decreased the expression of CD36 and lipid binding proteins. The expression of LDL-R and HMG CoA reductase was also investigated using real time PCR. The expression of both of these genes was increased after copper treatment of macrophages (Pmechanism for the association between copper and atherosclerosis. The effect of copper on cholesterogenic genes may also have implications for liver steatosis in early stages of Wilson's disease.

  8. Mechanics of the IL2RA gene activation revealed by modeling and atomic force microscopy.

    Directory of Open Access Journals (Sweden)

    Pascale Milani

    Full Text Available Transcription implies recruitment of RNA polymerase II and transcription factors (TFs by DNA melting near transcription start site (TSS. Combining atomic force microscopy and computer modeling, we investigate the structural and dynamical properties of the IL2RA promoter and identify an intrinsically negative supercoil in the PRRII region (containing Elf-1 and HMGA1 binding sites, located upstream of a curved DNA region encompassing TSS. Conformational changes, evidenced by time-lapse studies, result in the progressive positioning of curvature apex towards the TSS, likely facilitating local DNA melting. In vitro assays confirm specific binding of the General Transcription Factors (GTFs TBP and TFIIB over TATA-TSS position, where an inhibitory nucleosome prevented preinitiation complex (PIC formation and uncontrolled DNA melting. These findings represent a substantial advance showing, first, that the structural properties of the IL2RA promoter are encoded in the DNA sequence and second, that during the initiation process DNA conformation is dynamic and not static.

  9. The use of Optical Magnetic Twisting Cytometry and Flourescence Resonance Energy Transfer to quantify force-induced protein dissociation in the nucleus of a living cell

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Yeh-Chuin Poh & Ning Wang ### Abstract Mechanical forces are known to play a significant role in biological processes. These forces can be transmitted to the cell through the cytoskeletal filament network, inducing different biochemical responses within the cytoplasm. Although there have been ample reports showing that cytoplasmic enzymes can be directly activated by a local stress on the cell surface via integrins, there has been no evidence that mechanical forces can di...

  10. Fluoroquinolone-induced gene transfer in multidrug-resistant Salmonella

    Science.gov (United States)

    Fluoroquinolones are broad spectrum antibiotics that inhibit bacterial DNA gyrase and topoisomerase activity. Bacterial exposure to fluoroquinolones can cause DNA damage and induce a bacterial SOS response to stimulate repair of damaged DNA. Certain prophages (integrated in bacterial chromosomes) ...

  11. Orbitofrontal cortex action of 5-hydroxytryptamine and its receptor in an acute forced swimming stress-induced depression model

    Institute of Scientific and Technical Information of China (English)

    Huipeng Li; Fengli An; Shucheng An

    2009-01-01

    BACKGROUND: The orbitofrontal cortex (OFC) is a brain region closely associated with emotion.5-hydroxytryptamine (5-HT) has been shown to be involved in human depression.OBJECTIVE: To investigate OFC actions and mechanisms of 5-HT and 5-HT1A receptor (5-HT1AR)in stress-induced depression.DESIGN, TIME AND SEI-rlNG: A randomized, controlled, animal experiment was performed at Laboratory of Neurobiology, College of Life Science, Shaanxi Normal University between May 2006 and March 2008.MATERIALS: 5-HT, p-chlorophenylalanine (PCPA, an inhibitor to tryptophan hydroxylase) andspiperone (5-HT1AR antagonist) were provided by Sigma, USA; rabbit anti-rat 5-HT1AR antibody was provided by Tlanjin Haoyang Biological Manufacture.METHODS: A total of 40 male Sprague Dawley rats, aged 3 months, were randomly divided into five groups: control, model, 5-HT, spiperone+5-HT, and PCPA, with 8 rats in each group. Except for control group, rats in the other four groups were used to establish depression models by forced swimming for 15 minutes. At 30 minutes before forced swimming test, 0.5μL of 5-HT (12.5μg/pL),PCPA (20μg/μL), spiperone (1.3 μg/μL)+5-HT (12.5μg/μL, 10 minutes later), and saline were respectively injected into the OFC of 5-HT, PCPA, spiperone+5-HT, and model groups, respectively.The control group received a saline microinjection into the OFC.MAIN OUTCOME MEASURES: Forced swimming and open field tests were employed to measure animal behaviors, and immunohistochemistry was used to analyze 5-HT1AR expression in the OFC,cingulate cortex, and piriform cortex.RESULTS: (1) Compared with the model group, 5-HT microinjection into the OFC prominently reduced immobility time in the forced swimming test and rearing in open field test (P0.05). Furthermore, following PCPA microinjection into the OFC (PCPA + forced swimming stress),immobility time in forced swimming test increased dramatically (P<0.01), locomotion and rearing inopen field test declined (P<0.05 and P<0

  12. Radiation-induced gene expression in the nematode caenorhabditis elegans

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, G.A.; Jones, T.A.; Chesnut, A.; Smith, A.L. [Loma Linda Univ., CA (United States)

    2002-12-01

    We used the nematode C. elegans to characterize the genotoxic and cytotoxic effects of ionizing radiation in a simple animal model emphasizing the unique effects of charged particle radiation. Here we demonstrate by reverse transcription polymerase chain reaction (RT-PCR) differential display and whole genome microarray hybridization experiments that gamma rays, accelerated protons and iron ions at the same physical dose lead to unique transcription profiles. 599 of 17871 genes analyzed (3.4%) showed differential expression 3 hrs after exposure to 3 Gy of radiation. 193 were up-regulated, 406 were down-regulated and 90% were affected only by a single species of radiation. A novel statistical clustering technique identified the regulatory relationships between the radiation-modulated genes and showed that genes affected by each radiation species were associated with unique regulatory clusters. This suggests that independent homeostatic mechanisms are activated in response to radiation exposure as a function of track structure or ionization density. (author)

  13. Radiation-induced gene expression in the nematode Caenorhabditis elegans

    Science.gov (United States)

    Nelson, Gregory A.; Jones, Tamako A.; Chesnut, Aaron; Smith, Anna L.

    2002-01-01

    We used the nematode C. elegans to characterize the genotoxic and cytotoxic effects of ionizing radiation in a simple animal model emphasizing the unique effects of charged particle radiation. Here we demonstrate by RT-PCR differential display and whole genome microarray hybridization experiments that gamma rays, accelerated protons and iron ions at the same physical dose lead to unique transcription profiles. 599 of 17871 genes analyzed (3.4%) showed differential expression 3 hrs after exposure to 3 Gy of radiation. 193 were up-regulated, 406 were down-regulated and 90% were affected only by a single species of radiation. A novel statistical clustering technique identified the regulatory relationships between the radiation-modulated genes and showed that genes affected by each radiation species were associated with unique regulatory clusters. This suggests that independent homeostatic mechanisms are activated in response to radiation exposure as a function of track structure or ionization density.

  14. Substrate-induced gene expression screening: a method for high-throughput screening of metagenome libraries.

    Science.gov (United States)

    Uchiyama, Taku; Miyazaki, Kentaro

    2010-01-01

    The SIGEX (substrate-induced gene expression) method is a novel approach for the screening of gene (genome) libraries. In addition to the commonly used function- and sequence-driven approaches to screening, SIGEX provides a third option; in SIGEX, positives are identified using a reporter gene, and the library is constructed using an "operon-trap" vector. This vector contains the reporter gene immediately downstream of the cloning site for the genomic insert so that the expression of the inserted gene(s) is coupled with that of the reporter gene. This system is especially suitable for screening catabolic genes that are induced in response to metabolically relevant compounds, such as substrates. If expression of the inserted gene(s) is activated in response to the addition of these compounds, then positive clones can be identified based on the reporter signal. The most effective selection is obtained by the use of a FACS (fluorescence-activated cell sorter) in conjunction with a FACS-compatible fluorescent reporter protein, such as GFP (green fluorescent protein). Activity-based screening of metagenomic libraries often suffers from low sensitivity and low throughput. In contrast, the high throughput, high sensitivity, and versatility of SIGEX make it a particularly suitable method for screening metagenomic libraries.

  15. Modulation of Gene Expression Networks underlying Realgar-Induced Differentiation of Acute Promyelocytic Leukemia Cells

    Institute of Scientific and Technical Information of China (English)

    王怀宇; 刘陕西

    2002-01-01

    Objective: To elucidate the molecular mechanism of the differentiation of acute promyelocytic leukemia (APL) cell line NB4 induced by realgar. Methods: The response of NB4 cell to realgar was explored with a cDNA microarray representing 1003 different human genes. Results: The analysis of gene expression profiles indicated that 8 genes were up-regulated and 33 genes were down-regulated 48 hrs after realgar treatment. Among the 8 up-regulated genes, 2 genes were involved in ubiquitin proteasome degradation pathway. Some genes related to RNA processing, protein synthesis and signal transduction were down-regulated. Conclusion: The ubiquitin-proteasome degradation pathway may play an important role in the degradation of PML/RAR α fusion protein and the differentiation of NB4 cells.

  16. Resonance energy transfer (RET)-Induced intermolecular pairing force: a tunable weak interaction and its application in SWNT separation.

    Science.gov (United States)

    Pan, Xiaoyong; Chen, Hui; Wang, Wei Zhi; Ng, Siu Choon; Chan-Park, Mary B

    2011-07-21

    This paper explores evidence of an optically mediated interaction that is active in the separation mechanism of certain selective agents through consideration of the contrasting selective behaviors of two conjugated polymers with distinct optical properties. The involvement of a RET-induced intermolecular pairing force is implied by the different illumination response behaviors. The magnitude of this interaction scales with the external stimulus parameter, the illumination irradiance (I), and thus is tunable. This suggests a facile technique to modify the selectivity of polymers toward specific SWNT species by altering the polymer structure to adjust the corresponding intermolecular interaction. This is the first experimental verification and application of a RET-induced intermolecular pairing force to SWNT separation. With this kind of interaction taken into account, reasonable interpretation of some conflicting data, especially PLE maps, can be easily made. The above conclusion can be applied to other substances as long as they are electrically neutral and there is photon-induced RET between them. The significant magnitude of this interaction makes direct manipulation of molecules/particles possible and is expected to have applications in molecular engineering.

  17. Forced egg retention induced by diethyl-phenylacetamide diminishes the fecundity and longevity of dengue vectors

    Directory of Open Access Journals (Sweden)

    Thangaraj Seenivasagan

    2015-01-01

    Interpretation & conclusion: Presence of DEPA odor on water surface forced the gravid Aedes females to retain their eggs, affecting their fecundity and longevity. This indigenously developed topical repellent DEPA may be applied to container′s water that act as breeding grounds for dengue vectors to deter gravid females from oviposition in the vicinity of a household.

  18. Self-induced dipole force and filamentation instability of a matter wave

    DEFF Research Database (Denmark)

    Saffman, M.

    1998-01-01

    The interaction of copropagating electromagnetic and matter waves is described with a set of coupled higher-order nonlinear Schrodinger equations. Optical self-focusing modulates an initially planar wave leading to the generation of dipole forces on the atoms. Atomic channeling due to the dipole...

  19. "Atomic Force Masking" Induced Formation of Effective Hot Spots along Grain Boundaries of Metal Thin Films.

    Science.gov (United States)

    Kim, Kwang Hyun; Chae, Soo Sang; Jang, Seunghun; Choi, Won Jin; Chang, Hyunju; Lee, Jeong-O; Lee, Tae Il

    2016-11-30

    We present an interesting phenomenon, "atomic force masking", which is the deposition of a few-nanometer-thick gold film on ultrathin low-molecular-weight (LMW) polydimethylsiloxane (PDMS) engineered on a polycrystalline gold thin film, and demonstrated the formation of hot spot based on SERS. The essential principle of this atomic force masking phenomenon is that an LMW PDMS layer on a single crystalline grain of gold thin film would repel gold atoms approaching this region during a second cycle of evaporation, whereas new nucleation and growth of gold atoms would occur on LMW PDMS deposited on grain boundary regions. The nanostructure formed by the atomic force masking, denoted here as "hot spots on grain boundaries" (HOGs), which is consistent with finite-difference time-domain (FDTD) simulation, and the mechanism of atomic force masking were investigated by carrying out systematic experiments, and density functional theory (DFT) calculations were made to carefully explain the related fundamental physics. Also, to highlight the manufacturing advantages of the proposed method, we demonstrated the simple synthesis of a flexible HOG SERS, and we used this substrate in a swabbing test to detect a common pesticide placed on the surface of an apple.

  20. The effect of a single nucleotide polymorphism in the matrix metalloproteinase-1 (MMP-1) promoter on force-induced MMP-1 expression in human periodontal ligament cells

    NARCIS (Netherlands)

    Huang, Sheng-Fu; Li, Yu-Hong; Ren, Yi-Jin; Cao, Zheng-Guo; Long, Xing

    2008-01-01

    Matrix metalloproteinase-1 (MMP-1) 1G/2G (-1,607) polymorphisms have been identified and shown to influence the transcription of the MMP-1 gene. In order to compare the expression of MMP-1 with different MMP-1 gene promoter alleles after force loading, human periodontal ligament (PDL) cells were cul

  1. A finite element perturbation method for computing fluid-induced forces on a certrifugal impeller rotating and whirling in a volute casing

    NARCIS (Netherlands)

    Jonker, Jan B.; van Essen, T.G.; van Essen, T.G.

    1997-01-01

    A finite element based method has been developed for computing time-averaged fluid-induced radial excitation forces and rotor dynamic forces on a two-dimensional centrifugal impeller rotating and whirling in a volute casing. In this method potential flow theory is used, which implies the assumption

  2. A finite element perturbation method for computing fluid-induced forces on a centrifugal impeller rotating and whirling in a volute casing

    NARCIS (Netherlands)

    Jonker, J.B.; Essen, van T.G.

    1997-01-01

    A finite element based method has been developed for computing time-averaged fluid-induced radial excitation forces and rotor dynamic forces on a two-dimensional centrifugal impeller rotating and whirling in a volute casing. In this method potential flow theory is used, which implies the assumption

  3. Chitosan nanoparticles as non-viral gene delivery vehicles based on atomic force microscopy study

    Institute of Scientific and Technical Information of China (English)

    Yujing Yuan; Jieyi Tan; Yifei Wang; Chuiwen Qian; Meiying Zhang

    2009-01-01

    Chitosan (CS), a biocompatible and biodegradable material, can act as a non-viral delivery vehicle with low toxicity. In this study, plasmid DNA (pDNA) and siRNA were encapsulated in CS nanoparticles (NPs) to prepare CS-DNA and CS-siRNA NPs using a complex coacervation process. The CS-DNA particle size was within the range of 180-370 nm with a surface charge ranging from 0 to 18 mV at pH 5.5. The stability of pDNA in CS-DNA was investigated by pDNA release study and DNase I protection assay. The release of pDNA from NPs was studied in pH 7.4 phosphatebuffered saline at 37℃ and the CS-DNA NPs could delay the DNA release. Results of DNase I protection assay showed that CS-DNA NPs could protect the encapsulated pDNA from nuclease degradation. In the transfection study, it was found that the transfection efficiency in vitro was dependent on the molecular weight, charge ratio, and DNA concentration of the CS-DNA NP as well as the type of cell transfected. Moreover, the morphology of HeLa cells transfected with CS-siRNA complexes was studied using atomic force microscopy. The results suggest that CS may be more capable than liposome in delivering siRNA to target cells. In summary,our analysis suggests that pDNA and siRNA can be encapsulated in CS NPs without being damaged.

  4. Differentially Gene Expression Profile Related to Inflammation in Endometrial Cells Induce by Lipopolysaccharide

    Institute of Scientific and Technical Information of China (English)

    Li-hua QIN; Ruo-guang WANG; Sheng LI; Chun-mei LI

    2009-01-01

    Objective To investigate differentially expressed genes related to inflammation in endometrial cells induced by Lipopolysaccharide(LPS).Methods Normal endometrium in the proliferative phase of specimen from 3 cases for the experiment was collected. The LPS group were treated with 50 μg/ml LPS. Total RNA was extracted using Trizol reagent from cells. RNA quality was assessed by determining the OD260/280 ratio by agarose gel electrophoresis, the chip was scanned by laser scanner. The acquired was analyzed. Results A total of differentially expressed genes were found, these genes were relative to many aspects. Among them, the expression of genes involved in inflammation were up-regulated by LPS, such as overexpression of lL-lβ, 8, etc.Conclusion The results indicates that inflammation-related genes may be one of the mechanisms of abnormal uterine bleeding by LPS-induced.

  5. Identification of in vivo-induced genes during infection of chickens with Salmonella enterica serovar Enteritidis.

    Science.gov (United States)

    Geng, Shizhong; Liu, Zhicheng; Lin, Zhijie; Barrow, Paul; Pan, Zhiming; Li, Qiuchun; Jiao, Xinan

    2015-06-01

    Chickens are an important source of food worldwide and are often infected with food-poisoning serovars of Salmonella enterica, frequently Salmonella Enteritidis (SE), without exhibiting clinical signs of disease. Ivi (in vivo induced) genes identified using in vivo-induced antigen technology (IVIAT) are expressed only during bacterial infection and have the potential value of identifying epidemic strains and antigens which can form the basis for sub-unit vaccine development. We applied IVIAT to SE strain 50041 and identified 42 ivi genes. Eight representative ivi genes were further confirmed by qRT-PCR as being expressed only in vivo within 48 h of infection compared with that of in vitro-cultured. Although our results indicated that the identified ivi genes are expressed only in vivo, further research is needed to elucidate the exact roles of these genes during infection and pathogenesis.

  6. Coordinate gene regulation by fimbriae-induced signal transduction

    DEFF Research Database (Denmark)

    Schembri, Mark; Klemm, Per

    2001-01-01

    of Ag43 production. No effect was observed in an oxyR mutant. We conclude that fimbriae expression per se constitutes a signal transduction mechanism that affects a number of unrelated genes via the thiol-disulfide status of OxyR. Thus, phase variation in fimbrial expression is coordinated...

  7. Identification of genes induced by salt stress from Medicago ...

    African Journals Online (AJOL)

    hope&shola

    2010-11-08

    Nov 8, 2010 ... Local Alignment Search Tool (BLAST) analysis of deduced protein sequences revealed that 35 .... uni-ESTs may be either new genes or segments unique to .... Yang QC, Wu MS, Wang QP, Kang JM (2005). Cloning ... Page 7 ...

  8. Concise Review: The Dynamics of Induced Pluripotency and Its Behavior Captured in Gene Network Motifs

    NARCIS (Netherlands)

    Muraro, M.J.; Kempe, H.; Verschure, P.J.

    2013-01-01

    The flexibility of cellular identity is clearly demonstrated by the possibility to reprogram fully differentiated somatic cells to induced pluripotent stem (iPS) cells through forced expression of a set of transcription factors. The generation of iPS cells is of great interest as they provide a trem

  9. Mechanically induced silyl ester cleavage under acidic conditions investigated by AFM-based single-molecule force spectroscopy in the force-ramp mode.

    Science.gov (United States)

    Schmidt, Sebastian W; Pill, Michael F; Kersch, Alfred; Clausen-Schaumann, Hauke; Beyer, Martin K

    2014-01-01

    AFM-based dynamic single-molecule force spectroscopy was used to stretch carboxymethylated amylose (CMA) polymers, which have been covalently tethered between a silanized glass substrate and a silanized AFM tip via acid-catalyzed ester condensation at pH 2.0. Rupture forces were measured as a function of temperature and force loading rate in the force-ramp mode. The data exhibit significant statistical scattering, which is fitted with a maximum likelihood estimation (MLE) algorithm. Bond rupture is described with a Morse potential based Arrhenius kinetics model. The fit yields a bond dissociation energy De = 35 kJ mol(-1) and an Arrhenius pre-factor A = 6.6 × 10(4) s(-1). The bond dissociation energy is consistent with previous experiments under identical conditions, where the force-clamp mode was employed. However, the bi-exponential decay kinetics, which the force-clamp results unambiguously revealed, are not evident in the force-ramp data. While it is possible to fit the force-ramp data with a bi-exponential model, the fit parameters differ from the force-clamp experiments. Overall, single-molecule force spectroscopy in the force-ramp mode yields data whose information content is more limited than force-clamp data. It may, however, still be necessary and advantageous to perform force-ramp experiments. The number of successful events is often higher in the force-ramp mode, and competing reaction pathways may make force-clamp experiments impossible.

  10. Use of induced acceleration to quantify the (de)stabilization effect of external and internal forces on postural responses.

    Science.gov (United States)

    van Asseldonk, Edwin H F; Carpenter, Mark G; van der Helm, Frans C T; van der Kooij, Herman

    2007-12-01

    Due to the mechanical coupling between the body segments, it is impossible to see with the naked eye the causes of body movements and understand the interaction between movements of different body parts. The goal of this paper is to investigate the use of induced acceleration analysis to reveal the causes of body movements. We derive the analytical equations to calculate induced accelerations and evaluate its potential to study human postural responses to support-surface translations. We measured the kinematic and kinetic responses of a subject to sudden forward and backward translations of a moving platform. The kinematic and kinetics served as input to the induced acceleration analyses. The induced accelerations showed explicitly that the platform acceleration and deceleration contributed to the destabilization and restabilization of standing balance, respectively. Furthermore, the joint torques, coriolis and centrifugal forces caused by swinging of the arms, contributed positively to stabilization of the Center of Mass. It is concluded that induced acceleration analyses is a valuable tool in understanding balance responses to different kinds of perturbations and may help to identify the causes of movement in different pathologies.

  11. Cold Atom Physics Using Ultra-Thin Optical Fibers: Light-Induced Dipole Forces and Surface Interactions

    CERN Document Server

    Sagu'e, G; Meschede, D; Rauschenbeutel, A; Vetsch, E

    2007-01-01

    The strong evanescent field around ultra-thin unclad optical fibers bears a high potential for detecting, trapping, and manipulating cold atoms. Introducing such a fiber into a cold atom cloud, we investigate the interaction of a small number of cold Caesium atoms with the guided fiber mode and with the fiber surface. Using high resolution spectroscopy, we observe and analyze light-induced dipole forces, van der Waals interaction, and a significant enhancement of the spontaneous emission rate of the atoms. The latter can be assigned to the modification of the vacuum modes by the fiber.

  12. Ongoing characterization of the forced electron beam induced arc discharge ion source for the selective production of exotic species facility

    Energy Technology Data Exchange (ETDEWEB)

    Manzolaro, M., E-mail: mattia.manzolaro@lnl.infn.it; Andrighetto, A.; Monetti, A.; Scarpa, D.; Rossignoli, M.; Vasquez, J.; Corradetti, S.; Calderolla, M.; Prete, G. [INFN, Laboratori Nazionali di Legnaro, Viale dell’Universita’ 2 - 35020 Legnaro, Padova,Italy (Italy); Meneghetti, G. [Department of Industrial Engineering, University of Padova, Via Venezia 1 - 35131 Padova (Italy)

    2014-02-15

    An intense research and development activity to finalize the design of the target ion source system for the selective production of exotic species (SPES) facility (operating according to the isotope separation on line technique) is at present ongoing at Legnaro National Laboratories. In particular, the characterization of ion sources in terms of ionization efficiency and transversal emittance is currently in progress, and a preliminary set of data is already available. In this work, the off-line ionization efficiency and emittance measurements for the SPES forced electron beam induced arc discharge ion source in the case of a stable Ar beam are presented in detail.

  13. Isolation of the alkane inducible cytochrome P450 (P450alk) gene from the yeast Candida tropicalis

    Science.gov (United States)

    The gene for the alkane-inducible cytochrome P450, P450alk, has been isolated from the yeast Candida tropicalis by immunoscreening a λgt11 library. Isolation of the gene has been identified on the basis of its inducibility and partial DNA sequence. Transcripts of this gene were i...

  14. Isolation of the alkane inducible cytochrome P450 (P450alk) gene from the yeast Candida tropicalis

    Science.gov (United States)

    The gene for the alkane-inducible cytochrome P450, P450alk, has been isolated from the yeast Candida tropicalis by immunoscreening a λgt11 library. Isolation of the gene has been identified on the basis of its inducibility and partial DNA sequence. Transcripts of this gene were i...

  15. Development of Agrobacterium-mediated virus-induced gene silencing and performance evaluation of four marker genes in Gossypium barbadense.

    Directory of Open Access Journals (Sweden)

    Jinhuan Pang

    Full Text Available Gossypiumbarbadense is a cultivated cotton species and possesses many desirable traits, including high fiber quality and resistance to pathogens, especially Verticilliumdahliae (a devastating pathogen of Gossypium hirsutum, the main cultivated species. These elite traits are difficult to be introduced into G. hirsutum through classical breeding methods. In addition, genetic transformation of G. barbadense has not been successfully performed. It is therefore important to develop methods for evaluating the function and molecular mechanism of genes in G. barbadense. In this study, we had successfully introduced a virus-induced gene silencing (VIGS system into three cultivars of G. barbadense by inserting marker genes into the tobacco rattle virus (TRV vector. After we optimized the VIGS conditions, including light intensity, photoperiod, seedling age and Agrobacterium strain, 100% of plants agroinfiltrated with the GaPDS silencing vector showed white colored leaves. Three other marker genes, GaCLA1, GaANS and GaANR, were employed to further test this VIGS system in G. barbadense. The transcript levels of the endogenous genes in the silenced plants were reduced by more than 99% compared to control plants; these plants presented phenotypic symptoms 2 weeks after inoculation. We introduced a fusing sequence fragment of GaPDS and GaANR gene silencing vectors into a single plant, which resulted in both photobleaching and brownish coloration. The extent of silencing in plants agroinfiltrated with fusing two-gene-silencing vector was consistent with plants harboring a single gene silencing vector. The development of this VIGS system should promote analysis of gene function in G. barbadense, and help to contribute desirable traits for breeding of G. barbadense and G. hirsutum.

  16. Changes in gene expression induced by Micro-Immunotherapy

    Directory of Open Access Journals (Sweden)

    Maurice Jeaner

    2012-09-01

    Full Text Available Background: Metabolic syndrome (MS is a metabolic disorder associated with obesity, type-II diabetes, and “low grade inflammation”, with the concomitant increased risk of cardiovascular events. As a chronic inflammatory process, MS results in a dysregulation of the cytokine profile. 2L®INFLAM, a Micro-immunotherapy (MI medication formulated with highly diluted cytokines, is currently prescribed in Belgium for inflammatory diseases and potentially may be helpful for MS patients. Aims: To investigate the impact of 2L®INFLAM on selected gene expression markers (mRNA in patients suffering from MS, in addition to biological and clinical parameters. Methodology: Four well characterized MS adult patients with stabilized body-weight were advised to take one capsule of 2L®INFLAM per day (by sublingual-oral route for 6 months (composition in table 1. Concomitantly to biological and clinical examination, genes expression status was assessed by a DNA microarray technology (Oxygen™ comprising 200 genes involved mainly in oxidative stress and inflammation. Whole blood collection was performed before and after treatment (3-6 months and mRNA levels measured. Gene expression was classified in 3 series (normally expressed, up or down-regulated and genes related to diabetes predisposition were scored by using a proprietary Diascore (Probiox. Results: Before MI medication, a significant percentage of dysregulated genes (median: 16.3% as well as a positive Diascore (median: 1.6 were noticed. Impressive correction of dysregulated genes (reaching 90% for one patient was observed after 3 months of treatment (median: 2.3% in addition to an improvement of Diascore in 3 MS patients out of 4 (median: 0.5. During the same period, both clinical and biological parameters remained unchanged. Conclusions: MS patients showing a high level of gene dysregulation efficiently normalized after 3 months of 2L

  17. Force-induced dynamical properties of multiple cytoskeletal filaments are distinct from that of single filaments

    CERN Document Server

    Das, Dipjyoti; Padinhateeri, Ranjith

    2014-01-01

    How cytoskeletal filaments collectively undergo growth and shrinkage is an intriguing question. Collective properties of multiple bio-filaments (actin or microtubules) undergoing hydrolysis, have not been studied extensively earlier, within simple theoretical frameworks. In this paper, we show that collective properties of multiple filaments under force are very distinct from the properties of a single filament under similar conditions -- these distinctions manifest as follows: (i) the collapse time during collective catastrophe for a multifilament system is much larger than that of a single filament with the same average length, (ii) force-dependence of the cap-size distribution of multiple filaments are quantitatively different from that of single filament, (iii) the diffusion constant associated with the system length fluctuations is distinct for multiple filaments, (iv) switching dynamics of multiple filaments between capped and uncapped states and the fluctuations therein are also distinct. We build a un...

  18. Capillary-force-induced formation of luminescent polystyrene/(rare-earth-doped nanoparticle) hybrid hollow spheres.

    Science.gov (United States)

    Chen, Min; Xie, Lin; Li, Fuyou; Zhou, Shuxue; Wu, Limin

    2010-10-01

    This paper presents a "one-pot" procedure to synthesize polystyrene/(rare-earth-doped nanoparticles) (PS/REDNPs) hybrid hollow spheres via the in situ diffusion of organic core into inorganic shell under strong capillary force. In this approach, when carboxyl-capped PS colloids were deposited by different REDNPs in aqueous medium, such as LaF3:Eu3+, LaF3:Ce3+-Tb3+, and YVO4:Dy3+, PS/REDNPs inorganic-organic hybrid hollow spheres could be directly obtained via the in situ diffusion of core PS chains into the voids between rare-earth-doped nanoparticles through the strong capillary force. Not only is the synthetic procedure versatile and very simple, but also the obtained hybrid hollow spheres are hydrophilic and luminescent and could be directly used in chemical and biological fields.

  19. Wheel-rail dynamic forces induced by random vertical track irregularities

    Science.gov (United States)

    Spiroiu, M. A.

    2016-08-01

    The present paper investigates the wheel-rail dynamic forces produced by railway vehicles in motion, which are an important issue especially for the high-speed rail transport from the point of view of traffic safety, ride quality and undesirable effects on vehicles, on track and on the land in the vicinity of railways. The research is carried out on a model which includes track system, vehicle unsprung mass, vehicle primary suspension and the bogie sprung mass. The wheel-rail dynamic overloads are evaluated assuming random vertical irregularities of the track. The estimation of wheel-rail dynamic forces is made for a range of vehicle speeds up to 300 km/h and the influence of track and vehicle various parameters is investigated.

  20. Biomechanically inspired modelling of pedestrian-induced forces on laterally oscillating structures

    Science.gov (United States)

    Bocian, M.; Macdonald, J. H. G.; Burn, J. F.

    2012-07-01

    Despite considerable interest among engineers and scientists, bi-directional interaction between walking pedestrians and lively bridges has still not been well understood. In an attempt to bridge this gap a biomechanically inspired model of the human response to lateral bridge motion is presented and explored. The simple inverted pendulum model captures the key features of pedestrian lateral balance and the resulting forces on the structure. The forces include self-excited components that can be effectively modelled as frequency-dependent added damping and mass to the structure. The results of numerical simulations are in reasonable agreement with recent experimental measurements of humans walking on a laterally oscillating treadmill, and in very good agreement with measurements on full-scale bridges. In contrast to many other models of lateral pedestrian loading, synchronisation with the bridge motion is not involved. A parametric study of the model is conducted, revealing that as pedestrians slow down as a crowd becomes more dense, their resulting lower pacing rates generate larger self-excited forces. For typical pedestrian parameters, the potential to generate negative damping arises for any lateral bridge vibration frequency above 0.43 Hz, depending on the walking frequency. Stability boundaries of the combined pedestrian-structure system are presented in terms of the structural damping ratio and pedestrian-to-bridge mass ratio, revealing complex relations between damping demand and bridge and pedestrian frequencies, due to the added mass effect. Finally it is demonstrated that the model can produce simultaneous self-excited forces on multiple structural modes, and a realistic full simulation of a large number of pedestrians, walking randomly and interacting with a bridge, produces structural behaviour in very good agreement with site observations.

  1. Steady dynein forces induce flutter instability and propagating waves in mathematical models of flagella.

    Science.gov (United States)

    Bayly, P V; Dutcher, S K

    2016-10-01

    Cilia and flagella are highly conserved organelles that beat rhythmically with propulsive, oscillatory waveforms. The mechanism that produces these autonomous oscillations remains a mystery. It is widely believed that dynein activity must be dynamically regulated (switched on and off, or modulated) on opposite sides of the axoneme to produce oscillations. A variety of regulation mechanisms have been proposed based on feedback from mechanical deformation to dynein force. In this paper, we show that a much simpler interaction between dynein and the passive components of the axoneme can produce coordinated, propulsive oscillations. Steady, distributed axial forces, acting in opposite directions on coupled beams in viscous fluid, lead to dynamic structural instability and oscillatory, wave-like motion. This 'flutter' instability is a dynamic analogue to the well-known static instability, buckling. Flutter also occurs in slender beams subjected to tangential axial loads, in aircraft wings exposed to steady air flow and in flexible pipes conveying fluid. By analysis of the flagellar equations of motion and simulation of structural models of flagella, we demonstrate that dynein does not need to switch direction or inactivate to produce autonomous, propulsive oscillations, but must simply pull steadily above a critical threshold force.

  2. Gene expression induced by Toll-like receptors in macrophages requires the transcription factor NFAT5.

    Science.gov (United States)

    Buxadé, Maria; Lunazzi, Giulia; Minguillón, Jordi; Iborra, Salvador; Berga-Bolaños, Rosa; Del Val, Margarita; Aramburu, José; López-Rodríguez, Cristina

    2012-02-13

    Toll-like receptors (TLRs) engage networks of transcriptional regulators to induce genes essential for antimicrobial immunity. We report that NFAT5, previously characterized as an osmostress responsive factor, regulates the expression of multiple TLR-induced genes in macrophages independently of osmotic stress. NFAT5 was essential for the induction of the key antimicrobial gene Nos2 (inducible nitric oxide synthase [iNOS]) in response to low and high doses of TLR agonists but is required for Tnf and Il6 mainly under mild stimulatory conditions, indicating that NFAT5 could regulate specific gene patterns depending on pathogen burden intensity. NFAT5 exhibited two modes of association with target genes, as it was constitutively bound to Tnf and other genes regardless of TLR stimulation, whereas its recruitment to Nos2 or Il6 required TLR activation. Further analysis revealed that TLR-induced recruitment of NFAT5 to Nos2 was dependent on inhibitor of κB kinase (IKK) β activity and de novo protein synthesis, and was sensitive to histone deacetylases. In vivo, NFAT5 was necessary for effective immunity against Leishmania major, a parasite whose clearance requires TLRs and iNOS expression in macrophages. These findings identify NFAT5 as a novel regulator of mammalian anti-pathogen responses.

  3. System for stable β-estradiol-inducible gene expression in the moss Physcomitrella patens.

    Science.gov (United States)

    Kubo, Minoru; Imai, Akihiro; Nishiyama, Tomoaki; Ishikawa, Masaki; Sato, Yoshikatsu; Kurata, Tetsuya; Hiwatashi, Yuji; Reski, Ralf; Hasebe, Mitsuyasu

    2013-01-01

    Inducible transgene expression provides a useful tool to analyze gene function. The moss Physcomitrellapatens is a model basal land plant with well-developed research tools, including a high efficiency of gene targeting and substantial genomics resources. However, current systems for controlled transgene expression remain limited. Here we report the development of an estrogen receptor mediated inducible gene expression system, based on the system used in flowering plants. After identifying the appropriate promoters to drive the chimeric transducer, we succeeded in inducing transcription over 1,000-fold after 24 h incubation with β-estradiol. The P. patens system was also effective for high-level long-term induction of gene expression; transcript levels of the activated gene were maintained for at least seven days on medium containing β-estradiol. We also established two potentially neutral targeting sites and a set of vectors for reproducible expression of two transgenes. This β-estradiol-dependent system will be useful to test genes individually or in combination, allowing stable, inducible transgenic expression in P. patens.

  4. Interferon induced IFIT family genes in host antiviral defense

    Science.gov (United States)

    Secretion of interferons (IFNs) from virus-infected cells is a hallmark of host antiviral immunity and in fact, IFNs exert their antiviral activities through the induction of antiviral proteins. The IFN-induced protein with tetratricopeptide repeats (IFITs) family is among hundreds of IF stimulated ...

  5. Thermally induced osteocyte damage initiates pro-osteoclastogenic gene expression in vivo.

    Science.gov (United States)

    Dolan, Eimear B; Tallon, David; Cheung, Wing-Yee; Schaffler, Mitchell B; Kennedy, Oran D; McNamara, Laoise M

    2016-06-01

    Bone is often subject to harsh temperatures during orthopaedic procedures resulting in thermally induced bone damage, which may affect the healing response. Postsurgical healing of bone is essential to the success of surgery, therefore, an understanding of the thermally induced responses of bone cells to clinically relevant temperatures in vivo is required. Osteocytes have been shown to be integrally involved in the bone remodelling cascade, via apoptosis, in micro-damage systems. However, it is unknown whether this relationship is similar following thermal damage. Sprague-Dawley rat tibia were exposed to clinically relevant temperatures (47°C or 60°C) to investigate the role of osteocytes in modulating remodelling related factors. Immunohistochemistry was used to quantify osteocyte thermal damage (activated caspase-3). Thermally induced pro-osteoclastogenic genes (Rankl, Opg and M-csf), in addition to genes known to mediate osteoblast and osteoclast differentiation via prostaglandin production (Cox2), vascularization (Vegf) and inflammatory (Il1a) responses, were investigated using gene expression analysis. The results demonstrate that heat-treatment induced significant bone tissue and cellular damage. Pro-osteoclastogenic genes were upregulated depending on the amount of temperature elevation compared with the control. Taken together, the results of this study demonstrate the in vivo effect of thermally induced osteocyte damage on the gene expression profile.

  6. Virus-induced opposite effect on Bombyx mori gene transcriptions

    Directory of Open Access Journals (Sweden)

    Y Yin

    2016-09-01

    Full Text Available Bombyx mori bidensovirus (BmBDV and Bombyx mori nucleopolyhedrovirus (BmNPV are serious pathogens of Bombyx mori. In this study, we reported the changes of transcription level of several immune genes, including bmi, argo, dicer, cap1, cap3 and car, in Bombyx mori midgut after exposure to BmBDV or BmNPV. Silkworm strains 798 (anti-BmBDV and 306 (susceptible to BmBDV were subjected to BmBDV infection, and NB (anti-BmNPV and HUABA (35 (susceptible to BmNPV were subjected to BmNPV infection. The results showed that the transcription levels differ largely among different silkworm strains, and that the extent to which the gene transcriptions were affected by the viruses was different. However, both BmNPV and BmBDV viruses can reverse the transcription patterns of these genes when the silkworms were administered with the viruses compared with those control groups. The transcript levels of bmi and dicer were decreased in 798 and 306 strains that were inoculated with BmBDV compared with their respective controls, but were increased in NB and HUABA (35 inoculated with BmNPV. The transcript levels of argo and cap3 were risen in 798, 306 and NB strains when inoculated with their respective viruses, but were decreased in HUABA (35 strain. The transcript levels of cap1 were risen in all silkworm strains, while the levels of car were decreased in 798, 306 and HUABA (35 strains, and increased in NB strain when inoculated with their respective viruses. These findings may contribute to more in-depth understanding on functions of these genes in virus infection and proliferation.

  7. HBV X Gene Transfection Upregulates IL-1β and IL-6 Gene Expression and Induces Rat Glomerular Mesangial Cell Proliferation

    Institute of Scientific and Technical Information of China (English)

    Hongzhu LU; Jianhua ZHOU

    2008-01-01

    The X gene of HBV encodes a 17-KD protein, termed HBx, which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements. The aim of this study was to investigate the effect of HBx on gene expression of interleukin (IL)-1β and IL-6, and proliferation of rat mesangial cells in vitro. The X gene of HBV was amplified by PCR assay, and inserted into the eukaryotic expression vector pCI-neo. The structure of recombinant pCI-neo-X plasmid was proved by restrict endonuclease digestion and sequencing analysis. pCI-neo-X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfected mesangial cells was detected by Western blot. The IL-1β and IL-6 mRNA expression in those cells was assayed by semiquantitative RT-PCR. Mesangial cell proliferation was tested by MTT. The results showed that HBx was obviously expressed in cultured mesangial cell line at 36th and 48th h after transfection. The expression of IL-1β and IL-6 mRNA was simultaneously increased. The cell proliferation was also obvious at the same time. It was concluded that HBx gene transfection could induce IL-1β and IL-6 gene expression and mesangial cell proliferation. HBx may play a critical role in mesangial cell proliferation through upregulation of the IL-1β and IL-6 gene expression.

  8. Exploring Mycobacterium tuberculosis infection-induced alterations in gene expression in macrophage by microarray hybridization

    Institute of Scientific and Technical Information of China (English)

    XIE; Jianping; (谢建平); LI; Yao; (李; 瑶); YUE; Jun; (乐; 军); XU; Yongzhong; (徐永忠); HUANG; Daqiang; (黄达蔷); LIANG; Li; (梁; 莉); WANG; Honghai; (王洪海)

    2003-01-01

    Tuberculosis remains a serious threat to public health. Its causative agent Mycobacte- rium tuberculosis is an intracellular pathogen which survives and replicates within cells of the host immune system, primarily macrophages. Knowledge of the bacteria-macrophage interaction can help to develop novel measures to combat the disease. The global gene expression of macro- phage following invasion by and growth of M. tuberculosis was studied by cDNA microarray. Of the 12800 human genes analyzed, totally 473 (3.7%) macrophage genes were differentially expressed after being infected by M. tuberculosis, among which, only 25 (5.2%, corresponding to less than 0.2% of the 12800 genes) genes were up-regulated, while others (94.8%) were down-regulated against the control. Of the 473 genes, 376 genes are registered in the GenBank, and 97 are novel genes. Expression of 5 up-regulated genes has been induced by more than 3-fold. 25 genes were down-regulated by more than 3-fold. Syndecan binding protein has been down-regu- lated up to 12.5-fold. The data gave an insight into the early gene expression in macrophage ensuing M. tuberculosis infection and a basis for further study.

  9. Myostatin gene mutated mice induced with tale nucleases.

    Science.gov (United States)

    Zhou, Fangfang; Sun, Ruilin; Chen, Hongyan; Fei, Jian; Lu, Daru

    2015-01-01

    Myostain gene (MSTN) is expressed primarily in skeletal muscle, and negatively regulates skeletal muscle mass; it has been suggested that mice with MSTN inhibition have reduced adiposity and improved insulin sensitivity. Therefore, it is important to establish a fast and effective gene editing method. In this report, we established the myostatin mutated-mouse model by microinjection of Transcription Activator-Like Effector Nucleases (TALENs) mRNA within the mouse fertilized oocytes and achieved high rates of mutagenesis of the mouse MSTN in C57BL/6J. Six of 45 born mice carried target mutations and we appointed one as the parental mating with wild mouse to produce the F1 and backcross to produce the F2 generation. All the mutations of the mice were examined quickly and efficiently by high-resolution melting curve analysis (HRMA) and then verified by direct sequencing. We obtained the homozygous of the F2 generation which transmitted the mutant alleles to the progeny with 100% efficiency. Mutant mice exhibited increases in muscle mass comparable to those observed in wild-type mice. Therefore, combining TALEN-mediated gene targeting with HRMA technology is a superior method of constructing genetically modified mice through microinjection in the mouse fertilized oocytes with high efficiency and short time of selection.

  10. Isolation and characterization of a human apoptosis-inducing gene with yeast two-hybrid system

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    asy gene is a novel apoptosis-inducing gene,but its mechanism is unclear.To investigate the mechanism of asy inducing apoptosis,a novel gene encoding ASY interacting protein (asyip) is isolated from human lung cell line (WI-38) cDNA library with yeast two-hybrid system.The asyip gene is constitutively expressed as two mRNA transcripts with the size of 1.8 and 2.7 kb in various human tissues at different levels.Sequence analysis of full-length cDNA reveals that the two alternative transcripts of asyip gene contain common 5' end and different 3' end,and share a common open reading frame encoding a polypeptide of 236 amino acids.Two protein kinase C phosphorylation sites and two casein kinase II phosphorylation sites are found in ASYIP amino acid sequence.Two highly hydrophobic regions encoding potentially two transmembrane domains are present.The ASYIP protein contains a C-terminal endoplasmic reticulum retrieval signal (Lys-Lys-Lys-Ala-Glu).Immunoprecipitation assay confirmed the interaction of ASY and ASYIP in mammalian cells.Compared with asy gene,overexpression of asyip gene can inhibit growth of tumor cell Saos2 and induce cell apoptosis with a low efficiency.

  11. Cigarette smoke modulates expression of human rhinovirus-induced airway epithelial host defense genes.

    Directory of Open Access Journals (Sweden)

    David Proud

    Full Text Available Human rhinovirus (HRV infections trigger acute exacerbations of chronic obstructive pulmonary disease (COPD and asthma. The human airway epithelial cell is the primary site of HRV infection and responds to infection with altered expression of multiple genes, the products of which could regulate the outcome to infection. Cigarette smoking aggravates asthma symptoms, and is also the predominant risk factor for the development and progression of COPD. We, therefore, examined whether cigarette smoke extract (CSE modulates viral responses by altering HRV-induced epithelial gene expression. Primary cultures of human bronchial epithelial cells were exposed to medium alone, CSE alone, purified HRV-16 alone or to HRV-16+ CSE. After 24 h, supernatants were collected and total cellular RNA was isolated. Gene array analysis was performed to examine mRNA expression. Additional experiments, using real-time RT-PCR, ELISA and/or western blotting, validated altered expression of selected gene products. CSE and HRV-16 each induced groups of genes that were largely independent of each other. When compared to gene expression in response to CSE alone, cells treated with HRV+CSE showed no obvious differences in CSE-induced gene expression. By contrast, compared to gene induction in response to HRV-16 alone, cells exposed to HRV+CSE showed marked suppression of expression of a number of HRV-induced genes associated with various functions, including antiviral defenses, inflammation, viral signaling and airway remodeling. These changes were not associated with altered expression of type I or type III interferons. Thus, CSE alters epithelial responses to HRV infection in a manner that may negatively impact antiviral and host defense outcomes.

  12. A comparative study examining the cytotoxicity of inducible gene expression system ligands in different cell types.

    Science.gov (United States)

    Xie, Jinger; Nair, Ayyappan; Hermiston, Terry W

    2008-02-01

    Inducible gene expression systems are being used in many in vitro and in vivo applications for target discovery, target validation and as components in exploratory therapeutic agents. Ideally, the ligands, which activate the systems, are benign so that the effects can be strictly attributed to the induced protein. As a first step to defining the potential effects of these inducers, we tested three of them, doxycycline, muristerone A and mifepristone (for tet-, ecdysone- and progesterone antagonist-inducible systems respectively), for toxicity across a panel of normal cells and cancer cell lines. In contrast to both muristerone A and mifepristone that showed no significant toxicity on any of the tested cells, we observed that doxycycline induced cell death in selected cancer and primary cell lines. The different susceptibility of cell lines to the ligands commonly used in these inducible systems suggests that it is important to consider the effects of the inducers prior to their use in experimental in vitro cell culture systems.

  13. Inducible amplification of gene copy number and heterologous protein production in the yeast Kluyveromyces lactis.

    Science.gov (United States)

    Morlino, G B; Tizzani, L; Fleer, R; Frontali, L; Bianchi, M M

    1999-11-01

    Heterologous protein production can be doubled by increasing the copy number of the corresponding heterologous gene. We constructed a host-vector system in the yeast Kluyveromyces lactis that was able to induce copy number amplification of pKD1 plasmid-based vectors upon expression of an integrated copy of the plasmid recombinase gene. We increased the production and secretion of two heterologous proteins, glucoamylase from the yeast Arxula adeninivorans and mammalian interleukin-1beta, following gene dosage amplification when the heterologous genes were carried by pKD1-based vectors. The choice of the promoters for expression of the integrated recombinase gene and of the episomal heterologous genes are critical for the mitotic stability of the host-vector system.

  14. Regulation of endogenous human gene expression by ligand-inducible TALE transcription factors.

    Science.gov (United States)

    Mercer, Andrew C; Gaj, Thomas; Sirk, Shannon J; Lamb, Brian M; Barbas, Carlos F

    2014-10-17

    The construction of increasingly sophisticated synthetic biological circuits is dependent on the development of extensible tools capable of providing specific control of gene expression in eukaryotic cells. Here, we describe a new class of synthetic transcription factors that activate gene expression in response to extracellular chemical stimuli. These inducible activators consist of customizable transcription activator-like effector (TALE) proteins combined with steroid hormone receptor ligand-binding domains. We demonstrate that these ligand-responsive TALE transcription factors allow for tunable and conditional control of gene activation and can be used to regulate the expression of endogenous genes in human cells. Since TALEs can be designed to recognize any contiguous DNA sequence, the conditional gene regulatory system described herein will enable the design of advanced synthetic gene networks.

  15. Technical advances in trigger-induced RNA interference gene silencing in the parasite Entamoeba histolytica.

    Science.gov (United States)

    Khalil, Mohamed I; Foda, Bardees M; Suresh, Susmitha; Singh, Upinder

    2016-03-01

    Entamoeba histolytica has a robust endogenous RNA interference (RNAi) pathway. There are abundant 27 nucleotide (nt) anti-sense small RNAs (AS sRNAs) that target genes for silencing and the genome encodes many genes involved in the RNAi pathway such as Argonaute proteins. Importantly, an E. histolytica gene with numerous AS sRNAs can function as a "trigger" to induce silencing of a gene that is fused to the trigger. Thus, the amebic RNAi pathway regulates gene expression relevant to amebic biology and has additionally been harnessed as a tool for genetic manipulation. In this study we have further improved the trigger-induced gene silencing method. We demonstrate that rather than using the full-length gene, a short portion of the coding region fused to a trigger is sufficient to induce silencing; the first 537 bp of the E. histolytica rhomboid gene (EhROM1) fused in-frame to the trigger was sufficient to silence EhROM1. We also demonstrated that the trigger method could silence two amebic genes concomitantly; fusion of the coding regions of EhROM1 and transcription factor, EhMyb, in-frame to a trigger gene resulted in both genes being silenced. Alternatively, two genes can be silenced sequentially: EhROM1-silenced parasites with no drug selection plasmid were transfected with trigger-EhMyb, resulting in parasites with both EhROM1 and EhMyb silenced. With all approaches tested, the trigger-mediated silencing was substantive and silencing was maintained despite loss of the G418 selectable marker. All gene silencing was associated with generation of AS sRNAs to the silenced gene. We tested the reversibility of the trigger system using inhibitors of histone modifications but found that the silencing was highly stable. This work represents a technical advance in the trigger gene silencing method in E. histolytica. Approaches that readily silence multiple genes add significantly to the genetic toolkit available to the ameba research community. Copyright © 2016

  16. Knockdown of col22a1 gene in zebrafish induces a muscular dystrophy by disruption of the myotendinous junction.

    Science.gov (United States)

    Charvet, Benjamin; Guiraud, Alexandre; Malbouyres, Marilyne; Zwolanek, Daniela; Guillon, Emilie; Bretaud, Sandrine; Monnot, Catherine; Schulze, Jörg; Bader, Hannah L; Allard, Bruno; Koch, Manuel; Ruggiero, Florence

    2013-11-01

    The myotendinous junction (MTJ) is the major site of force transfer in skeletal muscle, and defects in its structure correlate with a subset of muscular dystrophies. Col22a1 encodes the MTJ component collagen XXII, the function of which remains unknown. Here, we have cloned and characterized the zebrafish col22a1 gene and conducted morpholino-based loss-of-function studies in developing embryos. We showed that col22a1 transcripts localize at muscle ends when the MTJ forms and that COLXXII protein integrates the junctional extracellular matrix. Knockdown of COLXXII expression resulted in muscular dystrophy-like phenotype, including swimming impairment, curvature of embryo trunk/tail, strong reduction of twitch-contraction amplitude and contraction-induced muscle fiber detachment, and provoked significant activation of the survival factor Akt. Electron microscopy and immunofluorescence studies revealed that absence of COLXXII caused a strong reduction of MTJ folds and defects in myoseptal structure. These defects resulted in reduced contractile force and susceptibility of junctional extracellular matrix to rupture when subjected to repeated mechanical stress. Co-injection of sub-phenotypic doses of morpholinos against col22a1 and genes of the major muscle linkage systems showed a synergistic gene interaction between col22a1 and itga7 (α7β1 integrin) that was not observed with dag1 (dystroglycan). Finally, pertinent to a conserved role in humans, the dystrophic phenotype was rescued by microinjection of recombinant human COLXXII. Our findings indicate that COLXXII contributes to the stabilization of myotendinous junctions and strengthens skeletal muscle attachments during contractile activity.

  17. Local Muscle Metabolic Demand Induced by Neuromuscular Electrical Stimulation and Voluntary Contractions at Different Force Levels: A NIRS Study.

    Science.gov (United States)

    Muthalib, Makii; Kerr, Graham; Nosaka, Kazunori; Perrey, Stephane

    2016-06-13

    Functional Muscle metabolic demand during contractions evoked by neuromuscular electrical stimulation (NMES) has been consistently documented to be greater than voluntary contractions (VOL) at the same force level (10-50% maximal voluntary contraction-MVC). However, we have shown using a near-infrared spectroscopy (NIRS) technique that local muscle metabolic demand is similar between NMES and VOL performed at MVC levels, thus controversy exists. This study therefore compared biceps brachii muscle metabolic demand (tissue oxygenation index-TOI and total hemoglobin volume-tHb) during a 10s isometric contraction of the elbow flexors between NMES (stimulation frequency of 30Hz and current level to evoke 30% MVC) and VOL at 30% MVC (VOL-30%MVC) and MVC (VOL-MVC) level in 8 healthy men (23-33-y). Greater changes in TOI and tHb induced by NMES than VOL-30%MVC confirm previous studies of a greater local metabolic demand for NMES than VOL at the same force level. The same TOI and tHb changes for NMES and VOL-MVC suggest that local muscle metabolic demand and intramuscular pressure were similar between conditions. In conclusion, these findings indicate that NMES induce a similar local muscle metabolic demand as that of maximal VOL.

  18. Identification of antiviralrelevant genes in the cultured fish cells induced by UV-inactivated virus

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    UV-inactivated grass carp hemorrhage virus (GCHV) can induce high titer of interferon in cultured CAB (crucian carp (Carassius auratus L.) blastulae) cells, and thus defend host cells against the virus invasion. The mechanism is proposed that an antiviral state should be established in the host cells by activating expression of a set of antivi-ral-relevant genes. In this study, suppressive subtractive hy-bridization is applied to constructing a subtracted cDNA library with mRNAs isolated from UV-inactivated GCHV infected and mock-infected CAB cells. 272 differential cDNA fragments are identified by both PCR and dot blot from the subtractive cDNA library. Sequencing analysis reveals 69 genes, including 46 known gene homologues, and 23 unknown putative genes. The known genes include the genes involved in interferon signaling pathways, such as Stat1 and Jak1, the antiviral genes, such as Mx and Viperin, and a set of interferon-stimulated genes observed in mammalian cells. Most of the unknown putative genes contain AU-rich ele-ment in their sequences. Differential expressions of these genes are further confirmed by virtual Northern blot and RT-PCR. The data imply that UV-inactivated GCHV is not only able to induce production of interferon in the infected CAB cells, but also leads to the expression of a series of antiviral-relevant genes or immune-relevant genes, and therefore reveals that the signaling pathway of interferon system and antiviral mechanism in fish are similar to those in mammals.

  19. Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis.

    Science.gov (United States)

    van der Linde, Karina; Kastner, Christine; Kumlehn, Jochen; Kahmann, Regine; Doehlemann, Gunther

    2011-01-01

    Infection of maize (Zea mays) plants with the corn smut fungus Ustilago maydis leads to the formation of large tumors on the stem, leaves and inflorescences. In this biotrophic interaction, plant defense responses are actively suppressed by the pathogen, and previous transcriptome analyses of infected maize plants showed massive and stage-specific changes in host gene expression during disease progression. To identify maize genes that are functionally involved in the interaction with U. maydis, we adapted a virus-induced gene silencing (VIGS) system based on the brome mosaic virus (BMV) for maize. Conditions were established that allowed successful U. maydis infection of BMV-preinfected maize plants. This set-up enabled quantification of VIGS and its impact on U. maydis infection using a quantitative real-time PCR (qRT-PCR)-based readout. In proof-of-principle experiments, an U. maydis-induced terpene synthase was shown to negatively regulate disease development while a protein involved in cell death inhibition was required for full virulence of U. maydis. The results suggest that this system is a versatile tool for the rapid identification of maize genes that determine compatibility with U. maydis.

  20. Differential expression of genes during aflatoxin B1-induced hepatocarcinogenesis in tree shrews

    Institute of Scientific and Technical Information of China (English)

    Yuan Li; Dan Luo; Hui-Fen Yue; Li-Sheng Zhang; Jian-Ren Gu; Da-Fang Wan; Jian-Jia Su; Ji Cao; Chao Ou; Xiao-Kun Qiu; Ke-Chen Ban; Chun Yang; Liu-Liang Qin

    2004-01-01

    AIM: Through exploring the regulation of gene expression during hepatocarcinogenesis induced by aflatoxin B1 (AFB1),to find out the responsible genes for hepatocellular carcinoma (HCC) and to further understand the underlying molecular mechanism.METHODS: Tree shrews ( 7upaia belangeri chinensis)were treated with or without AFB1 for about 90 weeks. Liver biopsies were performed regularly during the animal experiment. Eight shares of total RNA were respectively isolated from 2 HCC tissues, 2 HCC-surrounding noncancerous liver tissues, 2 biopsied tissues at the early stage (30th week) of the experiment from the same animals as above, 1 mixed sample of three liver tissues biopsied at the beginning (0th week) of the experiment, and another 1 mixed sample of two liver tissues from the untreated control animals biopsied at the 90th week of the experiment. The samples were then tested with the method of AtlasTM cDNA microarray assay. The levels of gene expression in these tissues taken at different time points during hepatocarcinogenesis were compared.RESULTS: The profiles of differently expressed genes were quite different in different ways of comparison. At the same period of hepatocarcinogenesis, the genes in the same function group usually had the same tendency for up- or down-regulation. Among the checked 588 genes that were known to be related to human cancer, 89 genes (15.1%) were recognized as "important genes" because they showed frequent changes in different ways of comparison. The differentially expressed genes during hepatocarcinogenesis could be classified into four categories: genes up-regulated in HCC tissue, genes with similar expressing levels in both HCC and HCC-surrounding liver tissues which were higher than that in the tissues prior to the development of HCC,genes down-regulated in HCC tissue, and genes up-regulated prior to the development of HCC but down-regulated after the development of HCC.CONCLUSION: A considerable number of genes could change

  1. Gene expression in rat skin induced by irritating chemicals.

    Science.gov (United States)

    Rogers, James V; Garrett, Carol M; McDougal, James N

    2003-01-01

    Occupational skin disease is the second most significant cause of occupational disease, after accidents. Irritation from occupational chemicals such as solvents, hydrocarbons, and surfactants are one cause of this disease. Gene expression studies provide useful information about normal processes in the skin and responses of the skin to exogenous chemicals. We exposed rats, cutaneously, to sodium lauryl sulfate (SLS, 1% and 10% aqueous solution), m-xylene (pure liquid), and d-limonene (pure liquid) for 1 h and measured transcriptional responses at the end of the exposure and 3 h later for comparison with untreated skin samples. Total skin RNA was isolated and analyzed using the Affymetrix RatTox U34 array. Using the Affymetrix software, we found that 234 of approximately 850 genes were detected as present in at least 80% of the normal skin samples. The largest number of these genes was related to metabolism, oxidative/cellular stress, and signal transduction. Limonene caused the largest change in mRNA levels with a total of 34 increased transcripts and 4 decreased transcripts. Xylene treatment resulted in 6 increased transcripts and 14 decreased transcripts, while 10% SLS caused 5 transcripts to increase and 17 to decrease. Only two transcripts were observed to change in skin following a 1% SLS exposure. Sodium lauryl sulfate transcript changes increased with dose and were maximum at 4 h. Limonene transcript changes were more numerous at 1 h than at 4 h. The observed differences may reflect different mechanisms of irritation. Copyright 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:123-137, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.10079

  2. Atomic Force Microscopy Studies on DNA Structural Changes Induced by Vincristine Sulfate and Aspirin

    Science.gov (United States)

    Zhu, Yi; Zeng, Hu; Xie, Jianming; Ba, Long; Gao, Xiang; Lu, Zuhong

    2004-04-01

    We report that atomic force microscopy (AFM) studies on structural variations of a linear plasmid DNA interact with various concentrations of vincristine sulfate and aspirin. The different binding images show that vincrinstine sulfate binding DNA chains caused some loops and cleavages of the DNA fragments, whereas aspirin interaction caused the width changes and conformational transition of the DNA fragments. Two different DNA structural alternations could be explained by the different mechanisms of the interactions with these two components. Our work indicates that the AFM is a powerful tool in studying the interaction between DNA and small molecules.

  3. Directed Current Induced by an Symmetrically ac Force Coexisting with a Time-Delayed Feedback

    Institute of Scientific and Technical Information of China (English)

    易述婷; 宋晖; 欧志娥; 艾保全; 熊建文

    2012-01-01

    We study the transport of overdamped Brownian particles in a symmetricaJly periodic potential in the presence of an asymmetrically ac driving force and a time-delayed feedback. It is found that for low frequencies, the average velocity can be negative by changing the driving amplitude, for high frequencies, there exists an optimized driving amplitude at which the average velocity takes its maximum value. Additionally, there is a threshold value of driving amplitude below which no directed transport can be obtained for high frequencies. For the large value of the delay time, the average velocity is independent of the delay time.

  4. Stable suspension and dispersion-induced transitions from repulsive Casimir forces between fluid separated eccentric cylinders

    Energy Technology Data Exchange (ETDEWEB)

    Dalvit, Diego A1 [Los Alamos National Laboratory; Rodriguez, Alejandro W [MASS INST OF TECH; Munday, J N [HARVARD UNIV; Joannopoulos, J D [MASS INST OF TECH

    2008-01-01

    Using accurate numerical methods for finite-size nonplanar objects, we demonstrate a stable mechanical suspension of a silica cylinder within a metallic cylinder separated by ethanol, via a repulsive Casimir force between the silica and the metal. We investigate cylinders with both circular and square cross sections, and show that the latter exhibit a stable orientation as well as a stable position, employing a new method to accurately compute Casimir torques for finite objects. Furthermore, the stable orientation of the square cylinder is shown to undergo an unusual 45 transition as a function of the separation lengthscale, and this transition is explained as a consequence of material dispersion.

  5. Ferromagnetic-resonance induced electromotive forces in Ni81Fe19 | p-type diamond

    Science.gov (United States)

    Fukui, Naoki; Morishita, Hiroki; Kobayashi, Satoshi; Miwa, Shinji; Mizuochi, Norikazu; Suzuki, Yoshishige

    2016-10-01

    We report on direct-current (DC) electromotive forces (emfs) in a nickel-iron alloy (Ni81 Fe19) | p-type diamond under the ferromagnetic resonance of the Ni81Fe19 layer at room temperature. The observed DC emfs take its maximum around the ferromagnetic resonant frequency of the Ni81Fe19, and their signs are reversed by reversing the direction of an externally-applied magnetic field; it shows that the observed DC emfs are spin-related emfs.

  6. Host-Induced Silencing of Pathogenicity Genes Enhances Resistance to Fusarium oxysporum Wilt in Tomato.

    Science.gov (United States)

    Bharti, Poonam; Jyoti, Poonam; Kapoor, Priya; Sharma, Vandana; Shanmugam, V; Yadav, Sudesh Kumar

    2017-08-01

    This study presents a novel approach of controlling vascular wilt in tomato by RNAi expression directed to pathogenicity genes of Fusarium oxysporum f. sp. lycopersici. Vascular wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici leads to qualitative and quantitative loss of the crop. Limitation in the existing control measures necessitates the development of alternative strategies to increase resistance in the plants against pathogens. Recent findings paved way to RNAi, as a promising method for silencing of pathogenicity genes in fungus and provided effective resistance against fungal pathogens. Here, two important pathogenicity genes FOW2, a Zn(II)2Cys6 family putative transcription regulator, and chsV, a putative myosin motor and a chitin synthase domain, were used for host-induced gene silencing through hairpinRNA cassettes of these genes against Fusarium oxysporum f. sp. lycopersici. HairpinRNAs were assembled in appropriate binary vectors and transformed into tomato plant targeting FOW2 and chsV genes, for two highly pathogenic strains of Fusarium oxysporum viz. TOFOL-IHBT and TOFOL-IVRI. Transgenic tomatoes were analyzed for possible attainment of resistance in transgenic lines against fungal infection. Eight transgenic lines expressing hairpinRNA cassettes showed trivial disease symptoms after 6-8 weeks of infection. Hence, the host-induced posttranscriptional gene silencing of pathogenicity genes in transgenic tomato plants has enhanced their resistance to vascular wilt disease caused by Fusarium oxysporum.

  7. Identification of Candida albicans genes induced during thrush offers insight into pathogenesis.

    Science.gov (United States)

    Cheng, Shaoji; Clancy, Cornelius J; Checkley, Mary Ann; Handfield, Martin; Hillman, Jeffrey D; Progulske-Fox, Ann; Lewin, Alfred S; Fidel, Paul L; Nguyen, M Hong

    2003-06-01

    Candida albicans causes a wide spectrum of diseases, ranging from mucocutaneous infections like oral thrush to disseminated candidiasis. Screening for C. albicans genes expressed within infected hosts might advance understanding of candidal pathogenesis, but is impractical using existing techniques. In this study, we used an antibody-based strategy to identify C. albicans genes expressed during thrush. We adsorbed sera from HIV-infected patients with thrush against candidal cells grown in vitro and screened a C. albicans genomic expression library. We identified 10 genes encoding immunogenic antigens and used reverse transcription-polymerase chain reaction to verify that they were induced within thrush pseudomembranes recovered from a patient. The in vivo induced genes are involved in diverse functions, including regulation of yeast-hyphal morphogenesis, adhesion to host cells, nutrient uptake, phospholipid biosynthesis and amino acid catabolism. Four genes encode known virulence determinants (HWP1, CST20, CPP1 and RBF1). Another gene, LPD1, for which a role in candidal pathogenesis is unknown, encodes a protein homologous to a bacterial virulence determinant. Most importantly, disruption of CaNOT5, a newly identified gene, conferred defects in morphogenesis, decreased adherence to human buccal epithelial cells and attenuated mortality during murine disseminated candidiasis, proving that our strategy can identify genes encoding novel virulence determinants.

  8. Identification of Differently Expressed Genes in Chemical Carcinogen-induced Rat Bladder Cancers

    Institute of Scientific and Technical Information of China (English)

    Guangfu CHEN; Franky L. CHAN; Xu ZHANG; Peter S.F. CHAN

    2009-01-01

    Possible altered gene expression patterns in bladder turnout carcinogenesis in rat bladder cancers induced by BBN [N-butyl-N-(4-hydroxybutyl)nitrosamine] was examined by cDNA microarray analysis of gene expression profiles.Thirty Sprague-Dawley rats were given drinking water containing 0.05% BBN ad libitum for 24 to 28-weeks.Equal numbers of control rats were given tap water without BBN.After treatment,the rat bladders were excised for RNA extraction and histopathological examinations.Total RNAs were extracted from rat transitional cell carcinoma (TCC) tissues and micro-dissected normal rat bladder epithelia.The atlas glass rat microarray was used,which included oligonucleotides of 1081 rat genes.Some of the up-regulated genes in rat bladder TCCs were further confirmed by Northern blotting.Our results showed that the transcriptions of 30 genes were significantly elevated in the rat bladder TCCs,and these included fly proto-oncogene,Lipocortin 2,COX Ⅳ,COX Ⅴ a,and cathepsin D.Also,15 genes were significantly down-regulated in the rat bladder TCCs and they included B7.1,TNFrl,APOAI and VHL.The resuits of cDNA microarray analysis demonstrated that normal rat bladder epithelia and bladder TCC exhibited different and specific gene statement profiles.The increased expressions of the identified genes may play an important role in the chemically induced bladder carcinogenesis.

  9. TIMP-1 gene deficiency increases tumour cell sensitivity to chemotherapy-induced apoptosis

    DEFF Research Database (Denmark)

    Davidsen, Marie Louise; Würts, S.Ø.; Rømer, Maria Unni Koefoed;

    2006-01-01

    in cancer. In this regard, several studies have demonstrated an antiapoptotic effect of TIMP-1 in a number of different cell types. Since chemotherapy works by inducing apoptosis in cancer cells, we raised the hypothesis that TIMP-1 promotes resistance against chemotherapeutic drugs. In order to investigate...... this hypothesis, we have established TIMP-1 gene-deficient and TIMP-1 wild-type fibrosarcoma cells from mouse lung tissue. We have characterised these cells with regard to TIMP-1 genotype, TIMP-1 expression, malignant transformation and sensitivity to chemotherapy-induced apoptosis. We show that TIMP-1 gene...... deficiency increases the response to chemotherapy considerably, confirming that TIMP-1 protects the cells from apoptosis. This is to our knowledge the first study investigating TIMP-1 and chemotherapy-induced apoptosis employing a powerful model system comprising TIMP-1 gene-deficient cells...

  10. The model of defense gene expression induced by signaling molecule β-ocimene

    Institute of Scientific and Technical Information of China (English)

    LIU Chunlin; RUAN Ying; GUAN Chunyun

    2004-01-01

    @@ β-ocimene, a kind of monoterpene, was found recently as a plant communication signal molecule[1]. It has two isomeric forms in nature: cis-β-ocimene and trans-β- ocimene. According to recent reports, all investigated plants, such as corn, cotton, lima bean, potato, tobacco, arabidopsis, and Mediterranean pine, could release the chemical component β-ocimene after fed by arthropod herbivores[2-5], suggesting thatβ-ocimene is an important functioal component in the herbivore-induced volatile. Nowadays, we know that β-ocimene can induce the expression of defense genes relative to salicylic acid in detatched leaves. But many problems of β-ocimene, for example, whether β-ocimene can induce the defense gene expression in intact plants, what role it can play in the expression model of defense genes, are elusive[1,6].

  11. Muscle Contraction Induces Acute Hydroxymethylation of the Exercise-Responsive Gene Nr4a3

    DEFF Research Database (Denmark)

    Pattamaprapanont, Pattarawan; Garde, Christian; Fabre, Odile

    2016-01-01

    promoters. Exercise induces dynamic DNA demethylation at gene promoters; however, the contribution of the demethylation precursor hydroxymethylcytosine is unknown. Given the evanescent nature of hydroxymethylcytosine, a muscle contraction model that allows for the collection of samples that are repeatedly...... stimulated over time is required to determine whether contraction-induced demethylation is preceded by changes in the hydroxymethylcytosine level. Here, we established an acute skeletal muscle contraction model to mimic the effects of acute exercise on gene expression. We used this model to investigate...... the effect of muscle contraction on DNA demethylation and hydroxymethylation. First, we performed an acute exercise study in healthy humans to identify an exercise-responsive gene that we could study in culture. We identified the nuclear receptor subfamily 4 group A member 3 (Nr4a3) gene with the highest...

  12. Nonimmunoglobulin target loci of activation-induced cytidine deaminase (AID) share unique features with immunoglobulin genes.

    Science.gov (United States)

    Kato, Lucia; Begum, Nasim A; Burroughs, A Maxwell; Doi, Tomomitsu; Kawai, Jun; Daub, Carsten O; Kawaguchi, Takahisa; Matsuda, Fumihiko; Hayashizaki, Yoshihide; Honjo, Tasuku

    2012-02-14

    Activation-induced cytidine deaminase (AID) is required for both somatic hypermutation and class-switch recombination in activated B cells. AID is also known to target nonimmunoglobulin genes and introduce mutations or chromosomal translocations, eventually causing tumors. To identify as-yet-unknown AID targets, we screened early AID-induced DNA breaks by using two independent genome-wide approaches. Along with known AID targets, this screen identified a set of unique genes (SNHG3, MALAT1, BCL7A, and CUX1) and confirmed that these loci accumulated mutations as frequently as Ig locus after AID activation. Moreover, these genes share three important characteristics with the Ig gene: translocations in tumors, repetitive sequences, and the epigenetic modification of chromatin by H3K4 trimethylation in the vicinity of cleavage sites.

  13. The Agricultural Antibiotic Carbadox Induces Phage-mediated Gene Transfer in Salmonella

    Directory of Open Access Journals (Sweden)

    Bradley L. Bearson

    2014-02-01

    Full Text Available Antibiotics are used for disease therapeutic or preventative effects in humans and animals, as well as for enhanced feed conversion efficiency in livestock. Antibiotics can also cause undesirable effects in microbial populations, including selection for antibiotic resistance, enhanced pathogen invasion, and stimulation of horizontal gene transfer. Carbadox is a veterinary antibiotic used in the U.S. during the starter phase of swine production for improved feed efficiency and control of swine dysentery and bacterial swine enteritis. Carbadox has been shown in vitro to induce phage-encoded Shiga toxin in Shiga toxin-producing Escherichia coli and a phage-like element transferring antibiotic resistance genes in Brachyspira hyodysenteriae, but the effect of carbadox on prophages in other bacteria is unknown. This study examined carbadox exposure on prophage induction and genetic transfer in Salmonella enterica serovar Typhimurium, a human foodborne pathogen that frequently colonizes swine without causing disease. S. Typhimurium LT2 exposed to carbadox induced prophage production, resulting in bacterial cell lysis and release of virions that were visible by electron microscopy. Carbadox induction of phage-mediated gene transfer was confirmed by monitoring the transduction of a sodCIII::neo cassette in the Fels-1 prophage from LT2 to a recipient Salmonella strain. Furthermore, carbadox frequently induced generalized transducing phages in multidrug-resistant phage type DT104 and DT120 isolates, resulting in the transfer of chromosomal and plasmid DNA that included antibiotic resistance genes. Our research indicates that exposure of Salmonella to carbadox induces prophages that can transfer virulence and antibiotic resistance genes to susceptible bacterial hosts. Carbadox-induced, phage-mediated gene transfer could serve as a contributing factor in bacterial evolution during animal production, with prophages being a reservoir for bacterial fitness

  14. Reduced Tyk2 gene expression in β-cells due to natural mutation determines susceptibility to virus-induced diabetes

    OpenAIRE

    Izumi, Kenichi; Mine, Keiichiro; Inoue, Yoshitaka; Teshima, Miho; Ogawa, Shuichiro; Kai, Yuji; Kurafuji, Toshinobu; Hirakawa, Kanako; Miyakawa, Daiki; Ikeda, Haruka; Inada, Akari; Hara, Manami; Yamada, Hisakata; Akashi, Koichi; Niho, Yoshiyuki

    2015-01-01

    Accumulating evidence suggests that viruses play an important role in the development of diabetes. Although the diabetogenic encephalomyocarditis strain D virus induces diabetes in restricted lines of inbred mice, the susceptibility genes to virus-induced diabetes have not been identified. We report here that novel Tyrosine kinase 2 (Tyk2) gene mutations are present in virus-induced diabetes-sensitive SJL and SWR mice. Mice carrying the mutant Tyk2 gene on the virus-resistant C57BL/6 backgrou...

  15. Stable oncogenic transformation induced by microcell-mediated gene transfer

    Institute of Scientific and Technical Information of China (English)

    吕有勇; Donald G.Blair

    1995-01-01

    Oncogenes have been identified using DNA-mediated transfection, but the size of the transferable and unrearranged DNA, gene rearrangement and amplification which occur during the transfection process limit the use of the techniques. We have evaluated microcell-mediated gene transfer techniques for the transfer and analysis of dominant oncogenes. MNNG-HOS, a transformed human cell line which contained the met oncogene mapping to human chromosome 7 was infected with retroviruses carrying drug resistance markers and used to optimize microcell preparation and transfer. Stable and drug-resistant hybrids containing single human chromosomes as well as the foci of the transformed cells containing the activated met oncogene and intact hitman chromosomes were obtained. Hybridization analysis with probes (i.e. collA2, pJ3.11) mapping up to 1 Mb away from met shows that the cells from the individual focr contain different amounts of apparently unrearranged human DNA associated with the oncogene, and the microcell-g

  16. Differential expression of genes during aflatoxin B1-induced hepatocarcinogenesis in tree shrews

    Science.gov (United States)

    Li, Yuan; Wan, Da-Fang; Su, Jian-Jia; Cao, Ji; Ou, Chao; Qiu, Xiao-Kun; Ban, Ke-Chen; Yang, Chun; Qin, Liu-Liang; Luo, Dan; Yue, Hui-Fen; Zhang, Li-Sheng; Gu, Jian-Ren

    2004-01-01

    AIM: Through exploring the regulation of gene expression during hepatocarcinogenesis induced by aflatoxin B1 (AFB1), to find out the responsible genes for hepatocellular carcinoma (HCC) and to further understand the underlying molecular mechanism. METHODS: Tree shrews (Tupaia belangeri chinensis) were treated with or without AFB1 for about 90 weeks. Liver biopsies were performed regularly during the animal experiment. Eight shares of total RNA were respectively isolated from 2 HCC tissues, 2 HCC-surrounding non-cancerous liver tissues, 2 biopsied tissues at the early stage (30th week) of the experiment from the same animals as above, 1 mixed sample of three liver tissues biopsied at the beginning (0th week) of the experiment, and another 1 mixed sample of two liver tissues from the untreated control animals biopsied at the 90th week of the experiment. The samples were then tested with the method of AtlasTM cDNA microarray assay. The levels of gene expression in these tissues taken at different time points during hepatocarcinogenesis were compared. RESULTS: The profiles of differently expressed genes were quite different in different ways of comparison. At the same period of hepatocarcinogenesis, the genes in the same function group usually had the same tendency for up- or down-regulation. Among the checked 588 genes that were known to be related to human cancer, 89 genes (15.1%) were recognized as “important genes” because they showed frequent changes in different ways of comparison. The differentially expressed genes during hepatocarcinogenesis could be classified into four categories: genes up-regulated in HCC tissue, genes with similar expressing levels in both HCC and HCC-surrounding liver tissues which were higher than that in the tissues prior to the development of HCC, genes down-regulated in HCC tissue, and genes up-regulated prior to the development of HCC but down-regulated after the development of HCC. CONCLUSION: A considerable number of genes could

  17. Reynolds-dependence of turbulent skin-friction drag reduction induced by spanwise forcing

    CERN Document Server

    Gatti, Davide

    2015-01-01

    This paper examines how increasing the value of the Reynolds number $Re$ affects the ability of spanwise-forcing techniques to yield turbulent skin-friction drag reduction. The control strategy is the streamwise-travelling waves of spanwise wall velocity (Quadrio {\\em et al. J. Fluid Mech.}, vol. 627, 2009, pp. 161--178). The study builds upon an extensive drag-reduction database created with Direct Numerical Simulation of a turbulent channel flow for two, 5-fold separated values of $Re$, namely $Re_\\tau=200$ and $Re_\\tau=1000$. The sheer size of the database, which for the first time systematically addresses the amplitude of the forcing, allows a comprehensive view of the drag-reducing characteristics of the traveling waves, and enables a detailed description of the changes occurring when $Re$ increases. The effect of using a viscous scaling based on the friction velocity of either the non-controlled flow or the drag-reduced flow is described. In analogy with other wall-based drag reduction techniques, like ...

  18. ohr, Encoding an Organic Hydroperoxide Reductase, Is an In Vivo-Induced Gene in Actinobacillus pleuropneumoniae

    OpenAIRE

    Shea, Robin J.; Mulks, Martha H.

    2002-01-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, a disease characterized by pulmonary necrosis and hemorrhage caused in part by neutrophil degranulation. In an effort to understand the pathogenesis of this disease, we have developed an in vivo expression technology (IVET) system to identify genes that are specifically up-regulated during infection. One of the genes that we have identified as being induced in vivo is ohr, encoding organic hydroperoxide reducta...

  19. Glucose ingestion during exercise blunts exercise-induced gene expression of skeletal muscle fat oxidative genes.

    Science.gov (United States)

    Civitarese, Anthony E; Hesselink, Matthijs K C; Russell, Aaron P; Ravussin, Eric; Schrauwen, Patrick

    2005-12-01

    Ingestion of carbohydrate during exercise may blunt the stimulation of fat oxidative pathways by raising plasma insulin and glucose concentrations and lowering plasma free fatty acid (FFA) levels, thereby causing a marked shift in substrate oxidation. We investigated the effects of a single 2-h bout of moderate-intensity exercise on the expression of key genes involved in fat and carbohydrate metabolism with or without glucose ingestion in seven healthy untrained men (22.7 +/- 0.6 yr; body mass index: 23.8 +/- 1.0 kg/m(2); maximal O(2) consumption: 3.85 +/- 0.21 l/min). Plasma FFA concentration increased during exercise (P glucose ingestion, whereas fat oxidation (indirect calorimetry) was higher in the fasted state vs. glucose feeding (P expression of pyruvate dehydrogenase kinase-4 (P glucose ingestion during exercise produced minimal effects on the expression of genes involved in carbohydrate utilization. However, glucose ingestion resulted in a decrease in the expression of genes involved in fatty acid transport and oxidation (CD36, carnitine palmitoyltransferase-1, uncoupling protein 3, and 5'-AMP-activated protein kinase-alpha(2); P glucose ingestion during exercise decreases the expression of genes involved in lipid metabolism rather than increasing genes involved in carbohydrate metabolism.

  20. Absence of missense mutations in activated c-myc genes in avian leukosis virus-induced B-cell lymphomas

    Energy Technology Data Exchange (ETDEWEB)

    Hahn, M.; Hayward, W.S.

    1988-06-01

    The authors determined the nucleotide sequences of two independent DNA clones which contained the activated c-myc genes from avian leukosis virus-induced B-cell lymphomas. Neither of these c-myce genes contained missense mutations. This strongly supports the notion that the c-myc photo-oncogene in avian leukosis virus-induced B-cell lymphomas can be oncogenically activated by altered expression of the gene without a change in the primary structure of the gene product.

  1. Integrated modelling of corrosion-induced deterioration in rein-forced concrete structures

    DEFF Research Database (Denmark)

    Michel, Alexander; Geiker, M.R.; Stang, Henrik

    2013-01-01

    An integrated finite element based modelling approach is presented, which allows for fully coupled simulation of reinforcement corrosion and corrosion-induced concrete damage. While a finite element method (FEM) based corrosion model was used to describe electrochemical processes at the reinforce...... use of the modelling approach, a numerical example is presented which illustrates full coupling of formation of corrosion cells, propagation of corrosion, and subsequent development of corrosion-induced concrete damage.......An integrated finite element based modelling approach is presented, which allows for fully coupled simulation of reinforcement corrosion and corrosion-induced concrete damage. While a finite element method (FEM) based corrosion model was used to describe electrochemical processes...... at the reinforcement surface, a FEM based me-chanical model was used to simulate corrosion-induced concrete damage. Both FEM models were fully coupled, i.e. information, such as corrosion current density, dam-age state of concrete cover, etc., were constantly exchanged between the models. To demonstrate the potential...

  2. Construction of an inducible cell-communication system that amplifies Salmonella gene expression in tumor tissue.

    Science.gov (United States)

    Dai, Yumei; Toley, Bhushan J; Swofford, Charles A; Forbes, Neil S

    2013-06-01

    Bacterial therapies have the potential to overcome resistances that cause chemotherapies to fail. When using bacteria to produce anticancer agents in tumors, triggering gene expression is necessary to prevent systemic toxicity. The use of chemical triggers, however, is hampered by poor delivery of inducing molecules, which reduces the number of activated bacteria. To solve this problem, we created a cell-communication system that enables activated bacteria to induce inactive neighbors. We hypothesized that introducing cell communication into Salmonella would improve direct triggering strategies by increasing protein production, increasing sensitivity to inducer molecules, and enabling expression in tumor tissue. To test these hypotheses we integrated the PBAD promoter into the quorum-sensing machinery from Vibrio fischeri. The expression of a fluorescent reporter gene was compared to expression from non-communicating controls. Function in three-dimensional tissue was tested in a tumor-on-a-chip device. Bacterial communication increased fluorescence 40-fold and increased sensitivity to inducer molecules more than 10,000-fold. The system enabled bacteria to activate neighbors and increased the time-scale of protein production. Gene expression was controllable and tightly regulated. At the optimal inducing signal, communicating bacteria produced 350 times more protein than non-communicating bacteria. The cell-communication system created in this study has uses beyond cancer therapy, including protein manufacturing, bioremediation and biosensing. It would enable amplified induction of gene expression in any environment that limits availability of inducer molecules. Ultimately, because inducible cellular communication enables gene expression in tissue, it will be a critical component of bacterial anticancer therapies.

  3. Importance of interferon inducible trans-membrane proteins and retinoic acid inducible gene I for influenza virus replication: A review.

    Science.gov (United States)

    Suo, Siqingaowa; Ren, Xiaofeng

    2016-01-01

    Understanding the interplay between Influenza viruses and host cells is key to elucidating the pathogenesis of these viruses. Several host factors have been identified that exert antiviral functions; however, influenza viruses continue to replicate utilizing host cell machinery. Herein, we review the mechanisms of action of two host-derived proteins on conferring cellular resistance to the influenza virus; (1) the interferon inducible trans-membrane proteins, 1, 2 and 3, a recently identified family of early restriction factors; and (2) retinoic acid inducible gene I, a key mediator of antiviral immunity. These data may contribute to the design of novel and efficient anti-influenza treatments.

  4. Gene expression in rats with Barrett's esophagus and esophageal adenocarcinoma induced by gastroduodenoesophageal reflux

    Institute of Scientific and Technical Information of China (English)

    Peng Cheng; Jun Gong; Tao Wang; Jie Chen; Gui-Sheng Liu; Ru Zhang

    2005-01-01

    AIM: To study the different gene expression profiles in rats with Barrett's esophagus (BE) and esophageal adenocarcinoma (EA) induced by gastro-duodenoesophageal reflux.METHODS: Esophagoduodenostomy was performed in 8-wk old Sprague-Dawley rats to induce gastro-duodenoesophageal reflux, and a group of rats that received sham operation served as control. Esophageal epithelial pathological tissues were dissected and frozen in liquid nitrogen immediately. The expression profiles of 4 096genes in EA and BE tissues were compared to normal esophagus epithelium in normal control (NC) by cDNA microarray.RESULTS: Four hundred and forty-eight genes in BE were more than three times different from those in NC, including 312 upregulated and 136 downregulated genes. Three hundred and seventy-seven genes in EA were more than three times different from those in NC, including 255upregulated and 142 downregulated genes. Compared to BE, there were 122 upregulated and 156 downregulated genes in EA. In the present study, the interested genes were those involved in carcinogenesis. Among them, the upregulated genes included cathepsin C, aminopeptidase M, arachidonic acid epoxygenase, tryptophan-2,3-dioxygenase, ubiquitin-conjugating enzyme, cyclic GMP-stimulated phosphodiesterase, tissue inhibitor of metalloproteinase-1, betaine-homocysteine methyltransferase, lysozyme, complement 4b binding protein,complement 9 protein, insulin-like growth factor binding protein, UDP-glucuronosyltransferase, tissue inhibitor of metalloproteinase-3, aldolase B, retinoid X receptor gamma, carboxylesterase and testicular cell adhesion molecule 1. The downregulated genes included glutathione synthetase, lecithin-cholesterol acyltransferase, p55CDC,heart fatty acid binding protein, cell adhesion regulator and endothelial cell selectin ligand.CONCLUSION: Esophageal epithelium exposed excessively to harmful ingredients of duodenal and gastric reflux may develop into BE and even EA gradually. The gene

  5. Spin and electric currents induced by a spin-motive force in two-dimensional Dirac semimetals protected by nonsymmorphic symmetries

    Science.gov (United States)

    Habe, Tetsuro

    2017-08-01

    We theoretically study the spin and charge currents induced by a spin-motive force in a two-dimensional Dirac semimetal protected by nonsymmorphic symmetries. Glide mirror plane symmetry, a nonsymmorphic symmetry, leads to a constraint to the induced current; the spin-motive force acting on out-of-plane spin (in-plane spin) induces the pure spin (charge) current. We calculate the response function to the spin-motive force in linear-response theory and find that the conductivity for the pure spin current remains nonzero even if the Fermi energy is crossing the node of linear dispersion. We also find that the conserved spin current is induced at the charge neutral point.

  6. Stimulation of primary osteoblasts with ATP induces transient vinculin clustering at sites of high intracellular traction force.

    Science.gov (United States)

    Tan, Toh Weng; Pfau, Bastian; Jones, David; Meyer, Thomas

    2014-02-01

    Adenosine 5'-triphosphate (ATP), released in response to mechanical and inflammatory stimuli, induces the dynamic and asynchronous protrusion and subsequent retraction of local membrane structures in osteoblasts. The molecular mechanisms involved in the ligand-stimulated herniation of the plasma membrane are largely unknown, which prompted us to investigate whether the focal-adhesion protein vinculin is engaged in the cytoskeletal alterations that underlie the ATP-induced membrane blebbing. Using time-lapse fluorescence microscopy of primary bovine osteoblast-like cells expressing green fluorescent protein-tagged vinculin, we found that stimulation of cells with 100 μM ATP resulted in the transient and rapid clustering of recombinant vinculin in the cell periphery, starting approximately 100 s after addition of the nucleotide. The ephemeral nature of the vinculin clusters was made evident by the brevity of their mean assembly and disassembly times (66.7 ± 13.3 s and 99.0 ± 6.6 s, respectively). Traction force vector maps demonstrated that the vinculin-rich clusters were localized predominantly at sites of high traction force. Intracellular calcium measurements showed that the ligand-induced increase in [Ca(2+)]i clearly preceded the clustering of vinculin, since [Ca(2+)]i levels returned to normal within 30 s of exposure to ATP, indicating that intracellular calcium transients trigger a cascade of signalling events that ultimately result in the incorporation of vinculin into membrane-associated focal aggregates.

  7. Effect of electro-acupuncture on gene expression in heart of rats with stress-induced pre-hypertension based on gene chip technology.

    Science.gov (United States)

    Guo, Yan; Xie, Xiaojia; Guo, Changqing; Wang, Zhaoyang; Liu, Qingguo

    2015-06-01

    To explore electro-acupuncture's (EA's) effect on gene expression in heart of rats with stress-induced pre-hypertension and try to reveal its biological mechanism based on gene chip technology. Twenty-seven Wistar male rats were randomly divided into 3 groups. The stress-induced hypertensive rat model was prepared by electric foot-shocks combined with generated noise. Molding cycle lasted for 14 days and EA intervene was applied,on rats in model + EA group during model preparation. Rat Gene 2.0 Sense Target Array technology was used for the determination of gene expression profiles and the screened key genes were verified by real-time quantitative polymerase chain reaction (RT-PCR) method. Compared with blank control group, 390 genes were changed in model group; compared with model control group, 330 genes were changed in model+EA group. Significance analysis of gene function showed that the differentially expressed genes are those involved in biological process, molecular function and cellular components. RT-PCR result of the screened key genes is consistent with that of gene chip test. EA could significantly lower blood pressure of stress-induced pre-hypertension rats and affect its gene expression profile in heart. Genes that related to the contraction of vascular smooth muscle may be involved in EA's anti-hypertensive mechanism.

  8. Functional connections and pathways of coenzyme Q10-inducible genes: an in-silico study.

    Science.gov (United States)

    Schmelzer, Constance; Lindner, Inka; Vock, Christina; Fujii, Kenji; Döring, Frank

    2007-10-01

    Coenzyme Q10 (CoQ10, ubiquinone) is an essential cofactor in the electron transport chain, serves as a potent antioxidant in mitochondria and lipid membranes, and is often used as a dietary supplement for a number of diseases including cardiovascular diseases. Recently, we obtained evidence that CoQ10 (Kaneka Q10) affects the expression of hundreds of human genes. To decipher the functional and regulatory connections of these genes, a literature search combined with transcription factor binding site analysis was performed using Genomatix BiblioSphere and MatInspector. This in-silico analysis revealed 17 CoQ10-inducible genes which are functionally connected by signalling pathways of G-protein coupled receptors, JAK/STAT, integrin, and beta-arrestin. Promoter analysis of these CoQ10-inducible genes showed one group of NF B-regulated genes, namely IL5, thrombin, vitronectin receptor and C-reactive protein (CRP). Furthermore, a common promoter framework containing binding sites of the transcription factor families EVI1, HOXF, HOXC, and CLOX was identified in the promoters of IL5, CRP, and vitronectin receptor. The identified CoQ10-inducible genes and pathways play an important role in inflammatory response. Since these effects are based on an in-vitro study, the effect of CoQ10 on vascular health in vivo needs to be addressed in further animal and/or human intervention studies.

  9. A CRISPR-Based Screen Identifies Genes Essential for West-Nile-Virus-Induced Cell Death.

    Science.gov (United States)

    Ma, Hongming; Dang, Ying; Wu, Yonggan; Jia, Gengxiang; Anaya, Edgar; Zhang, Junli; Abraham, Sojan; Choi, Jang-Gi; Shi, Guojun; Qi, Ling; Manjunath, N; Wu, Haoquan

    2015-07-28

    West Nile virus (WNV) causes an acute neurological infection attended by massive neuronal cell death. However, the mechanism(s) behind the virus-induced cell death is poorly understood. Using a library containing 77,406 sgRNAs targeting 20,121 genes, we performed a genome-wide screen followed by a second screen with a sub-library. Among the genes identified, seven genes, EMC2, EMC3, SEL1L, DERL2, UBE2G2, UBE2J1, and HRD1, stood out as having the strongest phenotype, whose knockout conferred strong protection against WNV-induced cell death with two different WNV strains and in three cell lines. Interestingly, knockout of these genes did not block WNV replication. Thus, these appear to be essential genes that link WNV replication to downstream cell death pathway(s). In addition, the fact that all of these genes belong to the ER-associated protein degradation (ERAD) pathway suggests that this might be the primary driver of WNV-induced cell death. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Comprehensive set of integrative plasmid vectors for copper-inducible gene expression in Myxococcus xanthus.

    Science.gov (United States)

    Gómez-Santos, Nuria; Treuner-Lange, Anke; Moraleda-Muñoz, Aurelio; García-Bravo, Elena; García-Hernández, Raquel; Martínez-Cayuela, Marina; Pérez, Juana; Søgaard-Andersen, Lotte; Muñoz-Dorado, José

    2012-04-01

    Myxococcus xanthus is widely used as a model system for studying gliding motility, multicellular development, and cellular differentiation. Moreover, M. xanthus is a rich source of novel secondary metabolites. The analysis of these processes has been hampered by the limited set of tools for inducible gene expression. Here we report the construction of a set of plasmid vectors to allow copper-inducible gene expression in M. xanthus. Analysis of the effect of copper on strain DK1622 revealed that copper concentrations of up to 500 μM during growth and 60 μM during development do not affect physiological processes such as cell viability, motility, or aggregation into fruiting bodies. Of the copper-responsive promoters in M. xanthus reported so far, the multicopper oxidase cuoA promoter was used to construct expression vectors, because no basal expression is observed in the absence of copper and induction linearly depends on the copper concentration in the culture medium. Four different plasmid vectors have been constructed, with different marker selection genes and sites of integration in the M. xanthus chromosome. The vectors have been tested and gene expression quantified using the lacZ gene. Moreover, we demonstrate the functional complementation of the motility defect caused by lack of PilB by the copper-induced expression of the pilB gene. These versatile vectors are likely to deepen our understanding of the biology of M. xanthus and may also have biotechnological applications.

  11. Cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene.

    Science.gov (United States)

    Kim, Tae Hyun; Park, Jong Ho; Kim, Moon Chul; Cho, Sung Ho

    2008-01-01

    Treatment with the cutin monomer 16-hydroxypalmitic acid (HPA), a major component of cutin, elicited the synthesis of hydrogen peroxide (H2O2) in rice leaves and induced the expression of the lipid transfer protein gene OsLTP5. Treatment with HPA also induced expression of OsLTP1, OsLTP2, and the pathogen-related PR-10 genes to a lesser extent. The OsLTP5 transcript was expressed prominently in stems and flowers, but was barely detectable in leaves. Expression of OsLTP5 was induced in shoots in response to ABA and salicylic acid. It is proposed that HPA is perceived by rice as a signal, inducing defense reactions.

  12. In Vivo Imaging of Local Gene Expression Induced by Magnetic Hyperthermia

    Science.gov (United States)

    Sandre, Olivier; Genevois, Coralie; Garaio, Eneko; Adumeau, Laurent; Mornet, Stéphane; Couillaud, Franck

    2017-01-01

    The present work aims to demonstrate that colloidal dispersions of magnetic iron oxide nanoparticles stabilized with dextran macromolecules placed in an alternating magnetic field can not only produce heat, but also that these particles could be used in vivo for local and noninvasive deposition of a thermal dose sufficient to trigger thermo-induced gene expression. Iron oxide nanoparticles were first characterized in vitro on a bio-inspired setup, and then they were assayed in vivo using a transgenic mouse strain expressing the luciferase reporter gene under transcriptional control of a thermosensitive promoter. Iron oxide nanoparticles dispersions were applied topically on the mouse skin or injected subcutaneously with Matrigel™ to generate so-called pseudotumors. Temperature was monitored continuously with a feedback loop to control the power of the magnetic field generator and to avoid overheating. Thermo-induced luciferase expression was followed by bioluminescence imaging 6 h after heating. We showed that dextran-coated magnetic iron oxide nanoparticle dispersions were able to induce in vivo mild hyperthermia compatible with thermo-induced gene expression in surrounding tissues and without impairing cell viability. These data open new therapeutic perspectives for using mild magnetic hyperthermia as noninvasive modulation of tumor microenvironment by local thermo-induced gene expression or drug release. PMID:28208731

  13. In Vivo Imaging of Local Gene Expression Induced by Magnetic Hyperthermia

    Directory of Open Access Journals (Sweden)

    Olivier Sandre

    2017-02-01

    Full Text Available The present work aims to demonstrate that colloidal dispersions of magnetic iron oxide nanoparticles stabilized with dextran macromolecules placed in an alternating magnetic field can not only produce heat, but also that these particles could be used in vivo for local and noninvasive deposition of a thermal dose sufficient to trigger thermo-induced gene expression. Iron oxide nanoparticles were first characterized in vitro on a bio-inspired setup, and then they were assayed in vivo using a transgenic mouse strain expressing the luciferase reporter gene under transcriptional control of a thermosensitive promoter. Iron oxide nanoparticles dispersions were applied topically on the mouse skin or injected subcutaneously with Matrigel™ to generate so-called pseudotumors. Temperature was monitored continuously with a feedback loop to control the power of the magnetic field generator and to avoid overheating. Thermo-induced luciferase expression was followed by bioluminescence imaging 6 h after heating. We showed that dextran-coated magnetic iron oxide nanoparticle dispersions were able to induce in vivo mild hyperthermia compatible with thermo-induced gene expression in surrounding tissues and without impairing cell viability. These data open new therapeutic perspectives for using mild magnetic hyperthermia as noninvasive modulation of tumor microenvironment by local thermo-induced gene expression or drug release.

  14. Degranulation of rat cerebellum induces selective variations in gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Eliyahu, D.; Soreq, H.

    1982-02-01

    Selective variations in the composition of poly(A)-containing mRNA were found to be induced in the rat cerebellum by X-irradiation. mRNA populations prepared from normal and X-irradiated rat cerebella at different stages of their development displayed equal efficiencies when translated in vitro in reticulocyte lysates. Specific differences were revealed, however, when the labeled translation products of both mRNA preparations were subjected to two-dimensional gel electrophoresis followed by fluorography of the dried gels. Of more than 100 polypeptide products, several showed marked intensity differences, indicating changes in the abundance of their directing mRNA species. These differences appear both in developing and in mature cerebellar mRNA, and the extent of modification in mRNA is much higher than the consequent changes in the composition of proteins in the irradiated cerebellum. The degranulation-induced modifications in levels of specific cerebellar mRNA species can be used to identify proteins whose biosynthesis depends on the presence of interneurons.

  15. Turbulence-induced bubble collision force modeling and validation in adiabatic two-phase flow using CFD

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, Subash L., E-mail: sharma55@purdue.edu [School of Nuclear Engineering, Purdue University, West Lafayette, IN 47907-1290 (United States); Hibiki, Takashi; Ishii, Mamoru [School of Nuclear Engineering, Purdue University, West Lafayette, IN 47907-1290 (United States); Brooks, Caleb S. [Department of Nuclear, Plasma, and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Schlegel, Joshua P. [Nuclear Engineering Program, Missouri University of Science and Technology, Rolla, MO 65409 (United States); Liu, Yang [Nuclear Engineering Program, Department of Mechanical Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States); Buchanan, John R. [Bechtel Marine Propulsion Corporation, Bettis Laboratory, West Mifflin, PA 15122 (United States)

    2017-02-15

    Highlights: • Void distribution in narrow rectangular channel with various non-uniform inlet conditions. • Modeling of void diffusion due to bubble collision force. • Validation of new modeling in adiabatic air–water two-phase flow in a narrow channel. - Abstract: The prediction capability of the two-fluid model for gas–liquid dispersed two-phase flow depends on the accuracy of the closure relations for the interfacial forces. In previous studies of two-phase flow Computational Fluid Dynamics (CFD), interfacial force models for a single isolated bubble has been extended to disperse two-phase flow assuming the effect in a swarm of bubbles is similar. Limited studies have been performed investigating the effect of the bubble concentration on the lateral phase distribution. Bubbles, while moving through the liquid phase, may undergo turbulence-driven random collision with neighboring bubbles without significant coalescence. The rate of these collisions depends upon the bubble approach velocity and bubble spacing. The bubble collision frequency is expected to be higher in locations with higher bubble concentrations, i.e., volume fraction. This turbulence-driven random collision causes the diffusion of the bubbles from high concentration to low concentration. Based on experimental observations, a phenomenological model has been developed for a “turbulence-induced bubble collision force” for use in the two-fluid model. For testing the validity of the model, two-phase flow data measured at Purdue University are utilized. The geometry is a 10 mm × 200 mm cross section channel. Experimentally, non-uniform inlet boundary conditions are applied with different sparger combinations to vary the volume fraction distribution across the wider dimension. Examining uniform and non-uniform inlet data allows for the influence of the volume fraction to be studied as a separate effect. The turbulence-induced bubble collision force has been implemented in ANSYS CFX. The

  16. Molecular cloning and functional analyses of low-temperature induced genes from Ammopiptanthus mongolicus

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Studies on the cold-responsive genes and cold signaling of woody species drop far behind in comparison to herbaceous plants.Due to similar lignified structure,perennial characteristic,and enhanced tolerance,it seems much easier to find strongly antifreeze genes and obtain effective results in transgenic woody plants.In this study,Ammopiptanthus mongolicus,an evergreen,broadleaf and cold-resist leguminous shrub growing in the desert of Inner Mongolia,was used as a material for low-temperature induced gene is...

  17. [Osmotic shock induces expression of Vibrio fischeri lux genes in Escherichia coli cells].

    Science.gov (United States)

    Zavil'gel'skiĭ, G B; Kotova, V Iu

    2003-04-01

    The effect of osmotic shock on the expression of genes in the lux regulon of marine bacteria Vibrio fischeri was studied in cells of Escherichia coli. Bioluminescence of cells was shown to drastically increase, when cells were exposed to osmotic shock at the early logarithmic growth phase. The expression of lux genes induced by osmotic shock is determined by the two-component regulatory system RcsC-RcsB. A nucleotide sequence in the regulatory region of the luxR gene homologous to the RcsB-box consensus of E. coli is assumed to be a primary site for this system.

  18. Elasticity-induced force reversal between active spinning particles in dense passive media.

    Science.gov (United States)

    Aragones, J L; Steimel, J P; Alexander-Katz, A

    2016-04-26

    The self-organization of active particles is governed by their dynamic effective interactions. Such interactions are controlled by the medium in which such active agents reside. Here we study the interactions between active agents in a dense non-active medium. Our system consists of actuated, spinning, active particles embedded in a dense monolayer of passive, or non-active, particles. We demonstrate that the presence of the passive monolayer alters markedly the properties of the system and results in a reversal of the forces between active spinning particles from repulsive to attractive. The origin of such reversal is due to the coupling between the active stresses and elasticity of the system. This discovery provides a mechanism for the interaction between active agents in complex and structured media, opening up opportunities to tune the interaction range and directionality via the mechanical properties of the medium.

  19. Serum induced degradation of 3D DNA box origami observed by high speed atomic force microscope

    DEFF Research Database (Denmark)

    Jiang, Zaixing; Zhang, Shuai; Yang, Chuanxu;

    2015-01-01

    3D DNA origami holds tremendous potential to encapsulate and selectively release therapeutic drugs. Observations of real-time performance of 3D DNA origami structures in physiological environment will contribute much to its further applications. Here, we investigate the degradation kinetics of 3D...... DNA box origami in serum using high-speed atomic force microscope optimized for imaging 3D DNA origami in real time. The time resolution allows characterizing the stages of serum effects on individual 3D DNA box origami with nanometer resolution. Our results indicate that the whole digest process...... is a combination of a rapid collapse phase and a slow degradation phase. The damages of box origami mainly happen in the collapse phase. Thus, the structure stability of 3D DNA box origami should be further improved, especially in the collapse phase, before clinical applications...

  20. Spatiotemporal variation of surface shortwave forcing from fire-induced albedo change in interior Alaska

    Science.gov (United States)

    Huang, Shengli; Dahal, Devendra; Liu, Heping; Jin, Suming; Young, Claudia J.; Liu, Shuang; Liu, Shu-Guang

    2015-01-01

    The albedo change caused by both fires and subsequent succession is spatially heterogeneous, leading to the need to assess the spatiotemporal variation of surface shortwave forcing (SSF) as a component to quantify the climate impacts of high-latitude fires. We used an image reconstruction approach to compare postfire albedo with the albedo assuming fires had not occurred. Combining the fire-caused albedo change from the 2001-2010 fires in interior Alaska and the monthly surface incoming solar radiation, we examined the spatiotemporal variation of SSF in the early successional stage of around 10 years. Our results showed that while postfire albedo generally increased in fall, winter, and spring, some burned areas could show an albedo decrease during these seasons. In summer, the albedo increased for several years and then declined again. The spring SSF distribution did not show a latitudinal decrease from south to north as previously reported. The results also indicated that although the SSF is usually largely negative in the early successional years, it may not be significant during the first postfire year. The annual 2005-2010 SSF for the 2004 fire scars was -1.30, -4.40, -3.31, -4.00, -3.42, and -2.47 Wm-2. The integrated annual SSF map showed significant spatial variation with a mean of -3.15 Wm-2 and a standard deviation of 3.26 Wm-2, 16% of burned areas having positive SSF. Our results suggest that boreal deciduous fires would be less positive for climate change than boreal evergreen fires. Future research is needed to comprehensively investigate the spatiotemporal radiative and non-radiative forcings to determine the effect of boreal fires on climate.

  1. Experimental studies in magnetically induced transverse force-frequency effect in thin quartz microresonators

    Energy Technology Data Exchange (ETDEWEB)

    Hatipoglu, Gokhan [Department of Electrical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802 (United States); Tadigadapa, Srinivas, E-mail: sat10@psu.edu [Department of Electrical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802 (United States); Department of Biomedical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802 (United States)

    2015-07-21

    In this work, the transverse force-frequency sensitivity of magnetostrictive Metglas{sup ®} (Fe{sub 85}B{sub 5}Si{sub 10}) thin film coated AT-cut thickness shear mode quartz thin plate microresonator (500 μm × 500 μm × 19 μm) is experimentally measured and modeled in Lagrangian formulation by coupling magnetostrictive deformation equations with the basic plate equations from the theory of small deformation. The quartz plate resonator is fabricated by micromachining techniques and released into fixed-free structure using focused ion beam milling. Application of a magnetic field results in the out-of-plane bending of the structure due to elastic coupling between the magnetostrictive Metglas{sup ®} and quartz resonator layers. As a result of the transverse loading and out-of-plane bending, the admittance characteristics of the resonator shifts, and these shifts are recorded in real time utilizing a network analyzer. The sensitivity is experimentally measured to be 162.3 mdeg/Oe for phase, corresponding to a frequency sensitivity of Δf/H = 11 Hz/Oe. The equivalent force-frequency sensitivity can then be calculated as 2.36 μN/Hz using the developed model. The coupled domain analysis fits well with the experimental data. Further reduction of quartz thickness and optimization of the thickness ratio of the magnetostrictive to quartz layers offers the possibility of exploiting the stress sensitivity of plate microresonators as sensitive magnetic field sensors capable of low nanoTesla to picoTesla level magnetic flux densities.

  2. Modeling of fluid-induced vibrations and identification of hydrodynamic forces on flow control valves

    Institute of Scientific and Technical Information of China (English)

    Samad Mehrzad; Ilgar Javanshir; Ahmad Rahbar Ranji; Seyyed Hadi Taheri

    2015-01-01

    Dynamics and vibration of control valves under flow-induced vibration are analyzed. Hydrodynamic load characteristics and structural response under flow-induced vibration are mainly influenced by inertia, damping, elastic, geometric characteristics and hydraulic parameters. The purpose of this work is to investigate the dynamic behavior of control valves in the response to self-excited fluid flow. An analytical and numerical method is developed to simulate the dynamic and vibrational behavior of sliding dam valves, in response to flow excitation. In order to demonstrate the effectiveness of proposed model, the simulation results are validated with experimental ones. Finally, to achieve the optimal valve geometry, numerical results for various shapes of valves are compared. Rounded valve with the least amount of flow turbulence obtains lower fluctuations and vibration amplitude compared with the flat and steep valves. Simulation results demonstrate that with the optimal design requirements of valves, vibration amplitude can be reduced by an average to 30%.

  3. Characterization of cell lysis in Pseudomonas putida induced upon expression of heterologous killing genes

    DEFF Research Database (Denmark)

    Ronchel, M.C.; Molina, L.; Witte, A.;

    1998-01-01

    Active biological containment systems are based on the controlled expression of killing genes. These systems are of interest for the Pseudomonadaceae because of the potential applications of these microbes as bioremediation agents and biopesticides, The physiological effects that lead to cell death...... upon the induction of expression of two different heterologous killing genes in nonpathogenic Pseudomonas putida KT2440 derivatives have been analyzed, P. putida CMC4 and CMC12 carry in their chromosomes a fusion of the PAl-04/03 promoter to the Escherichia coli gef gene and the phi X174 lysis gene E......, respectively. Expression of the killing genes is controlled by the LacI protein, whose expression is initiated from the XylS-dependent Pm promoter. Under induced conditions, killing of P. putida CMC12 cells mediated by phi X174 lysis protein E was faster than that observed for P. putida CMC4, for which the Gef...

  4. Identification of genes regulated during mechanical load-induced cardiac hypertrophy

    Science.gov (United States)

    Johnatty, S. E.; Dyck, J. R.; Michael, L. H.; Olson, E. N.; Abdellatif, M.; Schneider, M. (Principal Investigator)

    2000-01-01

    Cardiac hypertrophy is associated with both adaptive and adverse changes in gene expression. To identify genes regulated by pressure overload, we performed suppressive subtractive hybridization between cDNA from the hearts of aortic-banded (7-day) and sham-operated mice. In parallel, we performed a subtraction between an adult and a neonatal heart, for the purpose of comparing different forms of cardiac hypertrophy. Sequencing more than 100 clones led to the identification of an array of functionally known (70%) and unknown genes (30%) that are upregulated during cardiac growth. At least nine of those genes were preferentially expressed in both the neonatal and pressure over-load hearts alike. Using Northern blot analysis to investigate whether some of the identified genes were upregulated in the load-independent calcineurin-induced cardiac hypertrophy mouse model, revealed its incomplete similarity with the former models of cardiac growth. Copyright 2000 Academic Press.

  5. Identification of genes regulated during mechanical load-induced cardiac hypertrophy

    Science.gov (United States)

    Johnatty, S. E.; Dyck, J. R.; Michael, L. H.; Olson, E. N.; Abdellatif, M.; Schneider, M. (Principal Investigator)

    2000-01-01

    Cardiac hypertrophy is associated with both adaptive and adverse changes in gene expression. To identify genes regulated by pressure overload, we performed suppressive subtractive hybridization between cDNA from the hearts of aortic-banded (7-day) and sham-operated mice. In parallel, we performed a subtraction between an adult and a neonatal heart, for the purpose of comparing different forms of cardiac hypertrophy. Sequencing more than 100 clones led to the identification of an array of functionally known (70%) and unknown genes (30%) that are upregulated during cardiac growth. At least nine of those genes were preferentially expressed in both the neonatal and pressure over-load hearts alike. Using Northern blot analysis to investigate whether some of the identified genes were upregulated in the load-independent calcineurin-induced cardiac hypertrophy mouse model, revealed its incomplete similarity with the former models of cardiac growth. Copyright 2000 Academic Press.

  6. Gene Analysis of Arsenic Trioxide—induced Apoptosis of Lymphoma Cells

    Institute of Scientific and Technical Information of China (English)

    ZHANGZidong; LIWeiyu; 等

    2002-01-01

    Objective The effect of arsenic trioxide on apoptosis gene expression of Raji cell was explored when Raji cells were incubated with 0.5μmol/L of arsenic trioxide for 6h。Methods Cell culture,extraction and isolation of mRNA,preparation of probes labeled with fluorescence,hybridization technique of DNA chip(each chip containing 200 apoptosis genes,Chinese Shanghai Biostar,In.)were used.Results Arsenic trioxide induced significant changes in 10%(20/200 genes)of the apoptosis genes:18 genes were downregulated,only two upregulated.In particular,inhibitors of apoptosis protein,such as X-linked inhibitor of apoptosis protein,were significantly downregulated.P53 and the other apoptosis genes were also downregulatec.Of the upregulated genes,high expression of heat-shock protein could promote apoptosis of Raji cells.Conclusion The inhibitors of apoptosis protein play an important role in the process of arsenic trioxide-induced apoptosis of Raji cells.

  7. Forced diffusion via electrically induced crystallization for fabricating ZnO–Ti–Si structures

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Yen-Ting; Hung, Fei-Yi, E-mail: fyhung@mail.ncku.edu.tw

    2014-11-15

    Highlights: • ZnO–Ti–Si system is very important for the structural design. • The electrically induced crystallization method is useful to diffusion process. • Intermetallic compound characteristics have been presented using electrically induced crystallization. • Interface mechanism about diffusion of TZO–TiSi{sub x}–Si structure is presented. - Abstract: Electrically induced crystallization (EIC) is a recently developed process for material modification. This study is applied to EIC to fabricate ZnO–Ti–Si multi-layer structures of various thicknesses to dope Ti into ZnO thin film and to form TiSi{sub x} intermetallic compound (IMC) in a single step. The IMC layer was confirmed using transmission electron microscopy images. The Ti layer thickness was more than 40 nm, which enhanced electron transmission and decreased the total electrical resistance in the structure. Finally, the diffusion mechanisms of EIC and the annealing process were investigated. This study shows that the EIC process has potential for industrial applications.

  8. Moisture-induced surface crystallization of spray-dried amorphous lactose particles studied by atomic force microscopy.

    Science.gov (United States)

    Mahlin, Denny; Berggren, Jonas; Alderborn, Göran; Engström, Sven

    2004-01-01

    The aim of this study was to show that atomic force microscopy (AFM) can be used to obtain mechanistic and kinetic information about the process of moisture-induced surface crystallization of single particles of amorphous lactose. Completely amorphous lactose particles were prepared by spray-drying a solution of alpha-lactose monohydrate, and moisture-induced crystallization was monitored for a bed of particles by microcalorimetry and for single particles by AFM. From the AFM images it was found that crystallization of the surface of single particles can be described in terms of a sequence of three events: an initial smoothening of the surface, formation of crystalline nanostructures dispersed in amorphous material, and growth of these structures to a complete crystalline surface. The surface roughness parameter rugosity was used to estimate the fraction crystalline surface, and the growth kinetics were found to obey the JMAK equation. The fraction crystalline surface at different times could also be estimated by determining the growth rate of individual crystals. It was concluded that AFM offers a unique means of visualizing the process of moisture-induced surface crystallization of amorphous particles and enables mechanistic and kinetic information about the process to be extracted. Copyright 2004 Wiley-Liss, Inc.

  9. Aggregation-induced fluorescence behavior of triphenylamine-based Schiff bases: the combined effect of multiple forces.

    Science.gov (United States)

    Yang, Mingdi; Xu, Dongling; Xi, Wengang; Wang, Lianke; Zheng, Jun; Huang, Jing; Zhang, Jingyan; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2013-10-18

    Eight triphenylamine (TPA)-based Schiff bases that exhibit different aggregation-induced emission (AIE) or aggregation-caused quenching (ACQ) behavior in tetrahydrofuran (THF)/water mixtures have been synthesized and characterized. The photophysical properties in solution, aqueous suspension, film, and the crystalline state along with their relationships were comparatively investigated. The single-crystal structures of 1-8 indicate that compact π···π stacking or excimers induce fluorescence quenching of 1, 2, 5, and 7. However, the existence of J aggregates or multiple intra- and intermolecular interactions restrict the intramolecular vibration and rotation, enabling compounds 3, 4, 6, and 8 to exhibit good AIE character. The size and growth process of particles with different water fractions were studied using scanning electron microscopy, which demonstrated that smaller uniformly dispersed nanoparticles in the THF/water mixtures favor fluorescence emission. The above results suggest that the combined effects of multiple forces caused by structural variation have a great influence on their molecular packing, electronic structure, and aggregation-induced fluorescence properties. In addition, piezofluorochromic experiments verified the potential applications of 4 and 6.

  10. Forced running exercise attenuates hippocampal neurogenesis impairment and the neurocognitive deficits induced by whole-brain irradiation via the BDNF-mediated pathway

    Energy Technology Data Exchange (ETDEWEB)

    Ji, Jian-feng; Ji, Sheng-jun; Sun, Rui; Li, Kun; Zhang, Yuan; Zhang, Li-yuan; Tian, Ye, E-mail: dryetian@hotmail.com

    2014-01-10

    Highlights: •Forced exercise can ameliorate WBI induced cognitive impairment in our rat model. •Mature BDNF plays an important role in the effects of forced exercise. •Exercise may be a possible treatment of the radiation-induced cognitive impairment. -- Abstract: Cranial radiotherapy induces progressive and debilitating cognitive deficits, particularly in long-term cancer survivors, which may in part be caused by the reduction of hippocampal neurogenesis. Previous studies suggested that voluntary exercise can reduce the cognitive impairment caused by radiation therapy. However, there is no study on the effect of forced wheel exercise and little is known about the molecular mechanisms mediating the effect of exercise. In the present study, we investigated whether the forced running exercise after irradiation had the protective effects of the radiation-induced cognitive impairment. Sixty-four Male Sprague–Dawley rats received a single dose of 20 Gy or sham whole-brain irradiation (WBI), behavioral test was evaluated using open field test and Morris water maze at 2 months after irradiation. Half of the rats accepted a 3-week forced running exercise before the behavior detection. Immunofluorescence was used to evaluate the changes in hippocampal neurogenesis and Western blotting was used to assess changes in the levels of mature brain-derived neurotrophic factor (BDNF), phosphorylated tyrosine receptor kinase B (TrkB) receptor, protein kinase B (Akt), extracellular signal-regulated kinase (ERK), calcium-calmodulin dependent kinase (CaMKII), cAMP-calcium response element binding protein (CREB) in the BDNF–pCREB signaling. We found forced running exercise significantly prevented radiation-induced cognitive deficits, ameliorated the impairment of hippocampal neurogenesis and attenuated the down-regulation of these proteins. Moreover, exercise also increased behavioral performance, hippocampal neurogenesis and elevated BDNF–pCREB signaling in non

  11. Acute ozone-induced differential gene expression profiles in rat lung.

    Science.gov (United States)

    Nadadur, Srikanth S; Costa, Daniel L; Slade, Ralph; Silbjoris, Robert; Hatch, Gary E

    2005-12-01

    Ozone is an oxidant gas that can directly induce lung injury. Knowledge of the initial molecular events of the acute O3 response would be useful in developing biomarkers of exposure or response. Toward this goal, we exposed rats to toxic concentrations of O3 (2 and 5 ppm) for 2 hr and the molecular changes were assessed in lung tissue 2 hr postexposure using a rat cDNA expression array containing 588 characterized genes. Gene array analysis indicated differential expression in almost equal numbers of genes for the two exposure groups: 62 at 2 ppm and 57 at 5 ppm. Most of these genes were common to both exposure groups, suggesting common roles in the initial toxicity response. However, we also identified the induction of nine genes specific to 2-ppm (thyroid hormone-beta receptor c-erb-A-beta; and glutathione reductase) or 5-ppm exposure groups (c-jun, induced nitric oxide synthase, macrophage inflammatory protein-2, and heat shock protein 27). Injury markers in bronchoalveolar lavage fluid (BALF) were used to assess immediate toxicity and inflammation in rats similarly exposed. At 2 ppm, injury was marked by significant increases in BALF total protein, N-acetylglucosaminidase, and lavageable ciliated cells. Because infiltration of neutrophils was observed only at the higher 5 ppm concentration, the distinctive genes suggested a potential amplification role for inflammation in the gene profile. Although the specific gene interactions remain unclear, this is the first report indicating a dose-dependent direct and immediate induction of gene expression that may be separate from those genes involved in inflammation after acute O3 exposure.

  12. Resistance to the development of stress-induced behavioral despair in the forced swim test associated with elevated hippocampal Bcl-xl expression.

    Science.gov (United States)

    Shishkina, Galina T; Kalinina, Tatyana S; Berezova, Inna V; Bulygina, Veta V; Dygalo, Nikolay N

    2010-12-01

    Stress may predispose individuals toward depression through down-regulation of neurogenesis and increase in apoptosis in the brain. However, many subjects show high resistance to stress in relation to psychopathology. In the present study, we assessed the possibility that individual-specific patterns of gene expression associated with cell survival and proliferation may be among the molecular factors underlying stress resilience. Brain-derived neurotrophic factor (BDNF), anti-apoptotic B cell lymphoma like X (Bcl-xl) and pro-apoptotic bcl2-associated X protein (Bax) expression were determined in the hippocampus and frontal cortex of rats naturally differed in despair-like behavior in the forced swim test. In the hippocampus, BDNF messenger RNA (mRNA) level was significantly down-regulated 2h after the forced swim test exposure, and at this time point, Bcl-xl mRNA and protein levels were significantly higher in stressed than in untested animals. The ratios of hippocampal Bcl-xl to Bax mRNA negatively correlated with the total time spent immobile in the test. When animals were divided in two groups according to immobility responses in two consecutive swim sessions and designated as stress resilient if their immobility time did not increase in the second session as it did in stress sensitive rats, it was found that resilient rats had significantly higher Bcl-xl/Bax ratios in the hippocampus than stress sensitive animals. The data suggest that naturally occurring variations in the Bcl-xl/Bax ratio in the hippocampus may contribute to individual differences in vulnerability to stress-induced depression-like behaviors.

  13. Gene expression profiling of human neural progenitor cells following the serum-induced astrocyte differentiation.

    Science.gov (United States)

    Obayashi, Shinya; Tabunoki, Hiroko; Kim, Seung U; Satoh, Jun-ichi

    2009-05-01

    Neural stem cells (NSC) with self-renewal and multipotent properties could provide an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. However, the majority of transplanted NSC and neural progenitor cells (NPC) differentiate into astrocytes in vivo under pathological environments in the central nervous system, which potentially cause reactive gliosis. Because the serum is a potent inducer of astrocyte differentiation of rodent NPC in culture, we studied the effect of the serum on gene expression profile of cultured human NPC to identify the gene signature of astrocyte differentiation of human NPC. Human NPC spheres maintained in the serum-free culture medium were exposed to 10% fetal bovine serum (FBS) for 72 h, and processed for analyzing on a Whole Human Genome Microarray of 41,000 genes, and the microarray data were validated by real-time RT-PCR. The serum elevated the levels of expression of 45 genes, including ID1, ID2, ID3, CTGF, TGFA, METRN, GFAP, CRYAB and CSPG3, whereas it reduced the expression of 23 genes, such as DLL1, DLL3, PDGFRA, SOX4, CSPG4, GAS1 and HES5. Thus, the serum-induced astrocyte differentiation of human NPC is characterized by a counteraction of ID family genes on Delta family genes. Coimmunoprecipitation analysis identified ID1 as a direct binding partner of a proneural basic helix-loop-helix (bHLH) transcription factor MASH1. Luciferase assay indicated that activation of the DLL1 promoter by MASH1 was counteracted by ID1. Bone morphogenetic protein 4 (BMP4) elevated the levels of ID1 and GFAP expression in NPC under the serum-free culture conditions. Because the serum contains BMP4, these results suggest that the serum factor(s), most probably BMP4, induces astrocyte differentiation by upregulating the expression of ID family genes that repress the proneural bHLH protein-mediated Delta expression in human NPC.

  14. Regulation of mouse hepatic genes in response to diet induced obesity, insulin resistance and fasting induced weight reduction

    Directory of Open Access Journals (Sweden)

    Mantzoros Christos

    2005-06-01

    Full Text Available Abstract Background Obesity is associated with insulin resistance that can often be improved by caloric restriction and weight reduction. Although many physiological changes accompanying insulin resistance and its treatment have been characterized, the genetic mechanisms linking obesity to insulin resistance are largely unknown. We used DNA microarrys and RT-PCR to investigate significant changes in hepatic gene transcription in insulin resistant, diet-induced obese (DIO-C57/BL/6J mice and DIO-C57/BL/6J mice fasted for 48 hours, whose weights returned to baseline levels during these conditions. Results Transcriptional profiling of hepatic mRNA revealed over 1900 genes that were significantly perturbed between control, DIO, and fasting/weight reduced DIO mice. From this set, our bioinformatics analysis identified 41 genes that rigorously discriminate these groups of mice. These genes are associated with molecular pathways involved in signal transduction, and protein metabolism and secretion. Of particular interest are genes that participate in pathways responsible for modulating insulin sensitivity. DIO altered expression of genes in directions that would be anticipated to antagonize insulin sensitivity, while fasting/ weight reduction partially or completely normalized their levels. Among these discriminatory genes, Sh3kbp1 and RGS3, may have special significance. Sh3kbp1, an endogenous inhibitor of PI-3-kinase, was upregulated by high-fat feeding, but normalized to control levels by fasting/weight reduction. Because insulin signaling occurs partially through PI-3-kinase, increased expression of Sh3kbp1 by DIO mice may contribute to hepatic insulin resistance via inhibition of PI-3-kinase. RGS3, a suppressor of G-protein coupled receptor generation of cAMP, was repressed by high-fat feeding, but partially normalized by fasting/weight reduction. Decreased expression of RGS3 may augment levels of cAMP and thereby contribute to increased, cAMP-induced

  15. Bifidobacterium bifidum Actively Changes the Gene Expression Profile Induced by Lactobacillus acidophilus in Murine Dendritic Cells

    DEFF Research Database (Denmark)

    Weiss, Gudrun Margarethe; Rasmussen, Simon; Fink, Lisbeth Nielsen;

    2010-01-01

    Dendritic cells (DC) play a pivotal regulatory role in activation of both the innate as well as the adaptive immune system by responding to environmental microorganisms. We have previously shown that Lactobacillus acidophilus induces a strong production of the pro-inflammatory and Th1 polarizing...... cytokine IL-12 in DC, whereas bifidobacteria do not induce IL-12 but inhibit the IL-12 production induced by lactobacilli. In the present study, genome-wide microarrays were used to investigate the gene expression pattern of murine DC stimulated with Lactobacillus acidophilus NCFM and Bifidobacterium...

  16. Determination of forces induced by steam flow in turbines; Determinacion de fuerzas inducidas por flujo de vapor en turbinas

    Energy Technology Data Exchange (ETDEWEB)

    Garcia Castrejon, Juan Carlos

    2008-09-15

    The steam flow induced vibrations in turbines is a common problem during the operation of 300 MW turbines and it is one of the problems that requires solution to develop 1 GW steam turbines. The flow induced vibration is caused because there is an interaction between blades and the flow field. Blades are subjected to the forces caused by the flow field, but also the flow field is affected by the blades and its movement. The nozzle wakes cause uneven pressure field downstream and produces alternating forces on blades which lead to blade vibrations. Some of the vibrations originated in this way may damage the blades and affect the turbine performance. The forces acting on blades (and causing forced vibrations) as a result of the variations in the flow field in the axial rotor-stator clearance are studied in this project. These forces were determined applying computer fluid dynamics (CFD) in two turbine stages. The CFD analysis was carried out in the Curtis stage and in the last stage, using 2D and 3D models. An important part of the analysis is focused on discussing the pressure field variation, because this variation caused the forces acting on blades. The flow field was resolved using CFD and the computed pressure field was integrated around the blades to get the forces acting on blades. These computed dynamical forces were analyzed using a FFT analysis and the results were used in the blade useful life estimation and in the investigation of the failure causes of these blades. In every turbine stage the RNG - turbulence model and the sliding mesh method was used to deal with the blade motion. The 3D models were resolved using parallel computation in a cluster of 6 AMD 64 Opteron processors of 2412.36 MHz and 8 Gb of RAM. The results of the simulation in both stages get the pressure field behavior in the axial rotor-stator clearance and determine the force acting on the blades. These results showed that both, the pressure field and the force magnitude acting on

  17. Visual symptoms and G-induced loss of consciousness in 594 Chinese Air Force aircrew--a questionnaire survey.

    Science.gov (United States)

    Cao, Xin-Sheng; Wang, Yong-Chun; Xu, Li; Yang, Chang-Bing; Wang, Bing; Geng, Jie; Gao, Yuan; Wu, Yan-Hong; Wang, Xiao-Ying; Zhang, Shu; Sun, Xi-Qing

    2012-02-01

    A questionnaire survey was performed for the first time to assess the prevalence of visual symptoms and G-induced loss of consciousness (G-LOC) due to +Gz exposure in the Chinese Air Force (CAF) to determine the effectiveness of current G tolerance training. Responses were received from 594 individuals. Among them, 302 reported at least one episode of some sort of symptoms related to +Gz, including 110 (18.5%) with visual blurring, 231 (38.9%) with greyout, 111 (18.7%) with blackout, and 49 (8.2%) with G-LOC. Incidences were most common in aircrew with 250-1,000 flying hours (53.6%) and were more prevalent in those with fewer on type flying hours (p aircrew. There remains considerable scope for +Gz education, particularly in the early centrifuge training and selection of rational physical exercises.

  18. Energy transfer of surface wind-induced currents to the deep ocean via resonance with the Coriolis force

    Science.gov (United States)

    Ashkenazy, Yosef

    2017-03-01

    There are two main comparable sources of energy to the deep ocean-winds and tides. However, the identity of the most efficient mechanism that transfers wind energy to the deep ocean is still debated. Here we study, using oceanic general circulation model simulations and analytic derivations, the way that the wind directly supplies energy down to the bottom of the ocean when it is stochastic and temporally correlated or when it is periodic with a frequency that matches the Coriolis frequency. Basically, under these, commonly observed, conditions, one of the wind components resonates with the Coriolis frequency. Using reanalysis surface wind data and our simple model, we show that about one-third of the kinetic energy that is associated with wind-induced currents resides in the abyssal ocean, highlighting the importance of the resonance of the wind with the Coriolis force.

  19. Treatment of lip hemangioma using forced dehydration with induced photocoagulation via diode laser: report of three cases.

    Science.gov (United States)

    Jasper, Juliana; Camilotti, Renata Stifelman; Pagnoncelli, Rogério Miranda; Poli, Vladimir Dourado; da Silveira Gerzson, Alexandre; Gavin Zakszeski, Ana Maria

    2015-03-01

    Several vascular lesions are related to the lip area. There is no universally accepted protocol for the treatment of hemangiomas and vascular malformations. In the oral cavity, high-power lasers represent an excellent therapeutic option for this type of lesion. Their coagulative properties allow for the performance of procedures without the risk of bleeding, which promotes a better healing pattern and a differentiated postoperative appearance. This study describes three cases of lip hemangioma treated with forced dehydration with induced photocoagulation (FDIP) via diode laser. All the reported cases were followed up until complete healing of the operated area had total remission of lesions, with no complications or adverse effects. The findings of the present study suggest that FDIP is effective and useful in the treatment of hemangiomas in the oral cavity. Laser treatment of these lesions prevents their recurrence and is well tolerated by patients.

  20. Hypoxia-induced force increase (HIFI) is a novel mechanism underlying the strengthening of labor contractions, produced by hypoxic stresses

    Science.gov (United States)

    Alotaibi, Mohammed; Arrowsmith, Sarah; Wray, Susan

    2015-01-01

    For successful birth, contractions need to become progressively stronger. The underlying mechanisms are unknown, however. We have found that a novel mechanism, hypoxia-induced force increase (HIFI), is switched on selectively, at term, and is essential to strengthening contractions. HIFI is initiated as contractions cyclically reduce blood flow and produce repeated hypoxic stresses, with associated metabolic and transcriptomic changes. The increases in contractility are a long-lasting, oxytocin-independent, intrinsic mechanism present only in the full-term pregnant uterus. HIFI is inhibited by adenosine receptor antagonism and blockade of cyclooxygenase-2 signaling, and partially reproduced by brief episodes of acidic (but not alkalotic) pH. HIFI explains how labor can progress despite paradoxical metabolic challenge, and provides a new mechanistic target for the 1 in 10 women suffering dysfunctional labor because of poor contractions. PMID:26195731

  1. Effects of alpha-AMPK knockout on exercise-induced gene activation in mouse skeletal muscle

    DEFF Research Database (Denmark)

    Jørgensen, Sebastian Beck; Wojtaszewski, Jørgen; Viollet, Benoit

    2005-01-01

    We tested the hypothesis that 5'AMP-activated protein kinase (AMPK) plays an important role in regulating the acute, exercise-induced activation of metabolic genes in skeletal muscle, which were dissected from whole-body a2- and a1-AMPK knockout (KO) and wild-type (WT) mice at rest, after treadmi...

  2. Exercise induces transient transcriptional activation of the PGC-1a gene in human skeletal muscle

    DEFF Research Database (Denmark)

    Pilegaard, Henriette; Saltin, Bengt; Neufer, P. Darrell

    2003-01-01

    Endurance exercise training induces mitochondrial biogenesis in skeletal muscle. The peroxisome proliferator activated receptor co-activator 1a (PGC-1a) has recently been identified as a nuclear factor critical for coordinating the activation of genes required for mitochondrial biogenesis in cell...

  3. Transcriptomic sequencing reveals a set of unique genes activated by butyrate-induced histone modification

    Science.gov (United States)

    Butyrate is a nutritional element with strong epigenetic regulatory activity as an inhibitor of histone deacetylases (HDACs). Based on the analysis of differentially expressed genes induced by butyrate in the bovine epithelial cell using deep RNA-sequencing technology (RNA-seq), a set of unique gen...

  4. Abrogation of p53-induced apoptosis by the hepatitis B virus X gene.

    NARCIS (Netherlands)

    X.W. Wang (Xin Wei); M.K. Gibson (Michael); W. Vermeulen (Wim); H. Yeh; K. Forrester; H.-W. Stürzbecher; J.H.J. Hoeijmakers (Jan); C.C. Harris

    1996-01-01

    textabstractThe p53 tumor suppressor gene product is a transcriptional transactivator and a potent apoptotic inducer. The fact that many of the DNA tumor virus oncoproteins bind to p53 and affect these p53 functions indicates that this interaction is an important step in oncogenic transformation. We

  5. Gene expression profiling and pathway analysis of hepatotoxicity induced by triptolide in Wistar rats.

    Science.gov (United States)

    Wang, Jiaying; Jiang, Zhenzhou; Ji, Jinzi; Wang, Xinzhi; Wang, Tao; Zhang, Yun; Tai, Ting; Chen, Mi; Sun, Lixin; Li, Xia; Zhang, Luyong

    2013-08-01

    Triptolide (TP), a major component of TWHF, is widely used to treat rheumatoid arthritis, systemic lupus erythematosus, nephritis and leprosy. However, its clinical use is limited by hepatotoxicity. To further elucidate the underlying mechanism of its hepatotoxic effects, hepatic gene expression profiles were analyzed. TP (1000 and 300 μg/kg) was orally administered to Wistar rats for 14 days. Current study indicated that female rats were more sensitive to TP-induced hepatotoxicity than males. Genome-wide microarray analyses identified 3329 differentially expressed genes in liver of female rats. Analyses of these genes identified over-represented functions associated with insulin signaling pathway, glucose metabolism, cell cycle, oxidative stress and apoptosis, which were consistent with the results of significant increase of Caspase-3 activity and reduction of serum glucose, GSH/GSSG ratio, glucose-6-phosphatase and phosphoenolpyruvate carboxykinase activities, liver glycogen. In addition, it was observed for the first time that glucocorticoids and IGF1 might get involved in TP-induced hepatotoxicity. These data suggest that TP treatment could alter the hepatic redox status, reduce serum glucose and induce hepatocyte apoptosis, consistent with the differential expression of genes involved in insulin signaling pathway, glucose metabolism pathway and cell stress pathway, all of which might contribute to the overall TP-induced hepatotoxicity.

  6. An in planta induced gene of Phytophthora infestans codes for ubiquitin

    NARCIS (Netherlands)

    Pieterse, C.M.J.; Risseeuw, E.P.; Davidse, L.C.

    1991-01-01

    An in planta induced gene of Phytophthora infestans (the causal organism of potato late blight) was selected from a genomic library by differential hybridization using labelled cDNA derived from poly(A)+ RNA of P. infestans grown in vitro and labelled cDNA made from potato-P. infestans interaction

  7. An in planta induced gene of Phytophthora infestans codes for ubiquitin

    NARCIS (Netherlands)

    Pieterse, C.M.J.; Risseeuw, E.P.; Davidse, L.C.

    1991-01-01

    An in planta induced gene of Phytophthora infestans (the causal organism of potato late blight) was isolated from a genomic library by differential hybridization using labelled cDNA derived from poly(A)⁺ RNA of P. infestans grown in vitro and labelled cDNA made from potato-P,

  8. Gene expression array analyses predict increased proto-oncogene expression in MMTV induced mammary tumors.

    Science.gov (United States)

    Popken-Harris, Pamela; Kirchhof, Nicole; Harrison, Ben; Harris, Lester F

    2006-08-01

    Exogenous infection by milk-borne mouse mammary tumor viruses (MMTV) typically induce mouse mammary tumors in genetically susceptible mice at a rate of 90-95% by 1 year of age. In contrast to other transforming retroviruses, MMTV acts as an insertional mutagen and under the influence of steroid hormones induces oncogenic transformation after insertion into the host genome. As these events correspond with increases in adjacent proto-oncogene transcription, we used expression array profiling to determine which commonly associated MMTV insertion site proto-oncogenes were transcriptionally active in MMTV induced mouse mammary tumors. To verify our gene expression array results we developed real-time quantitative RT-PCR assays for the common MMTV insertion site genes found in RIII/Sa mice (int-1/wnt-1, int-2/fgf-3, int-3/Notch 4, and fgf8/AIGF) as well as two genes that were consistently up regulated (CCND1, and MAT-8) and two genes that were consistently down regulated (FN1 and MAT-8) in the MMTV induced tumors as compared to normal mammary gland. Finally, each tumor was also examined histopathologically. Our expression array findings support a model whereby just one or a few common MMTV insertions into the host genome sets up a dominant cascade of events that leave a characteristic molecular signature.

  9. Rescue Effects and Underlying Mechanisms of Intragland Shh Gene Delivery on Irradiation-Induced Hyposalivation.

    Science.gov (United States)

    Hai, Bo; Zhao, Qingguo; Qin, Lizheng; Rangaraj, Dharanipathy; Gutti, Veera R; Liu, Fei

    2016-05-01

    Irreversible hypofunction of salivary glands is common in head and neck cancer survivors treated with radiotherapy and can only be temporarily relieved with current treatments. We found in an inducible sonic hedgehog (Shh) transgenic mouse model that transient activation of the Hedgehog pathway after irradiation rescued salivary gland function in males by preserving salivary stem/progenitor cells and parasympathetic innervation. To translate these findings into feasible clinical application, we evaluated the effects of Shh gene transfer to salivary glands of wild-type mice on irradiation-induced hyposalivation. Shh or control GFP gene was delivered by noninvasive retrograde ductal instillation of corresponding adenoviral vectors. In both male and female mice, Shh gene delivery efficiently activated Hedgehog/Gli signaling, and significantly improved stimulated saliva secretion and preserved saliva-producing acinar cells after irradiation. In addition to preserving parasympathetic innervation through induction of neurotrophic factors, Shh gene delivery also alleviated the irradiation damage of the microvasculature, likely via inducing angiogenic factors, but did not expand the progeny of cells responsive to Hedgehog/Gli signaling. These data indicate that transient activation of the Hedgehog pathway by gene delivery is promising to rescue salivary function after irradiation in both sexes, and the Hedgehog/Gli pathway may function mainly in cell nonautonomous manners to achieve the rescue effect.

  10. Identification and characterization of the inducible murine mast cell gene, imc-415.

    Science.gov (United States)

    Cho, S H; Cho, J J; Kim, I S; Vliagoftis, H; Metcalfe, D D; Oh, C K

    1998-11-01

    Activation of mast cells results in the generation and release of bioactive mediators which in turn initiate allergic inflammation. Mast cell function is enhanced following stimulation in part because of the induction of specific genes and their products. To identify additional genes induced in mast cells that support this process, we thus constructed an activation-specific mast cell subtraction library. To date, we have isolated 26 novel inducible murine mast cell (imc) cDNA clones. Among them, a full-coding region of the murine gene imc-415 was found to have a greater than 90% nucleotide sequence homology and a 97.5% amino acid sequence homology to both a human beta4 integrin-binding protein (p27(BBP)) and a human translation initiation factor 6 (eIF6), which in turn are identical. In vitro translation of the imc-415 gene yielded a band of an approximately 26 kDa. This is the same as the calculated molecular weight of murine IMC-415 protein based on the predicted amino acid sequence and is the molecular weight of p27(BBP)/eIF6. Murine imc-415 message was also induced in inflamed lung tissues in a mouse model of asthma. These results suggest a role for murine imc-415 in allergic inflammation where it may enhance protein synthesis. Human eIF6/p27(BBP) may also play a role in allergic diseases based on the similarities in sequence and in gene expression patterns.

  11. Effect of bacoside extract from Bacopa monniera on physical fatigue induced by forced swimming.

    Science.gov (United States)

    Anand, T; Phani Kumar, G; Pandareesh, M D; Swamy, M S L; Khanum, Farhath; Bawa, A S

    2012-04-01

    The antifatigue effect of bacoside extract (BME) from Bacopa monniera (L.) Wettst. was investigated. Rats were subjected to weight-loaded forced swim test (WFST) every alternate day for 3 weeks. The BME at a dosage of 10 mg/kg body weight was administered orally to rats for 2 weeks in order to evaluate the following biomarkers of physical fatigue: swimming time, change in body weight, lipid peroxidation, lactic acid (LA), glycogen, antioxidant enzyme activities such as superoxide dismutase (SOD) and catalase (CAT) and blood parameters, namely blood urea nitrogen (BUN) and creatine kinase (CK). The exhaustive swimming time was increased by 3-fold in the BME supplemented group compared with that of the control group on day 13. The BME treatment lowered malondialdehyde (MDA) levels in brain, liver and muscle tissues by 11.2%, 16.2% and 37.7%, respectively, compared with the control exercised group (p < 0.05). The BME also reduced the LA, serum BUN and CK activities significantly compared with that of the control. Administration of BME significantly protected the depletion of SOD and CAT activities. The HSP-70 expression studies by western blot also confirmed the antifatigue property of BME. The present study thus indicates that BME ameliorates the various impairments associated with physical fatigue.

  12. Active noise control of forced and induced draft fans in power generating plants

    Energy Technology Data Exchange (ETDEWEB)

    Eldada, M.V.

    1985-05-01

    A study was carried out into active noise reduction of forced draft fans in power generation plants. Active noise reduction involves detecting noise, inverting the phase and re-introducing the anti-phase signal into the duct to cancel the noise through destructive interference. Acoustic pressure transfer functions were meaured in-situ along a cross section of a 15 ft diameter fan inlet. A computer modal analysis program was written and used to analyze the field data. It was found that in frequencies between ca 5 Hz and 45 Hz the energy propagates mainly in the plane wave mode, while in higher frequencies the energy was carried mainly by higher order propagation modes. The project objective was to cancel noise up to a frequency of 130 Hz, but current technology restricted active cancellation to plane waves only. Three alternatives were considered: install a feed forward active noise control system to cancel noise at frequencies below 45 Hz; conduct research on active noise control of higher order propagation modes in ducts; or install a feed back active noise control system and a duct splitter in order to cancel noise between 30 and 130 Hz. It was recommended that the third option be selected as the next phase of the research project, which would comprise a 20 ft duct splitter and microphones, filters, amplifiers, loudspekers and cabinets. 6 refs., 12 figs.

  13. Investigation of quercetin-induced HepG2 cell apoptosis-associated cellular biophysical alterations by atomic force microscopy.

    Science.gov (United States)

    Pi, Jiang; Li, Baole; Tu, Lvying; Zhu, Haiyan; Jin, Hua; Yang, Fen; Bai, Haihua; Cai, Huaihong; Cai, Jiye

    2016-01-01

    Quercetin, a wildly distributed bioflavonoid, has been proved to possess excellent antitumor activity on hepatocellular carcinoma (HCC). In the present study, the biophysical properties of HepG2 cells were qualitatively and quantitatively determined using high resolution atomic force microscopy (AFM) to understand the anticancer effects of quercetin on HCC cells at nanoscale. The results showed that quercetin could induce severe apoptosis in HepG2 cells through arrest of cell cycle and disruption of mitochondria membrane potential. Additionally, the nuclei and F-actin structures of HepG2 cells were destroyed by quercetin treatment as well. AFM morphological data showed some typical apoptotic characterization of HepG2 cells with increased particle size and roughness in the ultrastructure of cell surface upon quercetin treatment. As an important biophysical property of cells, the membrane stiffness of HepG2 cells was further quantified by AFM force measurements, which indicated that HepG2 cells became much stiffer after quercetin treatment. These results collectively suggest that quercetin can be served as a potential therapeutic agent for HCC, which not only extends our understanding of the anticancer effects of quercetin against HCC cells into nanoscale, but also highlights the applications of AFM for the investigation of anticancer drugs.

  14. Cloning of murine BRI3 gene and study on its function for inducing cell death

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    To understand the molecular mechanism of TNFα effects, the cDNA of murine BRI3 gene was cloned from the total RNA of murine brain endothelial cells (bEnd.3)treated with hTNFα by using the suppression subtractive hybridization (SSH) and the RT-PCR method. The fusion expression vector harbouring BRI3 gene and enhanced green fluorescence protein (EGFP) thus obtained were designated as pEGFP/I3. Then pEGFP/I3 was transiently transfected into L929 cells and the fusion protein EGFP/I3 was localized in cytoplasm. It is found that the expression of EGFP/I3 could induce cell death in L929 cells detected by TUNEL method and flow cytometry. And the overexpression of Bci-2 in L929 cells can block cell death induced by EGFP/I3, indicating that murine BRI3 gene might related to the TNFα mediated cytotoxicity.

  15. Gene-gun DNA vaccination aggravates respiratory syncytial virus-induced pneumonitis

    DEFF Research Database (Denmark)

    Bartholdy, Christina; Olszewska, Wieslawa; Stryhn, Anette

    2004-01-01

    elicited with recombinant vaccinia virus expressing the complete RSV M2 protein, but stronger than those induced by a similar DNA construct without the beta2m gene. DNA vaccination led to enhanced pulmonary disease after RSV challenge, with increased weight loss and cell recruitment to the lung. Depletion......A CD8+ T-cell memory response to respiratory syncytial virus (RSV) was generated by using a DNA vaccine construct encoding the dominant Kd-restricted epitope from the viral transcription anti-terminator protein M2 (M2(82-90)), linked covalently to human beta2-microglobulin (beta2m). Cutaneous gene......+ T-cell responses were not induced. Thus, in addition to specific CD8+ T cell-mediated immunopathology, gene-gun DNA vaccination causes non-specific enhancement of RSV disease without affecting virus clearance....

  16. Identification of LPS-inducible genes downregulated by ubiquinone in human THP-1 monocytes.

    Science.gov (United States)

    Schmelzer, Constance; Döring, Frank

    2010-01-01

    Coenzyme Q(10) (CoQ(10)) is an obligatory element in the respiratory chain and functions as a potent antioxidant of lipid membranes. More recently, anti-inflammatory effects as well as an impact of CoQ(10) on gene expression have been observed. To reveal putative effects of Q(10) on LPS-induced gene expression, whole genome expression analysis was performed in the monocytic cell line THP-1. Thousand one hundred twenty-nine and 710 probe sets have been identified to be significantly (P genes revealed a functional connection in the NFkappaB pathway and confirmed our applied in vitro stimulation model. Moreover, 33 LPS-sensitive genes have been identified to be significantly downregulated by Q(10)-treatment between a factor of 1.32 and 1.85. GeneOntology (GO) analysis revealed for the Q(10)-sensitve genes a primary involvement in protein metabolism (e.g., HERC1 and EPS15), cell proliferation (e.g., CCDC100 and SMURF1), and transcriptional processes (e.g., CNOT4 and STK4). Three genes were either related to NFkappaB transcription factor activity (ERC1), cytokinesis (DIAPH2), or modulation of oxidative stress (MSRA). In conclusion, our data provide evidence that Q(10) downregulates LPS-inducible genes in the monocytic cell line THP-1. Thus, the previously described effects of Q(10) on the reduction of proinflammatory mediators might be due to its antioxidant impact on gene expression.

  17. Gene expression in Barrett's esophagus and reflux esophagitis induced by gastroduodenoesophageal reflux in rats

    Institute of Scientific and Technical Information of China (English)

    Peng Cheng; Jun Gong; Tao Wang; Chen Jie; Gui-Sheng Liu; Ru Zhang

    2005-01-01

    AIM: To investigate the difference of gene expression profiles between Barrett's esophagus and reflux esophagitis induced by gastroduodenoesophageal reflux in rats.METHODS: Eight-week-old Sprague-Dawley rats were treated esophagoduodenostomy to produce gastroduodenoesophageal reflux, and another group received sham operation as control. Esophageal epithelial tissues were dissected and frozen in liquid nitrogen immediately for pathology 40 wk after surgery. The expression profiles of 4 096 genes in reflux esophagitis and Barrett's esophagus tissues were compared with normal esophageal epithelium by cDNA microarray.RESULTS: Four hundred and forty-eight genes in Barrett'sesophagus were more than three times different from those in normal esophageal epithelium, including 312 up regulated and 136 down-regulated genes. Two hundred and thirty-twogenes in RE were more than three times different from those in normal esophageal epithelium, 90up-regulated and 142 down-regulated genes. Compared to reflux esophagitis, there were 214 up-regulated and 142 down-regulated genes in Barrett's esophagus. CONCLUSION: Esophageal epithelium exposed excessively to harmful ingredients of duodenal and gastric reflux can develop esophagitis and Barrett's esophagus gradually.The gene expression level is different between reflux esophagitis and Barrett's esophagus and the differentially expressed genes might be related to the occurrence and development of Barrett's esophagus and the promotion or progression in adenocarcinoma.

  18. Combining Click Chemistry-Based Proteomics With Dox-Inducible Gene Expression.

    Science.gov (United States)

    Gebert, J; Schnölzer, M; Warnken, U; Kopitz, J

    2017-01-01

    Inactivating mutations in single genes can trigger, prevent, promote, or alleviate diseases. Identifying such disease-related genes is a main pillar of medical research. Since proteins play a crucial role in mediating these effects, their impact on the diseased cells' proteome including posttranslational modifications has to be elucidated for a detailed understanding of the role of these genes in the disease process. In complex disorders, like cancer, several genes contribute to the disease process, thereby hampering the assignment of a proteomic change to the corresponding causative gene. To enable comprehensive screening for the impact of inactivation of a gene, e.g., loss of a tumor suppressor in cancer, on the cellular proteome, we present a strategy based on combination of three technologies that is recombinase-mediated cassette exchange, click chemistry, and mass spectrometry. The methodology is exemplified by the analysis of the proteomic changes induced by the loss of a tumor suppressor gene in colorectal cancer cells. To demonstrate the applicability to screen for posttranslational modification changes, we also describe the analysis of protein glycosylation changes caused by the tumor suppressor inactivation. In principle, this strategy can be applied to analyze the effects of any gene of interest on protein expression as well as posttranslational modification by glycosylation. Moreover adaptation of the strategy to an appropriate cell culture model has the potential for application on a broad range of diseases where the disease-promoting mutations have been identified. © 2017 Elsevier Inc. All rights reserved.

  19. A virus-induced gene silencing approach to understanding alkaloid metabolism in Catharanthus roseus.

    Science.gov (United States)

    Liscombe, David K; O'Connor, Sarah E

    2011-11-01

    The anticancer agents vinblastine and vincristine are bisindole alkaloids derived from coupling vindoline and catharanthine, monoterpenoid indole alkaloids produced exclusively by the Madagascar periwinkle (Catharanthus roseus). Industrial production of vinblastine and vincristine currently relies on isolation from C. roseus leaves, a process that affords these compounds in 0.0003-0.01% yields. Metabolic engineering efforts to either improve alkaloid content or provide alternative sources of the bisindole alkaloids ultimately rely on the isolation and characterization of the genes involved. Several vindoline biosynthetic genes have been isolated, and the cellular and subcellular organization of the corresponding enzymes has been well studied. However, due to the leaf-specific localization of vindoline biosynthesis, and the lack of production of this precursor in cell suspension and hairy root cultures of C. roseus, further elucidation of this pathway demands the development of reverse genetics approaches to assay gene function in planta. The bipartite pTRV vector system is a Tobacco Rattle Virus-based virus-induced gene silencing (VIGS) platform that has provided efficient and effective means to assay gene function in diverse plant systems. A VIGS method was developed herein to investigate gene function in C. roseus plants using the pTRV vector system. The utility of this approach in understanding gene function in C. roseus leaves is demonstrated by silencing known vindoline biosynthetic genes previously characterized in vitro.

  20. Engineering Human Stem Cell Lines with Inducible Gene Knockout using CRISPR/Cas9.

    Science.gov (United States)

    Chen, Yuejun; Cao, Jingyuan; Xiong, Man; Petersen, Andrew J; Dong, Yi; Tao, Yunlong; Huang, Cindy Tzu-Ling; Du, Zhongwei; Zhang, Su-Chun

    2015-08-06

    Precise temporal control of gene expression or deletion is critical for elucidating gene function in biological systems. However, the establishment of human pluripotent stem cell (hPSC) lines with inducible gene knockout (iKO) remains challenging. We explored building iKO hPSC lines by combining CRISPR/Cas9-mediated genome editing with the Flp/FRT and Cre/LoxP system. We found that "dual-sgRNA targeting" is essential for biallelic knockin of FRT sequences to flank the exon. We further developed a strategy to simultaneously insert an activity-controllable recombinase-expressing cassette and remove the drug-resistance gene, thus speeding up the generation of iKO hPSC lines. This two-step strategy was used to establish human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC) lines with iKO of SOX2, PAX6, OTX2, and AGO2, genes that exhibit diverse structural layout and temporal expression patterns. The availability of iKO hPSC lines will substantially transform the way we examine gene function in human cells.

  1. UVB-induced gene expression in the skin of Xiphophorus maculatus Jp 163 B.

    Science.gov (United States)

    Yang, Kuan; Boswell, Mikki; Walter, Dylan J; Downs, Kevin P; Gaston-Pravia, Kimberly; Garcia, Tzintzuni; Shen, Yingjia; Mitchell, David L; Walter, Ronald B

    2014-06-01

    Xiphophorus fish and interspecies hybrids represent long-standing models to study the genetics underlying spontaneous and induced tumorigenesis. The recent release of the Xiphophorus maculatus genome sequence will allow global genetic regulation studies of genes involved in the inherited susceptibility to UVB-induced melanoma within select backcross hybrids. As a first step toward this goal, we report results of an RNA-Seq approach to identify genes and pathways showing modulated transcription within the skin of X. maculatus Jp 163 B upon UVB exposure. X. maculatus Jp 163 B were exposed to various doses of UVB followed by RNA-Seq analysis at each dose to investigate overall gene expression in each sample. A total of 357 genes with a minimum expression change of 4-fold (p-adjbasal expression level of each transcript for each skin sample, (2) the changes in expression levels for each gene in the transcriptome upon exposure to increasing doses of UVB, and (3) clusters of genes that exhibit similar patterns of change in expression upon UVB exposure. These data provide a foundation for understanding the molecular genetic response of fish skin to UVB exposure.

  2. Laparotomy in mice induces blood cell expression of inflammatory and stress genes.

    Science.gov (United States)

    Ko, Fred; Isoda, Fumiko; Mobbs, Charles

    2015-04-01

    Surgical trauma induces immune and stress responses although its effects on postsurgical inflammatory and stress gene expression remain poorly characterized. This study sought to improve current scientific knowledge by investigating the effects of laparotomy on mouse blood cell inflammatory and stress gene expression. Three-month-old male C57BL/6J mice were subjected to 2% isoflurane or 2% isoflurane with laparotomy and sacrificed 4 h postintervention. Blood was collected and blood cell expression of 158 genes central to inflammatory and stress responses was assayed using quantitative polymerase chain reaction arrays. Mice subjected to isoflurane with laparotomy, compared with mice receiving isoflurane alone, had >2-fold upregulation of genes in inflammation (Osm, IL1rn, IL1b, and Csf1), oxidative stress (Hmox1), heat shock (Hspa1b), growth arrest (Cdkn1a), and DNA repair (Ugt1a2). These genes demonstrated similar expression patterns by Pearson correlation and cluster analysis. Thus, laparotomy induces coordinated, postsurgical blood cell expression of unique inflammatory and stress genes whose roles in influencing surgical outcomes need further investigation.

  3. ADHESION INDUCES MATRIX METALLOPROTEINASE-9 GENE EXPRESSION IN OVARIAN CANCER CELLS

    Institute of Scientific and Technical Information of China (English)

    田方; 颜春洪; 薛红; 肖凤君

    2002-01-01

    Objective: To investigate the expression of matrix metalloproteinase-9 (MMP-9) gene in cancer cells induced by adhesion with fibronectin and the underlying mechanism of cell invasion. Methods: Following adhesion of ovarian cancer cells A2780 to fibronectin, MMP mRNA expression was assayed by using reverse transcription-polymerase chain reaction (RT-PCR). MMP-9 promoter was cloned from genomic DNA of HT1080 cells with PCR. The MMP-9-pGL2 reporter gene vector was constructed and then transiently transfected into A2780 cells. Results: Adhesion could induce the expression of MMP-9 gene in A2780 cells, but did not affect longer theexpression of MMP-2 or TIMP-1 gene. The induction was enhanced with longer adhesion time. When the transfected cells were allowed to adhere and spread on FN-coated surface, the promoter activity of MMP-9 gene was also enhanced dramatically. Conclusion: adhesion of cells with ECM may stimulate the expression of MMP-9 gene through stimulating the promoter activity, thereby enhancing cancer cell invasion and metastasis.

  4. Effect of induced shielding current transmission in longitudinal direction on levitation force of melt grown single-domain YBa2Cu3O7-x cylindrical superconductor

    Institute of Scientific and Technical Information of China (English)

    YANG Wanmin; ZHOU Lian; FENG Yong; ZHANG Pingxiang; R.Nicolsky; R.de Andrade Jr

    2004-01-01

    A novel layer deletion method is used to experimentally investigate the effect of induced shielding current transmission (ISCT) in the longitudinal direction on the levitation force of a single-domain YBa2Cu3O7-x (YBCO) cylindrical superconductor (φ30x7 mm). In the experiment the sample was gradually sliced into two equal sheets, at the middle height along a diameter with 5 mm every step. The experimental results show that the levitation force is closely related with the ISCT in the longitudinal direction. Any layer deletion, even a small piece of layer deletion can reduce the levitation force of the sample. After the whole layer was deleted the levitation force can diminish about 50%. It is also found that the levitation force is directly proportional to the effective factor of surface area, which is equal to the top surface area divided by the total surface area parallel to the top surface of the sample.

  5. Characterization of chemically induced liver injuries using gene co-expression modules.

    Directory of Open Access Journals (Sweden)

    Gregory J Tawa

    Full Text Available Liver injuries due to ingestion or exposure to chemicals and industrial toxicants pose a serious health risk that may be hard to assess due to a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific damage and clinical outcomes via biomarkers or biomarker panels will provide the foundation for highly specific and robust diagnostic tests. Here, we have used DrugMatrix, a toxicogenomics database containing organ-specific gene expression data matched to dose-dependent chemical exposures and adverse clinical pathology assessments in Sprague Dawley rats, to identify groups of co-expressed genes (modules specific to injury endpoints in the liver. We identified 78 such gene co-expression modules associated with 25 diverse injury endpoints categorized from clinical pathology, organ weight changes, and histopathology. Using gene expression data associated with an injury condition, we showed that these modules exhibited different patterns of activation characteristic of each injury. We further showed that specific module genes mapped to 1 known biochemical pathways associated with liver injuries and 2 clinically used diagnostic tests for liver fibrosis. As such, the gene modules have characteristics of both generalized and specific toxic response pathways. Using these results, we proposed three gene signature sets characteristic of liver fibrosis, steatosis, and general liver injury based on genes from the co-expression modules. Out of all 92 identified genes, 18 (20% genes have well-documented relationships with liver disease, whereas the rest are novel and have not previously been associated with liver disease. In conclusion, identifying gene co-expression modules associated with chemically induced liver injuries aids in generating testable hypotheses and has the potential to identify putative biomarkers of adverse health effects.

  6. Characterization of Chemically Induced Liver Injuries Using Gene Co-Expression Modules

    Science.gov (United States)

    Tawa, Gregory J.; AbdulHameed, Mohamed Diwan M.; Yu, Xueping; Kumar, Kamal; Ippolito, Danielle L.; Lewis, John A.; Stallings, Jonathan D.; Wallqvist, Anders

    2014-01-01

    Liver injuries due to ingestion or exposure to chemicals and industrial toxicants pose a serious health risk that may be hard to assess due to a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific damage and clinical outcomes via biomarkers or biomarker panels will provide the foundation for highly specific and robust diagnostic tests. Here, we have used DrugMatrix, a toxicogenomics database containing organ-specific gene expression data matched to dose-dependent chemical exposures and adverse clinical pathology assessments in Sprague Dawley rats, to identify groups of co-expressed genes (modules) specific to injury endpoints in the liver. We identified 78 such gene co-expression modules associated with 25 diverse injury endpoints categorized from clinical pathology, organ weight changes, and histopathology. Using gene expression data associated with an injury condition, we showed that these modules exhibited different patterns of activation characteristic of each injury. We further showed that specific module genes mapped to 1) known biochemical pathways associated with liver injuries and 2) clinically used diagnostic tests for liver fibrosis. As such, the gene modules have characteristics of both generalized and specific toxic response pathways. Using these results, we proposed three gene signature sets characteristic of liver fibrosis, steatosis, and general liver injury based on genes from the co-expression modules. Out of all 92 identified genes, 18 (20%) genes have well-documented relationships with liver disease, whereas the rest are novel and have not previously been associated with liver disease. In conclusion, identifying gene co-expression modules associated with chemically induced liver injuries aids in generating testable hypotheses and has the potential to identify putative biomarkers of adverse health effects. PMID:25226513

  7. Forced diuresis with the RenalGuard system: impact on contrast induced acute kidney injury.

    Science.gov (United States)

    Solomon, Richard

    2014-01-01

    Kidney injury following the administration of iodinated contrast media occurs particularly in patients with reduced kidney and cardiac function and when large doses of contrast are used. There is little compelling evidence that vasodilators and anti-oxidants prevent this injury. Most prevention trials have employed intravenous volume loading as a central strategy. However, the success of this approach depends upon maintaining euvolemia while producing a vigorous diuresis. A novel strategy for maintaining euvolemia and inducing a vigorous diuresis has been developed using the RenalGuard system. In this review; the mechanism of protective action is reviewed. The trials of the RenalGuard device are reviewed and future uses of the device are discussed.

  8. Effect of nifedipine on depolarization-induced force responses in skinned skeletal muscle fibres of rat and toad.

    Science.gov (United States)

    Posterino, G S; Lamb, G D

    1998-01-01

    The effect of the dihydropyridine, nifedipine, on excitation-contraction coupling was compared in toad and rat skeletal muscle, using the mechanically skinned fibre technique, in order to understand better the apparently disparate results of previous studies and to examine recent proposals on the importance of certain intracellular factors in determining the efficacy of dihydropyridines. In twitch fibres from the iliofibularis muscle of the toad, 10 microM nifedipine completely inhibited depolarization-induced force responses within 30 s, without interfering with direct activation of the Ca(2+)-release channels by caffeine application or reduction of myoplasmic [Mg2+]. At low concentrations of nifedipine, inhibition was considerably augmented by repeated depolarizations, with half-maximal inhibition occurring at < 0.1 microM nifedipine. In contrast, in rat extensor digitorum longus (EDL) fibres 1 microM nifedipine had virtually no effect on depolarization-induced force responses, and 10 microM nifedipine caused only approximately 25% reduction in the responses, even upon repeated depolarizations. In rat fibres, 10 microM nifedipine shifted the steady-state force inactivation curve to more negative potentials by < 11 mV, whereas in toad fibres the potent inhibitory effect of nifedipine indicated a much larger shift. The inhibitory effect of nifedipine in rat fibres was little, if at all, increased by the absence of Ca2+ in the transverse tubular (t-) system, provided that the Ca2+ was replaced with sufficient Mg2+. The presence of the reducing agents dithiothreitol (10 mM) or glutathione (10 mM) in the solution bathing a toad skinned fibre did not reduce the inhibitory effect of nifedipine, suggesting that the potency of nifedipine in toad skinned fibres was not due to the washout of intracellular reducing agents. The results are considered in terms of a model that can account for the markedly different effects of nifedipine on the two putative functions of the

  9. Dynamic gene expression in fish muscle during recovery growth induced by a fasting-refeeding schedule

    Directory of Open Access Journals (Sweden)

    Esquerré Diane

    2007-11-01

    Full Text Available Abstract Background Recovery growth is a phase of rapid growth that is triggered by adequate refeeding of animals following a period of weight loss caused by starvation. In this study, to obtain more information on the system-wide integration of recovery growth in muscle, we undertook a time-course analysis of transcript expression in trout subjected to a food deprivation-refeeding sequence. For this purpose complex targets produced from muscle of trout fasted for one month and from muscle of trout fasted for one month and then refed for 4, 7, 11 and 36 days were hybridized to cDNA microarrays containing 9023 clones. Results Significance analysis of microarrays (SAM and temporal expression profiling led to the segregation of differentially expressed genes into four major clusters. One cluster comprising 1020 genes with high expression in muscle from fasted animals included a large set of genes involved in protein catabolism. A second cluster that included approximately 550 genes with transient induction 4 to 11 days post-refeeding was dominated by genes involved in transcription, ribosomal biogenesis, translation, chaperone activity, mitochondrial production of ATP and cell division. A third cluster that contained 480 genes that were up-regulated 7 to 36 days post-refeeding was enriched with genes involved in reticulum and Golgi dynamics and with genes indicative of myofiber and muscle remodelling such as genes encoding sarcomeric proteins and matrix compounds. Finally, a fourth cluster of 200 genes overexpressed only in 36-day refed trout muscle contained genes with function in carbohydrate metabolism and lipid biosynthesis. Remarkably, among the genes induced were several transcriptional regulators which might be important for the gene-specific transcriptional adaptations that underlie muscle recovery. Conclusion Our study is the first demonstration of a coordinated expression of functionally related genes during muscle recovery growth

  10. Training-induced changes in muscle CSA, muscle strength, EMG, and rate of force development in elderly subjects after long-term unilateral disuse

    DEFF Research Database (Denmark)

    Suetta, Charlotte; Aagaard, Per; Rosted, Anna

    2004-01-01

    , the present study examined the effect of three types of training regimes after unilateral prolonged disuse and subsequent hip-replacement surgery on maximal muscle strength, rapid muscle force [rate of force development (RFD)], muscle activation, and muscle size. Thirty-six subjects (60-86 yr) were randomized......The ability to develop muscle force rapidly may be a very important factor to prevent a fall and to perform other tasks of daily life. However, information is still lacking on the range of training-induced neuromuscular adaptations in elderly humans recovering from a period of disuse. Therefore...

  11. Controlled Gene Expression Systems for Lactic Acid Bacteria : Transferable Nisin-Inducible Expression Cassettes for Lactococcus, Leuconostoc, and Lactobacillus spp.

    NARCIS (Netherlands)

    Kleerebezem, Michiel; Beerthuyzen, Marke M.; Vaughan, Elaine E.; Vos, Willem M. de; Kuipers, Oscar P.

    1997-01-01

    A transferable dual-plasmid inducible gene expression system for use in lactic acid bacteria that is based on the autoregulatory properties of the antimicrobial peptide nisin produced by Lactococcus lactis was developed. Introduction of the two plasmids allowed nisin-inducible gene expression in Lac

  12. Effects of modulation of calcium levels and calcium fluxes on ABA- induced gene expression in barley aleurone

    NARCIS (Netherlands)

    Meulen, R.M. van der; Visser, K.; Wang, M.

    1996-01-01

    We present data to elucidate the involvement of calcium ions in abscisic acid (ABA)-induced gene expression. Modulation of external calcium concentrations was able to affect ABA-induced specific RAB gene expression. At a constant ABA level with increasing extracellular calcium level, an increasing R

  13. Effects of modulation of calcium levels and calcium fluxes on ABA- induced gene expression in barley aleurone

    NARCIS (Netherlands)

    Meulen, R.M. van der; Visser, K.; Wang, M.

    1996-01-01

    We present data to elucidate the involvement of calcium ions in abscisic acid (ABA)-induced gene expression. Modulation of external calcium concentrations was able to affect ABA-induced specific RAB gene expression. At a constant ABA level with increasing extracellular calcium level, an increasing R

  14. Barcode Sequencing Screen Identifies SUB1 as a Regulator of Yeast Pheromone Inducible Genes

    Directory of Open Access Journals (Sweden)

    Anna Sliva

    2016-04-01

    Full Text Available The yeast pheromone response pathway serves as a valuable model of eukaryotic mitogen-activated protein kinase (MAPK pathways, and transcription of their downstream targets. Here, we describe application of a screening method combining two technologies: fluorescence-activated cell sorting (FACS, and barcode analysis by sequencing (Bar-Seq. Using this screening method, and pFUS1-GFP as a reporter for MAPK pathway activation, we readily identified mutants in known mating pathway components. In this study, we also include a comprehensive analysis of the FUS1 induction properties of known mating pathway mutants by flow cytometry, featuring single cell analysis of each mutant population. We also characterized a new source of false positives resulting from the design of this screen. Additionally, we identified a deletion mutant, sub1Δ, with increased basal expression of pFUS1-GFP. Here, in the first ChIP-Seq of Sub1, our data shows that Sub1 binds to the promoters of about half the genes in the genome (tripling the 991 loci previously reported, including the promoters of several pheromone-inducible genes, some of which show an increase upon pheromone induction. Here, we also present the first RNA-Seq of a sub1Δ mutant; the majority of genes have no change in RNA, but, of the small subset that do, most show decreased expression, consistent with biochemical studies implicating Sub1 as a positive transcriptional regulator. The RNA-Seq data also show that certain pheromone-inducible genes are induced less in the sub1Δ mutant relative to the wild type, supporting a role for Sub1 in regulation of mating pathway genes. The sub1Δ mutant has increased basal levels of a small subset of other genes besides FUS1, including IMD2 and FIG1, a gene encoding an integral membrane protein necessary for efficient mating.

  15. Performance of repetitive tasks induces decreased grip strength and increased fibrogenic proteins in skeletal muscle: role of force and inflammation.

    Directory of Open Access Journals (Sweden)

    Samir M Abdelmagid

    Full Text Available BACKGROUND: This study elucidates exposure-response relationships between performance of repetitive tasks, grip strength declines, and fibrogenic-related protein changes in muscles, and their link to inflammation. Specifically, we examined forearm flexor digitorum muscles for changes in connective tissue growth factor (CTGF; a matrix protein associated with fibrosis, collagen type I (Col1; a matrix component, and transforming growth factor beta 1 (TGFB1; an upstream modulator of CTGF and collagen, in rats performing one of two repetitive tasks, with or without anti-inflammatory drugs. METHODOLOGY/RESULTS: To examine the roles of force versus repetition, rats performed either a high repetition negligible force food retrieval task (HRNF, or a high repetition high force handle-pulling task (HRHF, for up to 9 weeks, with results compared to trained only (TR-NF or TR-HF and normal control rats. Grip strength declined with both tasks, with the greatest declines in 9-week HRHF rats. Quantitative PCR (qPCR analyses of HRNF muscles showed increased expression of Col1 in weeks 3-9, and CTGF in weeks 6 and 9. Immunohistochemistry confirmed PCR results, and also showed greater increases of CTGF and collagen matrix in 9-week HRHF rats than 9-week HRNF rats. ELISA, and immunohistochemistry revealed greater increases of TGFB1 in TR-HF and 6-week HRHF, compared to 6-week HRNF rats. To examine the role of inflammation, results from 6-week HRHF rats were compared to rats receiving ibuprofen or anti-TNF-α treatment in HRHF weeks 4-6. Both treatments attenuated HRHF-induced increases in CTGF and fibrosis by 6 weeks of task performance. Ibuprofen attenuated TGFB1 increases and grip strength declines, matching our prior results with anti-TNFα. CONCLUSIONS/SIGNIFICANCE: Performance of highly repetitive tasks was associated with force-dependent declines in grip strength and increased fibrogenic-related proteins in flexor digitorum muscles. These changes were

  16. Anoxia-Induced Suspended Animation in Budding Yeast as an Experimental Paradigm for Studying Oxygen-Regulated Gene Expression▿

    Science.gov (United States)

    Chan, Kin; Roth, Mark B.

    2008-01-01

    A lack of oxygen can force many organisms to enter into recoverable hypometabolic states. To better understand how organisms cope with oxygen deprivation, our laboratory previously had shown that when challenged with anoxia, both the nematode Caenorhabditis elegans and embryos of the zebrafish Danio rerio enter into suspended animation, in which all life processes that can be observed by light microscopy reversibly halt pending the restoration of oxygen (P. A. Padilla and M. B. Roth, Proc. Natl. Acad. Sci. USA 98:7331-7335, 2001, and P. A. Padilla, T. G. Nystul, R. A. Zager, A. C. Johnson, and M. B. Roth, Mol. Biol. Cell 13:1473-1483, 2002). Here, we show that both sporulating and vegetative cells of the budding yeast Saccharomyces cerevisiae also enter into a similar state of suspended animation when made anoxic on a nonfermentable carbon source. Transcriptional profiling using cDNA microarrays and follow-on quantitative real-time PCR analysis revealed a relative derepression of aerobic metabolism genes in carbon monoxide (CO)-induced anoxia when compared to nitrogen (N2) gas-induced anoxia, which is consistent with the known oxygen-mimetic effects of CO. We also found that mutants deleted for components of the mitochondrial retrograde signaling pathway can tolerate prolonged exposure to CO but not to N2. We conclude that the cellular response to anoxia is dependent on whether the anoxic gas is an oxygen mimetic and that the mitochondrial retrograde signaling pathway is functionally important for mediating this response. PMID:18708563

  17. Therapeutic angiogenesis induced by human hepatocyte growth factor (HGF) gene in rat myocardial ischemia models

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    In order to investigate the feasibility of myocardial ischemia gene therapy, we cloned human hepatocyte growth factor gene from human placenta cDNA library by the RT-PCR method. Recombination adenovirus Ad-HGF was constructed by the method of co-transfection and homologous recombination of plasmids in 293 cells. Ad-HGF was amplified in 293 cells and purified through CsCl density gradient centrifugation. Ad-HGF could be expressed in rat primary myocardial cells and HGF secreted into the culture media, which was tested by ELISA. The distribution and persistence of adenovirus in rat were investigated by green fluorescence protein as a report gene. In vivo we found that intramyocardial administration of Ad-HGF could induce angiogenesis in rat myocardium after ligation of coronary artery. The results suggested that Ad-HGF was effective in vitro and in vivo, and the data for designing human trial of gene therapy-- mediated cardiac angiogenesis were provided.

  18. Membrane fusion inducers, chloroquine and spermidine increase lipoplex-mediated gene transfection

    Energy Technology Data Exchange (ETDEWEB)

    Wong-Baeza, Carlos; Bustos, Israel; Serna, Manuel; Tescucano, Alonso; Alcantara-Farfan, Veronica; Ibanez, Miguel [Biochemistry Department, National Polytechnic Institute (IPN), Mexico City 11340 (Mexico); Montanez, Cecilia [Department of Genetics and Molecular Biology, Centre for Research and Advanced Studies (CINVESTAV), IPN, Mexico City 07360 (Mexico); Wong, Carlos [Biochemistry Department, National Polytechnic Institute (IPN), Mexico City 11340 (Mexico); Baeza, Isabel, E-mail: ibaeza@encb.ipn.mx [Biochemistry Department, National Polytechnic Institute (IPN), Mexico City 11340 (Mexico)

    2010-05-28

    Gene transfection into mammalian cells can be achieved with viral and non-viral vectors. Non-viral vectors, such as cationic lipids that form lipoplexes with DNA, are safer and more stable than viral vectors, but their transfection efficiencies are lower. Here we describe that the simultaneous treatment with a membrane fusion inducer (chlorpromazine or procainamide) plus the lysosomotropic agent chloroquine increases lipoplex-mediated gene transfection in human (HEK293 and C-33 A) and rat (PC12) cell lines (up to 9.2-fold), as well as in situ in BALB/c mice spleens and livers (up to 6-fold); and that the polyamine spermidine increases lipoplex-mediated gene transfection and expression in cell cultures. The use of these four drugs provides a novel, safe and relatively inexpensive way to considerably increase lipoplex-mediated gene transfection efficiency.

  19. Transcription activation of a UV-inducible Clostridium perfringens bacteriocin gene by a novel sigma factor.

    Science.gov (United States)

    Dupuy, Bruno; Mani, Nagraj; Katayama, Seiichi; Sonenshein, Abraham L

    2005-02-01

    Expression of the plasmid-encoded Clostridium perfringens gene for bacteriocin BCN5 was shown to depend in vivo and in vitro on the activity of UviA protein. UviA, also plasmid-encoded, proved to be an RNA polymerase sigma factor and was also partly autoregulatory. The uviA gene has two promoters; one provided a UviA-independent, basal level of gene expression while the stronger, UviA-dependent promoter was only utilized after the cell experienced DNA damage. As a result, BCN5 synthesis is induced by treatment with UV light or mitomycin C. UviA is related to a special class of sigma factors found to date only in Clostridium species and responsible for activating transcription of toxin genes in Clostridium difficile, Clostridium tetani, and Clostridium botulinum.

  20. Plant defense gene promoter enhances the reliability of shiva-1 gene-induced resistance to soft rot disease in potato.

    Science.gov (United States)

    Yi, Jung Yoon; Seo, Hyo Won; Yang, Moon Sik; Robb, E Jane; Nazar, Ross N; Lee, Shin Woo

    2004-11-01

    PAL5, a tomato (Lycopersicon esculentum Mill.) plant defense gene that encodes phenylalanine ammonia-lyase, is known to respond to a variety of environmental stresses including pathogen infection and wounding. A shiva-1 gene recombinant that encodes a small synthetic antibacterial peptide under the PAL5 gene promoter was transformed into potato (Solanum tuberosum L.) and its ability to induce resistance to Erwinia carotovora was compared with a construct under the control of the constitutive and widely used cauliflower mosaic virus (CaMV) 35S promoter. The shiva-1 peptide, an analog of natural cecropin B, was shown previously to have high bactericidal activity in vitro, but when expressed in vivo under the control of the CaMV 35S promoter, the effects were very inconsistent. As observed previously, in the present studies a few transformants with the CaMV 35S promoter were highly resistant when assayed for susceptibility to soft rot disease. In marked contrast the majority of transformants with the PAL5 gene promoter were highly resistant. More-detailed analyses of the incorporated DNA indicated that most of the transformants with the CaMV 35S promoter contained multiple copies of the transforming DNA while all of the PAL5 recombinants contained single copies. The highly resistant CaMV 35S recombinant also was present as a single copy. The results indicate that, at least in this instance, a constitutive promoter may not be ideal for the effective expression of a foreign gene and suggest that multiple insertions may have negative consequences.

  1. Visualization of Lipid Membrane Reorganization Induced by a Pore-Forming Toxin Using High-Speed Atomic Force Microscopy.

    Science.gov (United States)

    Yilmaz, Neval; Kobayashi, Toshihide

    2015-08-25

    We examined the effect of a sphingomyelin (SM)-binding pore-forming toxin (PFT), lysenin, on the dynamics of a phase-separated membrane of SM, where SM formed liquid-ordered (Lo) domains with cholesterol (Chol) within a phosphatidylcholine-rich liquid-disordered (Ld) phase. We visualized the lysenin-induced membrane reorganization using high-speed atomic force microscope (HS-AFM). Lysenin oligomerized on the SM-rich Lo domain and simultaneously its oligomers assembled into a hexagonal close-packed (hcp) structure. The phase boundary was stable during the assembling of lysenin on the SM-rich domain, indicating that lysenin did not affect the line tension between Lo and Ld phases. After the full coverage of the SM-rich domain by oligomers, their hcp assembly gradually expanded into the Ld phase and eventually covered the entire membrane. Our results suggest that pore formation, i.e., insertion of lysenin into the membrane in its oligomeric state, induced the exclusion of SM and Chol from the SM-rich domain, which was followed by further binding and oligomerization of lysenin.

  2. Atypical antipsychotics induce both proinflammatory and adipogenic gene expression in human adipocytes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Sárvári, Anitta K., E-mail: anittasarvari@med.unideb.hu [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Veréb, Zoltán, E-mail: jzvereb@gmail.com [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Uray, Iván P., E-mail: ipuray@mdanderson.org [Clinical Cancer Prevention Department, The University of Texas, MD Anderson Cancer Center, Houston, TX (United States); Fésüs, László, E-mail: fesus@med.unideb.hu [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); MTA DE Apoptosis, Genomics and Stem Cell Research Group of the Hungarian Academy of Sciences (Hungary); Balajthy, Zoltán, E-mail: balajthy@med.unideb.hu [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary)

    2014-08-08

    Highlights: • Antipsychotics modulate the expression of adipogenic genes in human adipocytes. • Secretion of proinflammatory cytokine IL8 and MCP-1 is induced by antipsychotics. • Adipocyte-dependent inflammatory abnormality could develop during chronic treatment. • Infiltrated macrophages would further enhance proinflammatory cytokine production. - Abstract: Schizophrenia requires lifelong treatment, potentially causing systemic changes in metabolic homeostasis. In the clinical setting, antipsychotic treatment may differentially lead to weight gain among individual patients, although the molecular determinants of such adverse effects are currently unknown. In this study, we investigated changes in the expression levels of critical regulatory genes of adipogenesis, lipid metabolism and proinflammatory genes during the differentiation of primary human adipose-derived stem cells (ADSCs). These cells were isolated from patients with body mass indices <25 and treated with the second-generation antipsychotics olanzapine, ziprasidone, clozapine, quetiapine, aripiprazole and risperidone and the first-generation antipsychotic haloperidol. We found that antipsychotics exhibited a marked effect on key genes involved in the regulation of cell cycle, signal transduction, transcription factors, nuclear receptors, differentiation markers and metabolic enzymes. In particular, we observed an induction of the transcription factor NF-KB1 and NF-KB1 target genes in adipocytes in response to these drugs, including the proinflammatory cytokines TNF-α, IL-1β, IL-8 and MCP-1. In addition, enhanced secretion of both IL8 and MCP-1 was observed in the supernatant of these cell cultures. In addition to their remarkable stimulatory effects on proinflammatory gene transcription, three of the most frequently prescribed antipsychotic drugs, clozapine, quetiapine and aripiprazole, also induced the expression of essential adipocyte differentiation genes and the adipocyte hormones leptin

  3. Mitochondrial Complex I Inhibitors and Forced Oxidative Phosphorylation Synergize in Inducing Cancer Cell Death

    Directory of Open Access Journals (Sweden)

    Roberta Palorini

    2013-01-01

    Full Text Available Cancer cells generally rely mostly on glycolysis rather than oxidative phosphorylation (OXPHOS for ATP production. In fact, they are particularly sensitive to glycolysis inhibition and glucose depletion. On the other hand mitochondrial dysfunctions, involved in the onset of the Warburg effect, are sometimes also associated with the resistance to apoptosis that characterizes cancer cells. Therefore, combined treatments targeting both glycolysis and mitochondria function, exploiting peculiar tumor features, might be lethal for cancer cells. In this study, we show that glucose deprivation and mitochondrial Complex I inhibitors synergize in inducing cancer cell death. In particular, our results reveal that low doses of Complex I inhibitors, ineffective on immortalized cells and in high glucose growth, become specifically cytotoxic on cancer cells deprived of glucose. Importantly, the cytotoxic effect of the inhibitors on cancer cells is strongly enhanced by forskolin, a PKA pathway activator, that we have previously shown to stimulate OXPHOS. Taken together, we demonstrate that induction in cancer cells of a switch from a glycolytic to a more respirative metabolism, obtained by glucose depletion or mitochondrial activity stimulation, strongly increases their sensitivity to low doses of mitochondrial Complex I inhibitors. Our findings might be a valuable approach to eradicate cancer cells.

  4. Exposure of mouse to high gravitation forces induces long-term potentiation in the hippocampus.

    Directory of Open Access Journals (Sweden)

    Ishii M

    2004-06-01

    Full Text Available The central nervous system is highly plastic and has been shown to undergo both transient and chronic adaptive changes in response to environmental influences. The purpose of this study was to investigate the effect of hypergravic field on long-term potentiation (LTP in the mouse hippocampus. Exposure of mice to 4G fields for 48 h had no effect on input-output coupling during extracellular stimulation of Schaffer collaterals and paired pulse facilitation, suggesting that the hypergravic exposure had no detrimental effect on basal neurotransmission in the hippocampus. However, the exposure to 4G fields for 48 h significantly induced LTP compared with the control mouse hippocampus. In contrast, no significant changes of late-phase LTP (L-LTP were found in the hippocampi of mice exposed to the hypergravic field. Exposure of mice to 4G fields for 48 h enhanced AMPA receptor phosphorylation but not cyclic AMP-responsive element binding protein (CREB phosphorylation. These results suggest that exposure to hyperdynamic fields influences the synaptic plasticity in the hippocampus.

  5. Identification of rice (Oryza sativa L.) signal factors capable of inducing Agrobacterium vir gene expression

    Institute of Scientific and Technical Information of China (English)

    许东晖; 李宝健; 刘煜; 黄志纾; 古练权

    1996-01-01

    Two kinds of signal factors capable of inducing Agrobaorerium vir gene expression were purified and identified from leaf extracts of panicle-differentiating to flowering stage of rice (Oryza saliva L. cv. IR 72) detected by Agrobacterium vir(?) lacZ. fusion genes. The induction was similar to that observed with 5 μm actosyringone (AS). Based on the comprehensive analysis of the data by UV, IR, NMR, MS, HMQC and HMBC, the structures of these two signal factors are identified as 5, 7, 4’-trihydroxy-3’, 5’-dimethoxy-flavone (named tricin) and 5, 4’ -dihydroxy-3’, 5’ -dimethoxy-7- (β-D-glucosyloxy) -flavone, respectively. These results demonstrate that monocotyledonous plants do contain highly efficient vir gene inducing factors of Agrobacterium, and the reason why monocotyledonous plants are difficult to transform by Ayrobacterium is not due to absence of vir gene inducing factors, but due to the signal factors only produced in specific stage and tissue of monocotyledonous plants

  6. Regulated gene insertion by steroid-induced PhiC31 integrase.

    Science.gov (United States)

    Sharma, Nynne; Moldt, Brian; Dalsgaard, Trine; Jensen, Thomas G; Mikkelsen, Jacob Giehm

    2008-06-01

    Nonviral integration systems are widely used genetic tools in transgenesis and play increasingly important roles in strategies for therapeutic gene transfer. Methods to efficiently regulate the activity of transposases and site-specific recombinases have important implications for their spatiotemporal regulation in live transgenic animals as well as for studies of their applicability as safe vectors for genetic therapy. In this report, strategies for posttranslational induction of a variety of gene-inserting proteins are investigated. An engineered hormone-binding domain, derived from the human progesterone receptor, hPR891, and specifically recognized by the synthetic steroid mifepristone, is fused to the Sleeping Beauty, Frog Prince, piggyBac and Tol2 transposases as well as to the Flp and PhiC31 recombinases. By analyzing mifepristone-directed inducibility of gene insertion in cultured human cells, efficient posttranslational regulation of the Flp recombinase and the PhiC31 integrase is documented. In addition, fusion of the PhiC31 integrase with the ER(T2) modified estrogen receptor hormone-binding domain results in a protein, which is inducible by a factor of 22-fold and retains 75% of the activity of the wild-type protein. These inducible PhiC31 integrase systems are important new tools in transgenesis and in safety studies of the PhiC31 integrase for gene therapy applications.

  7. Inducible gene silencing in podocytes: a new tool for studying glomerular function.

    Science.gov (United States)

    Bugeon, Laurence; Danou, Aliki; Carpentier, David; Langridge, Paul; Syed, Nelofer; Dallman, Margaret J

    2003-03-01

    Glomerular filtration is one of the primary functions of the kidney. Podocytes, a highly specialized cell type found in glomeruli, are believed to play a critical role in that function. Null mutations of genes expressed in podocytes like WT1, nephrin, and NEPH1 result in an embryo and perinatal lethal phenotype and therefore do not allow the functional analysis of these genes in the adult kidney. Here is describes the generation of a model that will allow such studies. We have engineered transgenic mice in which the disruption of targeted genes can be induced in a temporally controlled fashion in podocytes. For this, a transgene encoding the mutated estrogen receptor-Cre recombinase fusion protein was introduced into the mouse genome. Animals were crossed with Z/AP reporter mice to test for efficient and inducible recombination. We found that, after injection of inducer drug tamoxifen, Cre fusion protein translocates to the nuclei of podocytes, where it becomes active and mediates recombination of DNA carrying loxP target sequences. These animals provide for the first time a tool for silencing genes selectively in podocytes of adult animals.

  8. Oligonucleotide microarray identifies genes differentially expressed during tumorigenesis of DMBA-induced pancreatic cancer in rats.

    Directory of Open Access Journals (Sweden)

    Jun-Chao Guo

    Full Text Available The extremely dismal prognosis of pancreatic cancer (PC is attributed, at least in part, to lack of early diagnosis. Therefore, identifying differentially expressed genes in multiple steps of tumorigenesis of PC is of great interest. In the present study, a 7,12-dimethylbenzanthraene (DMBA-induced PC model was established in male Sprague-Dawley rats. The gene expression profile was screened using an oligonucleotide microarray, followed by real-time quantitative polymerase chain reaction (qRT-PCR and immunohistochemical staining validation. A total of 661 differentially expressed genes were identified in stages of pancreatic carcinogenesis. According to GO classification, these genes were involved in multiple molecular pathways. Using two-way hierarchical clustering analysis, normal pancreas, acute and chronic pancreatitis, PanIN, early and advanced pancreatic cancer were completely discriminated. Furthermore, 11 upregulated and 142 downregulated genes (probes were found by Mann-Kendall trend Monotone test, indicating homologous genes of rat and human. The qRT-PCR and immunohistochemistry analysis of CXCR7 and UBe2c, two of the identified genes, confirmed the microarray results. In human PC cell lines, knockdown of CXCR7 resulted in decreased migration and invasion. Collectively, our data identified several promising markers and therapeutic targets of PC based on a comprehensive screening and systemic validation.

  9. Effects of L-Theanine on Posttraumatic Stress Disorder Induced Changes in Rat Brain Gene Expression

    Directory of Open Access Journals (Sweden)

    Tomás Eduardo Ceremuga

    2014-01-01

    Full Text Available Posttraumatic stress disorder (PTSD is characterized by the occurrence of a traumatic event that is beyond the normal range of human experience. The future of PTSD treatment may specifically target the molecular mechanisms of PTSD. In the US, approximately 20% of adults report taking herbal products to treat medical illnesses. L-theanine is the amino acid in green tea primarily responsible for relaxation effects. No studies have evaluated the potential therapeutic properties of herbal medications on gene expression in PTSD. We evaluated gene expression in PTSD-induced changes in the amygdala and hippocampus of Sprague-Dawley rats. The rats were assigned to PTSD-stressed and nonstressed groups that received either saline, midazolam, L-theanine, or L-theanine + midazolam. Amygdala and hippocampus tissue samples were analyzed for changes in gene expression. One-way ANOVA was used to detect significant difference between groups in the amygdala and hippocampus. Of 88 genes examined, 17 had a large effect size greater than 0.138. Of these, 3 genes in the hippocampus and 5 genes in the amygdala were considered significant (P<0.05 between the groups. RT-PCR analysis revealed significant changes between groups in several genes implicated in a variety of disorders ranging from PTSD, anxiety, mood disorders, and substance dependence.

  10. Expression of putative expansin genes in phylloxera (Daktulosphaira vitifoliae Fitch) induced root galls of Vitis spp.

    Science.gov (United States)

    Lawo, N C; Griesser, M; Forneck, A

    Grape phylloxera (Daktulosphaira vitifoliae Fitch) is a serious global pest in viticulture. The insects are sedentary feeders and require a gall to feed and reproduce. The insects induce their feeding site within the meristematic zone of the root tip, where they stay attached, feeding both intra- and intercellularly, and causing damage by reducing plant vigour. Several changes in cell structure and composition, including increased cell division and tissue swelling close to the feeding site, cause an organoid gall called a nodosity to develop. Because alpha expansin genes are involved in cell enlargement and cell wall loosening in many plant tissues it may be anticipated that they are also involved in nodosity formation. To identify expansin genes in Vitis vinifera cv. Pinot noir, we mined for orthologues genes in a comparative analysis. Eleven putative expansin genes were identified and shown to be present in the rootstock Teleki 5C (V. berlandieri Planch. x V. riparia Michx.) using specific PCR followed by DNA sequencing. Expression analysis of young and mature nodosities and uninfested root tips were conducted via quantitative real time PCR (qRT-PCR). Up-regulation was measured for three putative expansin genes (VvEXPA15, -A17 and partly -A20) or down-regulation for three other putative genes (VvEXPA7, -A12, -A20) in nodosities. The present study clearly shows the involvement of putative expansin genes in the phylloxera-root interaction.

  11. Gene Signature of Human Oral Mucosa Fibroblasts: Comparison with Dermal Fibroblasts and Induced Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Keiko Miyoshi

    2015-01-01

    Full Text Available Oral mucosa is a useful material for regeneration therapy with the advantages of its accessibility and versatility regardless of age and gender. However, little is known about the molecular characteristics of oral mucosa. Here we report the first comparative profiles of the gene signatures of human oral mucosa fibroblasts (hOFs, human dermal fibroblasts (hDFs, and hOF-derived induced pluripotent stem cells (hOF-iPSCs, linking these with biological roles by functional annotation and pathway analyses. As a common feature of fibroblasts, both hOFs and hDFs expressed glycolipid metabolism-related genes at higher levels compared with hOF-iPSCs. Distinct characteristics of hOFs compared with hDFs included a high expression of glycoprotein genes, involved in signaling, extracellular matrix, membrane, and receptor proteins, besides a low expression of HOX genes, the hDFs-markers. The results of the pathway analyses indicated that tissue-reconstructive, proliferative, and signaling pathways are active, whereas senescence-related genes in p53 pathway are inactive in hOFs. Furthermore, more than half of hOF-specific genes were similarly expressed to those of hOF-iPSC genes and might be controlled by WNT signaling. Our findings demonstrated that hOFs have unique cellular characteristics in specificity and plasticity. These data may provide useful insight into application of oral fibroblasts for direct reprograming.

  12. Oligonucleotide microarray identifies genes differentially expressed during tumorigenesis of DMBA-induced pancreatic cancer in rats.

    Science.gov (United States)

    Guo, Jun-Chao; Li, Jian; Yang, Ying-Chi; Zhou, Li; Zhang, Tai-Ping; Zhao, Yu-Pei

    2013-01-01

    The extremely dismal prognosis of pancreatic cancer (PC) is attributed, at least in part, to lack of early diagnosis. Therefore, identifying differentially expressed genes in multiple steps of tumorigenesis of PC is of great interest. In the present study, a 7,12-dimethylbenzanthraene (DMBA)-induced PC model was established in male Sprague-Dawley rats. The gene expression profile was screened using an oligonucleotide microarray, followed by real-time quantitative polymerase chain reaction (qRT-PCR) and immunohistochemical staining validation. A total of 661 differentially expressed genes were identified in stages of pancreatic carcinogenesis. According to GO classification, these genes were involved in multiple molecular pathways. Using two-way hierarchical clustering analysis, normal pancreas, acute and chronic pancreatitis, PanIN, early and advanced pancreatic cancer were completely discriminated. Furthermore, 11 upregulated and 142 downregulated genes (probes) were found by Mann-Kendall trend Monotone test, indicating homologous genes of rat and human. The qRT-PCR and immunohistochemistry analysis of CXCR7 and UBe2c, two of the identified genes, confirmed the microarray results. In human PC cell lines, knockdown of CXCR7 resulted in decreased migration and invasion. Collectively, our data identified several promising markers and therapeutic targets of PC based on a comprehensive screening and systemic validation.

  13. RNA Interference (RNAi) Induced Gene Silencing: A Promising Approach of Hi-Tech Plant Breeding.

    Science.gov (United States)

    Younis, Adnan; Siddique, Muhammad Irfan; Kim, Chang-Kil; Lim, Ki-Byung

    2014-01-01

    RNA interference (RNAi) is a promising gene regulatory approach in functional genomics that has significant impact on crop improvement which permits down-regulation in gene expression with greater precise manner without affecting the expression of other genes. RNAi mechanism is expedited by small molecules of interfering RNA to suppress a gene of interest effectively. RNAi has also been exploited in plants for resistance against pathogens, insect/pest, nematodes, and virus that cause significant economic losses. Keeping beside the significance in the genome integrity maintenance as well as growth and development, RNAi induced gene syntheses are vital in plant stress management. Modifying the genes by the interference of small RNAs is one of the ways through which plants react to the environmental stresses. Hence, investigating the role of small RNAs in regulating gene expression assists the researchers to explore the potentiality of small RNAs in abiotic and biotic stress management. This novel approach opens new avenues for crop improvement by developing disease resistant, abiotic or biotic stress tolerant, and high yielding elite varieties.

  14. MOLECULAR ANALYSIS OF RADIATION-INDUCED MUTATION IN EXON 7/8 OF RAT HPRT GENE

    Institute of Scientific and Technical Information of China (English)

    任晓庆; 黄定九; 黄钢; 王利民

    2003-01-01

    Objective To investigate the relationship between the radiation dose and the HPRT gene locus mutation in rat smooth muscle cells, and provide the molecular basis for prevention of restenosis after percutaneous transluminal coronary angioplasty (PTCA).MethodsThe smooth muscle cells cultured in vitro were irradiated by radionuclide 188Re in different doses. HPRT gene mutation colonies were selected and isolated by 6 thioguanine. Analysis of mutation in exon 7/8 of HPRT gene were accomplished by polymerase chain reaction and single strand conformation polymorphism.ResultsThe HPRT gene mutation frequency of rat smooth muscle cells that were irradiated by radionuclide 188Re ranged from 5.5×10-6 to 13×10-6. Of 91 HPRT gene mutation colonies, 13(14.3%) contained exon 7/8 deletion and 15(16.5%) had point mutation. The exon 7/8 mutation frequency was 30.8%. There were significant relationships between radiation dose and mutation frequency of HPRT gene and exon 7/8.ConclusionThe DNA damage and gene mutation induced by radiation has positive relationship with radiation dose, and is a basis of proliferation inhibition and apoptosis of smooth muscle cells.

  15. The influence of hemodynamic forces on biomarkers in the walls of elastase-induced aneurysms in rabbits

    Energy Technology Data Exchange (ETDEWEB)

    Kadirvel, Ramanathan; Ding, Yong-Hong; Dai, Daying; Danielson, Mark A.; Lewis, Debra A.; Cloft, Harry J.; Kallmes, David F. [Mayo Clinic College of Medicine, Department of Radiology, Rochester, MN (United States); Zakaria, Hasballah; Robertson, Anne M. [University of Pittsburgh, Department of Mechanical Engineering, Pittsburgh, PA (United States)

    2007-12-15

    Biological and biophysical factors have been shown to play an important role in the initiation, progression, and rupture of intracranial aneurysms. The purpose of this study was to evaluate the association between hemodynamic forces and markers of vascular remodeling in elastase-induced saccular aneurysms in rabbits. Elastase-induced aneurysms were created at the origin of the right common carotid artery in rabbits. Hemodynamic parameters were estimated using computational fluid dynamic simulations based on 3-D-reconstructed models of the vasculature. Expression of matrix metalloproteinases (MMPs), their inhibitors (TIMPs) and markers of vascular remodeling were measured in different spatial regions within the aneurysms. Altered expression of biological markers relative to controls was correlated with the locations of subnormal time-averaged wall shear stress (WSS) but not with the magnitude of pressure. In the aneurysms, WSS was low and expression of biological markers was significantly altered in a time-dependent fashion. At 2 weeks, an upregulation of active-MMP-2, downregulation of TIMP-1 and TIMP-2, and intact endothelium were found in aneurysm cavities. However, by 12 weeks, endothelial cells were absent or scattered, and levels of pro- and active-MMP-2 were not different from those in control arteries, but pro-MMP-9 and both TIMPs were upregulated. These results reveal a strong, spatially localized correlation between diminished WSS and differential expression of biological markers of vascular remodeling in elastase-induced saccular aneurysms. The ability of the wall to function and maintain a healthy endothelium in a low shear environment appears to be significantly impaired by chronic exposure to low WSS. (orig.)

  16. Repeated Forced Swim Exacerbates Methamphetamine-Induced Neurotoxicity: Neuroprotective Effects of Nanowired Delivery of 5-HT3-Receptor Antagonist Ondansetron.

    Science.gov (United States)

    Lafuente, José Vicente; Sharma, Aruna; Muresanu, Dafin F; Ozkizilcik, Asya; Tian, Z Ryan; Patnaik, Ranjana; Sharma, Hari S

    2017-08-31

    The possibility that stress associated with chronic forced swim (FS) may exacerbate methamphetamine (METH) neurotoxicity was examined in a rat model. Rats were subjected to FS in a pool (30 °C) for 15 min daily for 8 days. Control rats were kept at room temperature. METH was administered (9 mg/kg, s.c.) in both control and FS rats and allowed to survive 4 h after the drug injection. METH in FS rats exacerbated BBB breakdown to Evans blue albumin (EBA) by 150 to 220% and ([131])-Iodine by 250 to 380% as compared to naive rats after METH. The METH-induced BBB leakage was most pronounced in the cerebral cortex followed by the hippocampus, cerebellum, thalamus, and hypothalamus in both FS and naive rats. The regional BBB changes were associated with a reduction in the local cerebral blood flow (CBF). Brain edema was also higher by 2 to 4% in FS rats after METH than in naive animals. Neuronal and glial cell injuries were aggravated by threefold to fivefold after METH in FS than the control group. Pretreatment with ondansetron (1 mg/kg, i.p.) 30 min before METH injection in naive rats reduced the brain pathology and improved the CBF. However, TiO2-nanowired delivery of ondansetron (1 mg/kg, i.p.) was needed to reduce METH-induced brain damage, BBB leakage, reduction in CBF, and edema formation in FS. Taken together, these observations are the first to show that METH exacerbates BBB breakdown leading to neurotoxicity in FS animals. This effect of METH-induced BBB breakdown and brain pathology in naive and FS rats is attenuated by ondansetron treatment indicating an involvement of 5-HT3 receptors, not reported earlier.

  17. An abrupt slowdown of Atlantic Meridional Overturning Circulation during 1915–1935 induced by solar forcing in a coupled GCM

    Directory of Open Access Journals (Sweden)

    P. Lin

    2014-06-01

    Full Text Available In this study, we explore an abrupt change of Atlantic Meridional Overturning Circulation (AMOC apparent in the historical run simulated by the second version of the Flexible Global Ocean–Atmosphere–Land System model – Spectral Version 2 (FGOALS-s2. The abrupt change is noted during the period from 1915 to 1935, in which the maximal AMOC value is weakened beyond 6 Sv (1 Sv = 106 m3 s−1. The abrupt signal first occurs at high latitudes (north of 46° N, then shifts gradually to middle latitudes (∼35° N three to seven years later. The weakened AMOC can be explained in the following. The weak total solar irradiance (TIS during early twentieth century decreases pole-to-equator temperature gradient in the upper stratosphere. The North polar vortex is weakened, which forces a negative North Atlantic Oscillation (NAO phase during 1905–1914. The negative phase of NAO induces anomalous easterly winds in 50–70° N belts, which decrease the release of heat fluxes from ocean to atmosphere and induce surface warming over these regions. Through the surface ice–albedo feedback, the warming may lead to continuously melting sea ice in Baffin Bay and Davis Strait, which results in freshwater accumulation. This can lead to salinity and density reductions and then an abrupt slowdown of AMOC. Moreover, due to increased TIS after 1914, the enhanced Atlantic northward ocean heat transport from low to high latitudes induces an abrupt warming of sea surface temperature or upper ocean temperature in mid–high latitudes, which can also weaken the AMOC. The abrupt change of AMOC also appears in the PiControl run, which is associated with the lasting negative NAO phases due to natural variability.

  18. Immobility stress induces depression-like behavior in the forced swim test in mice: effect of magnesium and imipramine.

    Science.gov (United States)

    Poleszak, Ewa; Wlaź, Piotr; Kedzierska, Ewa; Nieoczym, Dorota; Wyska, Elzbieta; Szymura-Oleksiak, Joanna; Fidecka, Sylwia; Radziwoń-Zaleska, Maria; Nowak, Gabriel

    2006-01-01

    Previously, we demonstrated antidepressant-like effect of magnesium (Mg) in the forced swim test (FST). Moreover, the joint administration of Mg and imipramine (IMI) at ineffective doses per se, resulted in a potent reduction in the immobility time in this test. In the present study, we examined the effect of immobility stress (IS), and Mg and/or IMI administration on FST behavior. IS induced enhancement of immobility time, which was reversed by Mg or IMI at doses ineffective in non-stressed mice (10 mg/kg and 15 mg/kg, respectively). The joint administration of Mg and IMI was effective in both IS and non-stressed animals in FST. IS did not significantly alter locomotor activity, while IMI or Mg + IMI treatment in IS mice reduced this activity. We also measured serum and brain Mg, IMI and its metabolite desipramine (DMI) concentration in mice subjected to FST and injected with Mg + IMI, both restrained and non-restrained. In the present study we demonstrated a significant increase (by 68%) in the brain IMI and a slight, non-significant reduction in DMI concentration in IS + Mg + IMI + FST vs. Mg + IMI + FST groups, which might indicate the reduction in brain IMI metabolism. The IS-induced reduction in brain IMI metabolism did not participate in the activity in FST, since no differences in such activity were noticed between IS + Mg + IMI + FST and Mg + IMI + FST groups. The present data suggest that IS-induced increase in immobility time in FST is more sensitive for detection antidepressant-like activity. However, further studies are needed to examine the effect of other antidepressants in such an experimental paradigm.

  19. Quantitative shear wave optical coherence elastography (SW-OCE) with acoustic radiation force impulses (ARFI) induced by phase array transducer

    Science.gov (United States)

    Song, Shaozhen; Le, Nhan Minh; Wang, Ruikang K.; Huang, Zhihong

    2015-03-01

    Shear Wave Optical Coherence Elastography (SW-OCE) uses the speed of propagating shear waves to provide a quantitative measurement of localized shear modulus, making it a valuable technique for the elasticity characterization of tissues such as skin and ocular tissue. One of the main challenges in shear wave elastography is to induce a reliable source of shear wave; most of nowadays techniques use external vibrators which have several drawbacks such as limited wave propagation range and/or difficulties in non-invasive scans requiring precisions, accuracy. Thus, we propose linear phase array ultrasound transducer as a remote wave source, combined with the high-speed, 47,000-frame-per-second Shear-wave visualization provided by phase-sensitive OCT. In this study, we observed for the first time shear waves induced by a 128 element linear array ultrasound imaging transducer, while the ultrasound and OCT images (within the OCE detection range) were triggered simultaneously. Acoustic radiation force impulses are induced by emitting 10 MHz tone-bursts of sub-millisecond durations (between 50 μm - 100 μm). Ultrasound beam steering is achieved by programming appropriate phase delay, covering a lateral range of 10 mm and full OCT axial (depth) range in the imaging sample. Tissue-mimicking phantoms with agarose concentration of 0.5% and 1% was used in the SW-OCE measurements as the only imaging samples. The results show extensive improvements over the range of SW-OCE elasticity map; such improvements can also be seen over shear wave velocities in softer and stiffer phantoms, as well as determining the boundary of multiple inclusions with different stiffness. This approach opens up the feasibility to combine medical ultrasound imaging and SW-OCE for high-resolution localized quantitative measurement of tissue biomechanical property.

  20. Oligonucleotide microarray analysis of dietary-induced hyperlipidemia gene expression profiles in miniature pigs.

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    Junko Takahashi

    Full Text Available BACKGROUND: Hyperlipidemia animal models have been established, but complete gene expression profiles of the transition from normal lipid levels have not been obtained. Miniature pigs are useful model animals for gene expression studies on dietary-induced hyperlipidemia because they have a similar anatomy and digestive physiology to humans, and blood samples can be obtained from them repeatedly. METHODOLOGY: Two typical dietary treatments were used for dietary-induced hyperlipidemia models, by using specific pathogen-free (SPF Clawn miniature pigs. One was a high-fat and high-cholesterol diet (HFCD and the other was a high-fat, high-cholesterol, and high-sucrose diet (HFCSD. Microarray analyses were conducted from whole blood samples during the dietary period and from white blood cells at the end of the dietary period to evaluate the transition of expression profiles of the two dietary models. PRINCIPAL FINDINGS: Variations in whole blood gene expression intensity within the HFCD or the HFCSD group were in the same range as the controls provide with normal diet at all periods. This indicates uniformity of dietary-induced hyperlipidemia for our dietary protocols. Gene ontology- (GO based functional analyses revealed that characteristics of the common changes between HFCD and HFCSD were involved in inflammatory responses and reproduction. The correlation coefficient between whole blood and white blood cell expression profiles at 27 weeks with the HFCSD diet was significantly lower than that of the control and HFCD diet groups. This may be due to the effects of RNA originating from the tissues and/or organs. CONCLUSIONS: No statistically significant differences in fasting plasma lipids and glucose levels between the HFCD and HFCSD groups were observed. However, blood RNA analyses revealed different characteristics corresponding to the dietary protocols. In this study, whole blood RNA analyses proved to be a useful tool to evaluate transitions in

  1. Glucose Oxidase Induces Cellular Senescence in Immortal Renal Cells through ILK by Downregulating Klotho Gene Expression

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    Nuria Troyano-Suárez

    2015-01-01

    Full Text Available Cellular senescence can be prematurely induced by oxidative stress involved in aging. In this work, we were searching for novel intermediaries in oxidative stress-induced senescence, focusing our interest on integrin-linked kinase (ILK, a scaffold protein at cell-extracellular matrix (ECM adhesion sites, and on the Klotho gene. Cultured renal cells were treated with glucose o