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Sample records for boldoa purpurascens cav

  1. Evaluación de la toxicidad aguda por el procedimiento de dosis fijas de un extracto de Boldoa purpurascens Cav. (Evaluation of the acute toxicity of the method fixed dose procedure of an extract of Boldoa prurpurascens Cav.)

    OpenAIRE

    Pérez Machín, Maykel; Monteagudo Jiménez, Emilio E; Boffill Cárdenas, María de los Ángeles; Méndez Triana, Rayza; Verdecía Machado, Belkys

    2008-01-01

    ResumenSe realizó un estudio experimental con el objetivo de evaluar el posible efecto tóxico de un extracto acuoso liofilizado de Boldoa purpurascen Cav. (nitro blanco). Para el desarrollo del mismo se utilizó el Procedimiento de Dosis Fijas (FDP), utilizando una dosis límite de 2000 mg/kg de peso corporal. Los animales seleccionados fueron ratas de la línea Sprague Dawley, con un peso comprendido entre 150 y 200 g. Los resultados demostraron la inocuidad de la planta al no observarse signo...

  2. Evaluación de la toxicidad aguda de un extracto de Boldoa purpurascens Cav. por el método de las clases

    OpenAIRE

    Pérez, Maykel; Jiménez, Emilio E.; Boffill Cárdenas, María de los Ángeles; González, Dulce M.; Verdecia Machado, Belkys; Blanco Machado, Freisman

    2008-01-01

    Se realizó un estudio experimental con el objetivo de evaluar el posible efecto tóxico de un extracto acuoso liofilizado de Boldoa purpurascens Cav. (nitro blanco). Para el desarrollo del mismo se utilizó el Método de las Clases de Toxicidad Aguda (CTA), utilizando una dosis límite de 2000 mg/kg de peso corporal. Los animales seleccionados fueron ratas de la línea Sprague Dawley, con un peso comprendido entre 180 y 220 g. El peso corporal se comportó acorde a la curva de crecimiento de la ...

  3. Evaluación de la toxicidad aguda por el procedimiento de dosis fijas de un extracto de Boldoa purpurascens Cav. (Evaluation of the acute toxicity of the method fixed dose procedure of an extract of Boldoa prurpurascens Cav.

    Directory of Open Access Journals (Sweden)

    Pérez Machín, Maykel

    2008-03-01

    Full Text Available ResumenSe realizó un estudio experimental con el objetivo de evaluar el posible efecto tóxico de un extracto acuoso liofilizado de Boldoa purpurascen Cav. (nitro blanco. Para el desarrollo del mismo se utilizó el Procedimiento de Dosis Fijas (FDP, utilizando una dosis límite de 2000 mg/kg de peso corporal. Los animales seleccionados fueron ratas de la línea Sprague Dawley, con un peso comprendido entre 150 y 200 g. Los resultados demostraron la inocuidad de la planta al no observarse signos ni síntomas de toxicidad. El peso corporal se comportó acorde a la curva de crecimiento de la especie y no se apreciaron alteraciones macroscópicas en los órganos estudiados, todo lo que permite afirmar que la DL50 se ubica por encima de 2000 mg/kg PC, considerándose el producto como No Clasificado (no tóxico. SummaryAn experimental study to assess the toxic effects of an aqueous and later lyophilised extract of Boldoa purpurascens Cav. (nitro blanco was carried out. It was employed the alternative Fixed Dose Procedure. There were used Sprague Dawley rats weighing between 150 and 200 g, which received a limit dose of 2000 mg/kg body weight and were kept under standardized experimental conditions. Results showed neither signs nor symptoms of toxicity. Bodyweight behaved consistent with the growing curve of this species and there were no alterations in macroscopic observations on main organs. The LD50 of this extract is located above 2000 mg/kg body weight, considering it as Non Classified (non toxic.

  4. Energioptimering VAV eller CAV?

    DEFF Research Database (Denmark)

    Kærn, Martin Ryhl; Barky, Tarek; Andersen, Dan Bomholt

    2008-01-01

    Når et HVAC-anlæg projekteres, vælges der oftest mellem fortrængnings- og opblandingsventilation. Herefter vælges hvorvidt luftfl owet skal være variabelt eller konstant – det vil sige, om der skal vælges. VAV eller CAV. Dette valg mellem VAV eller CAV bør foretages på baggrund af en vurdering af...

  5. The importance of arbuscular mycorrhiza for Cyclamen purpurascens subsp. immaculatum endemic in Slovakia.

    Science.gov (United States)

    Rydlová, Jana; Sýkorová, Zuzana; Slavíková, Renata; Turis, Peter

    2015-11-01

    At present, there is no relevant information on arbuscular mycorrhiza and the effect of the symbiosis on the growth of wild populations of cyclamens. To fill this gap, two populations of Cyclamen purpurascens subsp. immaculatum, endemic in Nízke Tatry (NT) mountains and Veľká Fatra (VF) mountains, Slovakia, were studied in situ as well as in a greenhouse pot experiment. For both populations, mycorrhizal root colonization of native plants was assessed, and mycorrhizal inoculation potential (MIP) of the soils at the two sites was determined in 3 consecutive years. In the greenhouse experiment, the growth response of cyclamens to cross-inoculation with arbuscular mycorrhizal fungi (AMF) was tested: plants from both sites were grown in their native soils and inoculated with a Septoglomus constrictum isolate originating either from the same or from the other plant locality. Although the MIP of soil at the NT site was significantly higher than at the VF site, the level of AMF root colonization of C. purpurascens subsp. immaculatum plants in the field did not significantly differ between the two localities. In the greenhouse experiment, inoculation with AMF generally accelerated cyclamen growth and significantly increased all growth parameters (shoot dry weight, leaf number and area, number of flowers, tuber, and root dry weight) and P uptake. The two populations of C. purpurascens subsp. immaculatum grown in their native soils, however, differed in their response to inoculation. The mycorrhizal growth response of NT plants was one-order higher compared to VF plants, and all their measured growth parameters were stimulated regardless of the fungal isolates' origin. In the VF plants, only the non-native (NT originating) isolate showed a significant positive effect on several growth traits. It can be concluded that mycorrhiza significantly increased fitness of C. purpurascens subsp. immaculatum, despite the differences between plant populations, implying that AMF

  6. Phytochemical Study of Condalia microphylla Cav.

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    M. A. Frontera

    2000-03-01

    Full Text Available From the petroleum ether extract of the aereal part of Condalia microphylla Cav, hydrocarbons, sterols, alcohols, and fatty acids were isolated. From fruits of the same plant anthocyanins were also isolated and characterized by chromatographic and spectroscopic methods.

  7. Curcuma purpurascens BI. rhizome accelerates rat excisional wound healing: involvement of Hsp70/Bax proteins, antioxidant defense, and angiogenesis activity

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    Rouhollahi E

    2015-10-01

    Full Text Available Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Fatemeh Hajiaghaalipour,3 Maryam Zahedifard,2 Faezeh Tayeby,2 Khalijah Awang,4 Mahmood Ameen Abdulla,3 Zahurin Mohamed1 1Pharmacogenomics Laboratory, Department of Pharmacology, Faculty of Medicine, 2Institute of Biological Sciences, Faculty of Science, 3Department of Biomedical Science, Faculty of Medicine, 4Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia Purpose: Curcuma purpurascens BI. is a member of Zingiberaceae family. The purpose of this study is to investigate the wound healing properties of hexane extract of C. purpurascens rhizome (HECP against excisional wound healing in rats.Materials and methods: Twenty four rats were randomly divided into 4 groups: A negative control (blank placebo, acacia gum, B low dose of HECP, C high dose of HECP, and D positive control, with 6 rats in each group. Full-thickness incisions (approximately 2.00 cm were made on the neck area of each rat. Groups 1–4 were treated two-times a day for 20 days with blank placebo, HECP (100 mg/kg, HECP (200 mg/kg, and intrasite gel as a positive control, respectively. After 20 days, hematoxylin and eosin and Masson’s trichrome stainings were employed to investigate the histopathological alterations. Protein expressions of Bax and Hsp70 were examined in the wound tissues using immunohistochemistry analysis. In addition, levels of enzymatic antioxidants and malondialdehyde representing lipid peroxidation were measured in wound tissue homogenates.Results: Macroscopic evaluation of wounds showed conspicuous elevation in wound contraction after topical administration of HECP at both doses. Moreover, histopathological analysis revealed noteworthy reduction in the scar width correlated with the enhanced collagen content and fibroblast cells, accompanied by a reduction of inflammatory cells in the granulation tissues. At the molecular level, HECP facilitates wound-healing process

  8. The role of CAV3 in long-QT syndrome

    DEFF Research Database (Denmark)

    Hedley, Paula L; Kanters, Jørgen K.; Dembic, Maja; Jespersen, Thomas; Skibsbye, Lasse; Aidt, Frederik H; Eschen, Ole; Graff, Claus; Behr, Elijah R; Schlamowitz, Sarah; Corfield, Valerie; McKenna, William J; Christiansen, Michael

    2013-01-01

    Mutations in CAV3, coding for caveolin-3, the major constituent scaffolding protein of cardiac caveolae, have been associated with skeletal muscle disease, cardiomyopathy, and most recently long-QT syndrome (LQTS) and sudden infant death syndrome. We examined the occurrence of CAV3 mutations in a...

  9. 9.3 kDa Components of the Injected Venom of Conus purpurascens Define a New 5-disulfide Conotoxin Framework

    OpenAIRE

    Möller, Carolina; Marí, Frank

    2011-01-01

    The 83-residue conopeptide (p21a) and its corresponding non-hydroxylated analog were isolated from the injected venom of Conus purpurascens. The complete conopeptide sequences were derived from Edman degradation sequencing of fragments from tryptic, chymotryptic and cyanogen bromide digestions. p21a has a unique, ten-cystine/5-disulfide 7-loop framework with extended 10-residue N-terminus and a 5-residue C-terminus tails, respectively. p21a has a 48% sequence homology with a recently describe...

  10. Essential Oil Content of the Rhizome of Curcuma purpurascens Bl. (Temu Tis) and Its Antiproliferative Effect on Selected Human Carcinoma Cell Lines

    OpenAIRE

    Sok-Lai Hong; Guan-Serm Lee; Syarifah Nur Syed Abdul Rahman; Omer Abdalla Ahmed Hamdi; Khalijah Awang; Nurfina Aznam Nugroho; Sri Nurestri Abd Malek

    2014-01-01

    Curcuma purpurascens Bl., belonging to the Zingiberaceae family, is known as temu tis in Yogyakarta, Indonesia. In this study, the hydrodistilled dried ground rhizome oil was investigated for its chemical content and antiproliferative activity against selected human carcinoma cell lines (MCF7, Ca Ski, A549, HT29, and HCT116) and a normal human lung fibroblast cell line (MRC5). Results from GC-MS and GC-FID analysis of the rhizome oil of temu tis showed turmerone as the major component, follow...

  11. Equal sensitivity of Cav1.2 and Cav1.3 channels to the opposing modulations of PKA and PKG in mouse chromaffin cells.

    Science.gov (United States)

    Mahapatra, Satyajit; Marcantoni, Andrea; Zuccotti, Annalisa; Carabelli, Valentina; Carbone, Emilio

    2012-10-15

    Mouse chromaffin cells (MCCs) express high densities of L-type Ca2+ channels (LTCCs), which control pacemaking activity and catecholamine secretion proportionally to their density of expression. In vivo phosphorylation of LTCCs by cAMP-PKA and cGMP–PKG, regulate LTCC gating in two opposing ways: the cAMP-PKA pathway potentiates while the cGMP–PKG cascade inhibits LTCCs. Despite this, no attempts have been made to answer three key questions related to the two Cav1 isoforms expressed in MCCs (Cav1.2 and Cav1.3): (i) how much are the two Cav1 channels basally modulated by PKA and PKG?, (ii) to what extent can Cav1.2 and Cav1.3 be further regulated by PKA or PKG activation?, and (iii) are the effects of both kinases cumulative when simultaneously active? Here, by comparing the size of L-type currents of wild-type (WT; Cav1.2+Cav1.3) and Cav1.3−/− KO (Cav1.2) MCCs, we provide new evidence that both PKA and PKG pathways affect Cav1.2 and Cav1.3 to the same extent either under basal conditions or induced stimulation. Inhibition of PKA by H89 (5 μM) reduced the L-type current in WT and KO MCCs by∼60%,while inhibition of PKG by KT 5823 (1 μM) increased by∼40% the same current in both cell types. Given that Cav1.2 and Cav1.3 carry the same quantity of Ca2+ currents, this suggests equal sensitivity of Cav1.2 and Cav1.3 to the two basal modulatory pathways. Maximal stimulation of cAMP–PKA by forskolin (100 μM) and activation of cGMP–PKG by pCPT-cGMP (1mM) uncovered a∼25% increase of L-type currents in the first case and∼65% inhibition in the second case in both WT and KO MCCs, suggesting equal sensitivity of Cav1.2 and Cav1.3 during maximal PKA or PKG stimulation. The effects of PKA and PKG were cumulative and most evident when one pathway was activated and the other was inhibited. The two extreme combinations(PKA activation–PKG inhibition vs. PKG activation-PKA inhibition) varied the size of L-type currents by one order of magnitude (from 180% to 18

  12. PREFACE: 9th International Symposium on Cavitation (CAV2015)

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    Farhat, M.; Müller, A.

    2015-12-01

    It is our pleasure and privilege to welcome all the participants of the 9th International Symposium on Cavitation (CAV2015) to Lausanne. Since its initiation in 1986 in Sendai, Japan, the CAV symposium has grown to become the world's foremost event dedicated to cavitation. Hosted by EPFL (Ecole Polytechnique Fédérale de Lausanne) and staged at the SwissTech Convention Center, CAV2015 is a unique opportunity to exchange with leading scientists and industry experts about the latest advances in theoretical modelling, numerical simulation and experimentation related to cavitation phenomena with a special emphasis on practical applications. The topics covered by CAV2015 include cavitation in ¬fluid machinery and fuel systems, bubble dynamics, cavitation erosion, advanced numerical simulation, sonochemistery, biomedicine and experimental techniques. CAV2015 will also host an exhibition of leading providers of state of the art measurement equipment, including high-speed imaging systems, non-intrusive velocimetry, pressure sensors, as well as numerical solvers. We have accepted over 190 papers, which will be presented in four parallel sessions. The proceedings will appear in the open access Journal of Physics: Conference Series (JPCS), which is part of the IOP Conference Series. All published papers are fully citable and upon publication will be free to download in perpetuity. We would like to thank all the reviewers for their great help during the selection process. We will also propose six plenary speakers to highlight cavitation issues in different fields. Finally, we would like to warmly thank our sponsors for their valuable support and the local Organizing Committee for the efforts in setting up this important event. We look forward to seeing you in Lausanne!

  13. High prevalence of antibodies against canine adenovirus (CAV) type 2 in domestic dog populations in South Africa precludes the use of CAV-based recombinant rabies vaccines.

    Science.gov (United States)

    Wright, N; Jackson, F R; Niezgoda, M; Ellison, J A; Rupprecht, C E; Nel, L H

    2013-08-28

    Rabies in dogs can be controlled through mass vaccination. Oral vaccination of domestic dogs would be useful in the developing world, where greater vaccination coverage is needed especially in inaccessible areas or places with large numbers of free-roaming dogs. From this perspective, recent research has focused on development of new recombinant vaccines that can be administered orally in a bait to be used as adjunct for parenteral vaccination. One such candidate, a recombinant canine adenovirus type 2 vaccine expressing the rabies virus glycoprotein (CAV2-RG), is considered a promising option for dogs, given host specificity and safety. To assess the potential use of this vaccine in domestic dog populations, we investigated the prevalence of antibodies against canine adenovirus type 2 in South African dogs. Blood was collected from 241 dogs from the Gauteng and KwaZulu-Natal provinces. Sampled dogs had not previously been vaccinated against canine adenovirus type 1 (CAV1) or canine adenovirus type 2 (CAV2). Animals from both provinces had a high percentage of seropositivity (45% and 62%), suggesting that CAV2 circulates extensively among domestic dog populations in South Africa. Given this finding, we evaluated the effect of pre-existing CAV-specific antibodies on the efficacy of the CAV2-RG vaccine delivered via the oral route in dogs. Purpose-bred Beagle dogs, which received prior vaccination against canine parvovirus, canine distemper virus and CAV, were immunized by oral administration of CAV2-RG. After rabies virus (RABV) infection all animals, except one vaccinated dog, developed rabies. This study demonstrated that pre-existing antibodies against CAV, such as naturally occurs in South African dogs, inhibits the development of neutralizing antibodies against RABV when immunized with a CAV-based rabies recombinant vaccine. PMID:23867013

  14. Zebrafish CaV2.1 Calcium Channels Are Tailored for Fast Synchronous Neuromuscular Transmission

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    Naranjo, David; Wen, Hua; Brehm, Paul

    2015-01-01

    The CaV2.2 (N-type) and CaV2.1 (P/Q-type) voltage-dependent calcium channels are prevalent throughout the nervous system where they mediate synaptic transmission, but the basis for the selective presence at individual synapses still remains an open question. The CaV2.1 channels have been proposed to respond more effectively to brief action potentials (APs), an idea supported by computational modeling. However, the side-by-side comparison of CaV2.1 and CaV2.2 kinetics in intact neurons failed to reveal differences. As an alternative means for direct functional comparison we expressed zebrafish CaV2.1 and CaV2.2 α-subunits, along with their accessory subunits, in HEK293 cells. HEK cells lack calcium currents, thereby circumventing the need for pharmacological inhibition of mixed calcium channel isoforms present in neurons. HEK cells also have a simplified morphology compared to neurons, which improves voltage control. Our measurements revealed faster kinetics and shallower voltage-dependence of activation and deactivation for CaV2.1. Additionally, recordings of calcium current in response to a command waveform based on the motorneuron AP show, directly, more effective activation of CaV2.1. Analysis of calcium currents associated with the AP waveform indicate an approximately fourfold greater open probability (PO) for CaV2.1. The efficient activation of CaV2.1 channels during APs may contribute to the highly reliable transmission at zebrafish neuromuscular junctions. PMID:25650925

  15. Zebrafish CaV2.1 calcium channels are tailored for fast synchronous neuromuscular transmission.

    Science.gov (United States)

    Naranjo, David; Wen, Hua; Brehm, Paul

    2015-02-01

    The CaV2.2 (N-type) and CaV2.1 (P/Q-type) voltage-dependent calcium channels are prevalent throughout the nervous system where they mediate synaptic transmission, but the basis for the selective presence at individual synapses still remains an open question. The CaV2.1 channels have been proposed to respond more effectively to brief action potentials (APs), an idea supported by computational modeling. However, the side-by-side comparison of CaV2.1 and CaV2.2 kinetics in intact neurons failed to reveal differences. As an alternative means for direct functional comparison we expressed zebrafish CaV2.1 and CaV2.2 α-subunits, along with their accessory subunits, in HEK293 cells. HEK cells lack calcium currents, thereby circumventing the need for pharmacological inhibition of mixed calcium channel isoforms present in neurons. HEK cells also have a simplified morphology compared to neurons, which improves voltage control. Our measurements revealed faster kinetics and shallower voltage-dependence of activation and deactivation for CaV2.1. Additionally, recordings of calcium current in response to a command waveform based on the motorneuron AP show, directly, more effective activation of CaV2.1. Analysis of calcium currents associated with the AP waveform indicate an approximately fourfold greater open probability (PO) for CaV2.1. The efficient activation of CaV2.1 channels during APs may contribute to the highly reliable transmission at zebrafish neuromuscular junctions. PMID:25650925

  16. The Low-Threshold Calcium Channel Cav3.2 Determines Low-Threshold Mechanoreceptor Function

    Directory of Open Access Journals (Sweden)

    Amaury François

    2015-01-01

    Full Text Available The T-type calcium channel Cav3.2 emerges as a key regulator of sensory functions, but its expression pattern within primary afferent neurons and its contribution to modality-specific signaling remain obscure. Here, we elucidate this issue using a unique knockin/flox mouse strain wherein Cav3.2 is replaced by a functional Cav3.2-surface-ecliptic GFP fusion. We demonstrate that Cav3.2 is a selective marker of two major low-threshold mechanoreceptors (LTMRs, Aδ- and C-LTMRs, innervating the most abundant skin hair follicles. The presence of Cav3.2 along LTMR-fiber trajectories is consistent with critical roles at multiple sites, setting their strong excitability. Strikingly, the C-LTMR-specific knockout uncovers that Cav3.2 regulates light-touch perception and noxious mechanical cold and chemical sensations and is essential to build up that debilitates allodynic symptoms of neuropathic pain, a mechanism thought to be entirely A-LTMR specific. Collectively, our findings support a fundamental role for Cav3.2 in touch/pain pathophysiology, validating their critic pharmacological relevance to relieve mechanical and cold allodynia.

  17. CAV-2--why a canine virus is a neurobiologist's best friend.

    Science.gov (United States)

    Junyent, Felix; Kremer, Eric J

    2015-10-01

    Canine adenovirus type 2 (CAV-2) vectors are powerful tools for fundamental and applied neurobiology due to their negligible immunogenicity, preferential transduction of neurons, widespread distribution via axonal transport, and duration of expression in the mammalian brain. CAV-2 vectors are internalized in neurons by the selective use of coxsackievirus and adenovirus receptor (CAR), which is located at the presynapse in neurons. Neuronal internalization and axonal transport is mediated by CAR, which potentiates vector biodistribution. The above characteristics, together with the ∼30kb cloning capacity of helper-dependent (HD) CAV-2 vectors, optimized CAV-2 vector creation, production and purification, is expanding the therapeutic and fundamental options for CNS gene transfer. PMID:26298516

  18. Protein kinase A modulation of CaV1.4 calcium channels

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    Sang, Lingjie; Dick, Ivy E.; Yue, David T.

    2016-07-01

    The regulation of L-type Ca2+ channels by protein kinase A (PKA) represents a crucial element within cardiac, skeletal muscle and neurological systems. Although much work has been done to understand this regulation in cardiac CaV1.2 Ca2+ channels, relatively little is known about the closely related CaV1.4 L-type Ca2+ channels, which feature prominently in the visual system. Here we find that CaV1.4 channels are indeed modulated by PKA phosphorylation within the inhibitor of Ca2+-dependent inactivation (ICDI) motif. Phosphorylation of this region promotes the occupancy of calmodulin on the channel, thus increasing channel open probability (PO) and Ca2+-dependent inactivation. Although this interaction seems specific to CaV1.4 channels, introduction of ICDI1.4 to CaV1.3 or CaV1.2 channels endows these channels with a form of PKA modulation, previously unobserved in heterologous systems. Thus, this mechanism may not only play an important role in the visual system but may be generalizable across the L-type channel family.

  19. Protein kinase A modulation of CaV1.4 calcium channels.

    Science.gov (United States)

    Sang, Lingjie; Dick, Ivy E; Yue, David T

    2016-01-01

    The regulation of L-type Ca(2+) channels by protein kinase A (PKA) represents a crucial element within cardiac, skeletal muscle and neurological systems. Although much work has been done to understand this regulation in cardiac CaV1.2 Ca(2+) channels, relatively little is known about the closely related CaV1.4 L-type Ca(2+) channels, which feature prominently in the visual system. Here we find that CaV1.4 channels are indeed modulated by PKA phosphorylation within the inhibitor of Ca(2+)-dependent inactivation (ICDI) motif. Phosphorylation of this region promotes the occupancy of calmodulin on the channel, thus increasing channel open probability (PO) and Ca(2+)-dependent inactivation. Although this interaction seems specific to CaV1.4 channels, introduction of ICDI1.4 to CaV1.3 or CaV1.2 channels endows these channels with a form of PKA modulation, previously unobserved in heterologous systems. Thus, this mechanism may not only play an important role in the visual system but may be generalizable across the L-type channel family. PMID:27456671

  20. The L-type calcium channel Cav1.3 is required for proper hippocampal neurogenesis and cognitive functions.

    Science.gov (United States)

    Marschallinger, Julia; Sah, Anupam; Schmuckermair, Claudia; Unger, Michael; Rotheneichner, Peter; Kharitonova, Maria; Waclawiczek, Alexander; Gerner, Philipp; Jaksch-Bogensperger, Heidi; Berger, Stefan; Striessnig, Jörg; Singewald, Nicolas; Couillard-Despres, Sebastien; Aigner, Ludwig

    2015-12-01

    L-type voltage gated Ca(2+) channels (LTCCs) are widely expressed within different brain regions including the hippocampus. The isoforms Cav1.2 and Cav1.3 have been shown to be involved in hippocampus-dependent learning and memory, cognitive functions that require proper hippocampal neurogenesis. In vitro, functional LTCCs are expressed on neuronal progenitor cells, where they promote neuronal differentiation. Expression of LTCCs on neural stem and progenitor cells within the neurogenic regions in the adult brain in vivo has not been examined so far, and a contribution of the individual isoforms Cav1.2 and Cav1.3 to adult neurogenesis remained to be clarified. To reveal the role of these channels we first evaluated the expression patterns of Cav1.2 and Cav1.3 in the hippocampal dentate gyrus and the subventricular zone (SVZ) in adult (2- and 3-month old) and middle-aged (15-month old) mice on mRNA and protein levels. We performed immunohistological analysis of hippocampal neurogenesis in adult and middle-aged Cav1.3(-/-) mice and finally addressed the importance of Cav1.3 for hippocampal function by evaluating spatial memory and depression-like behavior in adult Cav1.3(-/-) mice. Our results showed Cav1.2 and Cav1.3 expression at different stages of neuronal differentiation. While Cav1.2 was primarily restricted to mature NeuN(+) granular neurons, Cav1.3 was expressed in Nestin(+) neural stem cells and in mature NeuN(+) granular neurons. Adult and middle-aged Cav1.3(-/-) mice showed severe impairments in dentate gyrus neurogenesis, with significantly smaller dentate gyrus volume, reduced survival of newly generated cells, and reduced neuronal differentiation. Further, Cav1.3(-/-) mice showed impairment in the hippocampus dependent object location memory test, implicating Cav1.3 as an essential element for hippocampus-associated cognitive functions. Thus, modulation of LTCC activities may have a crucial impact on neurogenic responses and cognition, which should be

  1. Structures of CaV2 Ca2+/CaM-IQ Domain Complexes Reveal Binding Modes That Underlie Calcium-Dependent Inactivation And Facilitation

    International Nuclear Information System (INIS)

    Calcium influx drives two opposing voltage-activated calcium channel (CaV) self-modulatory processes: calcium-dependent inactivation (CDI) and calcium-dependent facilitation (CDF). Specific Ca2+/calmodulin (Ca2+/CaM) lobes produce CDI and CDF through interactions with the CaVα1 subunit IQ domain. Curiously, Ca2+/CaM lobe modulation polarity appears inverted between CaV1s and CaV2s. Here, we present crystal structures of CaV2.1, CaV2.2, and CaV2.3 Ca2+/CaM-IQ domain complexes. All display binding orientations opposite to CaV1.2 with a physical reversal of the CaM lobe positions relative to the IQ α-helix. Titration calorimetry reveals lobe competition for a high-affinity site common to CaV1 and CaV2 IQ domains that is occupied by the CDI lobe in the structures. Electrophysiological experiments demonstrate that the N-terminal CaV2 Ca2+/C-lobe anchors affect CDF. Together, the data unveil the remarkable structural plasticity at the heart of CaV feedback modulation and indicate that CaV1 and CaV2 IQ domains bear a dedicated CDF site that exchanges Ca2+/CaM lobe occupants

  2. Physicochemical properties of tamarillo (Solanum betaceum Cav.) hydrocolloid fractions.

    Science.gov (United States)

    Gannasin, Sri Puvanesvari; Adzahan, Noranizan Mohd; Hamzah, Mohd Yusof; Mustafa, Shuhaimi; Muhammad, Kharidah

    2015-09-01

    Tamarillo (Solanum betaceum Cav.) is an underutilised fruit in Malaysia. The fruit, however, contains good proportions of soluble fibre, protein, starch, anthocyanins and carotenoids. Amongst the fruits, only tamarillo mesocarp contains both polar (anthocyanins) and non-polar (carotenoids) pigments. The ability to retain both polar and non-polar pigments in the mesocarp could be related to the unique properties of its hydrocolloids. To understand the pigment-hydrocolloid interaction in the fruit, information on the physicochemical characteristics of the hydrocolloids is required. Therefore, hydrocolloids from the anthocyanin-rich seed mucilage fraction of the tamarillo and its carotenoid-rich pulp fraction were extracted and characterised. Water and 1% citric acid were used to extract the seed mucilage hydrocolloid while 72% ethanol and 20mM HEPES buffer were used for pulp hydrocolloid extraction. Seed mucilage hydrocolloid was primarily composed of arabinogalactan protein-associated pectin whereas pulp hydrocolloid was composed of hemicellulosic polysaccharides with some naturally interacting proteins and neutral polysaccharides. PMID:25842340

  3. Mechanism of direct Cav2.2 channel block by the κ-opioid receptor agonist U50488H.

    Science.gov (United States)

    Berecki, Géza; Motin, Leonid; Adams, David J

    2016-10-01

    U50488H is a benzeneacetamide κ-opioid receptor (κ-OR) agonist analgesic, widely used for investigating the pharmacology of G protein-coupled κ-ORs. However, U50488H is also known to directly block various voltage-gated ion channels in a G protein-independent manner. We investigated the direct actions of U50488H on various high voltage-activated (HVA) and low voltage-activated (LVA) neuronal Ca(2+) channels heterologously expressed in human embryonic kidney (HEK293) cells. U50488H inhibited HVA rat Cav1.3 (rCav1.3), human Cav2.1 (hCav2.1), hCav2.2, hCav2.3, and LVA hCav3.1 and hCav3.2 channels in a concentration-dependent manner, with similar potencies characterised with half-maximal inhibitory concentration (IC50) values of ∼30 μM. U50488H concentrations causing direct Cav inhibition are typically >100 times higher than those producing κ-OR activation. Investigation of the mechanism of U50488H block of the Cav2.2 channel revealed that U50488H interacted with all major kinetic states of the channel - resting, open, and inactivated. U50488H did not affect the voltage dependence of activation but shifted the steady-state inactivation curve by ∼11 mV to more hyperpolarized potentials. U50488H also increased the rate of Ba(2+) current inactivation during a step depolarization and significantly delayed recovery from slow inactivation, compared with control. Cav2.2 current inhibition was frequency dependent during repetitive step depolarization at 1 Hz and 3 Hz, consistent with use-dependent block. In summary, our results suggest that preferential interaction of U50488H with inactivated Cav2.2 channels significantly contributes to reduced Cav2.2 channel availability and slow recovery form inactivation. We conclude that U50488H non-selectively blocks heterologously expressed neuronal HVA and LVA Cav channels in the absence of κ-ORs. This cross-reactivity also suggests potentially common U50488H binding motifs across Cav channel targets. PMID:27245500

  4. Correlation Analysis Between Biomass, Active Components and Agronomic Traits of B. purpurascens%岩白菜的生物量和有效成分与农艺性状间的相关性分析

    Institute of Scientific and Technical Information of China (English)

    朱映安; 王仕玉; 李玉强; 赵桂茹; 陈骊君; 郭凤根

    2013-01-01

      为了探明不同种源岩白菜的农艺性状与生物量及有效成分间的相关性,为岩白菜的良种选育和永续利用提供理论依据。观测了24份岩白菜种源的7个农艺性状和3个生物量性状,并利用HPLC法测定岩白菜的有效成分含量。结果表明:株根状茎干重与株干重、株高、根状茎直径间均呈极显著正相关关系(P<0.01);株叶片干重与株高呈显著正相关关系(P<0.05),与根状茎直径极显著正相关关系(P<0.01)。岩白菜素单株产量与根状茎直径间呈显著正相关关系(P<0.05),与其他农艺性状间相关性不显著;叶片中熊果苷的含量与株高呈显著负相关(P<0.05),与其他农艺性状间呈负相关,但没有达到显著水平;而根状茎中岩白菜素的含量和熊果苷的单株产量与7个农艺性状间相关性均不强。株高和根状茎直径是岩白菜选育种时需重点关注的2个农艺性状。%In order to provide a theory basis for breeding and sustainable utilization of Bergenia purpurascens, the correlations between agronomic traits and biomass and effective components in different resources of B. purpurascens had been discussed. 7 agronomic characters and 3 biomass characters from 24 resources were observed, and the contents of effective component were determined in the method of HPLC. It was showed that there was a very significant positive correlation (P<0.01) between dry weight of rhizomes per plant, total dry weight of plant and plant height, diameter of rhizomes. A significant positive correlation (P<0.05) between dry weight of leaves per plant and plant height and a very significant positive correlation (P<0.01) between dry weight of leaves per plant and diameter of rhizomes per plant were found too. There also was a significant positive correlation (P<0.05) between the yield of bergenin per plant and diameter of rhizomes, although there was no significant correlation between the yield of

  5. Pharmacognostic standardization of stems of Thespesia lampas (Cav.) Dalz & Gibs

    Institute of Scientific and Technical Information of China (English)

    Chumbhale DS; Upasani CD

    2012-01-01

    Objective: To establish the standardization parameters for complete pharmacognostic evaluation of stems of Thespesia lampas (T. lampas ) (Cav.) Dalz & Gibs (Malvaceae), an important plant in the Indian system of medicine. Methods: Morphological, microscopical, physico-chemical evaluations, florescence analysis of T. lampas stems were investigated and preliminary phytochemical analysis, GC-MS analysis and HPTLC fingerprinting were carried out for qualitative phytochemical evaluation of various extracts of stems of T. lampas. Results: Chemo-microscopy revealed the presence of lignin, starch grains and calcium oxalate crystals. Physico-chemical evaluation used to determine numerical standards showed a result with total ash (9.03 ± 0.05) % w/w, acid insoluble ash (1.50 ± 0.01) % w/w, water soluble ash (2.51 ± 0.02) % w/w, sulphated ash (7.50 ± 0.01) % w/w, ethanol soluble extractive (0.24 ± 0.02) % w/w, water soluble extractive (0.08 ± 0.01) % w/w, moisture content (6.03 ± 0.05) % w/w and total crude fibre content of stem powder (47.36 ± 0.32) % w/w. Behavior characteristics of the stem powder showed presence of steroids, starch, alkaloid, flavonoids and proteins. Preliminary phytochemical analysis revealed presence of glycosides, phenolic compounds, tannins, steroids, saponins, flavonoids, carbohydrates and proteins. GC-MS analysis showed the presence of fatty acids such as dodecanoic acid, tetradecanoic acid, n- hexadecanoic acid, 9-tetradecenal and HPTLC fingerprinting revealed the presence of β– sitosterol and quercetin in stems of T. lampas. Conclusions: The pharmacognostic standardization of T. lampas is useful towards establishing standards for quality, purity and sample identification.

  6. A retrospective investigation of canine adenovirus (CAV infection in adult dogs in Turkey : article

    Directory of Open Access Journals (Sweden)

    S. Gur

    2009-05-01

    Full Text Available Canine adenovirus (CAV type 1 and 2, respectively, cause infectious canine hepatitis and infectious canine laryngotracheitis in members of the families Canidae and Ursidae worldwide. Both of these infections are acute diseases, especially in young dogs. The aim of this study was to conduct a serological investigation of canine adenovirus infection. For this purpose, serumsamples were collected from native pure-bred Kangal (n = 11, and Akbash dogs (n = 17 and Turkish Greyhounds (n=15 in Eskisehir and Konya provinces. None ofthe dogs were previously vaccinated against CAV types. Indirect ELISA detected 88.2 %, 93.3 % and 100 % prevalences in Akbash, Greyhound and Kangal dogs, respectively. The remainder of the samples (n = 51 were collected at the Afyonkarahisar Municipality Shelter. Fourty-two of these dogs (82.3 % were detected as seropositive. In total, 82 of 94 dogs (87.2 % were found to be positive for CAV serum antibodies.

  7. A retrospective investigation of canine adenovirus (CAV) infection in adult dogs in Turkey.

    Science.gov (United States)

    Gür, S; Acar, A

    2009-06-01

    Canine adenovirus (CAV) type 1 and 2, respectively, cause infectious canine hepatitis and infectious canine laryngotracheitis in members of the families Canidae and Ursidae worldwide. Both of these infections are acute diseases, especially in young dogs. The aim of this study was to conduct a serological investigation of canine adenovirus infection. For this purpose, serum samples were collected from native pure-bred Kangal(n = 11), and Akbash dogs (n = 17) and Turkish Greyhounds (n = 15) in Eskişehir and Konya provinces. None of the dogs were previously vaccinated against CAV types. Indirect ELISA detected 88.2%, 93.3% and 100% prevalences in Akbash, Greyhound and Kangal dogs, respectively. The remainder of the samples (n = 51) were collected at the Afyonkarahisar Municipality Shelter. Fourty-two of these dogs (82.3%) were detected as seropositive. In total, 82 of 94 dogs (87.2%) were found to be positive for CAV serum antibodies. PMID:19831268

  8. CaV channels and cancer: canonical functions indicate benefits of repurposed drugs as cancer therapeutics

    OpenAIRE

    Buchanan, Paul J.; McCloskey, Karen D.

    2016-01-01

    The importance of ion channels in the hallmarks of many cancers is increasingly recognised. This article reviews current knowledge of the expression of members of the voltage-gated calcium channel family (CaV) in cancer at the gene and protein level and discusses their potential functional roles. The ten members of the CaV channel family are classified according to expression of their pore-forming α-subunit; moreover, co-expression of accessory α2δ, β and γ confers a spectrum of biophysical c...

  9. The IQ Motif is Crucial for Ca(v)1.1 Function

    OpenAIRE

    Katarina Stroffekova

    2011-01-01

    Ca2+-dependent modulation via calmodulin, with consensus CaM-binding IQ motif playing a key role, has been documented for most high-voltage-activated Ca2+ channels. The skeletal muscle Cav1.1 also exhibits Ca2+-/CaM-dependent modulation. Here, whole-cell Ca2+ current, Ca2+ transient, and maximal, immobilization-resistant charge movement (Q max) recordings were obtained from cultured mouse myotubes, to test a role of IQ motif in function of Cav1.1. The effect of introducing mutation (IQ to AA)...

  10. Troponin T3 regulates nuclear localization of the calcium channel Cavβ1a subunit in skeletal muscle

    International Nuclear Information System (INIS)

    The voltage-gated calcium channel (Cav) β1a subunit (Cavβ1a) plays an important role in excitation–contraction coupling (ECC), a process in the myoplasm that leads to muscle-force generation. Recently, we discovered that the Cavβ1a subunit travels to the nucleus of skeletal muscle cells where it helps to regulate gene transcription. To determine how it travels to the nucleus, we performed a yeast two-hybrid screening of the mouse fast skeletal muscle cDNA library and identified an interaction with troponin T3 (TnT3), which we subsequently confirmed by co-immunoprecipitation and co-localization assays in mouse skeletal muscle in vivo and in cultured C2C12 muscle cells. Interacting domains were mapped to the leucine zipper domain in TnT3 COOH-terminus (160–244 aa) and Cavβ1a NH2-terminus (1–99 aa), respectively. The double fluorescence assay in C2C12 cells co-expressing TnT3/DsRed and Cavβ1a/YFP shows that TnT3 facilitates Cavβ1a nuclear recruitment, suggesting that the two proteins play a heretofore unknown role during early muscle differentiation in addition to their classical role in ECC regulation. - Highlights: • Previously, we demonstrated that Cavβ1a is a gene transcription regulator. • Here, we show that TnT3 interacts with Cavβ1a. • We mapped TnT3 and Cavβ1a interaction domain. • TnT3 facilitates Cavβ1a nuclear enrichment. • The two proteins play a heretofore unknown role during early muscle differentiation

  11. 基于CAV444的电容式液位传感器设计%Design of Capacitive Level Sensor Based on CAV444

    Institute of Scientific and Technical Information of China (English)

    周燕

    2012-01-01

    The paper introduced measuring principle of capacitive level sensor and proposed a design scheme of level sensor based on novel capacitance measuring chip CAV444. The testing result showed the sensor is stable with high precision and small error, which can meet requirements of water level monitoring for safety monitoring system of low-concentration gas transportation.%介绍了电容式液位传感器的测量原理,提出了一种基于新型电容测量集成电路芯片CAV444的电容式液位传感器的设计方案.测试结果表明,该传感器性能稳定,测量精度较高,误差较小,能够满足低浓度瓦斯输送安全监测系统的水位监测需求.

  12. Functional properties of the Cav1.2 calcium channel activated by calmodulin in the absence of α2δ subunits

    OpenAIRE

    Ravindran, Arippa; Kobrinsky, Evgeny; Lao, Qi Zong; Soldatov, Nikolai M.

    2009-01-01

    Voltage-activated Cav1.2 calcium channels require association of the pore-forming α1C subunit with accessory Cavβ and α2δ subunits. Binding of a single calmodulin (CaM) to α1C supports Ca2+-dependent inactivation (CDI). The human Cav1.2 channel is silent in the absence of Cavβ and/or α2δ. Recently, we found that coexpression of exogenous CaM (CaMex) supports plasma membrane targeting, gating facilitation and CDI of the channel in the absence of Cavβ. Here we discovered that CaMex and its Ca2+...

  13. TRANSCRIPTIONAL UPREGULATION OF α2δ-1 ELEVATES ARTERIAL SMOOTH MUSCLE CELL CAV1.2 CHANNEL SURFACE EXPRESSION AND CEREBROVASCULAR CONSTRICTION IN GENETIC HYPERTENSION

    OpenAIRE

    Bannister, John P.; Bulley, Simon; Narayanan, Damodaran; Thomas-Gatewood, Candice; Luzny, Patrik; Pachuau, Judith; Jaggar, Jonathan H.

    2012-01-01

    A hallmark of hypertension is an increase in arterial myocyte voltage-dependent Ca2+ (CaV1.2) currents that induces pathological vasoconstriction. CaV1.2 channels are heteromeric complexes comprising a pore forming CaV1.2α1 with auxiliary α2δ and β subunits. Molecular mechanisms that elevate CaV1.2 currents during hypertension and the potential contribution of CaV1.2 auxiliary subunits are unclear. Here, we investigated the pathological significance of α2δ subunits in vasoconstriction associa...

  14. CaV1.3 as pacemaker channels in adrenal chromaffin cells: specific role on exo- and endocytosis?

    Science.gov (United States)

    Comunanza, Valentina; Marcantoni, Andrea; Vandael, David H; Mahapatra, Satyajit; Gavello, Daniela; Carabelli, Valentina; Carbone, Emilio

    2010-01-01

    Voltage-gated L-type calcium channels (LTCCs) are expressed in adrenal chromaffin cells. Besides shaping the action potential (AP), LTCCs are involved in the excitation-secretion coupling controlling catecholamine release and in Ca (2+) -dependent vesicle retrieval. Of the two LTCCs expressed in chromaffin cells (CaV1.2 and CaV1.3), CaV1.3 possesses the prerequisites for pacemaking spontaneously firing cells: low-threshold, steep voltage-dependence of activation and slow inactivation. By using CaV1 .3 (-/-) KO mice and the AP-clamp it has been possible to resolve the time course of CaV1.3 pacemaker currents, which is similar to that regulating substantia nigra dopaminergic neurons. In mouse chromaffin cells CaV1.3 is coupled to fast-inactivating BK channels within membrane nanodomains and controls AP repolarization. The ability to carry subthreshold Ca (2+) currents and activate BK channels confers to CaV1.3 the unique feature of driving Ca (2+) loading during long interspike intervals and, possibly, to control the Ca (2+) -dependent exocytosis and endocytosis processes that regulate catecholamine secretion and vesicle recycling. PMID:21084859

  15. Ca2+ entry into neurons is facilitated by cooperative gating of clustered CaV1.3 channels

    Science.gov (United States)

    Moreno, Claudia M; Dixon, Rose E; Tajada, Sendoa; Yuan, Can; Opitz-Araya, Ximena; Binder, Marc D; Santana, Luis F

    2016-01-01

    CaV1.3 channels regulate excitability in many neurons. As is the case for all voltage-gated channels, it is widely assumed that individual CaV1.3 channels behave independently with respect to voltage-activation, open probability, and facilitation. Here, we report the results of super-resolution imaging, optogenetic, and electrophysiological measurements that refute this long-held view. We found that the short channel isoform (CaV1.3S), but not the long (CaV1.3L), associates in functional clusters of two or more channels that open cooperatively, facilitating Ca2+ influx. CaV1.3S channels are coupled via a C-terminus-to-C-terminus interaction that requires binding of the incoming Ca2+ to calmodulin (CaM) and subsequent binding of CaM to the pre-IQ domain of the channels. Physically-coupled channels facilitate Ca2+ currents as a consequence of their higher open probabilities, leading to increased firing rates in rat hippocampal neurons. We propose that cooperative gating of CaV1.3S channels represents a mechanism for the regulation of Ca2+ signaling and electrical activity. DOI: http://dx.doi.org/10.7554/eLife.15744.001 PMID:27187148

  16. Thermomechanical repository and shaft response analyses using the CAVS [Cracking And Void Strain] jointed rock model: Draft final report

    International Nuclear Information System (INIS)

    Numerical studies of the far-field repository and near-field shaft response for a nuclear waste repository in bedded salt have been performed with the STEALTH computer code using the CAVS model for jointed rock. CAVS is a constitutive model that can simulate the slip and dilatancy of fracture planes in a jointed rock mass. The initiation and/or propagation of fractures can also be modeled when stress intensity criteria are met. The CAVS models are based on the joint models proposed with appropriate modifications for numerical simulations. The STEALTH/CAVS model has been previously used to model (1) explosive fracturing of a wellbore, (2) earthquake effects on tunnels in a generic nuclear waste repository, (3) horizontal emplacement for a nuclear waste repository in jointed granite, and (4) tunnel response in jointed rock. The use of CAVS to model far-field repository and near-field shaft response was different from previous approaches because it represented a spatially oriented approach to rock response and failure, rather than the traditional stress invariant formulation for yielding. In addition, CAVS tracked the response of the joint apertures to the time-dependent stress changes in the far-field repository and near-field shaft regions. 28 refs., 21 figs., 11 tabs

  17. Non-contact wafer thickness measurement of capacitance sensor circuit based on CAV424

    Directory of Open Access Journals (Sweden)

    Yan You Jun

    2016-01-01

    Full Text Available Non-contact wafer thickness measurement with the CAV424 capacitance sensor special integrated circuit and arc pole plate capacitor sensor has good stability and linearity under low capacity of the bottom of sensor and low&entity; C condition. This method has a high technical advantages and practical value. Two capacitance sensors Cb, Ca measurement spacing 4mm install at the same axis which constitutes the size condition for measuring thickness. The static capacity of Ca and Cb is a constant value. The capacity of Cb and Ca will change when the silicon wafer is involved. This change is checked by the CAV424 capacitive sensor which has better linearity and higher thickness resolution.

  18. CAV3 mutations causing exercise intolerance, myalgia and rhabdomyolysis: Expanding the phenotypic spectrum of caveolinopathies.

    Science.gov (United States)

    Scalco, Renata Siciliani; Gardiner, Alice R; Pitceathly, Robert D S; Hilton-Jones, David; Schapira, Anthony H; Turner, Chris; Parton, Matt; Desikan, Mahalekshmi; Barresi, Rita; Marsh, Julie; Manzur, Adnan Y; Childs, Anne-Marie; Feng, Lucy; Murphy, Elaine; Lamont, Phillipa J; Ravenscroft, Gianina; Wallefeld, William; Davis, Mark R; Laing, Nigel G; Holton, Janice L; Fialho, Doreen; Bushby, Kate; Hanna, Michael G; Phadke, Rahul; Jungbluth, Heinz; Houlden, Henry; Quinlivan, Ros

    2016-08-01

    Rhabdomyolysis is often due to a combination of environmental trigger(s) and genetic predisposition; however, the underlying genetic cause remains elusive in many cases. Mutations in CAV3 lead to various neuromuscular phenotypes with partial overlap, including limb girdle muscular dystrophy type 1C (LGMD1C), rippling muscle disease, distal myopathy and isolated hyperCKemia. Here we present a series of eight patients from seven families presenting with exercise intolerance and rhabdomyolysis caused by mutations in CAV3 diagnosed by next generation sequencing (NGS) (n = 6). Symptoms included myalgia (n = 7), exercise intolerance (n = 7) and episodes of rhabdomyolysis (n = 2). Percussion-induced rapid muscle contractions (PIRCs) were seen in five out of six patients examined. A previously reported heterozygous mutation in CAV3 (p.T78M) and three novel variants (p.V14I, p.F41S, p.F54V) were identified. Caveolin-3 immunolabeling in muscle was normal in 3/4 patients; however, immunoblotting showed more than 50% reduction of caveolin-3 in five patients compared with controls. This case series demonstrates that exercise intolerance, myalgia and rhabdomyolysis may be caused by CAV3 mutations and broadens the phenotypic spectrum of caveolinopathies. In our series, immunoblotting was a more sensitive method to detect reduced caveolin-3 levels than immunohistochemistry in skeletal muscle. Patients presenting with muscle pain, exercise intolerance and rhabdomyolysis should be routinely tested for PIRCs as this may be an important clinical clue for caveolinopathies, even in the absence of other "typical" features. The use of NGS may expand current knowledge concerning inherited diseases, and unexpected/atypical phenotypes may be attributed to well-known human disease genes. PMID:27312022

  19. The first disease connection for Ca(v)2.2 channels

    Czech Academy of Sciences Publication Activity Database

    Weiss, Norbert

    2015-01-01

    Roč. 34, č. 3 (2015), s. 217-219. ISSN 0231-5882 R&D Projects: GA ČR GA15-13556S; GA MŠk 7AMB15FR015 Institutional support: RVO:61388963 Keywords : calcium channel * Ca(v)2.2 channel * channelopathies * myoclonus-dystonia syndrome Subject RIV: CE - Biochemistry Impact factor: 1.173, year: 2014

  20. DIFFERENT SUBSTRATA EFFECTS IN THE GERMINATION OF Ochroma pyramidale (CAV. EX LAM.) URB. (BOMBACACEAE)

    OpenAIRE

    Fabrízia de Oliveira Alvino; Breno Pinto Rayol

    2010-01-01

    The objective of this paper was to analyze the effect of different substrata in the germination of Ochroma pyramidale (Cav. ex Lam.) Urb. (Bombacaceae). The experiment was conducted in delineation randomized with three treatments (substratum) and four repetitions of 25 seeds. The following substrata had been tested: sand + vermiculite (1:1); vermiculite and, sand + shavings (1:1). The effect of substrata through the percentage of germination of the seeds, average time of germination and index...

  1. A retrospective investigation of canine adenovirus (CAV) infection in adult dogs in Turkey : article

    OpenAIRE

    Gur, S; A. Acar

    2009-01-01

    Canine adenovirus (CAV) type 1 and 2, respectively, cause infectious canine hepatitis and infectious canine laryngotracheitis in members of the families Canidae and Ursidae worldwide. Both of these infections are acute diseases, especially in young dogs. The aim of this study was to conduct a serological investigation of canine adenovirus infection. For this purpose, serumsamples were collected from native pure-bred Kangal (n = 11), and Akbash dogs (n = 17) and Turkish Greyhounds (n=15) in Es...

  2. Analysis of Volatile Compounds from Solanumbetaceum Cav. Fruits from Panama by Head-Space Micro Extraction

    OpenAIRE

    Durant, Armando A.; Candelario Rodríguez; Santana, Ana I; Carlos Herrero; Juan C. Rodríguez; Mahabir P. Gupta

    2012-01-01

    The characterization of the volatile compounds of two varieties of Solanum betaceum Cav. by means of headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry ( GC-MS) i s presented. The HS-SPME method for extraction of the volatiles compounds was optimized by using a 2 3 central composite design. Maximum extraction of volatile compounds was achieved by using a divinylbenzene-carboxen-polydimethylsiloxane (DVB/CAR/PDMS) fiber, extraction temperature 76°...

  3. Bupleurum virgatum Cav. [B. gerardii auct.; Apiaceae] im LSG "Harbke-Allertal"

    OpenAIRE

    Hofman, Constanze

    2013-01-01

    Im Rahmen der Untersuchungen an Trockenbiotopen im Allertal (Landkreis Börde, Sachsen- Anhalt) wurden im Jahr 2007 auf mehreren flächenhaften Naturdenkmalen vegetationskundliche Kartierarbeiten durchgeführt. Während dieser Kartierarbeiten wurde ein neuer Fundort von Bupleurum virgatum CAV. [Bupleurum gerardii auct.], dem Ruten-Hasenohr, nachgewiesen. Die Art gilt in Sachsen-Anhalt sowie in ganz Deutschland als vom Aussterben bedroht (FRANK et al. 2004, KORNECK et al. 1996). Das Ruten-Hasenohr...

  4. 基于CAV424的电容式液位检测系统%Capacitance Level Transducer Measurement System Based on CAV424

    Institute of Scientific and Technical Information of China (English)

    侯海岭; 解衍江

    2010-01-01

    介绍一种基于CAV424的电容式液位检测系统.根据电容式液位传感器的工作原理,传感器两个极板间的电容值会随液位的改变而改变,系统利用电容/电压转换芯片CAV424将电容值转化为电压值输出,然后使用16位高精度∑-△模数转换器AD7705再将电压值转换为数字信号,交由单片机对数据做进一步的处理,如数码管显示及电流输出.电流输出部分由12位串行数模转换器AD7943和4~20mA的电压/电流转换器组成.系统具有RS-485远传接口.

  5. Crystal structure of the CaV2 IQ domain in complex with Ca2+/calmodulin: High-resolution mechanistic implications for channel regulation by Ca2+

    OpenAIRE

    Mori, Masayuki X.; Vander Kooi, Craig W.; Leahy, Daniel J.; Yue, David T.

    2008-01-01

    Calmodulin (CaM) regulation of Ca2+ channels is central to Ca2+ signaling. CaV1 versus CaV2 classes of these channels exhibit divergent forms of regulation, potentially relating to customized CaM/IQ interactions among different channels. Here, we report the crystal structures for the Ca2+/CaM—IQ domains of both CaV2.1 and CaV2.3 channels. These highly similar structures emphasize that major CaM contacts with the IQ domain extend well upstream of traditional consensus residues. Surprisingly, u...

  6. Cooperative regulation of Cav1.2 channels by intracellular Mg2+, the proximal C-terminal EF-hand, and the distal C-terminal domain

    OpenAIRE

    Brunet, Sylvain; Scheuer, Todd; Catterall, William A.

    2009-01-01

    L-type Ca2+ currents conducted by Cav1.2 channels initiate excitation–contraction coupling in cardiac myocytes. Intracellular Mg2+ (Mgi) inhibits the ionic current of Cav1.2 channels. Because Mgi is altered in ischemia and heart failure, its regulation of Cav1.2 channels is important in understanding cardiac pathophysiology. Here, we studied the effects of Mgi on voltage-dependent inactivation (VDI) of Cav1.2 channels using Na+ as permeant ion to eliminate the effects of permeant divalent cat...

  7. Evaluation of the Functional Roles of Cav1.3 (α1D) Calcium Channel in Mouse Atrial and SA Node Cells

    Institute of Scientific and Technical Information of China (English)

    Nipavan; CHIAMVIMONVAT

    2005-01-01

    1 IntroductionMammalian α1 subunits of the voltage-gated Ca~(2+) channels are encoded by at least ten distinct genes. Previous data suggest that L-type Ca~(2+) channels containing Cav1.3 (α1D) subunit are expressed mainly in neurons and neuroendocrine cells, while those containing Cav1.2 (α1C) subunit are found in the brain, vascular smooth muscle and cardiac tissue. Recently, it has been shown that Cav1.3 Ca~(2+) channel is highly expressed in the cardiac tissue~([1]), especially in atrial tissue. Cav1.3 C...

  8. Characterization of the substituted N-triazole oxindole TROX-1, a small-molecule, state-dependent inhibitor of Ca(V)2 calcium channels.

    Science.gov (United States)

    Swensen, Andrew M; Herrington, James; Bugianesi, Randal M; Dai, Ge; Haedo, Rodolfo J; Ratliff, Kevin S; Smith, McHardy M; Warren, Vivien A; Arneric, Stephen P; Eduljee, Cyrus; Parker, David; Snutch, Terrance P; Hoyt, Scott B; London, Clare; Duffy, Joseph L; Kaczorowski, Gregory J; McManus, Owen B

    2012-03-01

    Biological, genetic, and clinical evidence provide validation for N-type calcium channels (Ca(V)2.2) as therapeutic targets for chronic pain. A state-dependent Ca(V)2.2 inhibitor may provide an improved therapeutic window over ziconotide, the peptidyl Ca(V)2.2 inhibitor used clinically. Supporting this notion, we recently reported that in preclinical models, the state-dependent Ca(V)2 inhibitor (3R)-5-(3-chloro-4-fluorophenyl)-3-methyl-3-(pyrimidin-5-ylmethyl)-1-(1H-1,2,4-triazol-3-yl)-1,3-dihydro-2H-indol-2-one (TROX-1) has an improved therapeutic window compared with ziconotide. Here we characterize TROX-1 inhibition of Cav2.2 channels in more detail. When channels are biased toward open/inactivated states by depolarizing the membrane potential under voltage-clamp electrophysiology, TROX-1 inhibits Ca(V)2.2 channels with an IC(50) of 0.11 μM. The voltage dependence of Ca(V)2.2 inhibition was examined using automated electrophysiology. TROX-1 IC(50) values were 4.2, 0.90, and 0.36 μM at -110, -90, and -70 mV, respectively. TROX-1 displayed use-dependent inhibition of Ca(V)2.2 with a 10-fold IC(50) separation between first (27 μM) and last (2.7 μM) pulses in a train. In a fluorescence-based calcium influx assay, TROX-1 inhibited Ca(V)2.2 channels with an IC(50) of 9.5 μM under hyperpolarized conditions and 0.69 μM under depolarized conditions. Finally, TROX-1 potency was examined across the Ca(V)2 subfamily. Depolarized IC(50) values were 0.29, 0.19, and 0.28 μM by manual electrophysiology using matched conditions and 1.8, 0.69, and 1.1 μM by calcium influx for Ca(V)2.1, Ca(V)2.2, and Ca(V)2.3, respectively. Together, these in vitro data support the idea that a state-dependent, non-subtype-selective Ca(V)2 channel inhibitor can achieve an improved therapeutic window over the relatively state-independent Ca(V)2.2-selective inhibitor ziconotide in preclinical models of chronic pain. PMID:22188924

  9. Transcriptomic Profiling of Virus-Host Cell Interactions following Chicken Anaemia Virus (CAV Infection in an In Vivo Model.

    Directory of Open Access Journals (Sweden)

    Efstathios S Giotis

    Full Text Available Chicken Anaemia Virus (CAV is an economically important virus that targets lymphoid and erythroblastoid progenitor cells leading to immunosuppression. This study aimed to investigate the interplay between viral infection and the host's immune response to better understand the pathways that lead to CAV-induced immunosuppression. To mimic vertical transmission of CAV in the absence of maternally-derived antibody, day-old chicks were infected and their responses measured at various time-points post-infection by qRT-PCR and gene expression microarrays. The kinetics of mRNA expression levels of signature cytokines of innate and adaptive immune responses were determined by qRT-PCR. The global gene expression profiles of mock-infected (control and CAV-infected chickens at 14 dpi were also compared using a chicken immune-related 5K microarray. Although in the thymus there was evidence of induction of an innate immune response following CAV infection, this was limited in magnitude. There was little evidence of a Th1 adaptive immune response in any lymphoid tissue, as would normally be expected in response to viral infection. Most cytokines associated with Th1, Th2 or Treg subsets were down-regulated, except IL-2, IL-13, IL-10 and IFNγ, which were all up-regulated in thymus and bone marrow. From the microarray studies, genes that exhibited significant (greater than 1.5-fold, false discovery rate <0.05 changes in expression in thymus and bone marrow on CAV infection were mainly associated with T-cell receptor signalling, immune response, transcriptional regulation, intracellular signalling and regulation of apoptosis. Expression levels of a number of adaptor proteins, such as src-like adaptor protein (SLA, a negative regulator of T-cell receptor signalling and the transcription factor Special AT-rich Binding Protein 1 (SATB1, were significantly down-regulated by CAV infection, suggesting potential roles for these genes as regulators of viral infection or

  10. Impact of virus load on immunocytological and histopathological parameters during clinical chicken anemia virus (CAV) infection in poultry.

    Science.gov (United States)

    Wani, Mohd Yaqoob; Dhama, Kuldeep; Malik, Yashpal Singh

    2016-07-01

    Chicken anemia virus (CAV) is one the important pathogen affecting commercial poultry sector globally by causing mortality, production losses, immunosuppression, aggravating co-infections and vaccination failures. Here, we describe the effects of CAV load on hematological, histopathological and immunocytochemical alterations in 1-day old infected chicks. The effects of CAV on cytokine expression profiles and generation of virus specific antibody titer were also studied and compared with viral clearance in various tissues. The results clearly confirmed that peak viral load was achieved mainly in lymphoid tissues between 10 and 20 days post infection (dpi), being highest in the blood (log1010.63 ±0.87/ml) and thymus (log1010.29 ±0.94/g) followed by spleen, liver, bone marrow and bursa. The histopathology and immunoflowcytometric analysis indicated specific degeneration of T lymphoid cells in the thymus, spleen and blood at 15 dpi. While the transcript levels of interleukin (IL)-1, IL-2, IL-12 decreased at all dpi, interferon (IFN)-γ increased (3-15 fold) during early stages of infection and the appearance of virus specific antibodies were found to be strongly associated with virus clearance in all the tissues. Our findings support the immunosuppressive nature of CAV and provide the relation between the virus load in the various body tissues and the immunopathological changes during clinical CAV infections. PMID:27165537

  11. The effects of S4 segments on the voltage-dependence of inactivation for Cav3.1 calcium channels

    Institute of Scientific and Technical Information of China (English)

    LI JunYing

    2007-01-01

    T-type calcium channels exhibit fast voltage-dependent inactivation,for which the underlying structure-function relationship still remains unclear.To investigate the roles of S4 segments in voltage-dependent inactivation of T-type calcium channels,we created S4 replacement chimeras between Cav3.1 calcium channels(fast voltage-dependent inactivation)and Cav1.2 calcium channels(little oltage-dependent inactivation)by replacing S4s in Cav3.1 with the corresponding regions in Cav1.2.Wild type and chimeric channels were expressed in Xenopus oocytes and channel currents were recorded with two-electrode voltage-clamp.We showed that replacing S4 region in domain I shifted voltage-dependence for inactivation of Cav3.1 to the left,and the V0.5 inact and kinact value were significantly changed.However replacing S4s in domains Ⅱ-Ⅳ had no effects on the voltage-dependent inactivation properties.These results suggest that the roles of S4 segments in domains Ⅰ-Ⅳ are different,and S4 in domain I is likely to be involved in voltage-dependent Inactivation process.Its movement during membrane depolarization may trigger a conformational change in the inactivation gate.

  12. Cell-type-specific tuning of Cav1.3 Ca2+-channels by a C-terminal automodulatory domain

    Science.gov (United States)

    Scharinger, Anja; Eckrich, Stephanie; Vandael, David H.; Schönig, Kai; Koschak, Alexandra; Hecker, Dietmar; Kaur, Gurjot; Lee, Amy; Sah, Anupam; Bartsch, Dusan; Benedetti, Bruno; Lieb, Andreas; Schick, Bernhard; Singewald, Nicolas; Sinnegger-Brauns, Martina J.; Carbone, Emilio; Engel, Jutta; Striessnig, Jörg

    2015-01-01

    Cav1.3 L-type Ca2+-channel function is regulated by a C-terminal automodulatory domain (CTM). It affects channel binding of calmodulin and thereby tunes channel activity by interfering with Ca2+- and voltage-dependent gating. Alternative splicing generates short C-terminal channel variants lacking the CTM resulting in enhanced Ca2+-dependent inactivation and stronger voltage-sensitivity upon heterologous expression. However, the role of this modulatory domain for channel function in its native environment is unkown. To determine its functional significance in vivo, we interrupted the CTM with a hemagglutinin tag in mutant mice (Cav1.3DCRDHA/HA). Using these mice we provide biochemical evidence for the existence of long (CTM-containing) and short (CTM-deficient) Cav1.3 α1-subunits in brain. The long (HA-labeled) Cav1.3 isoform was present in all ribbon synapses of cochlear inner hair cells. CTM-elimination impaired Ca2+-dependent inactivation of Ca2+-currents in hair cells but increased it in chromaffin cells, resulting in hyperpolarized resting potentials and reduced pacemaking. CTM disruption did not affect hearing thresholds. We show that the modulatory function of the CTM is affected by its native environment in different cells and thus occurs in a cell-type specific manner in vivo. It stabilizes gating properties of Cav1.3 channels required for normal electrical excitability. PMID:26379493

  13. Cell-type-specific tuning of Cav1.3 Ca2+-channels by a C-terminal automodulatory domain

    Directory of Open Access Journals (Sweden)

    Anja eScharinger

    2015-08-01

    Full Text Available Cav1.3 L-type Ca2+-channel function is regulated by a C-terminal automodulatory domain (CTM. It affects channel binding of calmodulin and thereby tunes channel activity by interfering with Ca2+- and voltage-dependent gating. Alternative splicing generates short C-terminal channel variants lacking the CTM resulting in enhanced Ca2+-dependent inactivation and stronger voltage-sensitivity upon heterologous expression. However, the role of this modulatory domain for channel function in its native environment is unkown. To determine its functional significance in vivo, we interrupted the CTM with a hemagglutinin tag in mutant mice (Cav1.3DCRDHA/HA. Using these mice we provide biochemical evidence for the existence of long (CTM-containing and short (CTM-deficient Cav1.3 α1-subunits in brain. The long (HA-labeled Cav1.3 isoform was present in all ribbon synapses of cochlear inner hair cells. CTM-elimination impaired Ca2+-dependent inactivation of Ca2+-currents in hair cells but increased it in chromaffin cells, resulting in hyperpolarized resting potentials and reduced pacemaking. CTM disruption did not affect hearing thresholds. We show that the modulatory function of the CTM is affected by its native environment in different cells and thus occurs in a cell-type specific manner in vivo. It is required to stabilize gating properties of Cav1.3 channels required for normal electrical excitability.

  14. Progress in the structural understanding of voltage-gated calcium channel (CaV) function and modulation.

    Science.gov (United States)

    Minor, Daniel L; Findeisen, Felix

    2010-01-01

    Voltage-gated calcium channels (CaVs) are large, transmembrane multiprotein complexes that couple membrane depolarization to cellular calcium entry. These channels are central to cardiac action potential propagation, neurotransmitter and hormone release, muscle contraction, and calcium-dependent gene transcription. Over the past six years, the advent of high-resolution structural studies of CaV components from different isoforms and CaV modulators has begun to reveal the architecture that underlies the exceptionally rich feedback modulation that controls CaV action. These descriptions of CaV molecular anatomy have provided new, structure-based insights into the mechanisms by which particular channel elements affect voltage-dependent inactivation (VDI), calcium‑dependent inactivation (CDI), and calcium‑dependent facilitation (CDF). The initial successes have been achieved through structural studies of soluble channel domains and modulator proteins and have proven most powerful when paired with biochemical and functional studies that validate ideas inspired by the structures. Here, we review the progress in this growing area and highlight some key open challenges for future efforts. PMID:21139419

  15. Transcriptional and electrophysiological consequences of KChIP2-mediated regulation of CaV1.2

    DEFF Research Database (Denmark)

    Thomsen, Morten B; Foster, Erika; Nguyen, Katherine H; Sosunov, Eugene A

    2009-01-01

    . Recently, we have reported that KChIP2 functionally modulates the cardiac Ca(V)1.2-governed L-type Ca(2+) current (I(Ca,L)) through a direct interaction between KChIP2 and the amino-terminus of Ca(V)1.2. Here, we show that KChIP2 and Ca(V)1.2 co-immunoprecipitate enhancing the biochemical support for our......IP2(-/-) mice, the action potential morphology is not altered. Furthermore, we show that the ventricular effective refractory period (VERP) is comparable in wild-type (53 +/- 5 ms) and KChIP2(-/-) mice (48 +/- 3 ms; p > 0.05). In summary, our findings document a novel function of KChIP2 and expand our...

  16. A Study on OBE Exceedance Criteria Using the CAV Concept for NPPs in Korea

    International Nuclear Information System (INIS)

    NPPs in Korea have so far complied with the US Regulatory Guide 1.12 Rev.1 (1974) for the seismic monitoring systems. However, researchers in Korea have been considering attempting to comply with Regulatory Guide 1.12 rev.2 (1997) when implementing all future seismic monitoring systems. To comply with the new regulations, Operating Basis Earthquake (OBE) Exceedance Criteria applied with the Cumulative Absolute Velocity (CAV) has to be developed for local use. The CAV was developed in the USA as one reasonable OBE exceedance criterion for NPPs; however, it has not been widely used outside the USA, not-withstanding its solid technical basis. It is, however, expected that the worldwide NPP construction boom will necessitate development and more frequent use of reasonable OBE exceedance criteria. As multi-units (four or more) are situated in one site in Korea, prevention of simultaneous shutdown of multi-unit caused by non damaging earthquakes is very important; otherwise, a serious economic loss could occur, and there could also be a great impact on the stability of the power grid that would arise due to the unnecessary shutdown. Actually, we have had the experience of observing a Peak Ground Acceleration (PGA)-value greater than 0.1g (the current OBE level for Korean NPPs) due to an M=4.8 earthquake (20th Jan. '07) that did not even cause any damage to the poorly-designed structures nearby. This experience gave a motivation to the KHNP should concern the needs of study on OBE exceedance criteria in view of application of the CAV to Korean seismic situation, which will be more practical. The present paper is intended to show the intermediate result of the research on Operating Basis Earthquake (OBE) exceedance criteria in Korea

  17. Mechanism of osthole inhibition of vascular Ca(v)1.2 current.

    Science.gov (United States)

    Fusi, Fabio; Sgaragli, Giampietro; Ha, Le Minh; Cuong, Nguyen Manh; Saponara, Simona

    2012-04-01

    Osthole is a coumarin extracted from Cnidium monnieri (L.) Cusson. The medicinal plant is widely used in Vietnamese as well as Chinese traditional medicine as a vasodilating and antihypertensive agent. Here we have tested the proposition that the block of Ca(v)1.2 channels is mainly responsible for its vascular activity. An in-depth analysis of the effect of osthole on Ca(v)1.2 current (I(Ca1.2)) was performed in rat tail artery myocytes using the whole-cell patch-clamp method. Osthole decreased I(Ca1.2) in a concentration- and voltage-dependent manner. At holding potentials of -50 and -80mV, the pIC(50) values were 4.78±0.07 and 4.36±0.08, respectively; the latter corresponded to the drug apparent dissociation constant for resting channels, K(R), of 47.8μM. Osthole speeded up the inactivation kinetics of I(Ca1.2) and shifted the voltage dependence of the inactivation curve to more negative potentials in a concentration-dependent manner, with an apparent dissociation constant for inactivated channels (K(I)) of 6.88μM. Block of I(Ca1.2) was frequency-dependent and the rate of recovery from inactivation was slowed down. In conclusion, osthole is a vascular Ca(v)1.2 channel antagonist stabilizing the channel in its inactivated state. This mechanism may account for the systolic blood pressure reduction induced by the drug in animal models of hypertension and points to osthole as a lead for the development of novel antihypertensive agents. PMID:22329900

  18. Spin-Peierls States of Quantum Antiferromagnets on the CaV4O9 Lattice

    International Nuclear Information System (INIS)

    We discuss the quantum paramagnetic phases of Heisenberg antiferromagnets on the 1/5-depleted square lattice found in CaV4O9. The possible phases of the quantum dimer model on this lattice are obtained by a mapping to a quantum-mechanical height model. In addition to the open-quote open-quote decoupled close-quote close-quote phases found earlier, we find a possible intermediate spin-Peierls phase with spontaneously broken lattice symmetry. Experimental signatures of the different quantum paramagnetic phases are discussed. copyright 1996 The American Physical Society

  19. Population genetic structure of Sisyrinchium micranthum Cav. (Iridaceae) in Itapuã State Park, Southern Brazil

    OpenAIRE

    Luana Olinda Tacuatiá; Lilian Eggers; Eliane Kaltchuk-Santos; Souza-Chies, Tatiana T.

    2012-01-01

    Sisyrinchium micranthum Cav. is a member of the family Iridaceae, which is distributed over the American continent. In Brazil, this species is found, not only in disturbed areas and coastal regions, but is also very common in urban centers, such as public parks, during the spring. Chromosome counts for North American specimens are 2n = 32 and 2n = 48, whereas in southern Brazil, there is a polyploidy series with three chromosome numbers, 2n = 16, 2n = 32, and 2n = 48. Population analyses usin...

  20. An automated electrophysiological assay for differentiating Ca(v)2.2 inhibitors based on state dependence and kinetics.

    Science.gov (United States)

    Swensen, Andrew M; Niforatos, Wende; Vortherms, Timothy A; Perner, Richard J; Li, Tao; Schrimpf, Michael R; Scott, Victoria E; Lee, Lance; Jarvis, Michael F; McGaraughty, Steve

    2012-12-01

    Ca(V)2.2 (N-type) calcium channels are key regulators of neurotransmission. Evidence from knockout animals and localization studies suggest that Ca(V)2.2 channels play a critical role in nociceptive transmission. Additionally, ziconotide, a selective peptide inhibitor of Ca(V)2.2 channels, is clinically used to treat refractory pain. However, the use of ziconotide is limited by its low therapeutic index, which is believed, at least in part, to be a consequence of ziconotide inhibiting Ca(V)2.2 channels regardless of the channel state. Subsequent efforts have focused on the discovery of state-dependent inhibitors that preferentially bind to the inactivated state of Ca(V)2.2 channels in order to achieve an improved safety profile relative to ziconotide. Much less attention has been paid to understanding the binding kinetics of these state-dependent inhibitors. Here, we describe a novel electrophysiology-based assay on an automated patch platform designed to differentiate Ca(V)2.2 inhibitors based on their combined state dependence and kinetics. More specifically, this assay assesses inactivated state block, closed state block, and monitors the kinetics of recovery from block when channels move between states. Additionally, a use-dependent assay is described that uses a train of depolarizing pulses to drive channels to a similar level of inactivation for comparison. This use-dependent protocol also provides information on the kinetics of block development. Data are provided to show how these assays can be utilized to screen for kinetic diversity within and across chemical classes. PMID:22428804

  1. A Restructuring of the CAV and FDI Package for the MIDAS Computer Code

    Energy Technology Data Exchange (ETDEWEB)

    Park, S. H.; Kim, D. H. [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of); Cho, S. W. [Korea Radiation Technology Institute Co., Taejon (Korea, Republic of)

    2006-07-01

    As one of the processes for a localized severe accident analysis code, KAERI is developing a severe accident code MIDAS. The MIDAS code is being developed based on MELCOR. The existing data saving method of MELCOR uses pointer variables for a fix-sized storage management, and it deteriorates the readability, maintainability and portability of the code. As a most important process for a localized severe accident analysis code, it is needed convenient method for data handling. So, it has been used the new features in FORTRAN90 such as a dynamic allocation for the restructuring. The restructuring of the data saving and transferring method of the existing code makes it easy to understand the code. Before an entire restructuring of the code, a restructuring template for a simple package was developed and tested. The target for the restructuring in this paper was the CAV and FDI packages. The CAV(cavity) package is responsible for modeling the attack on the basement concrete by hot core materials. The FDI(Fuel Dispersal Interactions) package is responsible for modeling both low and high pressure molten fuel ejection from the RPV into the reactor cavity, control volumes and surfaces. The verification was done through comparing the results before and after the restructuring.

  2. A Restructuring of the CAV and FDI Package for the MIDAS Computer Code

    International Nuclear Information System (INIS)

    As one of the processes for a localized severe accident analysis code, KAERI is developing a severe accident code MIDAS. The MIDAS code is being developed based on MELCOR. The existing data saving method of MELCOR uses pointer variables for a fix-sized storage management, and it deteriorates the readability, maintainability and portability of the code. As a most important process for a localized severe accident analysis code, it is needed convenient method for data handling. So, it has been used the new features in FORTRAN90 such as a dynamic allocation for the restructuring. The restructuring of the data saving and transferring method of the existing code makes it easy to understand the code. Before an entire restructuring of the code, a restructuring template for a simple package was developed and tested. The target for the restructuring in this paper was the CAV and FDI packages. The CAV(cavity) package is responsible for modeling the attack on the basement concrete by hot core materials. The FDI(Fuel Dispersal Interactions) package is responsible for modeling both low and high pressure molten fuel ejection from the RPV into the reactor cavity, control volumes and surfaces. The verification was done through comparing the results before and after the restructuring

  3. Disorder effects in the S=1 antiferromagnetic spin ladder CaV2O4

    Science.gov (United States)

    Guitarra, S. R.; Caneiro, A.; Niebieskikwiat, D.

    2015-10-01

    We study the physical properties of the antiferromagnetic spin ladder CaV2O4 (CVO) and the Y-doped related compound Ca0.9Y0.1V2O4. In the latter, X-ray diffraction demonstrates the segregation of a small amount of a vanadium-perovskite impurity phase, leading to the formation of V vacancies within the main CVO-type structure. The 1D character of this calcium-vanadite enhances the influence of the vacancies on the electric and magnetic properties of Ca0.9Y0.1V2O4. Electrical transport is characterized by a variable-range hopping mechanism determined by the charging energy of nm-sized segments of V chains delimited by V vacancies, i.e. a Coulomb gap is formed at the Fermi level. These vacancies also locally affect the magnetic correlations, breaking the long-range AFM order observed in CaV2O4 and producing exchange bias when the Y-doped sample is cooled with an applied magnetic field.

  4. Effect of structurally related flavonoids from Zuccagnia punctata Cav. on Caenorhabditis elegans.

    Science.gov (United States)

    D'Almeida, Romina E; Alberto, María R; Morgan, Phillip; Sedensky, Margaret; Isla, María I

    2014-03-01

    Zuccagnia punctata Cav. (Fabaceae), commonly called jarilla macho or pus-pus, is being used in traditional medicine as an antiseptic, anti-inflammatory and to relieve muscle and bone pain. The aim of this work was to study the anthelmintic effects of three structurally related flavonoids present in aerial parts of Z. punctata Cav. The biological activity of the flavonoids 7-hydroxyflavanone (HF), 3,7-dihydroxyflavone (DHF) and 2´,4´-dihydroxychalcone (DHC) was examined in the free-living nematode Caenorhabditis elegans. Our results showed that among the assayed flavonoids, only DHC showed an anthelmintic effect and alteration of egg hatching and larval development processes in C. elegans. DHC was able to kill 50% of adult nematodes at a concentration of 17 μg/mL. The effect on larval development was observed after 48 h in the presence of 25 and 50 μg/mL DHC, where 33.4 and 73.4% of nematodes remained in the L3 stage or younger. New therapeutic drugs with good efficacy against drug-resistant nematodes are urgently needed. Therefore, DHC, a natural compound present in Z. punctata, is proposed as a potential anthelmintic drug. PMID:26204036

  5. Graded Ca2+/calmodulin-dependent coupling of voltage-gated CaV1.2 channels

    Science.gov (United States)

    Dixon, Rose E; Moreno, Claudia M; Yuan, Can; Opitz-Araya, Ximena; Binder, Marc D; Navedo, Manuel F; Santana, Luis F

    2015-01-01

    In the heart, reliable activation of Ca2+ release from the sarcoplasmic reticulum during the plateau of the ventricular action potential requires synchronous opening of multiple CaV1.2 channels. Yet the mechanisms that coordinate this simultaneous opening during every heartbeat are unclear. Here, we demonstrate that CaV1.2 channels form clusters that undergo dynamic, reciprocal, allosteric interactions. This ‘functional coupling’ facilitates Ca2+ influx by increasing activation of adjoined channels and occurs through C-terminal-to-C-terminal interactions. These interactions are initiated by binding of incoming Ca2+ to calmodulin (CaM) and proceed through Ca2+/CaM binding to the CaV1.2 pre-IQ domain. Coupling fades as [Ca2+]i decreases, but persists longer than the current that evoked it, providing evidence for ‘molecular memory’. Our findings suggest a model for CaV1.2 channel gating and Ca2+-influx amplification that unifies diverse observations about Ca2+ signaling in the heart, and challenges the long-held view that voltage-gated channels open and close independently. DOI: http://dx.doi.org/10.7554/eLife.05608.001 PMID:25714924

  6. Block of CaV1.2 Channels by Gd3+ Reveals Preopening Transitions in the Selectivity Filter

    OpenAIRE

    Babich, Olga; Reeves, John; Shirokov, Roman

    2007-01-01

    Using the lanthanide gadolinium (Gd3+) as a Ca2+ replacing probe, we investigated the voltage dependence of pore blockage of CaV1.2 channels. Gd+3 reduces peak currents (tonic block) and accelerates decay of ionic current during depolarization (use-dependent block). Because diffusion of Gd3+ at concentrations used (

  7. Transcriptional Response of Human Neurospheres to Helper-Dependent CAV-2 Vectors Involves the Modulation of DNA Damage Response, Microtubule and Centromere Gene Groups.

    Directory of Open Access Journals (Sweden)

    Stefania Piersanti

    Full Text Available Brain gene transfer using viral vectors will likely become a therapeutic option for several disorders. Helper-dependent (HD canine adenovirus type 2 vectors (CAV-2 are well suited for this goal. These vectors are poorly immunogenic, efficiently transduce neurons, are retrogradely transported to afferent structures in the brain and lead to long-term transgene expression. CAV-2 vectors are being exploited to unravel behavior, cognition, neural networks, axonal transport and therapy for orphan diseases. With the goal of better understanding and characterizing HD-CAV-2 for brain therapy, we analyzed the transcriptomic modulation induced by HD-CAV-2 in human differentiated neurospheres derived from midbrain progenitors. This 3D model system mimics several aspects of the dynamic nature of human brain. We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes. Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways. We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.

  8. Ablation of Ca(V2.1 voltage-gated Ca²⁺ channels in mouse forebrain generates multiple cognitive impairments.

    Directory of Open Access Journals (Sweden)

    Robert Theodor Mallmann

    Full Text Available Voltage-gated Ca(V2.1 (P/Q-type Ca²⁺ channels located at the presynaptic membrane are known to control a multitude of Ca²⁺-dependent cellular processes such as neurotransmitter release and synaptic plasticity. Our knowledge about their contributions to complex cognitive functions, however, is restricted by the limited adequacy of existing transgenic Ca(V2.1 mouse models. Global Ca(V2.1 knock-out mice lacking the α1 subunit Cacna1a gene product exhibit early postnatal lethality which makes them unsuitable to analyse the relevance of Ca(V2.1 Ca²⁺ channels for complex behaviour in adult mice. Consequently we established a forebrain specific Ca(V2.1 knock-out model by crossing mice with a floxed Cacna1a gene with mice expressing Cre-recombinase under the control of the NEX promoter. This novel mouse model enabled us to investigate the contribution of Ca(V2.1 to complex cognitive functions, particularly learning and memory. Electrophysiological analysis allowed us to test the specificity of our conditional knock-out model and revealed an impaired synaptic transmission at hippocampal glutamatergic synapses. At the behavioural level, the forebrain-specific Ca(V2.1 knock-out resulted in deficits in spatial learning and reference memory, reduced recognition memory, increased exploratory behaviour and a strong attenuation of circadian rhythmicity. In summary, we present a novel conditional Ca(V2.1 knock-out model that is most suitable for analysing the in vivo functions of Ca(V2.1 in the adult murine forebrain.

  9. MiR-103 inhibits osteoblast proliferation mainly through suppressing Cav1.2 expression in simulated microgravity.

    Science.gov (United States)

    Sun, Zhongyang; Cao, Xinsheng; Hu, Zebing; Zhang, Lianchang; Wang, Han; Zhou, Hua; Li, Dongtao; Zhang, Shu; Xie, Manjiang

    2015-07-01

    Emerging evidence indicates that microRNAs (miRNAs) play important roles in modulating osteoblast function and bone formation. However, the influence of miRNA on osteoblast proliferation and the possible mechanisms underlying remain to be defined. In this study, we aimed to investigate whether miR-103 regulates osteoblast proliferation under simulated microgravity condition through regulating Cav1.2, the primary subunit of L-type voltage sensitive calcium channels (LTCCs). We first investigated the effect of simulated microgravity on osteoblast proliferation and the outcomes clearly demonstrated that the mechanical unloading inhibits MC3T3-E1 osteoblast-like cell proliferation. Using quantitative Real-Time PCR (qRT-PCR), we provided data showing that miR-103 was up-regulated in response to simulated microgravity. In addition, we observed that up-regulation of miR-103 inhibited and down-regulation of miR-103 promoted osteoblast proliferation under simulated microgravity condition. Furthermore, knocking-down or over-expressing miR-103, respectively, up- or down-regulated the level of Cav1.2 expression and LTCC currents, suggesting that miR-103 acts as an endogenous attenuator of Cav1.2 in osteoblasts under simulated microgravity condition. More importantly, we showed that the effect of miR-103 on osteoblast proliferation was diminished in simulated microgravity, when co-transfecting miR-103 mimic or inhibitor with Cav1.2 siRNA. Taken together, our data suggest that miR-103 inhibits osteoblast proliferation mainly through suppression of Cav1.2 expression under simulated microgravity condition. This work may provide a novel mechanism of microgravity-induced detrimental effects on osteoblast proliferation, identifying miR-103 as a novel possible therapeutic target in bone remodeling disorders in this mechanical unloading. PMID:25868801

  10. Alcohol Withdrawal-Induced Seizure Susceptibility is Associated with an Upregulation of CaV1.3 Channels in the Rat Inferior Colliculus

    Science.gov (United States)

    Akinfiresoye, Luli R.; Allard, Joanne S.; Lovinger, David M.

    2015-01-01

    Background: We previously reported increased current density through L-type voltage-gated Ca2+ (CaV1) channels in inferior colliculus (IC) neurons during alcohol withdrawal. However, the molecular correlate of this increased CaV1 current is currently unknown. Methods: Rats received three daily doses of ethanol every 8 hours for 4 consecutive days; control rats received vehicle. The IC was dissected at various time intervals following alcohol withdrawal, and the mRNA and protein levels of the CaV1.3 and CaV1.2 α1 subunits were measured. In separate experiments, rats were tested for their susceptibility to alcohol withdrawal–induced seizures (AWS) 3, 24, and 48 hours after alcohol withdrawal. Results: In the alcohol-treated group, AWS were observed 24 hours after withdrawal; no seizures were observed at 3 or 48 hours. No seizures were observed at any time in the control-treated rats. Compared to control-treated rats, the mRNA level of the CaV1.3 α1 subunit was increased 1.4-fold, 1.9-fold, and 1.3-fold at 3, 24, and 48 hours, respectively. In contrast, the mRNA level of the CaV1.2 α1 subunit increased 1.5-fold and 1.4-fold at 24 and 48 hours, respectively. At 24 hours, Western blot analyses revealed that the levels of the CaV1.3 and CaV1.2 α1 subunits increased by 52% and 32%, respectively, 24 hours after alcohol withdrawal. In contrast, the CaV1.2 and CaV1.3 α1 subunits were not altered at either 3 or 48 hours during alcohol withdrawal. Conclusions: Expression of the CaV1.3 α1 subunit increased in parallel with AWS development, suggesting that altered L-type CaV1.3 channel expression is an important feature of AWS pathogenesis. PMID:25556199

  11. THE ROLE OF MAP1A LIGHT CHAIN 2 IN SYNAPTIC SURFACE RETENTION OF CAV2.2 CHANNELS IN HIPPOCAMPAL NEURONS

    OpenAIRE

    Leenders, AG Miriam; Lin, Lin; Huang, Li-Dong; Gerwin, Claudia; Lu, Pei-Hua; Sheng, Zu-Hang

    2008-01-01

    Cav2.2 channels are localized at nerve terminals where they play a critical role in neurotransmission. However, the determinant that controls surface retention of these channels has not been identified. Here, we report that presynaptic surface localization of Cav2.2 is mediated through its interaction with light chain 2 (LC2) of microtubule-associated protein MAP1A. Deletion of a 23-residue binding-domain (BD) within the Cav2.2 C-terminus resulted in reduced synaptic distribution of the mutan...

  12. Synaptic Vesicle Tethering and the CaV2.2 Distal C-terminal

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    Fiona K Wong

    2014-03-01

    Full Text Available . Evidence that synaptic vesicles (SVs can be gated by a single voltage sensitive calcium channel (CaV2.2 predict a molecular linking mechanism or ‘tether’[Stanley 1993]. Recent studies have proposed that the SV binds to the distal C-terminal on the CaV2.2 calcium channel [Kaeser et al. 2011;Wong, Li, and Stanley 2013] while genetic analysis proposed a double tether mechanism via RIM: directly to the C terminus PDZ ligand domain or indirectly via a more proximal proline rich site [Kaeser et al. 2011]. Using a novel in vitro SV-PD binding assay, we reported that SVs bind to a fusion protein comprising the C-terminal distal third (C3, aa 2137-2357 [Wong, Li, and Stanley 2013]. Here we limit the binding site further to the last 58 aa, beyond the proline rich site, by the absence of SV capture by a truncated C3 fusion protein (aa 2137-2299. To test PDZ-dependent binding we generated two C terminus-mutant C3 fusion proteins and a mimetic blocking peptide (H-WC, aa 2349-2357 and validated these by elimination of MINT-1 or RIM binding. Persistence of SV capture with all three fusion proteins or with the full length C3 protein but in the presence of the blocking peptide, demonstrated that SVs can bind to the distal C-terminal via a PDZ-independent mechanism. These results were supported in situ by normal SV turnover in H-WC-loaded synaptosomes, as assayed by a novel peptide cryoloading method. Thus, SVs tether to the CaV2.2 C-terminal within a 49 aa region immediately prior to the terminus PDZ ligand domain. Long tethers that could reflect extended C termini were imaged by electron microscopy of synaptosome ghosts. To fully account for SV tethering we propose a model where SVs are initially captured, or ‘grabbed’, from the cytoplasm by a binding site on the distal region of the channel C-terminal and are then retracted to be ‘locked’ close to the channel by a second attachment mechanism in preparation for single channel domain gating.

  13. Effet du stress oxydant sur les cavéoles dans les cellules musculaires squelettiques

    OpenAIRE

    Mougeolle, Alexis

    2014-01-01

    La sarcopénie est une maladie dégénérative liée à l’âge qui se caractérise par une perte progressive et involontaire de la masse et de la force musculaire. Elle s’accompagne d’une atteinte de la régénération musculaire et d’une accumulation des espèces réactives de l’oxygène. Les cavéoles sont des invaginations de la membrane plasmique. Dans le muscle, elles jouent un rôle dans la différenciation des cellules satellites et dans le maintien de l’unité contractile dans le muscle différencié. Ce...

  14. Analysis of Volatile Compounds from Solanumbetaceum Cav. Fruits from Panama by Head-Space Micro Extraction

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    Armando A. Durant

    2012-11-01

    Full Text Available The characterization of the volatile compounds of two varieties of Solanum betaceum Cav. by means of headspace solid-phase microextraction (HS-SPME coupled with gas chromatography-mass spectrometry ( GC-MS i s presented. The HS-SPME method for extraction of the volatiles compounds was optimized by using a 2 3 central composite design. Maximum extraction of volatile compounds was achieved by using a divinylbenzene-carboxen-polydimethylsiloxane (DVB/CAR/PDMS fiber, extraction temperature 76° C, incubation time 44 min, and extraction time of 46 min. The main types of compounds detected in both varieties are terpenoids, followed by aromatics, esters, and aldehydes. Golden-yellow cultivars contained higher levels of esters and terpenes, while the reddish-purple variety contained a significant amount of aromatic compounds. The data structure of the chemical information obtained as well as the relationship between variables was evaluated by means of principal component analysis and cluster analysis.

  15. DIFFERENT SUBSTRATA EFFECTS IN THE GERMINATION OF Ochroma pyramidale (CAV. EX LAM. URB. (BOMBACACEAE

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    Fabrízia de Oliveira Alvino

    2010-08-01

    Full Text Available The objective of this paper was to analyze the effect of different substrata in the germination of Ochroma pyramidale (Cav. ex Lam. Urb. (Bombacaceae. The experiment was conducted in delineation randomized with three treatments (substratum and four repetitions of 25 seeds. The following substrata had been tested: sand + vermiculite (1:1; vermiculite and, sand + shavings (1:1. The effect of substrata through the percentage of germination of the seeds, average time of germination and index of germination speed (IGS were evaluated. There had been significant difference between the treatments in of all the observed variable. The seeds, when conditioned in vermiculite, had presented the biggest tax of germination, greatest speed and lowest germination time.

  16. Upper canopy pollinators of Eucryphia cordifolia Cav., a tree of South American temperate rain forest

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    Cecilia Smith-Ramírez

    2016-05-01

    Full Text Available Ecological processes in the upper canopy of temperate forests have been seldom studied because of the limited accessibility. Here, we present the results of the first survey of the pollinator assemblage and the frequency of insect visits to flowers in the upper branches of ulmo, Eucryphia cordifolia Cav., an emergent 30-40 m-tall tree in rainforests of Chiloé Island, Chile. We compared these findings with a survey of flower visitors restricted to lower branches of E. cordifolia 1- in the forest understory, 2- in lower branches in an agroforestry area. We found 10 species of pollinators in canopy, and eight, 12 and 15 species in understory, depending of tree locations. The main pollinators of E. cordifolia in the upper canopy differed significantly from the pollinator assemblage recorded in lower tree branches. We conclude that the pollinator assemblages of the temperate forest canopy and interior are still unknown.

  17. How “Pharmacoresistant” is Cav2.3, the Major Component of Voltage-Gated R-type Ca2+ Channels?

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    Jürgen Hescheler

    2013-05-01

    Full Text Available Membrane-bound voltage-gated Ca2+ channels (VGCCs are targets for specific signaling complexes, which regulate important processes like gene expression, neurotransmitter release and neuronal excitability. It is becoming increasingly evident that the so called “resistant” (R-type VGCC Cav2.3 is critical in several physiologic and pathophysiologic processes in the central nervous system, vascular system and in endocrine systems. However its eponymous attribute of pharmacologic inertness initially made in depth investigation of the channel difficult. Although the identification of SNX-482 as a fairly specific inhibitor of Cav2.3 in the nanomolar range has enabled insights into the channels properties, availability of other pharmacologic modulators of Cav2.3 with different chemical, physical and biological properties are of great importance for future investigations. Therefore the literature was screened systematically for molecules that modulate Cav2.3 VGCCs.

  18. Desiccation tolerance of seeds of Bunchosia armenica (Cav. DC.Tolerância à dessecação em sementes de Bunchosia armenica (Cav. DC.

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    Edson de Almeida Cardoso

    2012-08-01

    Full Text Available The seeds have different behaviors and tolerance to moisture loss. In order to determine desiccation tolerance in seeds of Bunchosia armenia (Cav. DC. (Ameixa-do-Pará was collected a lot of fruits of this species in Areia-PB. After the manual pulping the seeds were exposed to dry on paper towel, under laboratory conditions (22°C and 85% RH and greenhouse (24°C and 80% RH. Treatments consisted of sampling for the determination of water content and seedling emergence tests at baseline and after 24, 48, 72, 96 and 120 hours. The water content was measured by the oven method at 105 ± 3°C for 24 hours using four replicates of 10 seeds for each treatment. The seedling emergence test was conducted in a greenhouse with four replications of 25 seeds. The count of emergence has beenheld every two days, for a period of 35th day and calculated the IVE, length and dry mass of seedlings. The drying of the seeds of B.armenica, in both environments is viable for up to 24 hours; After 24 hours of drying the water content of seeds is reduced from 62.6% to 57%, a level considered critical to the desiccation of the seeds of Ameixa-do-Pará in both environments; Ameixa-do-Pará seeds nuts are dispersed with very high water content, and small reduction in physiological quality compromises their value, allowing the seeds to characterize this species as recalcitrant.As sementes têm comportamentos diferenciados quanto à tolerância à perda de umidade. Com o objetivo de determinar a tolerância à dessecação em sementes de Bunchosia armenica (Cav. DC. (ameixa-dopará foi coletado um lote de frutos desta espécie no município de Areia-PB. Após o despolpamento manual as sementes foram expostas para a secagem, sobre papel toalha, em condições de laboratório (22 ºC e 85% UR e casa de vegetação (24 ºC e 80% UR. Os tratamentos consistiram na amostragem para a determinação do teor de água e testes de emergência de plântulas no início do experimento e ap

  19. Isolation, synthesis and characterization of ω-TRTX-Cc1a, a novel tarantula venom peptide that selectively targets L-type Cav channels.

    Science.gov (United States)

    Klint, Julie K; Berecki, Géza; Durek, Thomas; Mobli, Mehdi; Knapp, Oliver; King, Glenn F; Adams, David J; Alewood, Paul F; Rash, Lachlan D

    2014-05-15

    Spider venoms are replete with peptidic ion channel modulators, often with novel subtype selectivity, making them a rich source of pharmacological tools and drug leads. In a search for subtype-selective blockers of voltage-gated calcium (CaV) channels, we isolated and characterized a novel 39-residue peptide, ω-TRTX-Cc1a (Cc1a), from the venom of the tarantula Citharischius crawshayi (now Pelinobius muticus). Cc1a is 67% identical to the spider toxin ω-TRTX-Hg1a, an inhibitor of CaV2.3 channels. We assembled Cc1a using a combination of Boc solid-phase peptide synthesis and native chemical ligation. Oxidative folding yielded two stable, slowly interconverting isomers. Cc1a preferentially inhibited Ba(2+) currents (IBa) mediated by L-type (CaV1.2 and CaV1.3) CaV channels heterologously expressed in Xenopus oocytes, with half-maximal inhibitory concentration (IC50) values of 825nM and 2.24μM, respectively. In rat dorsal root ganglion neurons, Cc1a inhibited IBa mediated by high voltage-activated CaV channels but did not affect low voltage-activated T-type CaV channels. Cc1a exhibited weak activity at NaV1.5 and NaV1.7 voltage-gated sodium (NaV) channels stably expressed in mammalian HEK or CHO cells, respectively. Experiments with modified Cc1a peptides, truncated at the N-terminus (ΔG1-E5) or C-terminus (ΔW35-V39), demonstrated that the N- and C-termini are important for voltage-gated ion channel modulation. We conclude that Cc1a represents a novel pharmacological tool for probing the structure and function of L-type CaV channels. PMID:24561180

  20. A CaV2.1 N-terminal fragment relieves the dominant-negative inhibition by an Episodic ataxia 2 mutant.

    Science.gov (United States)

    Dahimene, Shehrazade; Page, Karen M; Nieto-Rostro, Manuela; Pratt, Wendy S; D'Arco, Marianna; Dolphin, Annette C

    2016-09-01

    Episodic ataxia 2 (EA2) is an autosomal dominant disorder caused by mutations in the gene CACNA1A that encodes the pore-forming CaV2.1 calcium channel subunit. The majority of EA2 mutations reported so far are nonsense or deletion/insertion mutations predicted to form truncated proteins. Heterologous expression of wild-type CaV2.1, together with truncated constructs that mimic EA2 mutants, significantly suppressed wild-type calcium channel function, indicating that the truncated protein produces a dominant-negative effect (Jouvenceau et al., 2001; Page et al., 2004). A similar finding has been shown for CaV2.2 (Raghib et al., 2001). We show here that a highly conserved sequence in the cytoplasmic N-terminus is involved in this process, for both CaV2.1 and CaV2.2 channels. Additionally, we were able to interfere with the suppressive effect of an EA2 construct by mutating key N-terminal residues within it. We postulate that the N-terminus of the truncated channel plays an essential part in its interaction with the full-length CaV2.1, which prevents the correct folding of the wild-type channel. In agreement with this, we were able to disrupt the interaction between EA2 and the full length channel by co-expressing a free N-terminal peptide. PMID:27260834

  1. T-type calcium channel Cav3.2 deficient mice show elevated anxiety, impaired memory and reduced sensitivity to psychostimulants.

    Science.gov (United States)

    Gangarossa, Giuseppe; Laffray, Sophie; Bourinet, Emmanuel; Valjent, Emmanuel

    2014-01-01

    The fine-tuning of neuronal excitability relies on a tight control of Ca(2+) homeostasis. The low voltage-activated (LVA) T-type calcium channels (Cav3.1, Cav3.2 and Cav3.3 isoforms) play a critical role in regulating these processes. Despite their wide expression throughout the central nervous system, the implication of T-type Cav3.2 isoform in brain functions is still poorly characterized. Here, we investigate the effect of genetic ablation of this isoform in affective disorders, including anxiety, cognitive functions as well as sensitivity to drugs of abuse. Using a wide range of behavioral assays we show that genetic ablation of the cacna1h gene results in an anxiety-like phenotype, whereas novelty-induced locomotor activity is unaffected. Deletion of the T-type channel Cav3.2 also triggers impairment of hippocampus-dependent recognition memories. Acute and sensitized hyperlocomotion induced by d-amphetamine and cocaine are dramatically reduced in T-type Cav3.2 deficient mice. In addition, the administration of the T-type blocker TTA-A2 prevented the expression of locomotor sensitization observed in wildtype mice. In conclusion, our data reveal that physiological activity of this specific Ca(2+) channel is required for affective and cognitive behaviors. Moreover, our work highlights the interest of T-type channel blockers as therapeutic strategies to reverse drug-associated alterations. PMID:24672455

  2. T-type calcium channel Cav3.2 deficient mice show elevated anxiety, impaired memory and reduced sensitivity to psychostimulants.

    Directory of Open Access Journals (Sweden)

    Giuseppe Gangarossa

    2014-03-01

    Full Text Available The fine-tuning of neuronal excitability relies on a tight control of Ca2+ homeostasis. The low voltage-activated T-type calcium channels (Cav3.1, Cav3.2 and Cav3.3 isoforms play a critical role in regulating these processes. Despite their wide expression throughout the central nervous system, the implication of T-type Cav3.2 isoform in brain functions is still poorly characterized. Here we investigate the effect of genetic ablation of this isoform in affective disorders, including anxiety, cognitive functions as well as sensitivity to drugs of abuse. Using a wide range of behavioral assays we show that genetic ablation of the cacna1h gene results in an anxiety-like phenotype, whereas novelty-induced locomotor activity is unaffected. Deletion of the T-type channel Cav3.2 also triggers impairment of hippocampus-dependent recognition memories. Acute and sensitized hyperlocomotion induced by d-amphetamine and cocaine are dramatically reduced in T-type Cav3.2 deficient mice. In addition, the administration of the T-type blocker TTA-A2 prevented the expression of locomotor sensitization observed in wildtype mice. In conclusion, our data reveal that physiological activity of this specific Ca2+ channel is required for affective and cognitive behaviors. Moreover, our work highlights the interest of T-type channel blockers as therapeutic strategies to reverse drug-associated alterations.

  3. Observação da visita de morcegos (Chiroptera) às flores de Pseudobombax grandiflorum (Cav.) A. Robyns Observation of visit of bats (Chiroptera) to the flowers of Pseudobombax grandiflorum (Cav.) A. Robyns

    OpenAIRE

    Shirley S. Pereira da Silva; Adriano Lúcio Peracchi

    1995-01-01

    The visit of two species of bats to the flowers of Pseudobombax grandiflorum (Cav.) A. Robyns was observed in the Federal Rural University of Rio de Janeiro during the period of may and july of 1989. The flowers of this vegetal species present attributions in accordance to the Syndrome of Chiropterophily and receive the visit of Glossophaga soricina (Pallas. 1766) and Phyllostomus hastatus (Pallas. 1767). Glossophaga soricina has morfological caracteristes for nectarivory, Phyllostomus hastat...

  4. Thermal Hydrolysis Synthesis and Characterization of Monoclinic Metahewettite CaV6O16•3H2O

    Institute of Scientific and Technical Information of China (English)

    LI Lanjie; ZHENG Shili; WANG Shaona; DU Hao; ZHANG Yi

    2014-01-01

    Monoclinic metahewettite CaV6O16•3H2O has been fabricated via thermal hydrolysis of calcium vanadate (Ca10V6O25). High purity calcium vanadate precipitate, featuring column structure with surface area of 8.61 m2/g, can be obtained by reacting sodium orthovanadate (Na3VO4) with calcium oxide at 90℃for 2 h. By acidification of calcium vanadate in hot water at pH of 1.0-3.0, the monoclinic metahewettite crystals with uniform particle distribution, layered structure and nonporous structure can be fabricated. With the well crystallized layered structure, CaV6O16•3H2O may be a potential cathode material for secondary batteries as well as super capacitor materials.

  5. Karyotype Analysis of Malvaviscus arboreus Cav. (Malvaceae)%锦葵科悬铃花的核型分析

    Institute of Scientific and Technical Information of China (English)

    黄碧兰; 张玄兵; 范红霞

    2014-01-01

    The karyotype analysis of Malvaviscus arboreus Cav. was studied by using root-tip squashing method. The results showed that the number of chromosomes in somatic cells of Malvaviscus arboreus Cav. was 2n=2x=28, the relative length of chromosome was 2n=2x=28=2L+10M2+8M1+6S, and the karyotype asymmetry index was 0.5995. This species belongs to the 2B karyotypic symmetry class with a 20m+8sm karyotype formula. This study will provide the basis and reference for future work on karyotype analysis in genus of Malvaviscus arboreus Cav..%利用根尖压片法对悬铃花进行核型分析。结果表明:悬铃花体细胞的染色体数为2n=28,相对长度系数组成为:2L+10M2+8M1+6S,核型不对称系数为:59.95%;其核型公式为:20m+8sm,属2B 核型。为今后锦葵科悬铃花属植物核型分析研究提供一定的借鉴和依据。

  6. Cav1.4 L-Type Calcium Channels Contribute to Calpain Activation in Degenerating Photoreceptors of rd1 Mice

    Science.gov (United States)

    Schön, Christian; Paquet-Durand, François; Michalakis, Stylianos

    2016-01-01

    Retinitis pigmentosa is an inherited blinding disorder characterized by progressive degeneration and loss of photoreceptors. The exact mechanism of degeneration and cell death of photoreceptors is not known, but is thought to involve disturbed Ca2+—signaling. Ca2+ can enter the photoreceptor cell via outer segment cyclic nucleotide-gated (CNG) channels or synaptic Cav1.4 L-type voltage-gated calcium channels (VGCC). Previously, we have shown that genetic ablation of the Cngb1 gene encoding the B subunit of the rod CNG channel delays the fast progressing degeneration in the rd1 mutant mouse model of retinitis pigmentosa. In this study, we crossbred rd1 mice with the Cacna1f-deficient mouse lacking the Cav1.4 α1 subunit of the L-type VGCC. Longitudinal in vivo examinations of photoreceptor layer thickness by optical coherence tomography revealed a significant, but not sustained delay of retinal degeneration in Cacna1f x rd1 double mutant mice compared to rd1 mice. This was accompanied by a reduction of TUNEL positive cells in the early phase of rod degeneration. Remarkably, Cacna1f x rd1 double mutant mice displayed a strong decrease in the activation of the Ca2+-dependent protease calpain during photoreceptor loss. Our results show that genetic deletion of the synaptic Cav1.4 L-type VGCCs impairs calpain activation and leads to a short-term preservation of photoreceptors in the rd1 mouse. PMID:27270916

  7. Expression and Regulation of Cav3.2 T-Type Calcium Channels during Inflammatory Hyperalgesia in Mouse Dorsal Root Ganglion Neurons.

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    Masaya Watanabe

    Full Text Available The Cav3.2 isoform of the T-type calcium channel is expressed in primary sensory neurons of the dorsal root ganglion (DRG, and these channels contribute to nociceptive and neuropathic pain in rats. However, there are conflicting reports on the roles of these channels in pain processing in rats and mice. In addition, the function of T-type channels in persistent inflammatory hyperalgesia is poorly understood. We performed behavioral and comprehensive histochemical analyses to characterize Cav3.2-expressing DRG neurons and examined the regulation of T-type channels in DRGs from C57BL/6 mice with carrageenan-induced inflammatory hyperalgesia. We show that approximately 20% of mouse DRG neurons express Cav3.2 mRNA and protein. The size of the majority of Cav3.2-positive DRG neurons (69 ± 8% ranged from 300 to 700 μm2 in cross-sectional area and 20 to 30 μm in estimated diameter. These channels co-localized with either neurofilament-H (NF-H or peripherin. The peripherin-positive cells also overlapped with neurons that were positive for isolectin B4 (IB4 and calcitonin gene-related peptide (CGRP but were distinct from transient receptor potential vanilloid 1 (TRPV1-positive neurons during normal mouse states. In mice with carrageenan-induced inflammatory hyperalgesia, Cav3.2 channels, but not Cav3.1 or Cav3.3 channels, were upregulated in ipsilateral DRG neurons during the sub-acute phase. The increased Cav3.2 expression partially resulted from an increased number of Cav3.2-immunoreactive neurons; this increase in number was particularly significant for TRPV1-positive neurons. Finally, preceding and periodic intraplantar treatment with the T-type calcium channel blockers mibefradil and NNC 55-0396 markedly reduced and reversed mechanical hyperalgesia during the acute and sub-acute phases, respectively, in mice. These data suggest that Cav3.2 T-type channels participate in the development of inflammatory hyperalgesia, and this channel might play an

  8. Colocalization of insulin-like growth factor-1 receptor and T type Cav3.2 channel in dorsal root ganglia in chronic inflammatory pain mouse model.

    Science.gov (United States)

    Lin, Si-Fang; Yu, Xiao-Lu; Wang, Bing; Zhang, Ya-Jun; Sun, Yan-Gang; Liu, Xing-Jun

    2016-07-01

    Insulin-like growth factor-1 (IGF-1) is a neurotrophic factor and plays important roles in the nervous system. Increasing evidence supports that IGF-1 contributes to pain hypersensitivity through its insulin-like growth factor-1 receptor (IGF-1R) by activating IGF-1R/Akt or MAPK signaling pathways, whereas T-type Cav3.2 channel can facilitate and amplify pain signals originating from the sensory periphery. A recent study showed that activated IGF-1R can increase T-type Cav3.2 channel currents and further activate the G protein-dependent PKCα pathway to contribute to inflammatory pain sensitivity. However, the colocalization of IGF-1R and Cav3.2 in mouse dorsal root ganglion (DRG) under chronic inflammatory pain conditions remains elusive. In this study, we investigated changes in the expression of IGF-1R and the Cav3.2 channel, and their colocalization in mouse DRGs in chronic inflammatory pain condition (induced by complete Freund's adjuvant intraplanter injection) using real-time RT-PCR and immunohistochemistry approaches to confirm that Cav3.2 channel can mediate pain facilitation following IGF-1/IGF-1R signaling. We found that IGF-1R was expressed extensively in DRG neurons including small-, medium-, and large-sized neurons, whereas Cav3.2 channel was expressed exclusively in small-sized DRG neurons of naive mice. Expression of Cav3.2, but not IGF-1R, and colocalization of Cav3.2 and IGF-1R were increased in lumbar (L)4-L6 primary sensory neurons in DRGs of mice in chronic inflammatory pain. Moreover, the increased colocalization of IGF-1R and Cav3.2 is exclusively localized in small- and medium-sized primary sensory neurons. Our findings provided morphological evidence that T-type Cav3.2 channel, at least partially, mediates the pain facilitation of IGF-1/IGF-1R signaling in chronic inflammatory pain condition. PMID:27213932

  9. Spinal morphine but not ziconotide or gabapentin analgesia is affected by alternative splicing of voltage-gated calcium channel CaV2.2 pre-mRNA

    OpenAIRE

    Jiang, Yu-Qiu; Andrade, Arturo; Lipscombe, Diane

    2013-01-01

    Presynaptic voltage-gated calcium CaV2.2 channels play a privileged role in spinal level sensitization following peripheral nerve injury. Direct and indirect inhibitors of CaV2.2 channel activity in spinal dorsal horn are analgesic in chronic pain states. CaV2.2 channels represent a family of splice isoforms that are expressed in different combinations according to cell-type. A pair of mutually exclusive exons in the CaV2.2 encoding Cacna1b gene, e37a and e37b, differentially influence morphi...

  10. Alternative splicing at C terminus of Ca(V)1.4 calcium channel modulates calcium-dependent inactivation, activation potential, and current density.

    Science.gov (United States)

    Tan, Gregory Ming Yeong; Yu, Dejie; Wang, Juejin; Soong, Tuck Wah

    2012-01-01

    The Ca(V)1.4 voltage-gated calcium channel is predominantly expressed in the retina, and mutations to this channel have been associated with human congenital stationary night blindness type-2. The L-type Ca(V)1.4 channel displays distinct properties such as absence of calcium-dependent inactivation (CDI) and slow voltage-dependent inactivation (VDI) due to the presence of an autoinhibitory domain (inhibitor of CDI) in the distal C terminus. We hypothesized that native Ca(V)1.4 is subjected to extensive alternative splicing, much like the other voltage-gated calcium channels, and employed the transcript scanning method to identify alternatively spliced exons within the Ca(V)1.4 transcripts isolated from the human retina. In total, we identified 19 alternative splice variations, of which 16 variations have not been previously reported. Characterization of the C terminus alternatively spliced exons using whole-cell patch clamp electrophysiology revealed a splice variant that exhibits robust CDI. This splice variant arose from the splicing of a novel alternate exon (43*) that can be found in 13.6% of the full-length transcripts screened. Inclusion of exon 43* inserts a stop codon that truncates half the C terminus. The Ca(V)1.4 43* channel exhibited robust CDI, a larger current density, a hyperpolarized shift in activation potential by ∼10 mV, and a slower VDI. Through deletional experiments, we showed that the inhibitor of CDI was responsible for modulating channel activation and VDI, in addition to CDI. Calcium currents in the photoreceptors were observed to exhibit CDI and are more negatively activated as compared with currents elicited from heterologously expressed full-length Ca(V)1.4. Naturally occurring alternative splice variants may in part contribute to the properties of the native Ca(V)1.4 channels. PMID:22069316

  11. Functional and Preliminary Characterisation of Hydrocolloid from Tamarillo (Solanum betaceum Cav. Puree

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    Kharidah Muhammad

    2012-06-01

    Full Text Available Hydrocolloid from tamarillo (Solanum betaceum Cav. puree was extracted using water and characterised for the first time. Proximate compositions of the extracted hydrocolloid were also determined. Functional characteristics such as water-holding capacity, oil-holding capacity, emulsifying activity, emulsion stability, foaming capacity and stability of the hydrocolloid were evaluated in comparison to that of commercial hydrocolloids. Its functional groups and degree of esterification were determined using Fourier Transform Infrared (FT-IR spectroscopy. Monosaccharide profiling was done using reverse-phase high pressure liquid chromatography (RP-HPLC. Screening of various fruits for high hydrocolloid yield after water extraction resulted in tamarillo giving the highest yield. The yield on dry weight basis was 8.30%. The hydrocolloid constituted of 0.83% starch, 21.18% protein and 66.48% dietary fibre with 49.47% degree of esterification and the monosaccharides identified were mannose, ribose, rhamnose, galacturonic acid, glucose, galactose, xylose and arabinose. Higher oil-holding capacity, emulsifying activity and emulsion stability compared to commercial hydrocolloids propose its possible application as a food emulsifier and bile acid binder. Foaming capacity of 32.19% and good foam stabilisation (79.36% of initial foam volume after 2 h of foam formation suggest its promising application in frothy beverages and other foam based food products. These findings suggest that water-extracted tamarillo hydrocolloid can be utilised as an alternative to low methoxyl pectin.

  12. Population genetic structure of Sisyrinchium micranthum Cav. (Iridaceae) in Itapuã State Park, Southern Brazil.

    Science.gov (United States)

    Tacuatiá, Luana Olinda; Eggers, Lilian; Kaltchuk-Santos, Eliane; Souza-Chies, Tatiana T

    2012-01-01

    Sisyrinchium micranthum Cav. is a member of the family Iridaceae, which is distributed over the American continent. In Brazil, this species is found, not only in disturbed areas and coastal regions, but is also very common in urban centers, such as public parks, during the spring. Chromosome counts for North American specimens are 2n = 32 and 2n = 48, whereas in southern Brazil, there is a polyploidy series with three chromosome numbers, 2n = 16, 2n = 32, and 2n = 48. Population analyses using DNA molecular markers are inexistent for this species, in spite of its wide distribution and morphological variation. To study the genetic population structure of S. micranthum, five natural populations were accessed in a conservation park within the Atlantic Rain Forest Biome in southern Brazil. Here, the chromosome numbers 2n = 16 and 2n = 48 had already been described. Molecular analysis showed that the populations are highly structured with low gene flow among them. The population with 2n = 48 was genetically less variable than and distinct from the other populations. Population genetics in relation to cytogenetic data provided new insights regarding the genetic diversification and mating system of S. micranthum. PMID:22481881

  13. Agronomical and chemical characterisation of Thymbra capitata (L.) Cav. biotypes from Sicily, Italy.

    Science.gov (United States)

    Tuttolomondo, Teresa; Dugo, Giacomo; Leto, Claudio; Cicero, Nicola; Tropea, Alessia; Virga, Giuseppe; Leone, Raffaele; Licata, Mario; La Bella, Salvatore

    2015-01-01

    In this study, the agronomical and chemical characterisation of 13 Sicilian biotypes of Thymbra capitata (L.) Cav., grown under the same agricultural and environmental condition, are reported. The main morpho-productive parameters and quali-quantitative profile of essential oils (EOs) were determined. The EOs were analysed by gas chromatography-flame ionisation detector and gas chromatography-mass spectrometry. Hierarchical cluster analysis and principal component analysis statistical methods were used to group biotypes according to the EOs chemical composition. The EO yield ranged between 4.6 and 8.1 (v/w). A total of 38 EO compounds have been identified. The compounds mostly represented were α-pinene, myrcene, α-terpinene, p-cymene, γ-terpinene, borneol, carvacrol and β-caryophyllene. In all biotypes, the carvacrol (67.4-79.5%) was the main compound, confirming that T. capitata is a carvacrol chemotype. The results showed that all Sicilian Thymbra biotypes have a good adaptation to the climatic conditions of the test environment. PMID:25600887

  14. Antifungal activity of Zuccagnia punctata Cav.: evidence for the mechanism of action.

    Science.gov (United States)

    Svetaz, Laura; Agüero, María Belén; Alvarez, Sandra; Luna, Lorena; Feresin, Gabriela; Derita, Marcos; Tapia, Alejandro; Zacchino, Susana

    2007-08-01

    Petroleum ether and dichloromethane extracts of fruits, aerial parts and exudate of Zuccagnia punctata Cav. (Fabaceae) showed moderate antifungal activities against the yeasts C. albicans, S. cerevisiae and C. neoformans (MICs: 62.5 - 250 microg/mL) and very strong antifungal activities against the dermatophytes M. gypseum, T. rubrum and T. mentagrophytes (MICs: 8 - 16 microg/mL) thus supporting the ethnopharmacological use of this plant. Antifungal activity-directed fractionation of active extracts by using bioautography led to the isolation of 2',4'-dihydroxy-3'-methoxychalcone (1) and 2',4'-dihydroxychalcone (2) as the compounds responsible for the antifungal activity. Second-order studies included MIC (80), MIC (50) and MFC of both chalcones in an extended panel of clinical isolates of the most sensitive fungi, and also comprised a series of targeted assays. They showed that the most active chalcone 2 is fungicidal rather than fungistatic, does not disrupt the fungal membranes up to 4 x MFC and does not act by inhibiting the fungal cell wall. So, 2',4'-dihydroxychalcone would act by a different mechanism of action than the antifungal drugs in current clinical use, such as amphotericin B, azoles or echinocandins, and thus appears to be very promising as a novel antifungal agent. PMID:17628836

  15. Argentinean propolis from Zuccagnia punctata Cav. (Caesalpinieae) exudates: phytochemical characterization and antifungal activity.

    Science.gov (United States)

    Agüero, María Belén; Gonzalez, Mariela; Lima, Beatriz; Svetaz, Laura; Sánchez, Marianela; Zacchino, Susana; Feresin, Gabriela Egly; Schmeda-Hirschmann, Guillermo; Palermo, Jorge; Wunderlin, Daniel; Tapia, Alejandro

    2010-01-13

    This paper reports the in vitro antifungal activity of propolis extracts from the province of Tucuman (Argentina) as well as the identification of their main antifungal compounds and botanical origin. The antifungal activity was determined by the microdilution technique, using reference microorganisms and clinical isolates. All dermatophytes and yeasts tested were strongly inhibited by different propolis extracts (MICs between 16 and 125 microg mL(-1)). The most susceptible species were Microsporum gypseum, Trichophyton mentagrophytes, and Trichophyton rubrum. The main bioactive compounds were 2',4'-dihydroxy-3'-methoxychalcone 2 and 2',4'-dihydroxychalcone 3. Both displayed strong activity against clinical isolates of T. rubrum and T. mentagrophytes (MICs and MFCs between 1.9 and 2.9 microg mL(-1)). Additionally, galangin 5, pinocembrin 6, and 7-hydroxy-8-methoxyflavanone 9 were isolated from propolis samples and Zuccagnia punctata exudates, showing moderate antifungal activity. This is the first study matching the chemical profile of Z. punctata Cav. exudates with their corresponding propolis, giving strong evidence on the botanical origin of the studied propolis. PMID:19916546

  16. Observação da visita de morcegos (Chiroptera às flores de Pseudobombax grandiflorum (Cav. A. Robyns Observation of visit of bats (Chiroptera to the flowers of Pseudobombax grandiflorum (Cav. A. Robyns

    Directory of Open Access Journals (Sweden)

    Shirley S. Pereira da Silva

    1995-01-01

    Full Text Available The visit of two species of bats to the flowers of Pseudobombax grandiflorum (Cav. A. Robyns was observed in the Federal Rural University of Rio de Janeiro during the period of may and july of 1989. The flowers of this vegetal species present attributions in accordance to the Syndrome of Chiropterophily and receive the visit of Glossophaga soricina (Pallas. 1766 and Phyllostomus hastatus (Pallas. 1767. Glossophaga soricina has morfological caracteristes for nectarivory, Phyllostomus hastatus is omnivorous and starts its visils soon after the anthesis. That bats present landing behaviour during the visit.

  17. 基于CAV424的电容式液位传感器信号调理电路研究%The Research for a Signal Conditioning Circuits of Capacitive Level Sensor based on Chip CAV424

    Institute of Scientific and Technical Information of China (English)

    唐正茂

    2008-01-01

    介绍了一种电容式液位传感器信号调理电路设计的新方法,通过利用单片的电容/电压信号转换芯片CAV424实现了对电容量的简单、准确测量,同时通过单片机进行智能补偿以实现测量的高精度.该传感器具有测量精度高、可靠性好和较高的性价比等优点.

  18. Effects of Amyloid-β Peptides on Voltage-Gated L-Type CaV1.2 and CaV1.3 Ca2+ Channels

    OpenAIRE

    Kim, Sunoh; Rhim, Hyewhon

    2011-01-01

    Overload of intracellular Ca2+ has been implicated in the pathogenesis of neuronal disorders, such as Alzheimer’s disease. Various mechanisms produce abnormalities in intracellular Ca2+ homeostasis systems. L-type Ca2+ channels have been known to be closely involved in the mechanisms underlying the neurodegenerative properties of amyloid-β (Aβ) peptides. However, most studies of L-type Ca2+ channels in Aβ-related mechanisms have been limited to CaV1.2, and surprisingly little is known about t...

  19. Antidepressant-like effect of Tagetes lucida Cav. extract in rats: involvement of the serotonergic system.

    Science.gov (United States)

    Gabriela, Guadarrama-Cruz; Javier, Alarcón-Aguilar Francisco; Elisa, Vega-Avila; Gonzalo, Vázquez-Palacios; Herlinda, Bonilla-Jaime

    2012-01-01

    It has been demonstrated that the decoction of the aerial parts of Tagetes lucida Cav. produces an antidepressant effect during the forced swimming test (FST) in rats. The aim of this study was to evaluate the effect of different organic extracts and one aqueous extract of the aerial parts of T. lucida on the FST. In addition, the possible involvement of the serotonergic system in the antidepressant-like effect of T. lucida in the FST was evaluated, as was its potential toxicological effect. The different extracts of T. lucida (methanol, hexane, dichloromethane and aqueous, 10 and 50 mg/kg), as well as fluoxetine (FLX, 5 mg/kg), were administered per os (p.o.) to rats for 14 days. All animals were subjected to the FST. Only the aqueous extract of T. lucida at a dose of 50 mg/kg significantly reduced immobility behavior and increased swimming in the FST, similar to FLX. Later, the aqueous extract of T. lucida (50mg/kg) was administered for 1, 7 and 14 days. An antidepressant effect was observed after 7 days of treatment. To evaluate the participation of the serotoninergic system, the animals were pretreated with PCPA, an inhibitor of serotonin synthesis (100 mg/kg/day for 4 consecutive days). The animals were treated with the aqueous extract of T. lucida (50 mg/kg) and FLX (5 mg/kg) 24 h after the final injection and were then subjected to the FST. Pretreatment with PCPA inhibited the antidepressant effect of both T. lucida and FLX. Finally, T. lucida was administered p.o. and intraperitoneal route to evaluate its acute toxicological effect. The aqueous extract of T. lucida, administered p.o., did not produce lethality or any significant changes in behavior. In conclusion, the aqueous extract of T. lucida manifested an antidepressant-like effect in the FST mediated by the serotonergic system, with no adverse effects when administered p.o. PMID:22809029

  20. Investigation of biological activity of polar extracts isolated from Phlomis crinita Cav ssp. mauritanica Munby.

    Science.gov (United States)

    Limem-Ben Amor, Ilef; Skandrani, Ines; Boubaker, Jihed; Ben Sghaïer, Mohamed; Neffati, Aicha; Bhouri, Wissem; Bouhlel, Ines; Chouchane, Nabil; Kilani, Soumaya; Guedon, Emmanuel; Ghoul, Mohamed; Ghedira, Kamel; Chekir-Ghedira, Leila

    2009-01-01

    The lyophilized infusion, the methanol, the ethyl acetate, and the total oligomer flavonoid (TOF)-enriched extracts prepared from the dried leaves of Phlomis crinita Cav. ssp. mauritanica Munby were investigated for the contents of flavonoids, tannins, coumarines and steroids. Antibacterial activity was investigated toward five bacterial strains. An inhibitory effect was observed against Staphyllococcus aureus and Enterococcus feacalis, and the minimal inhibitory concentrations ranged from 2.5 to 5 mg/mL of extract. The tested extracts exhibit an important free radical scavenging activity toward the 1,1-diphenyl 2-picrylhydrazyl (DPPH) free radical; with IC(50) values of 30.5, 6, 32, and 31.5 microg/mL, respectively, in the presence of lyophilized infusion, the TOF, the methanol, and the ethyl acetate extracts. Genotoxic and antigenotoxic properties of the different extracts were studied by using the SOS chromotest with Escherichia coli PQ37. The lyophilized infusion and TOF extracts obtained from P. crinita ssp. mauritanica showed no genotoxicity, whereas methanol and ethyl acetate extracts are considered as marginally genotoxic. On the other hand, we showed that each extract inhibited the mutagenicity induced by aflatoxin B1 (AFB1) (10 microg/assay) and nifuroxazide (NF) (10 microg/assay). The ethyl acetate extract showed the strongest level of protection toward the genotoxicity induced by both directly and indirectly genotoxic NF and AFB1. These tests proved that the lyophilized infusion possesses an antiradical activity likewise, it showed no genotoxic effect; that is why we choose this extract to assess its antiulcerogenic activity by using an ethanol-induced ulcerogenesis model in the rat. This test demonstrates that 300 mg/kg of a P. crinita ssp. mauritanica lyophilized infusion was more effective than the reference compound, cimetidine. PMID:19514937

  1. Evaluation of genotoxic and antigenotoxic effects of hydroalcoholic extracts of Zuccagnia punctata Cav.

    Science.gov (United States)

    Zampini, Iris Catiana; Villarini, Milena; Moretti, Massimo; Dominici, Luca; Isla, María Inés

    2008-01-17

    Zuccagnia punctata Cav. (Fabaceae), a widely used plant species in Argentine folk medicine, has been shown to have a broad spectrum of antibacterial, antifungal, antioxidant and cytoprotective activities. In this study, the hydroalcoholic extract of Zuccagnia punctata and 2',4'-dihydroxychalcone isolated from it were investigated for genotoxicity/antigenotoxicity in the in vitro comet assay test on human hepatoma HepG2 cells. No acute toxicity of the extract could be determined. HepG2 cells were treated with three different concentrations (2.5, 5.0 and 10.0 microg/mL) or 2',4'-dihydroxychalcone (0.01, 0.10 and 1.00 microg/mL). To explore the potential mechanisms of action, two approaches were followed: co-treatment with 4-nitroquinoline-N-oxyde (4-NQO), a direct genotoxic compound, and a pre-treatment protocol with benzo[a]pyrene (B[a]P), an indirect genotoxic compound. The natural products neither affected cell viability nor induced DNA damage in the concentration range tested. Zuccagnia punctata tinctures were able to diminish the DNA damage induced in HepG2 cells by 4-NQO and B[a]P in 31% and 10%, respectively at 10 microg/mL. Pre-treatment of HepG2 cells with 2',4'-dihydroxychalcone was highly effective in decreasing B[a]P-induced DNA damage at a statistically significant level, with an almost clear dose-response relationship. The inhibition values were 28.2-43.9% for the tested concentrations of 0.01-1 microg/mL, respectively. The results clearly indicate that the phytoextract from Zuccagnia punctata, under the experimental conditions tested, is not genotoxic and that 2',4'-dihydroxychalcone contributes to a high degree to the antigenotoxic effects of Zuccagnia punctata tincture. PMID:18023546

  2. Conservação e vigor de sementes de pau-de-balsa Ochroma pyramidale (Cav. ex Lam.) Urban.

    OpenAIRE

    Pinto, Antonio Moçambite

    2013-01-01

    O experimento foi conduzido nos Laboratórios de Sementes do Departamento de Silvicultura e Manejo da Universidade Federal do Paraná e do Centro Nacional de Pesquisa de Florestas da Embrapa e teve como objetivo definir um método para o armazenamento de sementes de Ochroma pyramidale (Cav. ex Lam.) Urbano As sementes foram embaladas em sacos de papel tipo kraft e sacos de plástico ( 0,10 mm ) e armazenadas em ambiente de laboratório (22°C e 65% UR), câmara úmida (5°C e 86% UR) e câmara seca (15...

  3. EFEITO DE DIFERENTES SUBSTRATOS NA GERMINAÇÃO DE Ochroma pyramidale (CAV. EX LAM.) URB. (BOMBACACEAE)

    OpenAIRE

    Breno Pinto Rayol; Fabrízia de Oliveira Alvino

    2007-01-01

    O objetivo deste trabalho foi analisar o efeito de diferentes substratos na germinação de Ochroma pyramidale (Cav. ex Lam.) Urb. (Bombacaceae). O experimento foi conduzido em delineamento inteiramente casualizado com três tratamentos (substratos) e quatro repetições de 25 sementes. Foram testados os seguintes substratos: areia + vermiculita (1:1); vermiculita e, areia + serragem (1:1). Foi avaliado o efeito dos substratos pelo percentual de germinação das sementes, tempo médio de germinação e...

  4. Actividad repelente de formulaciones del aceite esencial de Melaleuca quinquenervia (Cav.) S.T. Blake (Myrtales: Myrtaceae) en mosquitos

    OpenAIRE

    Leyva, Maureen; Castex, Mayda; Montada, Domingo; Quintana, Felipe; Lezcano, Diosanys; Marquetti Fernández, María del Carmen; Companioni, Ariamys; Anaya, Jorge; González, Iván

    2012-01-01

    En este trabajo nos trazamos como objetivo evaluar la actividad repelente de diferentes formulaciones del aceite de Melaleuca quinquinervia Cav S.T. Blake en condiciones de laboratorio y terreno. Se prepararon formulaciones etanólicas, hidroalcohólicas, hidrofílicas y en base oleosa con aceite mineral y dipropilenglicol. Se trató el antebrazo con 1 ml de repelente y la mano fue cubierta con un guante de goma. El antebrazo fue expuesto por 3 minutos cada media hora. Se obt...

  5. Two Components of Voltage-Dependent Inactivation in Cav1.2 Channels Revealed by Its Gating Currents

    OpenAIRE

    Ferreira, Gonzalo; Ríos, Eduardo; Reyes, Nicolás

    2003-01-01

    Voltage-dependent inactivation (VDI) was studied through its effects on the voltage sensor in Cav1.2 channels expressed in tsA 201 cells. Two kinetically distinct phases of VDI in onset and recovery suggest the presence of dual VDI processes. Upon increasing duration of conditioning depolarizations, the half-distribution potential (V1/2) of intramembranous mobile charge was negatively shifted as a sum of two exponential terms, with time constants 0.5 s and 4 s, and relative amplitudes near 50...

  6. Comparison of a Constant Air Volume (CAV) and a Demand Controlled Ventilation (DCV) System in a Residential Building

    DEFF Research Database (Denmark)

    Mortensen, Dorthe Kragsig; Nielsen, Toke Rammer; Topp, Claus

    The aim of this paper was to compare the indoor climate and the energy performance of a Constant Air Volume (CAV) system of 0.5h-1 with a Demand Controlled Ventilation (DCV) system controlled by occupancy and relative humidity for a studio apartment. Furthermore the impact of building materials...... hygroscopic properties on indoor climate and energy consumption was investigated for the two systems. Dynamic simulations of the studio apartment were carried out in the program WUFI+ with weather data from Copenhagen including outside temperature end relative humidity. For the non-hygroscopic case it was...

  7. Expression of calcium channel CaV1.3 in cat spinal cord: light and electron microscopic immunohistochemical study

    DEFF Research Database (Denmark)

    Zhang, Mengliang; Møller, Morten; Broman, Jonas;

    2008-01-01

    in the cat spinal cord by light and electron microscopic immunohistochemistry. The results show that Ca(V)1.3-like immunoreactivity is widely distributed in all segments of the spinal cord but that the distribution in the different laminae of the spinal gray matter varies, with the highest density of labeled...... associated with the rough endoplasmic reticulum but some also with the plasma membrane. In dendrites, they were associated with both intracellular organelles, including microtubules and microchondria, and the plasma membrane. These results indicate that significant proportions of the neurons in cat spinal...

  8. Suppression of peripheral pain by blockade of Cav2.2 channels in nociceptors induces RANKL and impairs recovery from inflammatory arthritis

    Science.gov (United States)

    Baddack, Uta; Frahm, Silke; Antolin-Fontes, Beatriz; Grobe, Jenny; Lipp, Martin; Müller, Gerd; Ibañez-Tallon, Ines

    2015-01-01

    Objective A hallmark of rheumatoid arthritis (RA) is the chronic pain that accompanies the inflammation and joint deformation. Patients with RA rate pain relief with highest priority, however, few studies have addressed the efficacy and safety of therapies directed specifically towards pain pathways. The conotoxin MVIIA (Prialt/Ziconotide) is used in humans to alleviate persistent pain syndromes because it specifically blocks the CaV2.2 voltage-gated calcium channel, which mediates the release of neurotransmitters and proinflammatory mediators from peripheral nociceptor nerve terminals. The purpose of this study was to investigate whether block of CaV2.2 can suppress arthritic pain, and to examine the progression of induced arthritis during persistent CaV2.2 blockade. Methods Transgenic mice (Tg-MVIIA) expressing a membrane-tethered form of the ω-conotoxin MVIIA, under the control of a nociceptor-specific gene, were employed. These mice were subjected to unilateral induction of joint inflammation using the Antigen- and Collagen-Induced Arthritis (ACIA) model. Results We observed that CaV2.2-blockade mediated by t-MVIIA effectively suppressed arthritis-induced pain; however, in contrast to their wild-type littermates, which ultimately regained use of their injured joint as inflammation subsides, Tg-MVIIA mice showed continued inflammation with an up-regulation of the osteoclast activator RANKL and concomitant joint and bone destruction. Conclusion Altogether, our results indicate that alleviation of peripheral pain by blockade of CaV2.2- mediated calcium influx and signaling in nociceptor sensory neurons, impairs recovery from induced arthritis and point to the potentially devastating effects of using CaV2.2 channel blockers as analgesics during inflammation. PMID:25733371

  9. Inhibitory gene expression of the Cav3.1 T-type calcium channel to improve neuronal injury induced by lidocaine hydrochloride.

    Science.gov (United States)

    Wen, Xianjie; Xu, Shiyuan; Zhang, Qingguo; Li, Xiaohong; Liang, Hua; Yang, Chenxiang; Wang, Hanbing; Liu, Hongzhen

    2016-03-15

    Cav3.1 is a low-voltage-activated (LVA) calcium channel that plays a key role in regulating intracellular calcium ion levels. In this study, we observed the effects of lidocaine hydrochloride on the pshRNA-CACNA1G-SH-SY5Y cells that silenced Cav3.1 mRNA by RNA interference, and investigated the roles of p38 MAPK in these effects. We constructed the pNC-puro-CACNA1G-SH-SY5Y cells and pshRNA-CACNA1G -SH-SY5Y cells by the RNA interference. All the cells were cultured with or without 10mM lidocaine hydrochloride for 24h. The cell morphology, cell viability, Cav3.1 and p38 protein expression, cell apoptosis rate and intracellular calcium ion concentration were detected. We found that all cells treated with 10mM lidocaine hydrochloride for 24h showed cellular rounding, axonal regression, and cellular floating. Compared with the cells in SH-SY5Y+Lido group and NC+Lido group, those in the RNAi+Lido group showed similar changes, but of smaller magnitude. Additionally, following lidocaine hydrochloride all cells displayed increased Cav3.1 and p38 MAPK protein, apoptosis rate, and intracellular calcium ion levels; however,these changes in the RNAi+Lido group were less pronounced than in the SH-SY5Y+Lido and NC+Lido groups. The cell viability decreased following lidocaine hydrochloride treatment, but viability of the cells in the RNAi+Lido group was higher than in the SH-SY5Y+Lido and NC+Lido groups. The results showed that Cav3.1 may be involved in neuronal injury induced by lidocaine hydrochloride and that p38 MAPK phosphorylation was reduced upon Cav3.1 gene silencing. PMID:26852957

  10. Inhibitory and excitatory axon terminals share a common nano-architecture of their Cav2.1 (P/Q-type Ca2+ channels

    Directory of Open Access Journals (Sweden)

    Daniel eAlthof

    2015-08-01

    Full Text Available Tuning of the time course and strength of inhibitory and excitatory neurotransmitter release is fundamental for the precise operation of cortical network activity and is controlled by Ca2+ influx into presynaptic terminals through the high voltage-activated P/Q-type Ca2+ (Cav2.1 channels. Proper channel-mediated Ca2+-signaling critically depends on the topographical arrangement of the channels in the presynaptic membrane. Here, we used high-resolution SDS-digested freeze-fracture replica immunoelectron microscopy together with automatized computational analysis of Cav2.1 immunogold labeling to determine the precise subcellular organization of Cav2.1 channels in both inhibitory and excitatory terminals. Immunoparticles labeling the pore-forming α1 subunit of Cav2.1 channels were enriched over the active zone of the boutons with the number of channels (3-62 linearly correlated with the area of the synaptic membrane. Detailed analysis showed that Cav2.1 channels are non-uniformly distributed over the presynaptic membrane specialization where they are arranged in clusters of an average 5 channels per cluster covering a mean area with a diameter of about 70 nm. Importantly, clustered arrangement and cluster properties did not show any significant difference between GABAergic and glutamatergic terminals. Our data demonstrate a common nano-architecture of Cav2.1 channels in inhibitory and excitatory boutons in CA1 hippocampal neurons suggesting that the cluster arrangement is crucial for the precise release of transmitters from the axonal boutons.

  11. Inhibitory and excitatory axon terminals share a common nano-architecture of their Cav2.1 (P/Q-type) Ca(2+) channels.

    Science.gov (United States)

    Althof, Daniel; Baehrens, David; Watanabe, Masahiko; Suzuki, Noboru; Fakler, Bernd; Kulik, Ákos

    2015-01-01

    Tuning of the time course and strength of inhibitory and excitatory neurotransmitter release is fundamental for the precise operation of cortical network activity and is controlled by Ca(2+) influx into presynaptic terminals through the high voltage-activated P/Q-type Ca(2+) (Cav2.1) channels. Proper channel-mediated Ca(2+)-signaling critically depends on the topographical arrangement of the channels in the presynaptic membrane. Here, we used high-resolution SDS-digested freeze-fracture replica immunoelectron microscopy together with automatized computational analysis of Cav2.1 immunogold labeling to determine the precise subcellular organization of Cav2.1 channels in both inhibitory and excitatory terminals. Immunoparticles labeling the pore-forming α1 subunit of Cav2.1 channels were enriched over the active zone of the boutons with the number of channels (3-62) correlated with the area of the synaptic membrane. Detailed analysis showed that Cav2.1 channels are non-uniformly distributed over the presynaptic membrane specialization where they are arranged in clusters of an average five channels per cluster covering a mean area with a diameter of about 70 nm. Importantly, clustered arrangement and cluster properties did not show any significant difference between GABAergic and glutamatergic terminals. Our data demonstrate a common nano-architecture of Cav2.1 channels in inhibitory and excitatory boutons in stratum radiatum of the hippocampal CA1 area suggesting that the cluster arrangement is crucial for the precise release of transmitters from the axonal boutons. PMID:26321916

  12. Heparin/heparan sulfates bind to and modulate neuronal L-type (Cav1.2) voltage-dependent Ca2+ channels

    DEFF Research Database (Denmark)

    Garau, Gianpiero; Magotti, Paola; Heine, Martin;

    2015-01-01

    Our previous studies revealed that L-type voltage-dependent Ca2+ channels (Cav1.2 L-VDCCs) are modulated by the neural extracellular matrix backbone, polyanionic glycan hyaluronic acid. Here we used isothermal titration calorimetry and screened a set of peptides derived from the extracellular...... domains of Cav1.2α1 to identify putative binding sites between the channel and hyaluronic acid or another class of polyanionic glycans, such as heparin/heparan sulfates. None of the tested peptides showed detectable interaction with hyaluronic acid, but two peptides derived from the first pore...

  13. Value of the first post-transplant biopsy for predicting long-term cardiac allograft vasculopathy (CAV and graft failure in heart transplant patients.

    Directory of Open Access Journals (Sweden)

    Carlos A Labarrere

    Full Text Available BACKGROUND: Cardiac allograft vasculopathy (CAV is the principal cause of long-term graft failure following heart transplantation. Early identification of patients at risk of CAV is essential to target invasive follow-up procedures more effectively and to establish appropriate therapies. We evaluated the prognostic value of the first heart biopsy (median: 9 days post-transplant versus all biopsies obtained within the first three months for the prediction of CAV and graft failure due to CAV. METHODS AND FINDINGS: In a prospective cohort study, we developed multivariate regression models evaluating markers of atherothrombosis (fibrin, antithrombin and tissue plasminogen activator [tPA] and endothelial activation (intercellular adhesion molecule-1 in serial biopsies obtained during the first three months post-transplantation from 172 patients (median follow-up = 6.3 years; min = 0.37 years, max = 16.3 years. Presence of fibrin was the dominant predictor in first-biopsy models (Odds Ratio [OR] for one- and 10-year graft failure due to CAV = 38.70, p = 0.002, 95% CI = 4.00-374.77; and 3.99, p = 0.005, 95% CI = 1.53-10.40 and loss of tPA was predominant in three-month models (OR for one- and 10-year graft failure due to CAV = 1.81, p = 0.025, 95% CI = 1.08-3.03; and 1.31, p = 0.001, 95% CI = 1.12-1.55. First-biopsy and three-month models had similar predictive and discriminative accuracy and were comparable in their capacities to correctly classify patient outcomes, with the exception of 10-year graft failure due to CAV in which the three-month model was more predictive. Both models had particularly high negative predictive values (e.g., First-biopsy vs. three-month models: 99% vs. 100% at 1-year and 96% vs. 95% at 10-years. CONCLUSIONS: Patients with absence of fibrin in the first biopsy and persistence of normal tPA in subsequent biopsies rarely develop CAV or graft failure during the next 10 years and potentially could be monitored less invasively

  14. Simultaneous Determination of Nine Compounds in Bergenia purpurascens by HPLC-MS%液相色谱-质谱联用法同时测定岩白菜中熊果苷等9种成分的含量

    Institute of Scientific and Technical Information of China (English)

    任艳平; 曹亮; 常路; 支旭然; 苑霖; 生宁; 张兰桐

    2013-01-01

    目的 采用HPLC-MS测定岩白菜中熊果苷、没食子酸、岩白菜素、原儿茶酸、绿原酸、儿茶素、表儿茶素、表儿茶素没食子酸酯和阿魏酸9种成分的含量.方法采用Diamonsil C18柱(4.6 nn×150 mm,5μm);柱温为室温;流动相为甲醇体积分数为-0.2‰甲酸水,以0.8 mL· min-1的流速进行梯度洗脱,进样量10 μL;质谱条件:采用电喷雾离子源进行负离子模式检测,多反应监测模式(MRM)用于定量测定.结果 岩白菜中9种成分熊果苷、没食子酸、岩白菜素、原儿茶酸、绿原酸、儿茶素、表儿茶素、表儿茶素没食子酸酯和阿魏酸能完全分离;峰面积与浓度呈良好的线性关系.平均回收率(n=9)分别为100.2%,100.0%,99.8%,99.9%,97.9%,101.2%,99.8%,101.7%,102.4%. RSD分别为0.80%,3.06%,2.72%,3.28%,3.58%,1.44%,0.95%,0.96%,0.48%.结论 该方法简便,准确,重现性好,专属性高,可用于岩白菜中多种成分的含量测定和质量控制.%OBJECTIVE To develop a method for the determination of arbutin, gallic acid, bergenin, protocatechuic acid, chlo-rogenic acid, catechin, epicatechin, epicatechin gallate, and ferulic acid in Bergenia purpurascens by HPLC-MS/MS. METHODS The samples were separated on a Diamonsil C18 column (4. 6 mm × 150 mm, 5 μm) by gradient elution using methanol and 0. 2‰ ( V/V) formic acid aqueous solution as the mobile phase at a flow rate of 0. 8 mL · min-1. The column temperature was room temperature. Multiple-reaction monitoring (MRM) scanning was employed for quantification with switching electrospray ion source polarity in negative mode. RESULTS Complete separation was achieved within 10 min for the nine compounds (arbutin, gallic acid, bergenin, protocatechuic acid, chlorogenic acid, catechin, epicatechin, epicatechin gallate, ferulic acid). The nine regression equations showed linear relationships between peak areas and contents of each compound. The average

  15. Cadmium tolerance and accumulation of Althaea rosea Cav. and its potential as a hyperaccumulator under chemical enhancement.

    Science.gov (United States)

    Liu, Jia Nv; Zhou, Qi Xing; Wang, Song; Sun, Ting

    2009-02-01

    The role of ornamental plants has drawn much attention as the urban pollution levels exacerbate. Althaea rosea Cav. had showed its strong tolerance and accumulation ability of Cd in our previous work, thus, the effects of ethylenediamine triacetic acid (EDTA), ethylenegluatarotriacetic acid (EGTA) and sodium dodecyl sulfate (SDS) on its Cd phytoremediation capacity were further investigated in this work. It reconfirmed that the species had strong tolerance and accumulation ability of Cd. Particularly, the species can be regarded as a potential Cd-hyperaccumulator through applying chemical agents. However, different chelators and surfactants had great differences in affecting hyperaccumulating characteristics of the species. EGTA and SDS could not only increase the dry biomass of the plants, but also promote Cd accumulation in shoots and roots. On the contrary, EDTA was toxic to the species by restraining the growth of plants, although it could promote Cd accumulation in shoots and roots of the plants to a certain extent. Thus, EGTA and SDS were effective in enhancing phytoremediation with Althaea rosea Cav. for Cd contaminated soils, while EDTA is ineffective in this regard. PMID:18259884

  16. Voltage-independent inhibition of Cav2.2 channels is delimited to a specific region of the membrane potential in rat SCG neurons

    Institute of Scientific and Technical Information of China (English)

    Oscar Vivas; Isabel Arenas; David E.García

    2012-01-01

    Neurotransmitters and hormones regulate Cav2.2 channels through a voltage-independent pathway which is not well understood.It has been suggested that this voltageindependent inhibition is constant at all membrane voltages.However,changes in the percent of voltageindependent inhibition of Cav2.2 have not been tested within a physiological voltage range.Here,we used a double-pulse protocol to isolate the voltage-independent inhibition of Cav2.2 channels induced by noradrenaline in rat superior cervical ganglion neurons.To assess changes in the percent of the voltage-independent inhibition,the activation voltage of the channels was tested between -40 and +40 mV.We found that the percent of voltage-independent inhibition induced by noradrenaline changed with the activation voltage used.In addition,voltage-independent inhibition induced by oxo-M,a muscarinic agonist,exhibited thesame dependence on activation voltage,which supports that this pattern is not exclusive for adrenergic activation.Our results suggested that voltage-independent inhibition of Cav2.2 channels depends on the activation voltage of the channel in a physiological voltage range.This may have relevant implications in the understanding of the mechanism involved in voltage-independent inhibition.

  17. An autism-associated mutation in CaV1.3 channels has opposing effects on voltage- and Ca2+-dependent regulation

    Science.gov (United States)

    Limpitikul, Worawan B.; Dick, Ivy E.; Ben-Johny, Manu; Yue, David T.

    2016-01-01

    CaV1.3 channels are a major class of L-type Ca2+ channels which contribute to the rhythmicity of the heart and brain. In the brain, these channels are vital for excitation-transcription coupling, synaptic plasticity, and neuronal firing. Moreover, disruption of CaV1.3 function has been associated with several neurological disorders. Here, we focus on the de novo missense mutation A760G which has been linked to autism spectrum disorder (ASD). To explore the role of this mutation in ASD pathogenesis, we examined the effects of A760G on CaV1.3 channel gating and regulation. Introduction of the mutation severely diminished the Ca2+-dependent inactivation (CDI) of CaV1.3 channels, an important feedback system required for Ca2+ homeostasis. This reduction in CDI was observed in two major channel splice variants, though to different extents. Using an allosteric model of channel gating, we found that the underlying mechanism of CDI reduction is likely due to enhanced channel opening within the Ca2+-inactivated mode. Remarkably, the A760G mutation also caused an opposite increase in voltage-dependent inactivation (VDI), resulting in a multifaceted mechanism underlying ASD. When combined, these regulatory deficits appear to increase the intracellular Ca2+ concentration, thus potentially disrupting neuronal development and synapse formation, ultimately leading to ASD. PMID:27255217

  18. An autism-associated mutation in CaV1.3 channels has opposing effects on voltage- and Ca(2+)-dependent regulation.

    Science.gov (United States)

    Limpitikul, Worawan B; Dick, Ivy E; Ben-Johny, Manu; Yue, David T

    2016-01-01

    CaV1.3 channels are a major class of L-type Ca(2+) channels which contribute to the rhythmicity of the heart and brain. In the brain, these channels are vital for excitation-transcription coupling, synaptic plasticity, and neuronal firing. Moreover, disruption of CaV1.3 function has been associated with several neurological disorders. Here, we focus on the de novo missense mutation A760G which has been linked to autism spectrum disorder (ASD). To explore the role of this mutation in ASD pathogenesis, we examined the effects of A760G on CaV1.3 channel gating and regulation. Introduction of the mutation severely diminished the Ca(2+)-dependent inactivation (CDI) of CaV1.3 channels, an important feedback system required for Ca(2+) homeostasis. This reduction in CDI was observed in two major channel splice variants, though to different extents. Using an allosteric model of channel gating, we found that the underlying mechanism of CDI reduction is likely due to enhanced channel opening within the Ca(2+)-inactivated mode. Remarkably, the A760G mutation also caused an opposite increase in voltage-dependent inactivation (VDI), resulting in a multifaceted mechanism underlying ASD. When combined, these regulatory deficits appear to increase the intracellular Ca(2+) concentration, thus potentially disrupting neuronal development and synapse formation, ultimately leading to ASD. PMID:27255217

  19. Modified Ca(v1.4 expression in the Cacna1f(nob2 mouse due to alternative splicing of an ETn inserted in exon 2.

    Directory of Open Access Journals (Sweden)

    Clinton J Doering

    Full Text Available The Cacna1f(nob2 mouse is reported to be a naturally occurring null mutation for the Ca(v1.4 calcium channel gene and the phenotype of this mouse is not identical to that of the targeted gene knockout model. We found two mRNA species in the Cacna1f(nob2 mouse: approximately 90% of the mRNA represents a transcript with an in-frame stop codon within exon 2 of CACNA1F, while approximately 10% of the mRNA represents a transcript in which alternative splicing within the ETn element has removed the stop codon. This latter mRNA codes for full length Ca(v1.4 protein, detectable by Western blot analysis that is predicted to differ from wild type Ca(v1.4 protein in a region of approximately 22 amino acids in the N-terminal portion of the protein. Electrophysiological analysis with either mouse Ca(v1.4(wt or Ca(v1.4(nob2 cDNA revealed that the alternatively spliced protein does not differ from wild type with respect to activation and inactivation characteristics; however, while the wild type N-terminus interacted with filamin proteins in a biochemical pull-down experiment, the alternatively spliced N-terminus did not. The Cacna1f(nob2 mouse electroretinogram displayed reduced b-wave and oscillatory potential amplitudes, and the retina was morphologically disorganized, with substantial reduction in thickness of the outer plexiform layer and sprouting of bipolar cell dendrites ectopically into the outer nuclear layer. Nevertheless, the spatial contrast sensitivity (optokinetic response of Cacna1f(nob2 mice was generally similar to that of wild type mice. These results suggest the Cacna1f(nob2 mouse is not a CACNA1F knockout model. Rather, alternative splicing within the ETn element can lead to full-length Ca(v1.4 protein, albeit at reduced levels, and the functional Ca(v1.4 mutant may be incapable of interacting with cytoskeletal filamin proteins. These changes, do not alter the ability of the Cacna1f(nob2 mouse to detect and follow moving sine-wave gratings

  20. Loss of Cav1.3 channels reveals the critical role of L-type and BK channel coupling in pacemaking mouse adrenal chromaffin cells.

    Science.gov (United States)

    Marcantoni, Andrea; Vandael, David H F; Mahapatra, Satyajit; Carabelli, Valentina; Sinnegger-Brauns, Martina J; Striessnig, Joerg; Carbone, Emilio

    2010-01-13

    We studied wild-type (WT) and Cav1.3(-/-) mouse chromaffin cells (MCCs) with the aim to determine the isoform of L-type Ca(2+) channel (LTCC) and BK channels that underlie the pacemaker current controlling spontaneous firing. Most WT-MCCs (80%) were spontaneously active (1.5 Hz) and highly sensitive to nifedipine and BayK-8644 (1,4-dihydro-2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl)phenyl]-3-pyridinecarboxylic acid, methyl ester). Nifedipine blocked the firing, whereas BayK-8644 increased threefold the firing rate. The two dihydropyridines and the BK channel blocker paxilline altered the shape of action potentials (APs), suggesting close coupling of LTCCs to BK channels. WT-MCCs expressed equal fractions of functionally active Cav1.2 and Cav1.3 channels. Cav1.3 channel deficiency decreased the number of normally firing MCCs (30%; 2.0 Hz), suggesting a critical role of these channels on firing, which derived from their slow inactivation rate, sizeable activation at subthreshold potentials, and close coupling to fast inactivating BK channels as determined by using EGTA and BAPTA Ca(2+) buffering. By means of the action potential clamp, in TTX-treated WT-MCCs, we found that the interpulse pacemaker current was always net inward and dominated by LTCCs. Fast inactivating and non-inactivating BK currents sustained mainly the afterhyperpolarization of the short APs (2-3 ms) and only partially the pacemaker current during the long interspike (300-500 ms). Deletion of Cav1.3 channels reduced drastically the inward Ca(2+) current and the corresponding Ca(2+)-activated BK current during spikes. Our data highlight the role of Cav1.3, and to a minor degree of Cav1.2, as subthreshold pacemaker channels in MCCs and open new interesting features about their role in the control of firing and catecholamine secretion at rest and during sustained stimulations matching acute stress. PMID:20071512

  1. Role of PKC and CaV1.2 in detrusor overactivity in a model of obesity associated with insulin resistance in mice.

    Directory of Open Access Journals (Sweden)

    Luiz O Leiria

    Full Text Available Obesity/metabolic syndrome are common risk factors for overactive bladder. This study aimed to investigate the functional and molecular changes of detrusor smooth muscle (DSM in high-fat insulin resistant obese mice, focusing on the role of protein kinase C (PKC and Ca(v1.2 in causing bladder dysfunction. Male C57BL/6 mice were fed with high-fat diet for 10 weeks. In vitro functional responses and cystometry, as well as PKC and Ca(v1.2 expression in bladder were evaluated. Obese mice exhibited higher body weight, epididymal fat mass, fasting glucose and insulin resistance. Carbachol (0.001-100 µM, α,β-methylene ATP (1-10 µM, KCl (1-300 mM, extracellular Ca(2+ (0.01-100 mM and phorbol-12,13-dibutyrate (PDBu; 0.001-3 µM all produced greater DSM contractions in obese mice, which were fully reversed by the Ca(v1.2 blocker amlodipine. Cystometry evidenced augmented frequency, non-void contractions and post-void pressure in obese mice that were also prevented by amlodipine. Metformin treatment improved the insulin sensitivity, and normalized the in vitro bladder hypercontractility and cystometric dysfunction in obese mice. The PKC inhibitor GF109203X (1 µM also reduced the carbachol induced contractions. PKC protein expression was markedly higher in bladder tissues from obese mice, which was normalized by metformin treatment. The Ca(v1.2 channel protein expression was not modified in any experimental group. Our findings show that Ca(v1.2 blockade and improvement of insulin sensitization restores the enhanced PKC protein expression in bladder tissues and normalizes the overactive detrusor. It is likely that insulin resistance importantly contributes for the pathophysiology of this urological disorder in obese mice.

  2. The calmodulin-binding, short linear motif, NSCaTE is conserved in L-type channel ancestors of vertebrate Cav1.2 and Cav1.3 channels.

    Directory of Open Access Journals (Sweden)

    Valentina Taiakina

    Full Text Available NSCaTE is a short linear motif of (xWxxx(I or Lxxxx, composed of residues with a high helix-forming propensity within a mostly disordered N-terminus that is conserved in L-type calcium channels from protostome invertebrates to humans. NSCaTE is an optional, lower affinity and calcium-sensitive binding site for calmodulin (CaM which competes for CaM binding with a more ancient, C-terminal IQ domain on L-type channels. CaM bound to N- and C- terminal tails serve as dual detectors to changing intracellular Ca(2+ concentrations, promoting calcium-dependent inactivation of L-type calcium channels. NSCaTE is absent in some arthropod species, and is also lacking in vertebrate L-type isoforms, Cav1.1 and Cav1.4 channels. The pervasiveness of a methionine just downstream from NSCaTE suggests that L-type channels could generate alternative N-termini lacking NSCaTE through the choice of translational start sites. Long N-terminus with an NSCaTE motif in L-type calcium channel homolog LCav1 from pond snail Lymnaea stagnalis has a faster calcium-dependent inactivation than a shortened N-termini lacking NSCaTE. NSCaTE effects are present in low concentrations of internal buffer (0.5 mM EGTA, but disappears in high buffer conditions (10 mM EGTA. Snail and mammalian NSCaTE have an alpha-helical propensity upon binding Ca(2+-CaM and can saturate both CaM N-terminal and C-terminal domains in the absence of a competing IQ motif. NSCaTE evolved in ancestors of the first animals with internal organs for promoting a more rapid, calcium-sensitive inactivation of L-type channels.

  3. EFEITO DE DIFERENTES SUBSTRATOS NA GERMINAÇÃO DE Ochroma pyramidale (CAV. EX LAM. URB. (BOMBACACEAE

    Directory of Open Access Journals (Sweden)

    Breno Pinto Rayol

    2007-03-01

    Full Text Available O objetivo deste trabalho foi analisar o efeito de diferentes substratos na germinação de Ochroma pyramidale (Cav. ex Lam. Urb. (Bombacaceae. O experimento foi conduzido em delineamento inteiramente casualizado com três tratamentos (substratos e quatro repetições de 25 sementes. Foram testados os seguintes substratos: areia + vermiculita (1:1; vermiculita e, areia + serragem (1:1. Foi avaliado o efeito dos substratos pelo percentual de germinação das sementes, tempo médio de germinação e índice de velocidade de germinação (IVG. Houve diferença significativa entre os tratamentos em todas as variáveis observadas. As sementes, quando acondicionadas em vermiculita, apresentaram a maior taxa de germinação, maior velocidade e menor tempo de germinação.

  4. Singlet oxygen quenching and radical scavenging capacities of structurally-related flavonoids present in Zuccagnia punctata Cav.

    Science.gov (United States)

    Vieyra, Faustino E Morán; Boggetti, Héctor J; Zampini, Iris C; Ordoñez, Roxana M; Isla, María I; Alvarez, Rosa M S; De Rosso, Veridiana; Mercadante, Adriana Z; Borsarelli, Claudio D

    2009-06-01

    The singlet oxygen (1O2) quenching and free radical (DPPH(*), ABTS(* +) and O2(* -)) scavenging ability of three structurally-related flavonoids (7-hydroxyflavanone HF, 2',4'-dihydroxychalcone DHC and 3,7-dihydroxyflavone DHF) present in the Argentinean native shrub Zuccagnia punctata Cav. were studied in solution by combining electrochemical and kinetic measurements, mass spectroscopy, end-point antioxidant assays and computational calculations. The results showed that the antioxidant properties of these flavonoids depend on several factors, such as their electron- and hydrogen atom donor capacity, the ionization degree of the more acidic group, solvatation effects and electrostatic interactions with the oxidant species. The theoretical calculations for both the gas and solution phases at the B3LYP level of theory for the Osanger reaction field model agreed with the experimental findings, thus supporting the characterization of the antioxidant mechanism of the Z. punctata flavonoids. PMID:19431060

  5. GDF-15 enhances intracellular Ca2+ by increasing Cav1.3 expression in rat cerebellar granule neurons

    Science.gov (United States)

    Lu, Jun-Mei; Wang, Chang-Ying; Hu, Changlong; Fang, Yan-Jia; Mei, Yan-Ai

    2016-01-01

    GDF-15 (growth/differentiation factor 15) is a novel member of the TGF (transforming growth factor)-β superfamily that has critical roles in the central and peripheral nervous systems. We reported previously that GDF-15 increased delayed rectifier outward K+ currents and Kv2.1 α subunit expression through TβRII (TGF-β receptor II) to activate Src kinase and Akt/mTOR (mammalian target of rapamycin) signalling in rat CGNs (cerebellar granule neurons). In the present study, we found that treatment of CGNs with GDF-15 for 24 h increased the intracellular Ca2+ concentration ([Ca2+]i) in response to membrane depolarization, as determined by Ca2+ imaging. Whole-cell current recordings indicated that GDF-15 increased the inward Ca2+ current (ICa) without altering steady-state activation of Ca2+ channels. Treatment with nifedipine, an inhibitor of L-type Ca2+ channels, abrogated GDF-15-induced increases in [Ca2+]i and ICa. The GDF-15-induced increase in ICa was mediated via up-regulation of the Cav1.3 α subunit, which was attenuated by inhibiting Akt/mTOR and ERK (extracellular-signal-regulated kinase) pathways and by pharmacological inhibition of Src-mediated TβRII phosphorylation. Given that Cav1.3 is not only a channel for Ca2+ influx, but also a transcriptional regulator, our data confirm that GDF-15 induces protein expression via TβRII and activation of a non-Smad pathway, and provide novel insight into the mechanism of GDF-15 function in neurons. PMID:27114559

  6. Characterization of the triazine, T4, a representative from a novel series of CaV2 inhibitors with strong state-dependence, poor use-dependence, and distinctively fast kinetics.

    Science.gov (United States)

    Swensen, Andrew M; Niforatos, Wende; Lee, Chih-Hung; Jarvis, Michael F; McGaraughty, Steve

    2014-12-15

    There is strong pharmacological, biological, and genetic evidence supporting the role of N-type calcium channels (CaV2.2) in nociception. There is also human validation data from ziconotide, the CaV2.2-selective peptidyl inhibitor used clinically to treat refractory pain. Unfortunately, ziconotide utility is limited by its narrow therapeutic window and required intrathecal route of administration. A major focus has been placed on identifying state-dependent CaV2.2 inhibitors to improve safety margins. Much less attention, however, has been given to characterizing the kinetics of CaV2.2 inhibitors as a means to further differentiate compounds and maximize therapeutic potential. Here we provide a detailed characterization of the CaV2.2 inhibitor T4 in terms of its state-dependence, use-dependence, kinetics, and mechanism of inhibition. Compound T4 displayed a >20-fold difference in potency when measured under inactivating conditions (IC50=1.1 μM) as compared to closed-state conditions (IC50=25 μM). At 3 μM, T4 produced a 15-fold hyperpolarizing shift in the inactivation curve for CaV2.2 while having no effect on channel activation. To assess the kinetic properties of T4 in a more physiological manner, its inhibition kinetics were assessed at 32°C using 2 mM Ca(2+) as the charge carrier. Surprisingly, the repriming rate for CaV2.2 channels at hyperpolarized potentials was similar in both the presence and absence of T4. This was in contrast to other compounds which markedly delayed repriming. Furthermore, T4 inhibited CaV2.2 channels more potently when channel inactivation was driven through a tonic sub-threshold depolarization rather than through a use-dependent protocol, despite similar levels of inactivation. PMID:25446431

  7. Design and Optimization of Capacitive Liquid Level Sensor Based on CAV444%基于CAV444的电容式液位传感器设计与优化

    Institute of Scientific and Technical Information of China (English)

    赵明; 汤晓君; 张徐梁; 张涛; 寇福林

    2015-01-01

    The measurement principle of capacitive liquid level sensor was introduced at first .And then,that how to design and optimize such a sensor based on CAV444 was proposed.Finally,a liquid level sensor was fabricated and tested.According to the rang of capacitance ,which corresponds to the measured level , the output voltage of the conditioning circuit was converted to in-crease the sensitivity of the sensor ,and a low-pass digital filter was designed according to the bandwidth of the signal to improve the resistance of the sensor .Finally,the design and testing of the sensor system were completed .The testing results shovo that the max non-linearity error is 0.75%,maximum fiducial error is only 1.13%,and this sensor has good linearity ,high measurement accura-cy,and good stability .Therefore,this sensor meets the requirements of practical liquid level measurement .%文中介绍了电容式液位传感器的测量原理,提出了一种基于CAV444线性电容-电压转换芯片的电容式液位传感器设计与优化方案。该方案根据所测液位范围对应的电容值变化范围对调理电路的输出电压进行了变换,以提高传感器的灵敏度,并根据被测信号的带宽设计了低通数字滤波器,提高了传感器的抗干扰能力。测试结果表明,该传感器性能稳定,最大非线性误差为0.75%,测量结果线性度好;引用误差为1.13%,测量准确度高,能够满足大多数工业现场实际的液位测量需求。

  8. Suppression of peripheral pain by blockade of Cav2.2 channels in nociceptors induces RANKL and impairs recovery from inflammatory arthritis

    OpenAIRE

    Baddack, U.; Frahm, S.; Antolin-Fontes, B.; Grobe, J.; Lipp, M.; Mueller, G.; Ibanez-Tallon, I.

    2015-01-01

    Objective: A hallmark of rheumatoid arthritis (RA) is the chronic pain that accompanies the inflammation and joint deformation. Patients with RA rate pain relief with highest priority, however, few studies have addressed the efficacy and safety of therapies directed specifically towards pain pathways. The conotoxin MVIIA (Prialt/Ziconotide) is used in humans to alleviate persistent pain syndromes because it specifically blocks the CaV 2.2 voltage-gated calcium channel, which mediates the rele...

  9. Age-dependent impact of CaV 3.2 T-type calcium channel deletion on myogenic tone and flow-mediated vasodilatation in small arteries

    DEFF Research Database (Denmark)

    Mikkelsen, Miriam F; Björling, Karl; Jensen, Lars Jørn

    2016-01-01

    The myogenic response and flow-mediated vasodilatation are important regulators of local blood perfusion and total peripheral resistance, and are known to entail a calcium influx into vascular smooth muscle cells (VSMCs) and endothelial cells (ECs), respectively. CaV 3.2 T-type calcium channels a......-mediated vasomotor responses to prevent excess arterial tone, protect against cardiovascular disease. This article is protected by copyright. All rights reserved....

  10. Biophysical characterization of the honeybee DSC1 orthologue reveals a novel voltage-dependent Ca2+ channel subfamily: CaV4.

    Science.gov (United States)

    Gosselin-Badaroudine, Pascal; Moreau, Adrien; Simard, Louis; Cens, Thierry; Rousset, Matthieu; Collet, Claude; Charnet, Pierre; Chahine, Mohamed

    2016-08-01

    Bilaterian voltage-gated Na(+) channels (NaV) evolved from voltage-gated Ca(2+) channels (CaV). The Drosophila melanogaster Na(+) channel 1 (DSC1), which features a D-E-E-A selectivity filter sequence that is intermediate between CaV and NaV channels, is evidence of this evolution. Phylogenetic analysis has classified DSC1 as a Ca(2+)-permeable Na(+) channel belonging to the NaV2 family because of its sequence similarity with NaV channels. This is despite insect NaV2 channels (DSC1 and its orthologue in Blatella germanica, BSC1) being more permeable to Ca(2+) than Na(+) In this study, we report the cloning and molecular characterization of the honeybee (Apis mellifera) DSC1 orthologue. We reveal several sequence variations caused by alternative splicing, RNA editing, and genomic variations. Using the Xenopus oocyte heterologous expression system and the two-microelectrode voltage-clamp technique, we find that the channel exhibits slow activation and inactivation kinetics, insensitivity to tetrodotoxin, and block by Cd(2+) and Zn(2+) These characteristics are reminiscent of CaV channels. We also show a strong selectivity for Ca(2+) and Ba(2+) ions, marginal permeability to Li(+), and impermeability to Mg(2+) and Na(+) ions. Based on current ion channel nomenclature, the D-E-E-A selectivity filter, and the properties we have uncovered, we propose that DSC1 homologues should be classified as CaV4 rather than NaV2. Indeed, channels that contain the D-E-E-A selectivity sequence are likely to feature the same properties as the honeybee's channel, namely slow activation and inactivation kinetics and strong selectivity for Ca(2+) ions. PMID:27432995

  11. Association between Genetic Polymorphisms in Cav2.3 (R-type) Ca2+ Channels and Fentanyl Sensitivity in Patients Undergoing Painful Cosmetic Surgery

    OpenAIRE

    Soichiro Ide; Daisuke Nishizawa; Ken-ichi Fukuda; Shinya Kasai; Junko Hasegawa; Masakazu Hayashida; Masabumi Minami; Kazutaka Ikeda

    2013-01-01

    Individual differences in the sensitivity to fentanyl, a widely used opioid analgesic, lead to different proper doses of fentanyl, which can hamper effective pain treatment. Voltage-activated Ca(2+) channels (VACCs) play a crucial role in the nervous system by controlling membrane excitability and calcium signaling. Ca(v)2.3 (R-type) VACCs have been especially thought to play critical roles in pain pathways and the analgesic effects of opioids. However, unknown is whether single-nucleotide po...

  12. Calpain inhibition rescues troponin T3 fragmentation, increases Cav1.1, and enhances skeletal muscle force in aging sedentary mice.

    Science.gov (United States)

    Zhang, Tan; Pereyra, Andrea S; Wang, Zhong-Min; Birbrair, Alexander; Reisz, Julie A; Files, Daniel Clark; Purcell, Lina; Feng, Xin; Messi, Maria L; Feng, Hanzhong; Chalovich, Joseph; Jin, Jian-Ping; Furdui, Cristina; Delbono, Osvaldo

    2016-06-01

    Loss of strength in human and animal models of aging can be partially attributed to a well-recognized decrease in muscle mass; however, starting at middle-age, the normalized force (force/muscle cross-sectional area) in the knee extensors and single muscle fibers declines in a curvilinear manner. Strength is lost faster than muscle mass and is a more consistent risk factor for disability and death. Reduced expression of the voltage sensor Ca(2+) channel α1 subunit (Cav1.1) with aging leads to excitation-contraction uncoupling, which accounts for a significant fraction of the decrease in skeletal muscle function. We recently reported that in addition to its classical cytoplasmic location, fast skeletal muscle troponin T3 (TnT3) is fragmented in aging mice, and both full-length TnT3 (FL-TnT3) and its carboxyl-terminal (CT-TnT3) fragment shuttle to the nucleus. Here, we demonstrate that it regulates transcription of Cacna1s, the gene encoding Cav1.1. Knocking down TnT3 in vivo downregulated Cav1.1. TnT3 downregulation or overexpression decreased or increased, respectively, Cacna1s promoter activity, and the effect was ablated by truncating the TnT3 nuclear localization sequence. Further, we mapped the Cacna1s promoter region and established the consensus sequence for TnT3 binding to Cacna1s promoter. Systemic administration of BDA-410, a specific calpain inhibitor, prevented TnT3 fragmentation, and Cacna1s and Cav1.1 downregulation and improved muscle force generation in sedentary old mice. PMID:26892246

  13. "Slow" Voltage-Dependent Inactivation of CaV2.2 Calcium Channels Is Modulated by the PKC Activator Phorbol 12-Myristate 13-Acetate (PMA.

    Directory of Open Access Journals (Sweden)

    Lei Zhu

    Full Text Available CaV2.2 (N-type voltage-gated calcium channels (Ca2+ channels play key roles in neurons and neuroendocrine cells including the control of cellular excitability, neurotransmitter / hormone secretion, and gene expression. Calcium entry is precisely controlled by channel gating properties including multiple forms of inactivation. "Fast" voltage-dependent inactivation is relatively well-characterized and occurs over the tens-to- hundreds of milliseconds timeframe. Superimposed on this is the molecularly distinct, but poorly understood process of "slow" voltage-dependent inactivation, which develops / recovers over seconds-to-minutes. Protein kinases can modulate "slow" inactivation of sodium channels, but little is known about if/how second messengers control "slow" inactivation of Ca2+ channels. We investigated this using recombinant CaV2.2 channels expressed in HEK293 cells and native CaV2 channels endogenously expressed in adrenal chromaffin cells. The PKC activator phorbol 12-myristate 13-acetate (PMA dramatically prolonged recovery from "slow" inactivation, but an inactive control (4α-PMA had no effect. This effect of PMA was prevented by calphostin C, which targets the C1-domain on PKC, but only partially reduced by inhibitors that target the catalytic domain of PKC. The subtype of the channel β-subunit altered the kinetics of inactivation but not the magnitude of slowing produced by PMA. Intracellular GDP-β-S reduced the effect of PMA suggesting a role for G proteins in modulating "slow" inactivation. We postulate that the kinetics of recovery from "slow" inactivation could provide a molecular memory of recent cellular activity and help control CaV2 channel availability, electrical excitability, and neurotransmission in the seconds-to-minutes timeframe.

  14. Anion-sensitive regions of L-type CaV1.2 calcium channels expressed in HEK293 cells.

    Directory of Open Access Journals (Sweden)

    Norbert Babai

    Full Text Available L-type calcium currents (I(Ca are influenced by changes in extracellular chloride, but sites of anion effects have not been identified. Our experiments showed that CaV1.2 currents expressed in HEK293 cells are strongly inhibited by replacing extracellular chloride with gluconate or perchlorate. Variance-mean analysis of I(Ca and cell-attached patch single channel recordings indicate that gluconate-induced inhibition is due to intracellular anion effects on Ca(2+ channel open probability, not conductance. Inhibition of CaV1.2 currents produced by replacing chloride with gluconate was reduced from approximately 75%-80% to approximately 50% by omitting beta subunits but unaffected by omitting alpha(2delta subunits. Similarly, gluconate inhibition was reduced to approximately 50% by deleting an alpha1 subunit N-terminal region of 15 residues critical for beta subunit interactions regulating open probability. Omitting beta subunits with this mutant alpha1 subunit did not further diminish inhibition. Gluconate inhibition was unchanged with expression of different beta subunits. Truncating the C terminus at AA1665 reduced gluconate inhibition from approximately 75%-80% to approximately 50% whereas truncating it at AA1700 had no effect. Neutralizing arginines at AA1696 and 1697 by replacement with glutamines reduced gluconate inhibition to approximately 60% indicating these residues are particularly important for anion effects. Expressing CaV1.2 channels that lacked both N and C termini reduced gluconate inhibition to approximately 25% consistent with additive interactions between the two tail regions. Our results suggest that modest changes in intracellular anion concentration can produce significant effects on CaV1.2 currents mediated by changes in channel open probability involving beta subunit interactions with the N terminus and a short C terminal region.

  15. Low-threshold exocytosis induced by cAMP-recruited CaV3.2 (alpha1H) channels in rat chromaffin cells.

    Science.gov (United States)

    Giancippoli, A; Novara, M; de Luca, A; Baldelli, P; Marcantoni, A; Carbone, E; Carabelli, V

    2006-03-01

    We have studied the functional role of CaV3 channels in triggering fast exocytosis in rat chromaffin cells (RCCs). CaV3 T-type channels were selectively recruited by chronic exposures to cAMP (3 days) via an exchange protein directly activated by cAMP (Epac)-mediated pathway. Here we show that cAMP-treated cells had increased secretory responses, which could be evoked even at very low depolarizations (-50, -40 mV). Potentiation of exocytosis in cAMP-treated cells did not occur in the presence of 50 microM Ni2+, which selectively blocks T-type currents in RCCs. This suggests that the "low-threshold exocytosis" induced by cAMP is due to increased Ca2+ influx through cAMP-recruited T-type channels, rather than to an enhanced secretion downstream of Ca2+ entry, as previously reported for short-term cAMP treatments (20 min). Newly recruited T-type channels increase the fast secretory response at low voltages without altering the size of the immediately releasable pool. They also preserve the Ca2+ dependence of exocytosis, the initial speed of vesicle depletion, and the mean quantal size of single secretory events. All this indicates that cAMP-recruited CaV3 channels enhance the secretory activity of RCCs at low voltages by coupling to the secretory apparatus with a Ca2+ efficacy similar to that of already existing high-threshold Ca2+ channels. Finally, using RT-PCRs we found that the fast inactivating low-threshold Ca2+ current component recruited by cAMP is selectively associated to the alpha1H (CaV3.2) channel isoform. PMID:16361341

  16. Association between Genetic Polymorphisms in Cav2.3 (R-type) Ca2+ Channels and Fentanyl Sensitivity in Patients Undergoing Painful Cosmetic Surgery

    OpenAIRE

    Ide, Soichiro; Nishizawa, Daisuke; Fukuda, Ken-ichi; Kasai, Shinya; Hasegawa, Junko; Hayashida, Masakazu; Minami, Masabumi; Ikeda, Kazutaka

    2013-01-01

    Individual differences in the sensitivity to fentanyl, a widely used opioid analgesic, lead to different proper doses of fentanyl, which can hamper effective pain treatment. Voltage-activated Ca2+ channels (VACCs) play a crucial role in the nervous system by controlling membrane excitability and calcium signaling. Cav2.3 (R-type) VACCs have been especially thought to play critical roles in pain pathways and the analgesic effects of opioids. However, unknown is whether single-nucleotide polymo...

  17. THE VOLTAGE DEPENDENCE OF GATING CURRENTS OF THE NEURONAL CAV3.3 CHANNEL IS DETERMINED BY THE GATING BRAKE IN THE I-II LOOP

    Science.gov (United States)

    Karmažínová, Mária; Baumgart, Joel; Perez-Reyes, Edward; Lacinová, L'ubica

    2012-01-01

    Low-voltage activated CaV3 Ca2+ channels have an activation threshold around −60 mV, which is lower than the activation threshold of other voltage-dependent calcium channels (VDCC). The kinetics of their activation at membrane voltages just above the activation threshold is much slower than the activation kinetics of other VDCCs. It was demonstrated recently that the intracellular loop connecting repeats I and II of all three CaV3 channels contains a so-called “gating brake.” Disruption of this brake yields channels that activate at even more hyperpolarized potentials with significantly accelerated kinetics. We have compared gating of a wild type CaV3.3 channel and a mutated ID12 channel, in which the putative gating brake at the proximal part of the I-II loop was removed. Voltage dependence of the gating current activation was shifted by 34.6 mV towards more hyperpolarized potentials in ID12 channel. Kinetics of the on-charge activation was significantly accelerated, while kinetics of the off-charge was not altered. We conclude that the putative gating brake in I-II loop hinders not only the opening of the conducting pore but also the activating movement of voltage sensing S4 segments, stabilizing the channel in its closed state. PMID:21340458

  18. Apo-states of calmodulin and CaBP1 control CaV1 voltage-gated calcium channel function through direct competition for the IQ domain

    Science.gov (United States)

    Findeisen, Felix; Rumpf, Christine; Minor, Daniel L.

    2013-01-01

    In neurons, binding of calmodulin (CaM) or calcium-binding protein 1 (CaBP1) to the CaV1 (L-type) voltage-gated calcium channel IQ domain endows the channel with diametrically opposed properties. CaM causes calcium-dependent inactivation (CDI) and limits calcium entry, whereas CaBP1 blocks CDI and allows sustained calcium influx. Here, we combine isothermal titration calorimetry (ITC) with cell-based functional measurements and mathematical modeling to show that these calcium sensors behave in a competitive manner that is explained quantitatively by their apo-state binding affinities for the IQ domain. This competition can be completely blocked by covalent tethering of CaM to the channel. Further, we show that Ca2+/CaM has a sub-picomolar affinity for the IQ domain that is achieved without drastic alteration of calcium binding properties. The observation that the apo-forms of CaM and CaBP1 compete with each other demonstrates a simple mechanism for direct modulation of CaV1 function and suggests a means by which excitable cells may dynamically tune CaV activity. PMID:23811053

  19. Combined loss of INK4a and caveolin-1 synergistically enhances cell proliferation and oncogene-induced tumorigenesis: role of INK4a/CAV-1 in mammary epithelial cell hyperplasia.

    Science.gov (United States)

    Williams, Terence M; Lee, Hyangkyu; Cheung, Michelle W-C; Cohen, Alex W; Razani, Babak; Iyengar, Puneeth; Scherer, Philipp E; Pestell, Richard G; Lisanti, Michael P

    2004-06-01

    Tumorigenesis is a multistep process that involves a series of genetic changes or "multiple hits," leading to alterations in signaling, proliferation, immortalization, and transformation. Many of the molecular factors that govern tumor initiation and progression remain unknown. Here, we evaluate the transformation suppressor potential of caveolin-1 (Cav-1) and its ability to cooperate with a well established tumor suppressor, the INK4a locus. To study the effects of loss of caveolin-1 on cellular transformation, we established immortalized primary mouse embryonic fibroblasts (MEFs) expressing and lacking caveolin-1 by interbreeding Cav-1 (+/+) and Cav-1 (-/-) mice with INK4a (-/-) mice. Analysis of these cells reveals that loss of caveolin-1 confers a significant growth advantage, as measured via cellular proliferation and cell cycle analysis. Loss of caveolin-1 in the INK4a (-/-) genetic background results in constitutive hyperactivation of the p42/44 MAP kinase cascade, decreased expression of p21(Cip1), as well as cyclin D1 and PCNA overexpression, consistent with their hyperproliferative phenotype. Importantly, in cells lacking Cav-1 expression, transformation by activated oncogenes (H-Ras(G12V) or v-Src) results in increased tumor growth in vivo (up to >40-fold). Finally, INK4a (-/-)/Cav-1 (-/-) mice demonstrate disturbed mammary epithelial ductal morphology, with hyperplasia, increased side-branching, and fibrosis. Our results provide important new evidence for the transformation suppressor properties of Cav-1 and the first molecular genetic evidence that Cav-1 cooperates with a tumor suppressor, namely the INK4a genetic locus. PMID:15044451

  20. Two heterozygous Cav3.2 channel mutations in a pediatric chronic pain patient: recording condition-dependent biophysical effects.

    Science.gov (United States)

    Souza, Ivana A; Gandini, Maria A; Wan, Miranda M; Zamponi, Gerald W

    2016-04-01

    We report expression system-dependent effects of heterozygous mutations (P769L and A1059S) in the Cav3.2 CACNA1H gene identified in a pediatric patient with chronic pain and absence seizures. The mutations were introduced individually into recombinant channels and then analyzed by means of electrophysiology. When both mutants were co-expressed in tsA-201 cells, we observed a loss of channel function, with significantly smaller current densities across a wide range of voltages (-40 to +20 mV). In addition, when both mutant channels were co-expressed, the channels opened at a more depolarizing potential with a ~5-mV right shift in the half-activation potential, with no changes in half-inactivation potential and the rate of recovery from inactivation. Interestingly, when both mutants were co-expressed in the neuronal-derived CAD cells in a different extracellular milieu, the effect was remarkably different. Although not statistically significant (p replacement of extracellular sodium and potassium with tetraethylammonium chloride. Our results show that experimental conditions can be a confounding factor in the biophysical effects of T-type calcium channel mutations found in certain neurological disorders. PMID:26706850

  1. RAPD analysis on dwarf mutant of Dahlia pinnata Cav induced by 80 MeV/u 12C6+ ion

    International Nuclear Information System (INIS)

    Young shoots of Dahlia pinnata Cav cultivar were irradiated by 80 MeV/u 12C6+ ions. The differences at DNA level between dwarf mutant and wild type were detected by using RAPD (Random amplified polymorphic DNA) method. The results showed that there were different polymorphic bands induced by 18 primers among the tested 25 primers, and the ratio of polymorphic bands and total bands accounted for 19.57% after 1.80 x 108C6+/cm2 irradiation. Meanwhile, after 1.08 x l08 C6+/cm2 irradiation the polymorphic bands were induced by only 6 primers and the ratio accounted for 5.76%. Using the Jaccard formula, statistic analysis of amplified bands were obtained and it showed that after 1.80 x 108 C6+/cm2 and 1.08 x 108 C6+/cm2 irradiation the similarity index were 0.65 and 0.92 respectively. The results suggested that DNA should prone to mutant at high doses, and high dose irradiation might be more effective in corn improving and mutagenic breeding. (authors)

  2. Effect of 1-aminocyclopropane-1-carboxylic acid on the production of ethylene in senescing flowers of Ipomoea tricolor Cav

    International Nuclear Information System (INIS)

    Application of 1-aminocyclopropane-1-carboxylic acid (ACC) to rib segments excised from flowers of Ipomoea tricolor Cav. resulted in the formation of C2H4 in greater quantities than produced under natural conditions. The ability of ACC to enhance C2H4 production was independent of the physiological age of the tissue and its capacity to synthesize C2H4 without applied ACC. When ACC was fed to rib segments that had been treated with [14C]methionine, incorporation of radioactivity into C2H4 was reduced by 80%. Aminoethoxyvinylglycine and aminooxyacetic acid inhibited C2H4 production in rib segments of I. tricolor but had no effect on ACC-enhanced C2H4 production. Protoplasts obtained from flower tissue of I. tricolor did not form C2H4, even when incubated with methionine or selenomethionine. They produced C2H4 upon incubation with ACC, however. ACC-dependent C2H4 production in protoplasts was inhibited by n-propyl gallate, AgCl, CoCl2, KCN, Na2S, and NaN3. ACC-dependent C2H4 synthesis in rib segments and protoplasts was dependent on O2, the K/sub m/ for O2 being 1.0 to 1.4% (v/v). These results confirm the following pathway for C2H4 biosynthesis in I. tricolor: methionine [selenomethionine] →S-adenosylmethionine [selenoadenosylmethionine] → ACC → C2H4

  3. Techno-functional properties and in vitro bile acid-binding capacities of tamarillo (Solanum betaceum Cav.) hydrocolloids.

    Science.gov (United States)

    Gannasin, Sri Puvanesvari; Adzahan, Noranizan Mohd; Mustafa, Shuhaimi; Muhammad, Kharidah

    2016-04-01

    Hydrocolloids were extracted from seed mucilage and the pulp fractions from red tamarillo (Solanum betaceum Cav.) mesocarp, and characterisation of their techno-functional properties and in vitro bile acid-binding capacities was performed. The seed mucilage hydrocolloids that were extracted, using either 1% citric acid (THC) or water (THW), had a good foaming capacity (32-36%), whereas the pulp hydrocolloids that were extracted, using 72% ethanol (THE) or 20mM HEPES buffer (THH), had no foaming capacity. The pulp hydrocolloid, however, possessed high oil-holding and water-holding capacities in the range of 3.3-3.6 g oil/g dry sample and 25-27 g water/g dry sample, respectively. This enabled the pulp hydrocolloid to entrap more bile acids (35-38% at a hydrocolloid concentration of 2%) in its gelatinous network in comparison to commercial oat fibre and other hydrocolloids studied. The exceptional emulsifying properties (80-96%) of both hydrocolloids suggest their potential applications as food emulsifiers and bile acid binders. PMID:26593571

  4. Use of sewage sludge and organic residues in the growth of seedlings Sesbania virgata (Cav. Pers. = Uso de lodo de esgoto e resíduos orgânicos no crescimento de mudas de Sesbania virgata (Cav. Pers.

    Directory of Open Access Journals (Sweden)

    William Macedo Delarmelina

    2013-08-01

    Full Text Available During the production of seedlings, the substrate has a significant influence on plant growth, which can be used in an original or combined form. This has made necessary studies to obtain substrates able to ensure adequate growth of the seedlings grown in nurseries. With to contribute to the knowledge of the specie Sesbania virgata (Cav. Pers, this study determined the best ratio of the components for the formation of a suitable growth medium for theses seedlings. The treatments were formulated using sewage sludge (LE, organic compost (CO, coffee straw (PC in natura, and the commercial substrate (SC. Seedlings were grown in plastic pots with a capacity of 120 cm3. The experiment consisted of fourteen treatments withthree replicates of five seedlings each. After 150 days old the following variables were measured: plant height, stem diameter,ratio between plant height and stem diameter, dry mass of shoot, dry mass of root system, total dry mass, dry mass ratio of shoot/root dry mass, and Dickson quality index. The results indicated that the treatments containing sewage sludge and organic compost in its composition, especially the treatment T7 (40% LE + 60% CO provided the demonstrated the best morphological characteristics of Sesbania virgata seedlings. = Na fase de produção de mudas, o substrato exerce influência significativa no crescimento das plantas, e sua utilizaçãopode ser feita de forma original ou combinados, tornando necessário, estudos voltados para obtenção de substratos capazes de garantir adequado crescimento e qualidade das mudas produzidas em viveiro. Visando contribuir para o conhecimento da espécie Sesbania virgata (Cav. Pers, objetivou-se com este trabalho avaliar a melhor proporção entre componentes para formação de substrato para mudas. Os tratamentos foram formulados utilizando lodo de esgoto (LE, palha de café in natura (PC in natura, composto orgânico (CO e o substrato comercial (SC em diferentes propor

  5. Significance of CaV3.2 T-type Ca2+ channels for pressure- and flow-dependent vasomotor responses in rat and mouse mesenteric small arteries

    DEFF Research Database (Denmark)

    Jensen, Lars Jørn; Björling, K.; Hansen, Pernille B. Lærkegaard;

    We investigated the potential significance of CaV3.2 channels in the myogenic response (MR) and flow-mediated vasodilatation (FMVD). CaV3.2 channels were immunolocalized to EC and VSMC in rat and mouse small mesenteric arteries. The myogenic tone at pressures of 40-120 mmHg was significantly larger...... not seen (N=4). In young and old CaV3.2-/- mice no effects of NiCl2 were observed. The FMVD response in rat mesenteric arteries was not blocked by L-NAME, but was almost abolished by the SKCa/IKCa channel blockers apamin/TRAM-34 (50 nM/1 µM) (P<0.01; N=6). Interestingly the vessels constricted to flow...

  6. Functional properties of the CaV1.2 calcium channel activated by calmodulin in the absence of alpha2delta subunits.

    Science.gov (United States)

    Ravindran, Arippa; Kobrinsky, Evgeny; Lao, Qi Zong; Soldatov, Nikolai M

    2009-01-01

    Voltage-activated CaV1.2 calcium channels require association of the pore-forming alpha1C subunit with accessory CaVbeta and alpha2delta subunits. Binding of a single calmodulin (CaM) to alpha1C supports Ca2+-dependent inactivation (CDI). The human CaV1.2 channel is silent in the absence of CaVbeta and/or alpha2delta. Recently, we found that coexpression of exogenous CaM (CaMex) supports plasma membrane targeting, gating facilitation and CDI of the channel in the absence of CaVbeta. Here we discovered that CaMex and its Ca2+-insensitive mutant (CaM1234) rendered active alpha1C/CaVbeta channel in the absence of alpha2delta. Coexpression of CaMex with alpha1C and beta2d in calcium-channel-free COS-1 cells recovered gating of the channel and supported CDI. Voltage-dependence of activation was shifted by approximately +40 mV to depolarization potentials. The calcium current reached maximum at +40 mV (20 mM Ca2+) and exhibited approximately 3 times slower activation and 5 times slower inactivation kinetics compared to the wild-type channel. Furthermore, both CaMex and CaM1234 accelerated recovery from inactivation and induced facilitation of the calcium current by strong depolarization prepulse, the properties absent from the human vascular/neuronal CaV1.2 channel. The data suggest a previously unknown action of CaM that in the presence of CaVbeta; translates into activation of the alpha2delta-deficient calcium channel and alteration of its properties. PMID:19106618

  7. Two mechanistically distinct effects of dihydropyridine nifedipine on CaV1.2 L-type Ca²⁺ channels revealed by Timothy syndrome mutation.

    Science.gov (United States)

    Sheng, Xiaona; Nakada, Tsutomu; Kobayashi, Motohiro; Kashihara, Toshihide; Shibazaki, Toshihide; Horiuchi-Hirose, Miwa; Gomi, Simmon; Hirose, Masamichi; Aoyama, Toshifumi; Yamada, Mitsuhiko

    2012-06-15

    Dihydropyridine Ca(2+) channel antagonists (DHPs) block Ca(V)1.2 L-type Ca(2+) channels (LTCCs) by stabilizing their voltage-dependent inactivation (VDI); however, it is still not clear how DHPs allosterically interact with the kinetically distinct (fast and slow) VDI. Thus, we analyzed the effect of a prototypical DHP, nifedipine on LTCCs with or without the Timothy syndrome mutation that resides in the I-II linker (L(I)-(II)) of Ca(V)1.2 subunits and impairs VDI. Whole-cell Ba(2+) currents mediated by rabbit Ca(V)1.2 with or without the Timothy mutation (G436R) (analogous to the human G406R mutation) were analyzed in the presence and absence of nifedipine. In the absence of nifedipine, the mutation significantly impaired fast closed- and open-state VDI (CSI and OSI) at -40 and 0 mV, respectively, but did not affect channels' kinetics at -100 mV. Nifedipine equipotently blocked these channels at -80 mV. In wild-type LTCCs, nifedipine promoted fast CSI and OSI at -40 and 0 mV and promoted or stabilized slow CSI at -40 and -100 mV, respectively. In LTCCs with the mutation, nifedipine resumed the impaired fast CSI and OSI at -40 and 0 mV, respectively, and had the same effect on slow CSI as in wild-type LTCCs. Therefore, nifedipine has two mechanistically distinct effects on LTCCs: the promotion of fast CSI/OSI caused by L(I-II) at potentials positive to the sub-threshold potential and the promotion or stabilization of slow CSI caused by different mechanisms at potentials negative to the sub-threshold potential. PMID:22554770

  8. CaBP1 regulates voltage-dependent inactivation and activation of Ca(V)1.2 (L-type) calcium channels.

    Science.gov (United States)

    Oz, Shimrit; Tsemakhovich, Vladimir; Christel, Carl J; Lee, Amy; Dascal, Nathan

    2011-04-22

    CaBP1 is a Ca(2+)-binding protein that regulates the gating of voltage-gated (Ca(V)) Ca(2+) channels. In the Ca(V)1.2 channel α(1)-subunit (α(1C)), CaBP1 interacts with cytosolic N- and C-terminal domains and blunts Ca(2+)-dependent inactivation. To clarify the role of the α(1C) N-terminal domain in CaBP1 regulation, we compared the effects of CaBP1 on two alternatively spliced variants of α(1C) containing a long or short N-terminal domain. In both isoforms, CaBP1 inhibited Ca(2+)-dependent inactivation but also caused a depolarizing shift in voltage-dependent activation and enhanced voltage-dependent inactivation (VDI). In binding assays, CaBP1 interacted with the distal third of the N-terminal domain in a Ca(2+)-independent manner. This segment is distinct from the previously identified calmodulin-binding site in the N terminus. However, deletion of a segment in the proximal N-terminal domain of both α(1C) isoforms, which spared the CaBP1-binding site, inhibited the effect of CaBP1 on VDI. This result suggests a modular organization of the α(1C) N-terminal domain, with separate determinants for CaBP1 binding and transduction of the effect on VDI. Our findings expand the diversity and mechanisms of Ca(V) channel regulation by CaBP1 and define a novel modulatory function for the initial segment of the N terminus of α(1C). PMID:21383011

  9. Selección preliminar de plantas de arroz con resistencia vertical A pyricularia oryzae Cav por inducción de mutaciones (ING

    Directory of Open Access Journals (Sweden)

    A Cruz

    2016-03-01

    Full Text Available Seeds of rice Oryza sativa L. cv. CR-1113 susceptible to blast disease Pyricularia oryzae Cav. Were subjected to gamma radiation with 60Co using doses of 15 and 20 krads to obtain and select mutant plants with vertical resistance to blast. A total of 80.000 rice plants per treatment were individually evaluated in the second mutant generation (M2 at tillering and panicle emergence. Vertical resistance was observed in 47 and 8 plants from the 15 and 20 krads treatments respectively.

  10. Modified Cav1.4 Expression in the Cacna1fnob2 Mouse Due to Alternative Splicing of an ETn Inserted in Exon 2

    OpenAIRE

    Doering, Clinton J.; Rehak, Renata; Bonfield, Stephan; Peloquin, Jean B.; Stell, William K.; Mema, Silvina C.; Sauvé, Yves; McRory, John E.

    2008-01-01

    The Cacna1fnob2 mouse is reported to be a naturally occurring null mutation for the Cav1.4 calcium channel gene and the phenotype of this mouse is not identical to that of the targeted gene knockout model. We found two mRNA species in the Cacna1fnob2 mouse: approximately 90% of the mRNA represents a transcript with an in-frame stop codon within exon 2 of CACNA1F, while approximately 10% of the mRNA represents a transcript in which alternative splicing within the ETn element has removed the st...

  11. Familial hemiplegic migraine type-1 mutated cav2.1 calcium channels alter inhibitory and excitatory synaptic transmission in the lateral superior olive of mice.

    Science.gov (United States)

    Inchauspe, Carlota González; Pilati, Nadia; Di Guilmi, Mariano N; Urbano, Francisco J; Ferrari, Michel D; van den Maagdenberg, Arn M J M; Forsythe, Ian D; Uchitel, Osvaldo D

    2015-01-01

    CaV2.1 Ca(2+) channels play a key role in triggering neurotransmitter release and mediating synaptic transmission. Familial hemiplegic migraine type-1 (FHM-1) is caused by missense mutations in the CACNA1A gene that encodes the α1A pore-forming subunit of CaV2.1 Ca(2+) channels. We used knock-in (KI) transgenic mice harbouring the pathogenic FHM-1 mutation R192Q to study inhibitory and excitatory neurotransmission in the principle neurons of the lateral superior olive (LSO) in the auditory brainstem. We tested if the R192Q FHM-1 mutation differentially affects excitatory and inhibitory synaptic transmission, disturbing the normal balance between excitation and inhibition in this nucleus. Whole cell patch-clamp was used to measure neurotransmitter elicited excitatory (EPSCs) and inhibitory (IPSCs) postsynaptic currents in wild-type (WT) and R192Q KI mice. Our results showed that the FHM-1 mutation in CaV2.1 channels has multiple effects. Evoked EPSC amplitudes were smaller whereas evoked and miniature IPSC amplitudes were larger in R192Q KI compared to WT mice. In addition, in R192Q KI mice, the release probability was enhanced compared to WT, at both inhibitory (0.53 ± 0.02 vs. 0.44 ± 0.01, P = 2.10(-5), Student's t-test) and excitatory synapses (0.60 ± 0.03 vs. 0.45 ± 0.02, P = 4 10(-6), Student's t-test). Vesicle pool size was diminished in R192Q KI mice compared to WT mice (68 ± 6 vs 91 ± 7, P = 0.008, inhibitory; 104 ± 13 vs 335 ± 30, P = 10(-6), excitatory, Student's t-test). R192Q KI mice present enhanced short-term plasticity. Repetitive stimulation of the afferent axons caused short-term depression (STD) of E/IPSCs that recovered significantly faster in R192Q KI mice compared to WT. This supports the hypothesis of a gain-of-function of the CaV2.1 channels in R192Q KI mice, which alters the balance of excitatory/inhibitory inputs and could also have implications in the altered cortical excitability responsible for FHM

  12. Microencapsulación de tomate de árbol rojo (solanum betaceum cav.), mediante spray drying para aplicación en productos lácteos

    OpenAIRE

    Revelo Vargas, Danilo Andrés

    2014-01-01

    El proceso de microencapsulación por spray drying de extractos acuosos de tomate de árbol rojo (Solanum betaceum Cav.) se evaluó utilizando como material de partida el endocarpio (SE) de la fruta y la combinación de mesocarpio y endocarpio (ME) como una alternativa de generación de productos con valor agregado a partir de esta fruta para su utilización en la industria de alimentos. La encapsulación se realizó en un spray-dryer Lab Plant a condiciones optimizadas y con variación...

  13. Tolerância à inundação: aspectos da anatomia ecológica e do desenvolvimento de Sesbania virgata (Cav.) Pers. (Fabaceae)

    OpenAIRE

    Viviane M. Davanso-Fabro; Moacyr E. Medri; Edmilson Bianchini; José A. Pimenta

    1998-01-01

    Plantas de Sebania virgata (Cav.) Pers. (Fabaceae) cultivadas em casa de vegetação foram alagadas por 40 dias. Plantas alagadas apresentaram aumento do peso da matéria seca e comprimento, acentuadas rachaduras corticais e intumescimento cortical esponjoso em caules e raízes, raízes superficiais e raízes adventícias. É possível que a plasticidade morfo-anatômica apresentada por esta espécie esteja contribuindo para o seu estabelecimento em solos hipóxicos.Twenty days seedlings of Sesbania virg...

  14. Das Vorkommen des Ruten-Hasenohrs (Bupleurum virgatum Cav.) im Selketal : Anmerkungen zu Gefährdungsursachen sowie zu Schutz- und Erhaltungsmöglichkeiten der Art

    OpenAIRE

    Herrmann, Nick; Kison, Hans-Ulrich

    2013-01-01

    Die bedeutendste Gefährdung für das in Deutschland vom Aussterben bedrohte Ruten-Hasenohr (Bupleurum virgatum CAV.) geht derzeit vom Menschen aus. Durch touristische Übernutzung wurden am bislang letzten Standort der Art (Selketal: „Alter Falkenstein“) Ruderalisierungs- und Eutrophierungsprozesse initiiert, durch die sich der für B. virgatum verfügbare Lebensraum ständig verkleinert. Darüber hinaus werden jährlich zahlreiche Individuen der annuellen Art durch menschliche Tritteinwirkung gesch...

  15. A Single Amino Acid Deletion (ΔF1502 in the S6 Segment of CaV2.1 Domain III Associated with Congenital Ataxia Increases Channel Activity and Promotes Ca2+ Influx.

    Directory of Open Access Journals (Sweden)

    Maria Isabel Bahamonde

    Full Text Available Mutations in the CACNA1A gene, encoding the pore-forming CaV2.1 (P/Q-type channel α1A subunit, result in heterogeneous human neurological disorders, including familial and sporadic hemiplegic migraine along with episodic and progressive forms of ataxia. Hemiplegic Migraine (HM mutations induce gain-of-channel function, mainly by shifting channel activation to lower voltages, whereas ataxia mutations mostly produce loss-of-channel function. However, some HM-linked gain-of-function mutations are also associated to congenital ataxia and/or cerebellar atrophy, including the deletion of a highly conserved phenylalanine located at the S6 pore region of α1A domain III (ΔF1502. Functional studies of ΔF1502 CaV2.1 channels, expressed in Xenopus oocytes, using the non-physiological Ba2+ as the charge carrier have only revealed discrete alterations in channel function of unclear pathophysiological relevance. Here, we report a second case of congenital ataxia linked to the ΔF1502 α1A mutation, detected by whole-exome sequencing, and analyze its functional consequences on CaV2.1 human channels heterologously expressed in mammalian tsA-201 HEK cells, using the physiological permeant ion Ca2+. ΔF1502 strongly decreases the voltage threshold for channel activation (by ~ 21 mV, allowing significantly higher Ca2+ current densities in a range of depolarized voltages with physiological relevance in neurons, even though maximal Ca2+ current density through ΔF1502 CaV2.1 channels is 60% lower than through wild-type channels. ΔF1502 accelerates activation kinetics and slows deactivation kinetics of CaV2.1 within a wide range of voltage depolarization. ΔF1502 also slowed CaV2.1 inactivation kinetic and shifted the inactivation curve to hyperpolarized potentials (by ~ 28 mV. ΔF1502 effects on CaV2.1 activation and deactivation properties seem to be of high physiological relevance. Thus, ΔF1502 strongly promotes Ca2+ influx in response to either single or

  16. Coexpression of voltage-dependent calcium channels Cav1.2, 2.1a, and 2.1b in vascular myocytes

    DEFF Research Database (Denmark)

    Andreasen, Ditte; Friis, Ulla Glenert; Uhrenholt, Torben Rene;

    2006-01-01

    Voltage-dependent Ca2+ channels Cav1.2 (L type) and Cav2.1 (P/Q type) are expressed in vascular smooth muscle cells (VSMCs) and are important for the contraction of renal resistance vessels. In the present study we examined whether native renal VSMCs coexpress L-, P-, and Q-type Ca2+ currents. The...... microscopy revealed expression of both channels in all of the smooth muscle cells. Whole-cell patch clamp on single preglomerular VSMCs from mice showed L-, P-, and Q-type currents. Blockade of the L-type currents by calciseptine (20 nmol/L) inhibited 35.6+/-3.9% of the voltage-dependent Ca2+ current, and...... blocking P-type currents (omega-agatoxin IVA 10 nmol/L) led to 20.2+/-3.0% inhibition, whereas 300 nmol/L of omega agatoxin IVA (blocking P/Q-type) inhibited 45.0+/-7.3%. In rat aortic smooth muscle cells (A7r5), blockade of L-type channels resulted in 28.5+/-6.1% inhibition, simultaneous blockade of L...

  17. Cav-3 and Its Relationship with Human Muscular Disease%窖蛋白-3及其与人类肌肉疾病的关系

    Institute of Scientific and Technical Information of China (English)

    洪美亚; 钟文涛; 龚兴国

    2005-01-01

    窖蛋白-3(caveolin-3,Cav-3)是整合在窖上的肌细胞特异性蛋白质.人cav-3基因定位于3p25,其主要突变包括跨膜区的错义突变及支架区的染色体微缺失,所导致的表现型包括肢带型肌营养不良(limb-girdle muscular dystrophy-1C,LGMD-1C)、杜氏肌营养不良(Duchenne muscular dystrophy,DMD)、自发性和家族性高CK血症(hypeCKemia,HCK)、末端肌病(distal myopathy,DM)和波形肌肉疾病(rippling muscle disease,RMD)等.Cav-3不仅与肌肉营养不良症相关,也是维持心脏正常功能的必要因素.

  18. Tolerância à inundação: aspectos da anatomia ecológica e do desenvolvimento de Sesbania virgata (Cav. Pers. (Fabaceae

    Directory of Open Access Journals (Sweden)

    Viviane M. Davanso-Fabro

    1998-08-01

    Full Text Available Plantas de Sebania virgata (Cav. Pers. (Fabaceae cultivadas em casa de vegetação foram alagadas por 40 dias. Plantas alagadas apresentaram aumento do peso da matéria seca e comprimento, acentuadas rachaduras corticais e intumescimento cortical esponjoso em caules e raízes, raízes superficiais e raízes adventícias. É possível que a plasticidade morfo-anatômica apresentada por esta espécie esteja contribuindo para o seu estabelecimento em solos hipóxicos.Twenty days seedlings of Sesbania virgata (Cav. Pers. (Fabaceae cultivated in green house were flooded by forty days. Flooded plants presented increasing of dry weight and lenght, evident cortical fissures and spongy cortical swelling on the stem basis and root, superficial roots on the soil and adventitious roots. It is possible that the morpho-anatomic plasticity presented by this specie is contribuiting for its hipoxic soils stablishment.

  19. The exogenous particles of heavy metals and/or radionuclide interaction with cellular organelles in Phragmites australis (Cav.) Steudel leaf

    Science.gov (United States)

    Corneanu, Gabriel; Corneanu, Mihaela; Craciun, Constantin; Tripon, Septimiu

    2013-04-01

    Phragmites australis (Cav.) Steudel (reed), is a phytoremediatory species, meet in the swampy areas, being a hypperaccumulator for chromium (Calheiros et al., 2008; Ait Ali et al., 2004, a/o). In nature there are cytotypes with a different somatic chromosome number (6x - 16x), with a good adaptation at various environmental conditions. Weis and Weis (2004) consider that reed is an invasive species, sequester more metals than some native species and recommended to use it, in wetlands, for phytoremediation and marsh restoration. Researches performed by Hakmaoui et al. (2007) regarding the ultrastructural effect of cadmium and cooper on reed, evidenced the presence of the ferritin aggregates in the chloroplast stroma, as well as some reversible modifications in chloroplast. In this paper, the ultrastructural features of the leaf in three Phragmites australis genotypes, from the Middle Jiu river valley (Gorj county, Romania), were analyzed: Control (Ţânţăreni village); a population from neighbourhood of TEPP-Turceni; and other population developed at the basis a sterile waste dump of 40 years-old (near Cocoreni village). The heavy metal and radionuclide content of the soil was different in the three sites, with the lowest values in Control and the highest values for many heavy metals (Zn, Mn, Ni, Co, Cd) and radionuclide's (U-238, Ra-226, Pb-210, Bi-214, Pb-214, U-235, Ac-228, Pb-212, Cs-137) on the sterile waste dump. The analysis of the ultrastructural features of the leaf in mature plants revealed some differences between the three Phragmites australis genotypes. The ultrastructural investigations underlined the adaptation of this species against the stress factors (heavy metals and radionuclides). The exogenous particles penetrated the foliar tissue through the epidermis and stomata, being spread in the cells, at the plasmodesmata level, through endoplasmic reticulum, and through the vascular system. The exogenous particles were present on the endoplasmic

  20. Synaptic gain-of-function effects of mutant Cav2.1 channels in a mouse model of familial hemiplegic migraine are due to increased basal [Ca2+]i

    NARCIS (Netherlands)

    M.N. Di Guilmi (Mariano); T. Wang (Teng); C.G. Inchauspe (Carlota Gonzalez); I.D. Forsythe (Ian); M.D. Ferrari (Michel); A.M.J.M. van der Maagdenberg (Arn M. J.); J.G.G. Borst (Gerard); O.D. Uchitel (Osvaldo)

    2014-01-01

    textabstractSpecific missense mutations in the CACNA1A gene, which encodes a subunit of voltage-gated CaV2.1 channels, are associated with familial hemiplegic migraine type 1 (FHM1), a rare monogenic subtype of common migraine with aura. We used transgenic knock-in (KI) mice harboring the human path

  1. Spinal morphine but not ziconotide or gabapentin analgesia is affected by alternative splicing of voltage-gated calcium channel CaV2.2 pre-mRNA.

    Science.gov (United States)

    Jiang, Yu-Qiu; Andrade, Arturo; Lipscombe, Diane

    2013-01-01

    Presynaptic voltage-gated calcium Ca(V)2.2 channels play a privileged role in spinal level sensitization following peripheral nerve injury. Direct and indirect inhibitors of Ca(V)2.2 channel activity in spinal dorsal horn are analgesic in chronic pain states. Ca(V)2.2 channels represent a family of splice isoforms that are expressed in different combinations according to cell-type. A pair of mutually exclusive exons in the Ca(V)2.2 encoding Cacna1b gene, e37a and e37b, differentially influence morphine analgesia. In mice that lack exon e37a, which is enriched in nociceptors, the analgesic efficacy of intrathecal morphine against noxious thermal stimuli is reduced. Here we ask if sequences unique to e37a influence: the development of abnormal thermal and mechanical sensitivity associated with peripheral nerve injury; and the actions of two other classes of analgesics that owe part or all of their efficacy to Ca(V)2.2 channel inhibition. We find that: i) the analgesic efficacy of morphine, but not ziconotide or gabapentin, is reduced in mice lacking e37a, ii) the induction and maintenance of behaviors associated with sensitization that accompany peripheral nerve injury, do not require e37a-specific sequence, iii) intrathecal morphine, but not ziconotide or gabapentin analgesia to thermal stimuli is significantly lower in wild-type mice after peripheral nerve injury, iv) the analgesic efficacy of ziconotide and gabapentin to mechanical stimuli is reduced following nerve injury, and iv) intrathecal morphine analgesia to thermal stimuli in mice lacking e37a is not further reduced by peripheral nerve injury. Our findings show that the analgesic action of morphine, but not ziconotide or gabapentin, to thermal stimuli is linked to which Cacna1b exon, e37a or e37b, is selected during alternative pre-mRNA splicing. PMID:24369063

  2. Constructos teóricos sobre la atención en crisis: atención de victimas del CAV

    OpenAIRE

    Mejía Rivera, Susana

    2015-01-01

    Se pretende con este artículo entender a través de una revisión documental de los diferentes paradigmas teóricos sobre el concepto de atención en crisis, enfocándolo en la atención diaria de las víctimas en el CAV (centro de atención a víctimas) y la aplicación de los constructos teóricos en este lugar -- Se busca de manera clara y puntual una delimitación de aquellos paradigmas que se utilizan y que realmente no funcionan en la intervención misma, sin aras de demeritar el trabajo de ningún p...

  3. In vitro Antioxidant, Lipoxygenase and Xanthine Oxidase Inhibitory Activities of Fractions from Cienfuegosia digitata Cav., Sida alba L. and Sida acuta Burn f. (Malvaceae

    Directory of Open Access Journals (Sweden)

    J. Millogo-Rasolodimby

    2010-01-01

    Full Text Available In this study polyphenol content, antioxidant activity, lipoxygenase (LOX and Xanthine Oxidase (XO inhibitory effects of n-hexane, dichloromethane, ethyl acetate and n-butanol fractions of aqueous acetone extracts from S. alba L., S. acuta Burn f. and Cienfuegosia digitata Cav. were investigated. The total phenolics, flavonoids, flavonols and total tannins were determined by spectrophotometric methods using Folin-ciocalteu, AlCl3 reagents and tannic acid, respectively. The antioxidant potential was evaluated using three methods: inhibition of free radical 2,2-diphenyl-1-picrylhydramzyl (DPPH, ABTS radical cation decolorization assay and Iron (III to iron (II reduction activity (FRAP. For enzymatic activity, lipoxygenase and xanthine oxidase inhibitory activities were used. This study shows a relationship between polyphenol contents, antioxidant and enzymatic activities. Present results showed that ethyl acetate and dichloromethane fractions elicit the highest polyphenol content, antioxidant and enzymatic activities.

  4. EFICIÊNCIA DE FUNGICIDAS NO CONTROLE DE BRUSONE (Pyricularia oryzae DO ARROZ EFFICIENCY OF FUNGICIDES IN THE CONTROL OF RICE BLAST (Pyricularia oryzae Cav.

    Directory of Open Access Journals (Sweden)

    Wilson Ferreira de Oliveira

    2007-09-01

    Full Text Available

    A eficiência dos fungicidas ethiltrianol e edifenphos, em diferentes dosagens, foi avaliada na cultura de arroz (Var. IAC 165, visando ao controle da Brusone (Pyricularia oryzae Cav. a nível de campo. Os fungicidas foram pulverizados uma ou duas vezes, na fase do emborrachamento e 15 dias após a 1ª aplicação. Os resultados demonstraram que, quanto ao nível de infecção das folhas bandeiras, das panículas e lesões nos grãos, houve diferença estatística significativa entre a testemunha e a maioria dos tratamentos com ethiltrianol e uns poucos com edifenphos. Os melhores resultados foram conseguidos com ethiltrianol na maior concentração em duas épocas de aplicação. Com relação à produtividade não houve diferença significativa entre os tratamentos.

    Efficiency of fungicides ethiltrianol and edifenphos, at different concentrations, was evaluated in rice culture (Var. IAC-165, aiming control of rice blast (Pyricularia oryzae Cav. at field situation. Fungicides were pulverized one or two times, at seasoning phase and 15 days after first treatment. The results showed that, as to infection level of banner leaves, panicles and grain lesions there was significant statistical difference between testimony and majority of treatments with ethiltrianol and a few ones with edifenphos. The best results were obtained with ethiltrianol at maximum concentration in two treatments phases. Referring to productiveness, there was no significant difference between treatments.

  5. Cytoplasmic location of α1A voltage-gated calcium channel C-terminal fragment (Cav2.1-CTF) aggregate is sufficient to cause cell death.

    Science.gov (United States)

    Takahashi, Makoto; Obayashi, Masato; Ishiguro, Taro; Sato, Nozomu; Niimi, Yusuke; Ozaki, Kokoro; Mogushi, Kaoru; Mahmut, Yasen; Tanaka, Hiroshi; Tsuruta, Fuminori; Dolmetsch, Ricardo; Yamada, Mitsunori; Takahashi, Hitoshi; Kato, Takeo; Mori, Osamu; Eishi, Yoshinobu; Mizusawa, Hidehiro; Ishikawa, Kinya

    2013-01-01

    The human α1A voltage-dependent calcium channel (Cav2.1) is a pore-forming essential subunit embedded in the plasma membrane. Its cytoplasmic carboxyl(C)-tail contains a small poly-glutamine (Q) tract, whose length is normally 4∼19 Q, but when expanded up to 20∼33Q, the tract causes an autosomal-dominant neurodegenerative disorder, spinocerebellar ataxia type 6 (SCA6). A recent study has shown that a 75-kDa C-terminal fragment (CTF) containing the polyQ tract remains soluble in normal brains, but becomes insoluble mainly in the cytoplasm with additional localization to the nuclei of human SCA6 Purkinje cells. However, the mechanism by which the CTF aggregation leads to neurodegeneration is completely elusive, particularly whether the CTF exerts more toxicity in the nucleus or in the cytoplasm. We tagged recombinant (r)CTF with either nuclear-localization or nuclear-export signal, created doxycyclin-inducible rat pheochromocytoma (PC12) cell lines, and found that the CTF is more toxic in the cytoplasm than in the nucleus, the observations being more obvious with Q28 (disease range) than with Q13 (normal-length). Surprisingly, the CTF aggregates co-localized both with cAMP response element-binding protein (CREB) and phosphorylated-CREB (p-CREB) in the cytoplasm, and Western blot analysis showed that the quantity of CREB and p-CREB were both decreased in the nucleus when the rCTF formed aggregates in the cytoplasm. In human brains, polyQ aggregates also co-localized with CREB in the cytoplasm of SCA6 Purkinje cells, but not in other conditions. Collectively, the cytoplasmic Cav2.1-CTF aggregates are sufficient to cause cell death, and one of the pathogenic mechanisms may be abnormal CREB trafficking in the cytoplasm and reduced CREB and p-CREB levels in the nuclei. PMID:23505410

  6. Cytoplasmic location of α1A voltage-gated calcium channel C-terminal fragment (Cav2.1-CTF aggregate is sufficient to cause cell death.

    Directory of Open Access Journals (Sweden)

    Makoto Takahashi

    Full Text Available The human α1A voltage-dependent calcium channel (Cav2.1 is a pore-forming essential subunit embedded in the plasma membrane. Its cytoplasmic carboxyl(C-tail contains a small poly-glutamine (Q tract, whose length is normally 4∼19 Q, but when expanded up to 20∼33Q, the tract causes an autosomal-dominant neurodegenerative disorder, spinocerebellar ataxia type 6 (SCA6. A recent study has shown that a 75-kDa C-terminal fragment (CTF containing the polyQ tract remains soluble in normal brains, but becomes insoluble mainly in the cytoplasm with additional localization to the nuclei of human SCA6 Purkinje cells. However, the mechanism by which the CTF aggregation leads to neurodegeneration is completely elusive, particularly whether the CTF exerts more toxicity in the nucleus or in the cytoplasm. We tagged recombinant (rCTF with either nuclear-localization or nuclear-export signal, created doxycyclin-inducible rat pheochromocytoma (PC12 cell lines, and found that the CTF is more toxic in the cytoplasm than in the nucleus, the observations being more obvious with Q28 (disease range than with Q13 (normal-length. Surprisingly, the CTF aggregates co-localized both with cAMP response element-binding protein (CREB and phosphorylated-CREB (p-CREB in the cytoplasm, and Western blot analysis showed that the quantity of CREB and p-CREB were both decreased in the nucleus when the rCTF formed aggregates in the cytoplasm. In human brains, polyQ aggregates also co-localized with CREB in the cytoplasm of SCA6 Purkinje cells, but not in other conditions. Collectively, the cytoplasmic Cav2.1-CTF aggregates are sufficient to cause cell death, and one of the pathogenic mechanisms may be abnormal CREB trafficking in the cytoplasm and reduced CREB and p-CREB levels in the nuclei.

  7. (S)-Lacosamide Binding to Collapsin Response Mediator Protein 2 (CRMP2) Regulates CaV2.2 Activity by Subverting Its Phosphorylation by Cdk5.

    Science.gov (United States)

    Moutal, Aubin; François-Moutal, Liberty; Perez-Miller, Samantha; Cottier, Karissa; Chew, Lindsey Anne; Yeon, Seul Ki; Dai, Jixun; Park, Ki Duk; Khanna, May; Khanna, Rajesh

    2016-04-01

    The neuronal circuit remodels during development as well as in human neuropathologies such as epilepsy. Neurite outgrowth is an obligatory step in these events. We recently reported that alterations in the phosphorylation state of an axon specification/guidance protein, the collapsin response mediator protein 2 (CRMP2), play a major role in the activity-dependent regulation of neurite outgrowth. We also identified (S)-LCM, an inactive stereoisomer of the clinically used antiepileptic drug (R)-LCM (Vimpat®), as a novel tool for preferentially targeting CRMP2-mediated neurite outgrowth. Here, we investigated the mechanism by which (S)-LCM affects CRMP2 phosphorylation by two key kinases, cyclin-dependent kinase 5 (Cdk5) and glycogen synthase kinase 3β (GSK-3β). (S)-LCM application to embryonic cortical neurons resulted in reduced levels of Cdk5- and GSK-3β-phosphorylated CRMP2. Mechanistically, (S)-LCM increased CRMP2 binding to both Cdk5- and GSK-3β without affecting binding of CRMP2 to its canonical partner tubulin. Saturation transfer difference nuclear magnetic resonance (STD NMR) and differential scanning fluorimetry (DSF) experiments demonstrated direct binding of (S)-LCM to CRMP2. Using an in vitro luminescent kinase assay, we observed that (S)-LCM specifically inhibited Cdk5-mediated phosphorylation of CRMP2. Cross-linking experiments and analytical ultracentrifugation showed no effect of (S)-LCM on the oligomerization state of CRMP2. The increased association between Cdk5-phosphorylated CRMP2 and CaV2.2 was reduced by (S)-LCM in vitro and in vivo. This reduction translated into a decrease of calcium influx via CaV2.2 in (S)-LCM-treated neurons compared to controls. (S)-LCM, to our knowledge, is the first molecule described to directly inhibit CRMP2 phosphorylation and may be useful for delineating CRMP2-facilitated functions. PMID:25846820

  8. Parasuicide and drug self-poisoning: analysis of the epidemiological and clinical variables of the patients admitted to the Poisoning Treatment Centre (CAV, Niguarda General Hospital, Milan

    Directory of Open Access Journals (Sweden)

    Manfré Sergio

    2005-04-01

    Full Text Available Abstract Epidemiological knowledge of parasuicides and drug self-poisoning is still limited by a lack of data. A number of preliminary studies, which require further analysis, evidenced that parasuicidal acts occur more often among females, that the peak rate is generally recorded between the ages of 15 and 34 years and psychotropic medications seems to be the most frequently used. The aim of this study was to describe the demographic and clinical variables of a sample of subjects admitted to the Posisoning Treatment Centre (CAV, Niguarda General Hospital, Milan, following drug self-poisoning. Furthermore, this study is aimed to identify the risk factors associated to parasuicidal gestures, with special care for the used drugs, the presence of psychiatric or organic disorders, alcoholism and drug addiction. The study included the 201 patients attending the CAV in 1999 and 2000 who satisfied the criteria of self-poisoning attempts: 106 cases in 1999 and 95 in 2000. The sample had a prevalence of females (64%. The peak rates of parasuicides from drug self-poisoning were reached between 21 and 30 years among the females, and 31 and 40 years among the males. 81.6% of the patients used one or more psychoactive drugs, the most frequent being the benzodiazepines (58.7%, classic neuroleptics (16.9% and new-generation antidepressants (SSRIs, SNRIs, NARIs (12.9%. The prevalence of mood disorders was higher among females (64% vs 42%, whereas schizophrenia was more frequently diagnosed in males (22% vs 10%. 61% (33% had a history of previous attempted suicides. The presence of clinically relevant organic diseases was observed in 24.9% of the sample.

  9. In vitro Antioxidant, Lipoxygenase and Xanthine Oxidase Inhibitory Activities of Fractions from Cienfuegosia digitata Cav., Sida alba L. and Sida acuta Burn f. (Malvaceae)

    OpenAIRE

    J. Millogo-Rasolodimby; A. Lamien-Meda; M. Kiendrebeogo; N.T.R. Meda; A.Y. Coulibaly; De Souza, A.; K. Konate; M. Lamidi; O. G. NACOULMA

    2010-01-01

    In this study polyphenol content, antioxidant activity, lipoxygenase (LOX) and Xanthine Oxidase (XO) inhibitory effects of n-hexane, dichloromethane, ethyl acetate and n-butanol fractions of aqueous acetone extracts from S. alba L., S. acuta Burn f. and Cienfuegosia digitata Cav. were investigated. The total phenolics, flavonoids, flavonols and total tannins were determined by spectrophotometric methods using Folin-ciocalteu, AlCl3 reagents and tannic acid, respectively. The antioxidant poten...

  10. Association between genetic polymorphisms in Ca(v2.3 (R-type Ca2+ channels and fentanyl sensitivity in patients undergoing painful cosmetic surgery.

    Directory of Open Access Journals (Sweden)

    Soichiro Ide

    Full Text Available Individual differences in the sensitivity to fentanyl, a widely used opioid analgesic, lead to different proper doses of fentanyl, which can hamper effective pain treatment. Voltage-activated Ca(2+ channels (VACCs play a crucial role in the nervous system by controlling membrane excitability and calcium signaling. Ca(v2.3 (R-type VACCs have been especially thought to play critical roles in pain pathways and the analgesic effects of opioids. However, unknown is whether single-nucleotide polymorphisms (SNPs of the human CACNA1E (calcium channel, voltage-dependent, R type, alpha 1E subunit gene that encodes Cav2.3 VACCs influence the analgesic effects of opioids. Thus, the present study examined associations between fentanyl sensitivity and SNPs in the human CACNA1E gene in 355 Japanese patients who underwent painful orofacial cosmetic surgery, including bone dissection. We first conducted linkage disequilibrium (LD analyses of 223 SNPs in a region that contains the CACNA1E gene using genomic samples from 100 patients, and a total of 13 LD blocks with 42 Tag SNPs were observed within and around the CACNA1E gene region. In the preliminary study using the same 100 genomic samples, only the rs3845446 A/G SNP was significantly associated with perioperative fentanyl use among these 42 Tag SNPs. In a confirmatory study using the other 255 genomic samples, this SNP was also significantly associated with perioperative fentanyl use. Thus, we further analyzed associations between genotypes of this SNP and all of the clinical data using a total of 355 samples. The rs3845446 A/G SNP was associated with intraoperative fentanyl use, 24 h postoperative fentanyl requirements, and perioperative fentanyl use. Subjects who carried the minor G allele required significantly less fentanyl for pain control compared with subjects who did not carry this allele. Although further validation is needed, the present findings show the possibility of the involvement of CACNA1E

  11. Influência da temperatura e da quantidade de água no substrato sobre a germinação de sementes de Ochroma pyramidale (Cav. ex Lam.) Urban (pau-de-balsa) Influence of temperature and substrate moisture on seed germination of Ochroma pyramidale (Cav. ex Lam.) Urban (balsa wood)

    OpenAIRE

    Michele Braule Pinto Ramos; Vania Palmeira Varela; Maria de Fátima Figueiredo Melo

    2006-01-01

    O presente trabalho foi conduzido com o objetivo de avaliar diferentes volumes de água no substrato e temperaturas na germinação de sementes de Ochroma pyramidale (Cav. ex Lam.) Urban. Antes da instalação dos testes de germinação, as sementes foram tratadas com imersão em água quente a 80º C até o resfriamento, para superar a dormência. A semeadura foi realizada em rolos de papel germitest, umedecidos com volumes (mL) de água equivalentes a 1,5; 2,0; 2,5 e 3,0 vezes o peso do substrato nas te...

  12. Indução do enraizamento em estacas de Malvaviscus arboreus Cav. com diferentes concentrações de ácido indol–butírico (AIB = Induction of rooting in cuttings of Malvaviscus arboreus Cav. with different concentrations of Indolbutiric Acid (IBA

    Directory of Open Access Journals (Sweden)

    Arcângelo Loss

    2009-04-01

    Full Text Available Foi avaliado o efeito de três concentrações de auxina (0, 2.000 e 6.000 ppm (ácido indol-bultírico - AIB para induzir a formação de raízes adventícias em três tipos de estacas caulinares (herbáceas, semilenhosas e lenhosas de malvavisco (Malvaviscus arboreusCav.. As estacas foram cortadas em bisel, com aproximadamente 12 cm cada, tratadas com o fungicida benomyl e, posteriormente, com AIB. As avaliações foram feitas 30 dias após o plantio, sendo avaliado o aparecimento ou não de calos, raízes e brotações. Entre as estacas,as lenhosas apresentaram melhor capacidade regenerativa, atingindo o máximo de tecido caloso, enraizamento e brotação, independentemente da concentração de AIB utilizada. No tratamento com AIB na concentração de 2.000 ppm, em estacas herbáceas e lenhosas,obtiveram-se melhores resultados para enraizamento.The study evaluated the effect of three concentrations of auxin (0; 2,000 and 6,000 ppm (indolbutiric acid - IBA in inducingthe formation of adventitious roots in three types of stem stakes (herbaceous, woody and semi-woody of wax mallow (Malvaviscus arboreus Cav.. The stakes were cut into a bezel angle, with approximately 12 cm each, treated with the fungicide Benlate and later with IBA. The assessments were made 30 days after planting, evaluating the appearance or absence of calluses, roots and shoots. Among the stakes, the woody ones showed the best regenerative capacity, reaching the maximum corpus callosum, rooting and shooting,regardless of IBA concentration. In the treatment with 2000 ppm IBA concentration, herbaceous and woody stakes showed the best results for rooting.

  13. Development and validation of a liquid chromatography method for quantification of xanthorrhizol in roots of Iostephane heterophylla (Cav.) Benth ex Hemsl.

    Science.gov (United States)

    Aguilar, María Isabel; Osorio, Nadia; Bernal, Israel; Navarrete, Andrés; Bye, Robert

    2007-01-01

    Roots of Iostephane heterophylla (Cav.) Benth ex Hemsl are used mainly in Mexican traditional medicine to heal skin problems. The development of a column high-performance liquid chromatography (LC)-UV detector method for the determination of xanthorrhizol, the major and active component of the roots of I. heterophylla, is described in this paper. The content of this compound was quantitatively determined employing a Symmetry C18 5 microm particle size column with the isocratic mobile phase acetonitrile-water (85 + 15). The flow rate was 1.0 mL/min, and UV detection was at 230 nm. The limits of detection and quantitation were 0.2 and 0.5 microg/mL, respectively. Quantities of xanthorrhizol measured by this method ranged between 1.8 to 10.94 mg/g of root of the plant in 11 different samples of I. heterophylla. Xanthorrhizol was not detected in a sample of I. madrensis, so xanthorrhizol could be used as a marker compound of I. heterophylla. The LC method described here was shown to be reliable, reproducible, and accurate. PMID:17760325

  14. Effects of high Zn and Pb concentrations on Phragmites australis (Cav.) Trin. Ex. Steudel: Photosynthetic performance and metal accumulation capacity under controlled conditions.

    Science.gov (United States)

    Bernardini, A; Salvatori, E; Guerrini, V; Fusaro, L; Canepari, S; Manes, F

    2016-01-01

    The response of Phragmites australis (Cav.) Trin. Ex. Steudel to zinc (Zn) and lead (Pb) was studied separately in two hydroponic tests, during a three weeks experiment. The effects on ecophysiology and biomass partitioning were evaluated during the metal treatments and at the recovery, and total metal content and accumulation capacity in different plant organs were assessed. Zn and Pb had different effects on the overall measured parameters, highlighting different mechanism of action. In particular, Zn concentration was higher in roots and, being a micronutrient, it was translocated into leaves, producing a reduction of assimilation rate, stomatal conductance (-71.9 and -81.3% respect to the control plant respectively), and a strong down regulation of photosystems functionality both at PSII and PSI level. Otherwise, Pb was accumulated mainly in the more lignified tissue such as rhizomes, with slightly effect on gas exchange. Chlorophyll a fluorescence highlighted that Pb inhibits the electron transfer process at the PSI donor side, without recovery after the removal of the metal stress. Despite these physiological limitations, P. australis showed a high capacity to accumulate both metals, and only slight reduction of biomass, being therefore a suitable species for phytoremediation interventions. PMID:26114607

  15. Uptake of radionuclides by a common reed (Phragmites australis (Cav.) Trin. ex Steud.) grown in the vicinity of the former uranium mine at Zirovski vrh

    International Nuclear Information System (INIS)

    From uranium mining areas, in particular, the radionuclides are usually discharged to the environment during the mining and milling process. At the former uranium mine Zirovski vrh, Slovenia, mine waste and mill tailings were deposited at the Jazbec site and the Borst site, respectively. Plants grown in soils contaminated with the seepage waters from tailings may represent radiological concern if radionuclides from the uranium decay chain are transferred into the food chain. Uranium is usually accumulated in the roots and translocated to the shoots in limited amounts. Uranium plant accumulators are usually plants from Brassicaceae and Poaceae families. A common reed (Phragmites australis (Cav.) Trin. ex Steud.), a tall perennial grass, growing in a wetland habitats, accumulates metals in the above-ground parts. It may be used for phytoremediation of uranium-contaminated soils, because of high biomass production and high metal-accumulation potential. Preliminary results of radionuclide contents measured in such plants, growing on the deposit tailings are presented. A common reed, that was grown on the Borst tailings pile accumulated 8.6 ± 8 mBq/g dry weight (d.w.) and 2.4 ± 2 mBq/g dry weight (d.w.) of 238U in leaves and stems, respectively. In the paper, activity concentrations of other nuclides, i.e. 226Ra, 210Pb and 40K are also shown and discussed.

  16. Uptake of radionuclides by a common reed (Phragmites australis (Cav.) Trin. ex Steud.) grown in the vicinity of the former uranium mine at Zirovski vrh

    Energy Technology Data Exchange (ETDEWEB)

    Cerne, Marko, E-mail: marko.cerne@ijs.s [Jozef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana (Slovenia); Smodis, Borut, E-mail: borut.smodis@ijs.s [Jozef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana (Slovenia); Strok, Marko, E-mail: marko.strok@ijs.s [Jozef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana (Slovenia)

    2011-04-15

    From uranium mining areas, in particular, the radionuclides are usually discharged to the environment during the mining and milling process. At the former uranium mine Zirovski vrh, Slovenia, mine waste and mill tailings were deposited at the Jazbec site and the Borst site, respectively. Plants grown in soils contaminated with the seepage waters from tailings may represent radiological concern if radionuclides from the uranium decay chain are transferred into the food chain. Uranium is usually accumulated in the roots and translocated to the shoots in limited amounts. Uranium plant accumulators are usually plants from Brassicaceae and Poaceae families. A common reed (Phragmites australis (Cav.) Trin. ex Steud.), a tall perennial grass, growing in a wetland habitats, accumulates metals in the above-ground parts. It may be used for phytoremediation of uranium-contaminated soils, because of high biomass production and high metal-accumulation potential. Preliminary results of radionuclide contents measured in such plants, growing on the deposit tailings are presented. A common reed, that was grown on the Borst tailings pile accumulated 8.6 {+-} 8 mBq/g dry weight (d.w.) and 2.4 {+-} 2 mBq/g dry weight (d.w.) of {sup 238}U in leaves and stems, respectively. In the paper, activity concentrations of other nuclides, i.e. {sup 226}Ra, {sup 210}Pb and {sup 40}K are also shown and discussed.

  17. Antidepressant-like activity of Tagetes lucida Cav. is mediated by 5-HT(1A) and 5-HT(2A) receptors.

    Science.gov (United States)

    Bonilla-Jaime, H; Guadarrama-Cruz, G; Alarcon-Aguilar, F J; Limón-Morales, O; Vazquez-Palacios, G

    2015-10-01

    It has been demonstrated that the aqueous extract of Tagetes lucida Cav. shows an antidepressant-like effect on the forced swimming test (FST) in rats. The aim of this study was to analyze the participation of the serotoninergic system in the antidepressant-like effect of the aqueous extract of T. lucida. Different doses of the extract of T. lucida were administered at 72, 48, 24, 18 and 1 h before FST. The animals were pretreated with a 5-HT1A receptor antagonist (WAY-100635, 0.5 mg/kg), a 5-HT2A receptor antagonist (ketanserin, 5 mg/kg), a β-noradrenergic receptor antagonist (propranolol, 200 mg/kg), and with a α2-noradrenergic receptor antagonist (yohimbine, 1 mg/kg) alone or combined with the extract and pretreated with a serotonin synthesis inhibitor (PCPA) before treatment with 8-OH-DPAT + the extract of T. lucida. In addition, suboptimal doses of the 5-HT1A agonist (8-OH-DPAT) + non-effective dose of extract was analyzed in the FST. To determine the presence of flavonoids, the aqueous extract of T. lucida (20 µl, 4 mg/ml) was injected in HPLC; however, a quercetin concentration of 7.72 mg/g of extract weight was detected. A suboptimal dose of 8-OH-DPAT + extract of T. lucida decreased immobility and increased swimming and climbing. An antidepressant-like effect with the aqueous extract of T. lucida at doses of 100 and 200 mg/kg was observed on the FST with decreased immobility behavior and increased swimming; however, this effect was blocked by WAY-100635, ketanserin and PCPA but not by yohimbine and propranolol, suggesting that the extract of T. lucida could be modulating the release/reuptake of serotonin. PMID:26062718

  18. Ion fluxes through KCa2 (SK and Cav1 (L-type channels contribute to chronoselectivity of adenosine A1 receptor-mediated actions in spontaneously beating rat atria

    Directory of Open Access Journals (Sweden)

    Paulo eCorreia-De-Sá

    2016-03-01

    Full Text Available Impulse generation in supraventricular tissue is inhibited by adenosine and acetylcholine via the activation of A1 and M2 receptors coupled to inwardly rectifying GIRK/KIR3.1/3.4 channels, respectively. Unlike M2 receptors, bradycardia produced by A1 receptors activation predominates over negative inotropy. Such difference suggests that other ion currents may contribute to adenosine chronoselectivity. In isolated spontaneously beating rat atria, blockade of KCa2/SK channels with apamin and Cav1 (L-type channels with nifedipine or verapamil, sensitized atria to the negative inotropic action of the A1 agonist, R-PIA, without affecting the nucleoside negative chronotropy. Patch-clamp experiments in the whole-cell configuration mode demonstrate that adenosine, via A1 receptors, activates the inwardly-rectifying GIRK/KIR3.1/KIR3.4 current resulting in hyperpolarization of atrial cardiomyocytes, which may slow down heart rate. Conversely, the nucleoside inactivates a small conductance Ca2+-activated KCa2/SK outward current, which eventually reduces the repolarizing force and thereby prolong action potentials duration Ca2+ influx into cardiomyocytes. Immunolocalization studies showed that differences in A1 receptors distribution between the sinoatrial node and surrounding cardiomyocytes do not afford a rationale for adenosine chronoselectivity. Immunolabelling of KIR3.1, KCa2.2, KCa2.3 and Cav1 was also observed throughout the right atrium. Functional data indicate that while both A1 and M2 receptors favor the opening of GIRK/KIR3.1/3.4 channels modulating atrial chronotropy, A1 receptors may additionally restrain KCa2/SK activation thereby compensating atrial inotropic depression by increasing the time available for Ca2+ influx through Cav1 (L-type channels.

  19. Ion Fluxes through KCa2 (SK) and Cav1 (L-type) Channels Contribute to Chronoselectivity of Adenosine A1 Receptor-Mediated Actions in Spontaneously Beating Rat Atria.

    Science.gov (United States)

    Bragança, Bruno; Oliveira-Monteiro, Nádia; Ferreirinha, Fátima; Lima, Pedro A; Faria, Miguel; Fontes-Sousa, Ana P; Correia-de-Sá, Paulo

    2016-01-01

    Impulse generation in supraventricular tissue is inhibited by adenosine and acetylcholine via the activation of A1 and M2 receptors coupled to inwardly rectifying GIRK/KIR3.1/3.4 channels, respectively. Unlike M2 receptors, bradycardia produced by A1 receptors activation predominates over negative inotropy. Such difference suggests that other ion currents may contribute to adenosine chronoselectivity. In isolated spontaneously beating rat atria, blockade of KCa2/SK channels with apamin and Cav1 (L-type) channels with nifedipine or verapamil, sensitized atria to the negative inotropic action of the A1 agonist, R-PIA, without affecting the nucleoside negative chronotropy. Patch-clamp experiments in the whole-cell configuration mode demonstrate that adenosine, via A1 receptors, activates the inwardly-rectifying GIRK/KIR3.1/KIR3.4 current resulting in hyperpolarization of atrial cardiomyocytes, which may slow down heart rate. Conversely, the nucleoside inactivates a small conductance Ca(2+)-activated KCa2/SK outward current, which eventually reduces the repolarizing force and thereby prolong action potentials duration and Ca(2+) influx into cardiomyocytes. Immunolocalization studies showed that differences in A1 receptors distribution between the sinoatrial node and surrounding cardiomyocytes do not afford a rationale for adenosine chronoselectivity. Immunolabelling of KIR3.1, KCa2.2, KCa2.3, and Cav1 was also observed throughout the right atrium. Functional data indicate that while both A1 and M2 receptors favor the opening of GIRK/KIR3.1/3.4 channels modulating atrial chronotropy, A1 receptors may additionally restrain KCa2/SK activation thereby compensating atrial inotropic depression by increasing the time available for Ca(2+) influx through Cav1 (L-type) channels. PMID:27014060

  20. Synaptic Gain-of-Function Effects of Mutant Cav2.1 Channels in a Mouse Model of Familial Hemiplegic Migraine Are Due to Increased Basal [Ca2+]i

    OpenAIRE

    Di Guilmi, Mariano N.; Wang, Tiantian; Inchauspe, Carlota Gonzalez; Forsythe, Ian D; Ferrari, Michel D; van den Maagdenberg, Arn M. J. M.; Borst, J. Gerard G.; Uchitel, Osvaldo D.

    2014-01-01

    Specific missense mutations in the CACNA1A gene, which encodes a subunit of voltage-gated CaV2.1 channels, are associated with familial hemiplegic migraine type 1 (FHM1), a rare monogenic subtype of common migraine with aura. We used transgenic knock-in (KI) mice harboring the human pathogenic FHM1 mutation S218L to study presynaptic Ca2+ currents, EPSCs, and in vivo activity at the calyx of Held synapse. Whole-cell patch-clamp recordings of presynaptic terminals from S218L KI mice showed a s...

  1. Cortical synaptic transmission in CaV2.1 knockin mice with the S218L missense mutation which causes a severe familial hemiplegic migraine syndrome in humans.

    Directory of Open Access Journals (Sweden)

    Dania eVecchia

    2015-02-01

    Full Text Available Familial hemiplegic migraine type 1 (FHM1 is caused by gain-of-function mutations in CaV2.1 (P/Q-type Ca2+ channels. Knockin (KI mice carrying the FHM1 R192Q missense mutation show enhanced cortical excitatory synaptic transmission at pyramidal cell synapses but unaltered cortical inhibitory neurotransmission at fast-spiking interneuron synapses. Enhanced cortical glutamate release was shown to cause the facilitation of cortical spreading depression (CSD in R192Q KI mice. It, however, remains unknown how other FHM1 mutations affect cortical synaptic transmission. Here, we studied neurotransmission in cortical neurons in microculture from KI mice carrying the S218L mutation, which causes a severe FHM syndrome in humans and an allele-dosage dependent facilitation of experimental CSD in KI mice, which is larger than that caused by the R192Q mutation. We show gain-of-function of excitatory neurotransmission, due to increased action-potential evoked Ca2+ influx and increased probability of glutamate release at pyramidal cell synapses, but unaltered inhibitory neurotransmission at multipolar interneuron synapses in S218L KI mice. In contrast with the larger gain-of-function of neuronal CaV2.1 current in homozygous than heterozygous S218L KI mice, the gain-of-function of evoked glutamate release, the paired-pulse ratio and the Ca2+ dependence of the EPSC were all similar in homozygous and heterozygous S218L KI mice, suggesting compensatory changes in the homozygous mice. Furthermore, we reveal a unique feature of S218L KI cortical synapses which is the presence of a fraction of mutant CaV2.1 channels being open at resting potential. Our data suggest that, while the gain-of-function of evoked glutamate release may explain the facilitation of CSD in heterozygous S218L KI mice, the further facilitation of CSD in homozygous S218L KI mice is due to other CaV2.1-dependent mechanisms, that likely include Ca2+ influx at voltages sub-threshold for action

  2. Optimalisation de l'extraction d'ADN génomique de la morelle jaune (Solanum elaeagnifolium Cav., une plante invasive des milieux cultivés en région méditerranéenne

    Directory of Open Access Journals (Sweden)

    Ripoll, J.

    2011-01-01

    Full Text Available Optimization of the genomic DNA extraction method of silverleaf nightshade (Solanum elaeagnifolium Cav., an invasive plant in the cultivated areas within the Mediterranean region. The geographical origin of an invasive plant in the cultivated area within the Mediterranean region, silverleaf nightshade, Solanum elaeagnifolium Cav. (Solanaceae should be identified through the analysis of genetic similarities between native and introduced populations using microsatellite markers. Beforehand, an effective and less costly method for extracting genomic DNA from dehydrated and polysaccharide-rich leaves was investigated. The original Doyle's protocol based on the use of a cationic detergent, CTAB, was successfully optimized in order to extract high quality DNA (A260/280 ratio of 1.8; A260/230 ratio of 1.7 to 2. The DNA yields obtained with this new protocol were higher than those obtained with a commercial kit, although quality of the DNA extracted was comparable. The improvement of the protocol was further proven by the amplification of several microsatellite loci first developed in eggplant, Solanum melongena L.

  3. Evaluation of furcation perforation repair by MTA and Cav-Hycal in beagle dog%两种材料修复比格犬髓室底穿孔的效果评价

    Institute of Scientific and Technical Information of China (English)

    侯惠敏; 侯本祥; 卫书盛

    2011-01-01

    Objective To investigate the effect of MTA and Cav-Hycal for the repair of beagle dogs'furcation perforations. Methods Thirty-six premolars of three adult beagle dogs with the root canals filled were randomly divided into two groups. Furcation perforations were made and repaired with either MTA or Cav-Hycal. Twenty-four molars served as the positive and negative contros. Fureation perforations were made and covered with polyvinychlorid in positive control and no furcation perforations were made in negative control The periodontal status were evaluated 1, 4 and 12 weeks after treatment. Results At one week, all experimental teeth showed normal. At 4 weeks, minor to severe bone loss were observed in the two experimental groups. The teeth in positive group showed periodontal pockets with severe bone loss. At 12 weeks, the bone defects were entirely or partly repaired in 12 cases of either MTA or Cav-Hycal group with no periodontal pockets. The bone defects in the positive control group became bigger and the periodontal pockets became deeper. The teeth in negative control group showed nomal. There was no significant difference between MTA and Cav-Hycal group( P >0. 05). Conclusion The effect of MTA and Cav-Hycal on the repair of furcation perforations in beagle dogs was similar.%目的:评价MTA和光固化氢氧化钙修复比格犬牙齿髓室底穿孔的效果.方法:3只成年比格犬的36颗前磨牙根管治疗后做髓室底穿孔模型,随机分为两组,分别采用MTA和光固化氢氧化钙修复穿孔.另外24颗磨牙根管治疗后随机分为阳性和阴性对照组,阳性对照组做髓室底穿孔模型后以聚氯乙烯膜覆盖穿孔,阴性对照组不穿孔.分别于术后1,4,12周进行牙周检查及X线影像学评价.结果:术后1周,各组牙周及X线片检查未见异常.术后4周,两实验组各12例牙周未见异常,各6例牙周破坏,所有样本不同程度骨质破坏;阳性对照组12例牙周破坏伴重度骨质破坏.术后12

  4. Dose and time-dependent sub-chronic toxicity study of hydroethanolic leaf extract of Flabellaria paniculata Cav. (Malpighiaceae in rodents

    Directory of Open Access Journals (Sweden)

    Abidemi James Akindele

    2014-04-01

    Full Text Available Flabellaria paniculata Cav. (Malpighiaceae is a climbing shrub, the preparations of which are used in the treatment of wounds and ulcers in Nigeria and Ghana. This study investigated the sub-chronic toxicity profile of the hydroethanolic leaf extract of Flabellaria paniculata (HLE-FP. HLE-FP was administered p.o. (20, 100 and 500 mg/kg for 30 and 60 days to different groups of rats. Control animals received 10 ml/kg distilled water. In the group of animals for reversibility study, HLE-FP administration ceased on the 60th day and animals were monitored for a further 15 days. Results showed that oral treatment with HLE-FP for 30 days caused significant (p0.05 differences in relative organ weights between control and treatment groups were observed. HLE-FP-treated rats showed significant (p< 0.05 increases in Hb, PCV and RBC on day 30 and significant (p< 0.05 increases in MCV and MCH indices on day 60 compared to control. There were significant (p< 0.05 elevations in serum K+, urea and creatinine compared to control. The liver function tests showed slightly but non-significant alterations when compared to control. Biochemical findings were supported by histopathological observations of vital organs including the kidney and liver. Toxicities observed in respect of kidney function were irreversible at 15 days of stoppage of treatment. In the acute toxicity study, HLE-FP given p.o. caused no lethality at 5000 mg/kg but behavioural manifestations like restlessness, generalized body tremor, feed and water refusal were observed. The i.p. LD50 was estimated to be 2951.2 mg/kg. Findings in this study showed that HLE-FP is relatively non-toxic on acute exposure and generally safe on sub-chronic administration, but could be deleterious on the kidneys on prolonged oral exposure at a high dose. Thus, caution should be exercised with i

  5. Nitric oxide inhibits neuroendocrine CaV1 L-channel gating via cGMP-dependent protein kinase in cell-attached patches of bovine chromaffin cells

    Science.gov (United States)

    Carabelli, Valentina; D'Ascenzo, Marcello; Carbone, Emilio; Grassi, Claudio

    2002-01-01

    Nitric oxide (NO) regulates the release of catecholamines from the adrenal medulla but the molecular targets of its action are not yet well identified. Here we show that the NO donor sodium nitroprusside (SNP, 200 μM) causes a marked depression of the single CaV1 L-channel activity in cell-attached patches of bovine chromaffin cells. SNP action was complete within 3-5 min of cell superfusion. In multichannel patches the open probability (NPo) decreased by ∼60 % between 0 and +20 mV. Averaged currents over a number of traces were proportionally reduced and showed no drastic changes to their time course. In single-channel patches the open probability (Po) at +10 mV decreased by the same amount as that of multichannel patches (∼61 %). Such a reduction was mainly associated with an increased probability of null sweeps and a prolongation of mean shut times, while first latency, mean open time and single-channel conductance were not significantly affected. Addition of the NO scavenger carboxy-PTIO or cell treatment with the guanylate cyclase inhibitor ODQ prevented the SNP-induced inhibition. 8-Bromo-cyclicGMP (8-Br-cGMP; 400 μM) mimicked the action of the NO donor and the protein kinase G blocker KT-5823 prevented this effect. The depressive action of SNP was preserved after blocking the cAMP-dependent up-regulatory pathway with the protein kinase A inhibitor H89. Similarly, the inhibitory action of 8-Br-cGMP proceeded regardless of the elevation of cAMP levels, suggesting that cGMP/PKG and cAMP/PKA act independently on L-channel gating. The inhibitory action of 8-Br-cGMP was also independent of the G protein-induced inhibition of L-channels mediated by purinergic and opiodergic autoreceptors. Since Ca2+ channels contribute critically to both the local production of NO and catecholamine release, the NO/PKG-mediated inhibition of neuroendocrine L-channels described here may represent an important autocrine signalling mechanism for controlling the rate of

  6. Nitric oxide inhibits neuroendocrine Ca(V)1 L-channel gating via cGMP-dependent protein kinase in cell-attached patches of bovine chromaffin cells.

    Science.gov (United States)

    Carabelli, Valentina; D'Ascenzo, Marcello; Carbone, Emilio; Grassi, Claudio

    2002-06-01

    Nitric oxide (NO) regulates the release of catecholamines from the adrenal medulla but the molecular targets of its action are not yet well identified. Here we show that the NO donor sodium nitroprusside (SNP, 200 microM) causes a marked depression of the single Ca(V)1 L-channel activity in cell-attached patches of bovine chromaffin cells. SNP action was complete within 3-5 min of cell superfusion. In multichannel patches the open probability (NP(o)) decreased by approximately 60 % between 0 and +20 mV. Averaged currents over a number of traces were proportionally reduced and showed no drastic changes to their time course. In single-channel patches the open probability (P(o)) at +10 mV decreased by the same amount as that of multichannel patches (approximately 61 %). Such a reduction was mainly associated with an increased probability of null sweeps and a prolongation of mean shut times, while first latency, mean open time and single-channel conductance were not significantly affected. Addition of the NO scavenger carboxy-PTIO or cell treatment with the guanylate cyclase inhibitor ODQ prevented the SNP-induced inhibition. 8-Bromo-cyclicGMP (8-Br-cGMP; 400 microM) mimicked the action of the NO donor and the protein kinase G blocker KT-5823 prevented this effect. The depressive action of SNP was preserved after blocking the cAMP-dependent up-regulatory pathway with the protein kinase A inhibitor H89. Similarly, the inhibitory action of 8-Br-cGMP proceeded regardless of the elevation of cAMP levels, suggesting that cGMP/PKG and cAMP/PKA act independently on L-channel gating. The inhibitory action of 8-Br-cGMP was also independent of the G protein-induced inhibition of L-channels mediated by purinergic and opiodergic autoreceptors. Since Ca(2+) channels contribute critically to both the local production of NO and catecholamine release, the NO/PKG-mediated inhibition of neuroendocrine L-channels described here may represent an important autocrine signalling mechanism

  7. Influência da temperatura e da quantidade de água no substrato sobre a germinação de sementes de Ochroma pyramidale (Cav. ex Lam.) Urban (pau-de-balsa)

    OpenAIRE

    Ramos Michele Braule Pinto; Varela Vania Palmeira; Melo Maria de Fátima Figueiredo

    2006-01-01

    O presente trabalho foi conduzido com o objetivo de avaliar diferentes volumes de água no substrato e temperaturas na germinação de sementes de Ochroma pyramidale (Cav. ex Lam.) Urban. Antes da instalação dos testes de germinação, as sementes foram tratadas com imersão em água quente a 80º C até o resfriamento, para superar a dormência. A semeadura foi realizada em rolos de papel germitest, umedecidos com volumes (mL) de água equivalentes a 1,5; 2,0; 2,5 e 3,0 vezes o peso do substrato nas te...

  8. Effects of natural phenolic compounds from a desert dominant shrub Larrea divaricata Cav. on toxicity and survival in mice Efectos de los compuestos fenólicos naturales de un arbusto dominante del desierto, Larrea divaricata Cav. sobre la toxicidad y sobrevida en ratones

    Directory of Open Access Journals (Sweden)

    J.M. RÍOS

    2008-06-01

    Full Text Available It is known that generalist herbivores may circumvent intoxication by ingesting small quantities from a mixture of plant secondary metabolites. However a single chemical, a highly toxic one or the most abundant in the mixture could cause toxicity. Survivorship and toxicity in Rockland male mice were measured to determine if the toxic effects of the phenolic resin of creosote bush (Larrea divaricata Cav. is due to its major constituent, the nordihydroguaiaretic acid (NDGA or to the total concentration of phenolic compounds in this resin. This objective was accomplished by exposing mice to voluntary feeding on resin-treated rat chow and by oral gavaging of mice with the following doses and compounds: Resin-100 mg (n = 7, NDGA-15 mg (n = 8, and NDGA-100 mg (n = 10 and Control (n = 6. Our hypothesis was that NDGA is responsible for the toxicity of Larrea divaricata's phenolic resin. Voluntary resin intake by mice had a pronounced toxic effect, producing body mass loss and significant reduction of food intake. Mice gavaged with Resin-100 mg, NDGA-100 mg, and NDGA-15 mg showed a significant reduction in survival probability compared to mice under Control conditions. Animáis exposed to NDGA-15 mg had a higher survivorship compared to the NDGA-100 mg animáis, and equivalent survivorship to the Resin-100 mg (containing 15 mg of NDGA animáis. No significant differences in detoxification, measured as glucuronic acid conjugates in urine, were detected among gavage treatments. Therefore, given that just 15 mg of NDGA were enough to produce the same effect as the whole resin, we suggest that NDGA is the main constituent of Larrea divaricata's resin responsible for the toxic effect of the phenolic resin of this plant.Los herbívoros generalistas pueden evitar la intoxicación ingiriendo pequeñas cantidades de una mezcla de metabolitos secundarios de plantas. Sin embargo, un solo compuesto, uno altamente tóxico o el más abundante en la mezcla es el que

  9. The importance of arbuscular mycorrhiza for Cyclamen purpurascens subsp. immaculatum endemic in Slovakia

    Czech Academy of Sciences Publication Activity Database

    Rydlová, Jana; Sýkorová, Zuzana; Slavíková, Renata; Turis, P.

    2015-01-01

    Roč. 25, č. 8 (2015), s. 599-609. ISSN 0940-6360 Institutional support: RVO:67985939 ; RVO:61388971 Keywords : wild cyclamens * Septoglomus constrictum * growth response Subject RIV: EF - Botanics; EE - Microbiology, Virology (MBU-M) Impact factor: 3.459, year: 2014

  10. Cav 1.3 L-type Ca ( 2+) channels mediate long-term adaptation in dopamine D2L-mediated GluA1 trafficking in the dorsal striatum following cocaine exposure.

    Science.gov (United States)

    Schierberl, Kathryn; Giordano, Thomas; Satpute, Shirish; Hao, Jin; Kaur, Gagandeep; Hofmann, Franz; Moosmang, Sven; Striessnig, Joerg; Rajadhyaksha, Anjali

    2012-01-01

    AMPA receptor (AMPAR) plasticity at glutamatergic synapses in the mesostriatal dopaminergic pathway has been implicated in persistent cocaine-induced behavioral responses; however, the precise mechanism underlying these changes remains unknown. Utilizing cocaine psychomotor sensitization in mice we find that repeated cocaine results in a basal reduction of Ser 845 GluA1 and cell surface GluA1 levels in the dorsal striatum (dStr) following a protracted withdrawal period, an adaptation that is dependent on Cav 1.3 channels but not those expressed in the VTA. We find that the basally-induced decrease in this phosphoprotein is the result of recruitment of the striatal dopamine D2 pathway, as evidenced by enhanced levels of D2 receptor (D2R) mRNA expression and D2R function as examined using the D2R antagonist, eticlopride, as well as alterations in the phosphorylation status of several downstream molecular targets of D2R's, including CREB, DARPP-32, Akt and GSK3β. Taken together with our recently published findings examining similar phenomena in the nucleus accumbens (NAc), these results underscore the utilization of divergent molecular mechanisms in the dStr, in mediating cocaine-induced persistent behavioral changes. PMID:22419037

  11. NMR Studies of the Vanadium Spin Dynamics and Spin Structure in LiV2O4, CaV2O4, and (LixV1-x)3BO5 (x is almost equal to 0.33, 0.40)

    International Nuclear Information System (INIS)

    Strong electron correlation is believed to be an essential and unifying factor in diverse properties of condensed matter systems. Ground states that can arise due to electron correlation effects include Mott insulators, heavy fermion, ferromagnetism and antiferromagnetism, spin glasses, and high-temperature superconductivity. The electronic systems in transition metal oxide compounds are often highly correlated. In this thesis, the author presents experimental studies on three strongly correlated vanadium oxide compounds: LiV2O4, (LixV1-x)3BO5, and CaV2O4, which have completely different ground states

  12. Influência da temperatura e da quantidade de água no substrato sobre a germinação de sementes de Ochroma pyramidale (Cav. ex Lam. Urban (pau-de-balsa

    Directory of Open Access Journals (Sweden)

    Ramos Michele Braule Pinto

    2006-01-01

    Full Text Available O presente trabalho foi conduzido com o objetivo de avaliar diferentes volumes de água no substrato e temperaturas na germinação de sementes de Ochroma pyramidale (Cav. ex Lam. Urban. Antes da instalação dos testes de germinação, as sementes foram tratadas com imersão em água quente a 80º C até o resfriamento, para superar a dormência. A semeadura foi realizada em rolos de papel germitest, umedecidos com volumes (mL de água equivalentes a 1,5; 2,0; 2,5 e 3,0 vezes o peso do substrato nas temperaturas de 25, 30 e 35º C. O delineamento experimental adotado foi o inteiramente casualizado com os tratamentos em esquema fatorial 4 x 3. Para cada tratamento, utilizaram-se quatro repetições de 25 sementes. Após o encerramento do experimento (aos 21 dias, foi avaliada a porcentagem, o índice de velocidade de germinação e os comprimentos da raiz primária e do hipocótilo. A porcentagem de germinação final não foi influenciada pelos fatores estudados. O índice de velocidade de germinação foi favorecido pelas maiores temperaturas, 30 e 35º C. A temperatura de 30º C e a quantidade de água de 1,5 vezes o peso do papel, e 35º C e a quantidade de 3,0 vezes, foram as mais indicadas para o desenvolvimento da raiz primária e do hipocótilo da espécie. Para a germinação das sementes de O. pyramidale recomenda-se a temperatura de 30º C com a quantidade de água de 1,5 vezes o peso do papel, e a temperatura de 35º C com a quantidade de água de 3,0 vezes.

  13. Effects of Lactic Acid Bacteria Inoculant on the Fermentation Quality of Reed Grass (Phragmites australis Cav. Trin. ex Sterd. ) at Low Temperature%低温条件下混合乳酸菌制剂对芦苇发酵品质的影响

    Institute of Scientific and Technical Information of China (English)

    王鹏; 白春生; 刘林; 曹兵海

    2011-01-01

    为探讨低温条件下混合乳酸菌制剂对芦苇(Phragmites australis Cav.Trin.ex Sterd.)发酵品质的影响,对含水量为40%的芦苇添加混合乳酸菌制,分别设计4℃和0℃的发酵温度,密封发酵,分别取3,4,5,6,7和8周的发酵样品分析.结果表明:在0℃条件下,发酵后第5周pH值出现显著下降(P<0.05),第8周NDF含量显著降低(P<0.05).在4℃条件下,发酵第3周起pH值一直保持在4.20以下.在相同发酵时间内,4℃条件比0℃条件时的pH值显著降低(P<0.05),乳酸菌数量、乳酸产量显著提高(P<0.05),乙酸产量显著降低(P<0.05).在低温条件下,对每克干物质添加6.63(lg CFU·g-1DM)的混合乳酸菌,可使含水量为40%的芦苇启动发酵.为保证得到优良发酵质量,发酵时间应在6周以上.%Effects of lactic acid bacteria inoculant (LABs) on the fermentation quality of the reed grass silage (Phragmites australis Cav. Trin ex Sterd. ) at a lower temperature level was studied. The reed grass which had 40% water was ensiled with LABs. The silages were stored at 4℃ and 0℃, and then sampled at 3, 4, 5, 6, 7 and 8 weeks for analyzing respectively. Results showed that the pH of the silages stored at 0C were decreased significantly (P<0. 05) at 5th week. The contents of NDF (neutral detergent fiber) were decreased significantly (P<0.05) at 8th week. The pH of the silages stored at 4℃ dropped to be below 4.20 from 5th week. At the same ensiling time, the pH and the acetic acid content of the silages stored at 4℃ were significantly lower (P<0. 05), and the content of lactic acid were significantly higher (P< 0.05) than those stored at 0℃. At low temperature level, the reed grass silage of which moisture is 40% could start fermentation process if the 6.63 (lg CFU · g-1 DM) LABs were added to every gram of DM. For getting good silage, the ensiling time should be at least 6 weeks.

  14. Antibacterial activity of Zuccagnia punctata Cav. ethanolic extracts.

    Science.gov (United States)

    Zampini, Iris C; Vattuone, Marta A; Isla, Maria I

    2005-12-01

    The present study was conducted to investigate antibacterial activity of Zuccagnia punctata ethanolic extract against 47 strains of antibiotic-resistant Gram-negative bacteria and to identify bioactive compounds. Inhibition of bacterial growth was investigated using agar diffusion, agar macrodilution, broth microdilution and bioautographic methods. Zuccagnia punctata extract was active against all assayed bacteria (Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, Morganella morganii, Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia) with minimal inhibitory concentration (MIC) values ranging from 25 to 200 microg/mL. Minimal bactericidal concentration (MBC) values were identical or two-fold higher than the corresponding MIC values. Contact bioautography, indicated that Zuccagnia punctata extracts possess one major antibacterial component against Pseudomonas aeruginosa and at least three components against. Klebsiella pneumoniae and Escherichia coli. Activity-guided fractionation of 1he ethanol extract on a silica gel column yielded a compound (2',4'-dihydroxychalcone), which exhibited strong antibacterial activity with MIC values between 0.10 and 1.00 microg/mL for Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, Morganella morganii, Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia. These values are lower than imipenem (0.25-16 microg/mL). Zuccagnia punctata might provide promising therapeutic agents against infections with multi-resistant Gram-negative bacteria. PMID:16137849

  15. Genetics Home Reference: CAV3-related distal myopathy

    Science.gov (United States)

    ... gene causes a peculiar form of distal myopathy. Neurology. 2002 Jan 22;58(2):323-5. Erratum in: Neurology 2002 Mar 12;58(5):839. Itoyoma Y [ ... 3 cause four distinct autosomal dominant muscle diseases. Neurology. 2004 Feb 24;62(4):538-43. Review. ...

  16. High Frequency Pulsatile Electromagnetic Fields and Ultrasound Pulsatile Fields Impact on Germination Dynamic at Ocimum basilicum L. and O. basilicum var. purpurascens Benth., Observed with Open Source Software’s

    Directory of Open Access Journals (Sweden)

    Ioan ONAC

    2016-06-01

    Full Text Available Basil (Ocimum sp., generally knowned as “king of herbs” due to its pharmaceutical and culinary properties, is considered to be a native plant derived from Africa and Asia. Ocimum sp. grows in tropical and sub-tropical regions with more than 50 species knowned till present time. Germination percent of Ocimum sp. is considered to be between 85-95%. Even though Ocimum sp. are considered to have fast germination velocity index (GVI, short mean germination time (MGT and increased seedling vigor index (SVI values, their experimental monitoring is done with specialized software’s that are expensive. Low cost scientific solutions are keen on open source software germination protocols. For observing the accuracy of open source C.A.D. software’s Ocimum sp. seeds where submitted to high frequency pulsatile electromagnetic fields (300 impulses/3 peak penetrance - 293 W and ultrasound pulsatile fields (1/2 pulses by 0.5 W/cm2. All sequence images where taken using a positive/negative photo filter enhancing differences in seed development. At the end of the experiment all images where transformed into vector formats (dwg. On the dwg extension, selective free plug-ins such as Face Centroid and Area Properties help to collect data like seed development on X/Y scale, area, perimeter, no. of germinated seeds, length of seedling root, hypocotyl length.

  17. High Frequency Pulsatile Electromagnetic Fields and Ultrasound Pulsatile Fields Impact on Germination Dynamic at Ocimum basilicum L. and O. basilicum var. purpurascens Benth., Observed with Open Source Software’s

    OpenAIRE

    Ioan ONAC; Valentin SINGUREANU; Gelu MOLDOVAN; Rodica UNGUR

    2016-01-01

    Basil (Ocimum sp.), generally knowned as “king of herbs” due to its pharmaceutical and culinary properties, is considered to be a native plant derived from Africa and Asia. Ocimum sp. grows in tropical and sub-tropical regions with more than 50 species knowned till present time. Germination percent of Ocimum sp. is considered to be between 85-95%. Even though Ocimum sp. are considered to have fast germination velocity index (GVI), short mean germination time (MGT) and increased seedling vigor...

  18. Effect of Flabellaria paniculata Cav. extracts on gastric ulcer in rats

    Directory of Open Access Journals (Sweden)

    Sofidiya Margaret O

    2012-10-01

    Full Text Available Abstract Background The leaves and root of Flabellaria paniculata (Malpighiaceae are frequently used in the treatment of wounds and ulcers in Nigerian folk medicine. The purpose of this study was to compare the effect of ethanolic extracts from the leaves (FPL and root (FPR of F. paniculata on gastric ulcers in rats. Methods The effect of FPL and FPR (100, 200 and 400 mg/kg was evaluated in ethanol and indomethacin gastric ulcer models. Control groups for FPL and FPR were orally treated with 3% Tween 20 and distilled water respectively. FPL was further investigated in pylorus ligation model. Misoprostol and cimetidine were used as reference. Results FPL significantly (P 50 of FPL was estimated to be 4570 mg/kg while that of FPR was 2754 mg/kg. The LD50 in intraperitoneal injection was estimated to be 1202.26 and 1380.38 mg/kg for FPL and FPR respectively. The phytochemical investigation showed that both extracts possess triterpenoids and saponin, while the presence of flavonoid was detected only in FPL. Conclusions The results of this study indicated that FPL and not FPR is effective against experimentally induced gastric ulcers. The presence of varied phytochemical constituents probably influenced the pharmacological differences between the two extracts.

  19. Opaque closed chambers underestimate methane fluxes of Phragmites australis (Cav.) Trin. ex Steud.

    Science.gov (United States)

    Günther, Anke; Jurasinski, Gerald; Huth, Vytas; Glatzel, Stephan

    2014-04-01

    Closed chamber measurements for methane emission estimation are often carried out with opaque chambers to avoid heating of the headspace. However, mainly in wetlands, some plants possess an internal convective gas transport which quickly responds to changes in irradiation. These plants have also been found to often channel a large part of the released methane in temperate fens. We compare methane fluxes derived from transparent versus opaque chambers on Carex-, Phragmites-, and Typha-dominated stands of a temperate fen. Transparent chamber fluxes almost doubled opaque chamber fluxes in the convective transporting Phragmites stand. In Typha, a trend of higher fluxes determined with the transparent chambers was detectable, whereas in Carex, transparent and opaque chamber fluxes did not differ significantly. Thus, opaque chambers bias the outcome of methane measurements, depending on dominant vegetation. We recommend the use of transparent chambers when determining emissions of convective plants or extrapolating fluxes to larger scales. PMID:24213640

  20. Directrices académico-docentes en el grado de CAV: Un estudio exploratorio multifuente

    OpenAIRE

    Romeo Delgado, Marina; Yepes i Baldó, Montserrat; Sánchez, Lydia; Burset Burillo, Sílvia; García Asensio, Ma. Ángeles (María Ángeles); González Argüello, Ma. Vicenta (María Vicenta); Gustems Carnicer, Josep; Martín Piñol, Carolina; Bosch, Emma; Berger, Rita; Aguilar, Carlos

    2014-01-01

    En este artículo se presenta una experiencia de investigación en el marco de los estudios de Comunicación Audiovisual de la Universidad de Barcelona. El objetivo ha sido diseñar las directrices académico-docentes de la materia Proyectos del Grado de Comunicación Audiovisual. Durante los meses de octubre y noviembre de 2012, se realizó un estudio cualitativo descriptivo, de carácter exploratorio y con metodología multifuente para identificar aquellos aspectos que permiten el desarrollo de una ...

  1. cav-p60 expression in rat muscle tissues. Distribution of caveolar proteins

    DEFF Research Database (Denmark)

    Voldstedlund, M.; Vinten, Jørgen; Tranum-Jensen, Jørgen

    Caveolae, Caveolin, muscle, endothel, electron microscopy, immunofluorescence microscopy, rat (Wistar)......Caveolae, Caveolin, muscle, endothel, electron microscopy, immunofluorescence microscopy, rat (Wistar)...

  2. Directrices académico-docentes en el grado de CAV: Un estudio exploratorio multifuente

    Directory of Open Access Journals (Sweden)

    Marina Romeo

    2014-01-01

    Full Text Available En este artículo se presenta una experiencia de investigación en el marco de los estudios de Comunicación Audiovisual de la Universidad de Barcelona. El objetivo ha sido diseñar las directrices académico-docentes de la materia Proyectos del Grado de Comunicación Audiovisual. Durante los meses de octubre y noviembre de 2012, se realizó un estudio cualitativo descriptivo, de carácter exploratorio y con metodología multifuente para identificar aquellos aspectos que permiten el desarrollo de una propuesta didáctica caracterizada por el desarrollo de las competencias del estudiante. Se optó por realizar dos seminarios prospectivos como técnica de recogida de datos. Esta metodología es considerada especialmente útil para estudiar la generación de representaciones sociales, dado que a partir del debate grupal emerge el significado compartido acerca del tema de discusión. Los informantes fueron identificados como clave por el hecho de ser docentes, egresados y/o empleadores de Comunicación Audiovisual. El análisis de contenido muestra un cierto grado de consenso entre profesores, profesionales y egresados con respecto a los conocimientos/habilidades requeridos por los estudiantes y docentes para el desarrollo de un proyecto audiovisual. Se evidencia la conceptualización del proyecto como un proceso y un producto que requiere la puesta en práctica de conocimientos y habilidades, así como el papel relevante que estudiantes y profesores tienen frente a los empleadores. Así mismo se observa el gap existente entre las competencias descritas en la titulación y las expresadas en los seminarios prospectivos. Mientras la titulación apuesta principalmente por el desarrollo de las competencias relativas al “saber hacer”, los empleadores, docentes y egresados focalizan principalmente las relativas al “saber ser”. La principal contribución de esta experiencia es reflexionar sobre los mecanismos que permiten al profesorado diseñar de forma colaborativa, junto con egresados y profesionales del sector, pautas y estrategias de enseñanza-aprendizaje en el marco del Espacio Europeo de la Educación Superior (EEES.

  3. Chemical Composition and Antibacterial Activity of Essential Oil of Cosmos bipinnatus Cav. Leaves from South Africa

    OpenAIRE

    Olajuyigbe, Olufunmiso; Ashafa, Anofi

    2014-01-01

    The chemical composition of essential oils isolated from the leaves of Cosmos bipinnatus and its antibacterial activity were analyzed by GC-MS and microbroth dilution assay respectively. The essential oil extracted from this plant was predominantly composed of monoterpenes (69.62%) and sesquiterpenes (22.73%). The antibacterial assay showed that the oil had significant inhibitory effects against both Gram-negative and Gram-positive bacteria isolates. The MIC of Gram-positive strains ranged be...

  4. Variation of volatiles in Tunisian populations of Thymbra capitata (L.) Cav. (Lamiaceae).

    Science.gov (United States)

    Ali, Imen Ben El Hadj; Guetat, Arbi; Boussaid, Mohamed

    2012-07-01

    Volatiles from 14 wild Tunisian populations of Thymbra capitata (=Thymus capitatus Hoffmanns. et Link=Coridothymus capitatus Rchb.f.), sampled in five ecological areas (sub-humid, upper semi-arid, mean semi-arid, lower semi-arid, and upper arid areas) were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) techniques. Thirty-nine constituents representing 94.2 to 99.5% of the total oil were identified. Carvacrol (38; 51.1-75.9%), p-cymene (13; 3.7-15%), γ-terpinene (12; 1.4-11.9%), and trans-β-caryophyllene (22; 2.9-4.6%) are the major compounds. A significant variation among populations and population's bioclimatic stage for the majority of compounds was shown. The chemical population structure, estimated using a principal-component analysis (PCA) and an unweighted pair group method with arithmetic mean (UPGMA) cluster analysis performed on all populations and compounds and based on Euclidean distances among populations, was high. Both methods allowed separation of the populations into distinct groups defined rather by minor than by major compounds. The spatial compound distribution is linked to ecological factors, indicating that local selective environmental factors influence chemical-composition diversity. Conservation strategies should involve all populations because of their low size and their high level of destruction. Populations exhibiting particular compounds other than the major ones should first be protected. In situ conservation of populations should be accomplished appropriately according to bioclimate. PMID:22782875

  5. Chromosome studies in some Stevia. Cav. (Compositae) species from Southern Brazil

    OpenAIRE

    Adriana P. Frederico; Paulo M. Ruas; Marin-Morales, Maria A.; Claudete F. Ruas; Jimi N. Nakajima

    1996-01-01

    Karyotypes of six species of the genus Stevia from Southern Brazil were studied, utilizing root tip metaphases. All species were diploid with 2n = 22 chromosomes. It was possible to identify each species by chromosome morphology. The basic chromosome number for Brazilian species of Stevia is X = 11. This number is also found in almost all South American species. We suggest that in Stevia there is an evolutionary trend toward chromosomal rearrangement, caused mainly by pericentric inversions. ...

  6. Chromosome studies in some Stevia. Cav. (Compositae species from Southern Brazil

    Directory of Open Access Journals (Sweden)

    Adriana P. Frederico

    1996-01-01

    Full Text Available Karyotypes of six species of the genus Stevia from Southern Brazil were studied, utilizing root tip metaphases. All species were diploid with 2n = 22 chromosomes. It was possible to identify each species by chromosome morphology. The basic chromosome number for Brazilian species of Stevia is X = 11. This number is also found in almost all South American species. We suggest that in Stevia there is an evolutionary trend toward chromosomal rearrangement, caused mainly by pericentric inversions. It was found that, in addition to aneuploidy and polyploidy, chromosomal rearrangements are common in the tribe Eupatorieae.

  7. Porcellium conspersum, een in Nederland zeer zeldzame landpissebed van vochtige bossen (Crustacea: Isopoda: Oniscoidea)

    NARCIS (Netherlands)

    Soesbergen, M.

    1999-01-01

    The influence of recent inundations on the distribution pattern of the isopod Eluma purpurascens in the province of Zeeland (Crustacea: Isopoda: Oniscoidea) Small scale mapping of Eluma purpurascens Budde-Lund, 1885 in the province of Zeeland revealed a remarkable distribution pattern. The species p

  8. The Calmodulin-Binding, Short Linear Motif, NSCaTE Is Conserved in L-Type Channel Ancestors of Vertebrate Cav1.2 and Cav1.3 Channels

    OpenAIRE

    Taiakina, Valentina; Boone, Adrienne N.; Fux, Julia; Senatore, Adriano; Weber-Adrian, Danielle; Guillemette, J. Guy; Spafford, J. David

    2013-01-01

    NSCaTE is a short linear motif of (xWxxx(I or L)xxxx), composed of residues with a high helix-forming propensity within a mostly disordered N-terminus that is conserved in L-type calcium channels from protostome invertebrates to humans. NSCaTE is an optional, lower affinity and calcium-sensitive binding site for calmodulin (CaM) which competes for CaM binding with a more ancient, C-terminal IQ domain on L-type channels. CaM bound to N- and C- terminal tails serve as dual detectors to changing...

  9. POTENCIAL DE PROPAGACIÓN in vitro PARA EL TOMATE DE ÁRBOL PARTENOCÁRPICO Cyphomandra betacea Cav. (Sendt POTENTIAL OF in vitro PROPAGATION FOR THE TOMATO TREE PARTENOCARPIC Cyphomandra betacea Cav. (Sendt

    Directory of Open Access Journals (Sweden)

    John Alexander Espinosa Orrego

    2005-06-01

    Full Text Available Se evaluaron 10 medios de cultivo para el establecimiento y la multiplicación in vitro de tomate de árbol partenocárpico, los cuales contenían sales de MS, BAP de 0,17 a 5,82 mg/l, y AIA de 0,19 a 2,31 mg/l y una mezcla de tres sustancias antioxidantes: cisteína, ácido ascórbico y caseína hidrolizada (100 mg/l de cada una. Los explantes utilizados fueron esquejes de nudo de 1,5 a 2,0 cm de longitud. Para el análisis estadístico se empleó un diseño central compuesto triplicado con dos repeticiones del punto central. Los resultados fueron: en el material proveniente del altiplano Norte de Antioquia, Colombia, se determinó el punto óptimo para la variable longitud de brotes con reguladores de crecimiento (0,17 mg/l de BAP y 0,19 mg/l de AIA; en el material procedente del Oriente de Antioquia-Colombia, no se pudo determinar el punto óptimo. Las sustancias antioxidantes no fueron efectivas para evitar la oxidación fenólica, la cual fue mediana a alta. Las tasas de multiplicación fueron bajas (0 % y 20 % para los genotipos del Oriente y Norte de Antioquia, respectivamente y el desarrollo de los brotes en el material del Norte fue muy lento (cuatro meses.Ten tissue culture medias were evaluated to establish and multiply in vitro the tomato tree partenocarpic, that contained MS salts, BAP from 0,17 to 5,82 mg/l, and IAA from 0,17 to 2,31 mg/l and a mix of three antiozidant substances : cistein, ascorbic acid, and hydrolized casein (100 mg/l each. The explants employed were nodal stems of from 1, 5 to 2,0 cm in length. For the statistical analyses, a triplicated central composite design was employed with two repetitions of the central point. The results were: in the material from the high plains of northern Antioquia, Colombia , the optimal point was determined for the variable shoot length with growth regulators (0,17 mg/l of BAP and 0,19 mg/l of IAA; for the material from eastern Antioquia-Colombia, it was not possible to determine the optimum point. The antioxidizing substances were not effective for preventing fenolic oxidation, which was intermediate to high. Multiplication rates were low (0 and 20 % for the eastern and northern genotypes, respectively and shoot development in the material from the north was very slow (four months.

  10. DETECCIÓN DE LOS VIRUS AMV, CMV Y PLRV EN CULTIVOS DE TOMATE DE ÁRBOL (SOLANUM BETACEUM CAV.) EN ANTIOQUIA, COLOMBIA DETECTION OF VIRUSES AMV, CMV AND PLRV IN TAMARILLO (SOLANUM BETACEUM CAV.) ORCHARDS IN ANTIOQUIA, COLOMBIA

    OpenAIRE

    Margarita Jaramillo Zapata; Pablo Andrés Gutiérrez Sánchez; José Miguel Cotes Torres; Elena Paola González Jaimes; Mauricio Marín Montoya

    2011-01-01

    El tomate de árbol es uno de los principales frutales cultivados en la región andina de Colombia. Durante los últimos años las áreas dedicadas a su plantación se han reducido drásticamente, debido entre otras razones, al efecto detrimental de la enfermedad conocida como Virosis. A pesar de la importancia económica de esta enfermedad, el nivel de conocimiento que se tiene en el país, sobre la identidad taxonómica de los virus asociados a su ocurrencia es precario y por tanto, se carece de buen...

  11. DETECCIÓN DE LOS VIRUS AMV, CMV Y PLRV EN CULTIVOS DE TOMATE DE ÁRBOL (SOLANUM BETACEUM CAV. EN ANTIOQUIA, COLOMBIA DETECTION OF VIRUSES AMV, CMV AND PLRV IN TAMARILLO (SOLANUM BETACEUM CAV. ORCHARDS IN ANTIOQUIA, COLOMBIA

    Directory of Open Access Journals (Sweden)

    Margarita Jaramillo Zapata

    2011-06-01

    Full Text Available El tomate de árbol es uno de los principales frutales cultivados en la región andina de Colombia. Durante los últimos años las áreas dedicadas a su plantación se han reducido drásticamente, debido entre otras razones, al efecto detrimental de la enfermedad conocida como Virosis. A pesar de la importancia económica de esta enfermedad, el nivel de conocimiento que se tiene en el país, sobre la identidad taxonómica de los virus asociados a su ocurrencia es precario y por tanto, se carece de buena parte de la información necesaria para el diseño de estrategias de manejo efectivas contra esta enfermedad. En este trabajo se evaluó la presencia e incidencia de los virus isométricos AMV, CMV y PLRV en cultivos de tomate de árbol establecidos en el departamento de Antioquia, mediante el empleo de pruebas de ELISA, RT-PCR y secuenciación de la cápside viral. Las pruebas serológicas permitieron detectar la presencia de los tres virus, aunque las secuencias generadas sólo fueron confirmatorias para el virus PLRV, lo que posiblemente sugiere que existen diferencias en las secuencias dirigidas a los sitios de alineamiento de los cebadores para AMV y CMV. La incidencia de los virus CMV y PLRV en las muestras analizadas fue de un 57 y 41% respectivamente, mientras que para AMV ésta solo llegó a un nivel del 2%. Los hallazgos obtenidos permiten confirmar la presencia de un complejo viral asociado a la enfermedad de la Virosis del tomate de árbol en el territorio mencionado.Tamarillo is one of the main fruit crops in the Colombian andean region. However, during the past years, there has been a drastic reduction in the total crop area due to the detrimental effect of a disease known as Virosis. In spite of the economic impact of this disease, there is little knowledge about the taxonomical identity of its associated viruses; therefore, no information is available for the design of appropriate control strategies. In this work, was evaluated the presence and incidence levels of AMV, CMV and PLRV viruses in tamarillo crops in the department of Antioquia by ELISA, RT-PCR and DNA sequencing of the viral capsid. Even though serological tests detected the presence of all three viruses, DNA sequencing could only confirm the presence of PRLV. The failure to detect AMV and CMV at the DNA level suggests differences in the target sequences used for primer design. CMV and PLRV were detected in 57 and 41% of all the analysed samples in contrast to AMV whose incidence level was only 2%. These findings confirm the presence of viral complex associated to the Virosis disease of tamarillo orchards in the mentioned territory.

  12. Naamlijst van de Nederlandse landpissebedden (Crustacea: Isopoda: Oniscidea)

    NARCIS (Netherlands)

    Berg, M.P.

    1997-01-01

    Checklist of Dutch woodlice (Crustacea: Isopoda: Oniscidea). An updated checklist for the terrestrial isopods of The Netherlands is presented. Four species, Armadillo officinalis, Eluma purpurascens, Miktoniscus patiencei, and Trichoniscoides sarsi are new to the woodlice fauna of The Netherlands co

  13. Induction of resistance to blast disease (Pyricularia oryzae Cav.) in the high yielding variety, Ratna (IR 8 x TKM 6)

    International Nuclear Information System (INIS)

    The popular high yielding variety, Ratna (IR 8 x TKM 6), with a duration of 110 days, is susceptible to blast disease, (Pyricularia oryzae). Dehusked seeds of Ratna were pre-soaked in distilled water for 4 hours and then subjected to 0.1% and 0.2% EMS for 6 hours. The percentage of germination was lower in 0.2% EMS but survival higher. Seedlings of M1 were raised in pots and panicles of individual plants were harvested separately for M2 generation. The seeds of M2 generation were grown in ''Uniform Blast Nursery'' for observing blast disease reaction. Seeds of resistant plants were harvested and grown in M3 generation. Some susceptible plants were also carried forward to M3 generation. Ratna control (untreated) developed ''B'', ''C'' and ''D'' type spots. The EMS treated population (8000 plants) developed in addtion, ''O'', ''A'' as well as ''E'' type spots, showing a wider range of variability than the untreated control regarding resistance and susceptibility. Three hundred resistant plants (with ''O, ''A'' and ''B'' type of spots) from M2 were carried forward to M3 generation. These also segregated giving a range of reaction from 0-E spots as in the case of M2. The population of resistant plants was higher in M3 (31.65%) than in M2 (23.6%). The progenies of a few plants (50 plants) found susceptible in M2 also segregated in M3 towards resistance, but the percentage of resistant plants was low (7.6%)

  14. Microscopic analysis and histochemical observations of the medicinal root of Iostephane heterophylla (Cav.) Benth. ex Hemsl. (Asteraceae)

    OpenAIRE

    Estela Sandoval; Bye, Robert A.; Griselda Ríos; María Isabel Aguilar

    2005-01-01

    The roots of Iostephane heterophylla are popular in Mexican traditional medicine and as such are a good candidate to develop herbal drug preparations to be used as phytomedicine. International criteria for validation and standardization of a herbal product as phytomedicine include, among others, the integration of microscopic and histochemical characteristics of the raw material, as in this case the herbal drug, to guarantee its authenticity. As an original contribution to the knowledge of th...

  15. Cultivo in vitro del tomate de árbol (Cyphomandra betacea (Cav. Sendt. (Fenotipo naranja proveniente de Costa Rica

    Directory of Open Access Journals (Sweden)

    Randall Chacón-Cerdas

    2014-06-01

    Se determinó que la desinfección con 2,5% de Ca(ClO2 fue la mejor para el establecimiento in vitro; el mejor medio de cultivo para la micropropagación del material fue el M1, compuesto por sales MS(1962 al 100%, sacarosa al 3%, phytagel® 1,8g/L, 0,5mg/L de AG3, 0,25mg/L de BAP y 2,0 mg/L de PaCa, el cual presentó el mejor balance entre el número promedio de brotación/explante y el número promedio de entrenudos/explante, sin formación excesiva de callo, mientras que el medio de cultivo E5, constituido por sales MS(1962 al 100%, agar 6,0g/L y sacarosa al 3%, sin reguladores del crecimiento y el medio de cultivo E6, que incluía las sales MS(1962 al 100%, agar 8,0g/L y sacarosa al 3%, sin reguladores del crecimiento, mostraron el periodo más corto para la formación de raíces, el mayor número promedio de raíces y la mayor longitud promedio de raíz y tallo.

  16. Growth of Phragmites australis (Cav.) Trin ex. Steudel in mine water treatment wetlands: effects of metal and nutrient uptake

    International Nuclear Information System (INIS)

    The abandoned mine of Shilbottle Colliery, Northumberland, UK is an example of acidic spoil heap discharge that contains elevated levels of many metals. Aerobic wetlands planted with the common reed, Phragmites australis, were constructed at the site to treat surface runoff from the spoil heap. The presence of a perched water table within the spoil heap resulted in the lower wetlands receiving acidic metal contaminated water from within the spoil heap while the upper wetland receives alkaline, uncontaminated surface runoff from the revegetated spoil. This unique situation enabled the comparison of metal uptake and growth of plants used in treatment schemes in two cognate wetlands. Results indicated a significant difference in plant growth between the two wetlands in terms of shoot height and seed production. Analyses of metal and nutrient concentrations within plant tissues provided the basis for three hypotheses to explain these differences: (i) the toxic effects of high levels of metals in shoot tissues (ii) the inhibition of Ca (an essential nutrient) uptake by the presence of metals and H+ ions, and (iii) low concentrations of bioavailable nitrogen sources resulting in nitrogen deficiency. This has important implications for the engineering of constructed wetlands in terms of the potential success of plant establishment and vegetation development

  17. Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

    OpenAIRE

    Warnke, Elisabeth; Pietsch, Jessica; Wehland, Markus; Bauer, Johann; Infanger, Manfred; Görög, Mark; Hemmersbach, Ruth; Braun, Markus; Ma, Xiao; Sahana, Jayashree; Grimm, Daniela

    2014-01-01

    Background: Multicellular tumor spheroids (MCTS) formed scaffold-free under microgravity are of high interest for research and medicine. Their formation mechanism can be studied in space in real microgravity or on Earth using ground-based facilities (GBF), which simulate microgravity. On Earth, these experiments are more cost-efficient and easily performable. However, each GBF might exert device-specific and altered superimposingly gravity-dependent effects on the cells. Result...

  18. Effect of the essential volatile oils isolated from Thymbra capitata (L.) Cav. on olive and sunflower oils

    OpenAIRE

    Miguel, Maria Graça; Figueiredo, A. C.; M.M. Costa; Martins, D; Barroso, J.G.; Pedro, L. G.

    2003-01-01

    The chemical composition of the volatile constituents of the oils isolated from different parts of Thymbra capitata collected at different developmental stages were analysed by GC and GC/MS. The antioxidant ability of the oils isolated from T capitata was evaluated determining the peroxide values, on olive and sunflower oils, stored at 60 degreesC. These peroxide values were compared with those obtained when BHT, carvacrol and control (without adding antioxidants) were used and subjected to t...

  19. Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) andOriganum vulgare L.

    Science.gov (United States)

    Faleiro, Leonor; Miguel, Graça; Gomes, Sónia; Costa, Ludmila; Venâncio, Florencia; Teixeira, Adriano; Figueiredo, A Cristina; Barroso, José G; Pedro, Luis G

    2005-10-19

    Antilisterial activities of Thymbra capitata and Origanum vulgare essential oils were tested against 41 strains of Listeria monocytogenes. The oil of T. capitata was mainly constituted by one component, carvacrol (79%), whereas for O. vulgare three components constituted 70% of the oil, namely, thymol (33%), gamma-terpinene (26%), and p-cymene (11%). T. capitata essential oil had a significantly higher antilisterial activity in comparison to O. vulgare oil and chloramphenicol. No significant differences in L. monocytogenes susceptibilities to the essential oils tested were registered. The minimum inhibitory concentration values of T. capitata essential oil and of carvacrol were quite similar, ranging between 0.05 and 0.2 microL/mL. Antioxidant activity was also tested, the essential oil of T. capitata showing significantly higher antioxidant activity than that of O. vulgare. Use of T. capitata and O. vulgare essential oils can constitute a powerful tool in the control of L. monocytogenes in food and other industries. PMID:16218659

  20. Effect of the essential volatile oils isolated from Thymbra capitata (L. Cav. on olive and sunflower oils

    Directory of Open Access Journals (Sweden)

    Pedro, L.

    2003-09-01

    Full Text Available The chemical composition of the volatile constituents of the oils isolated from different parts of Thymbra capitata collected at different developmental stages were analysed by GC and GC/MS. The antioxidant ability of the oils isolated from T. capitata was evaluated determining the peroxide values, on olive and sunflower oils, stored at 60 ºC. These peroxide values were compared with those obtained when BHT, carvacrol and control (without adding antioxidants were used and subjected to the same conditions. The best yield oil was obtained from the whole aerial part of T. capitata collected during the flowering phase. The major component of the oils was carvacrol. Relative high amounts of p-cymene, γ-terpinene and β-caryophyllene were also found. BHT revealed to be the best antioxidant when the olive oil was used. On sunflower oil, the antioxidant ability of BHT was not so evident, being the carvacrol-rich essential oils of T. capitata or carvacrol more important antioxidants.Se analizaron, mediante GC y GC/MS, los componentes volátiles de aceites aislados de las distintas partes de la Thymbra capitata, recogida en diferentes etapas de desarrollo. Se evaluó la actividad antioxidante de estos aceites de la T. capitata, midiendo el índice de peróxidos, en aceites de oliva y girasol, almacenados a 60 ºC. Estos índices de peróxidos se compararon con los obtenidos cuando no se agregó ningún antioxidante (control y cuando se utilizó BHT o carvacrol, en las mismas condiciones de almacenamiento. El mayor rendimiento en aceite se obtuvo de la parte aérea de T. capitata recogida durante la etapa de floración. El componente mayoritario de los aceites fue el carvacrol. También se encontraron, cantidades relativamente elevadas, de p-cimeno, γ-terpineno y β-cariofileno. El mejor antioxidante para el aceite de oliva resultó ser el BHT. En el aceite del girasol, la actividad antioxidante del BHT no fue tan evidente, mientras que el carvacrol y los aceites esenciales de T. capitata ricos en carvacrol mostraron una mayor actividad antioxidante.

  1. Molecular endpoints of Ca2+/calmodulin- and voltage-dependent inactivation of Cav1.3 channels

    OpenAIRE

    Tadross, Michael R.; Johny, Manu Ben; Yue, David T.

    2010-01-01

    Ca2+/calmodulin- and voltage-dependent inactivation (CDI and VDI) comprise vital prototypes of Ca2+ channel modulation, rich with biological consequences. Although the events initiating CDI and VDI are known, their downstream mechanisms have eluded consensus. Competing proposals include hinged-lid occlusion of channels, selectivity filter collapse, and allosteric inhibition of the activation gate. Here, novel theory predicts that perturbations of channel activation should alter inactivation i...

  2. Growth of Phragmites australis (Cav.) Trin ex. Steudel in mine water treatment wetlands: effects of metal and nutrient uptake

    Energy Technology Data Exchange (ETDEWEB)

    Lesley C. Batty; Paul L. Younger [University of Newcastle, Newcastle upon Tyne (United Kingdom). School of Civil Engineering and Geosciences

    2004-11-01

    The abandoned mine of Shilbottle Colliery, Northumberland, UK is an example of acidic spoil heap discharge that contains elevated levels of many metals. Aerobic wetlands planted with the common reed, Phragmites australis, were constructed at the site to treat surface runoff from the spoil heap. The presence of a perched water table within the spoil heap resulted in the lower wetlands receiving acidic metal contaminated water from within the spoil heap while the upper wetland receives alkaline, uncontaminated surface runoff from the revegetated spoil. This unique situation enabled the comparison of metal uptake and growth of plants used in treatment schemes in two cognate wetlands. Results indicated a significant difference in plant growth between the two wetlands in terms of shoot height and seed production. Analyses of metal and nutrient concentrations within plant tissues provided the basis for three hypotheses to explain these differences: (i) the toxic effects of high levels of metals in shoot tissues, (ii) the inhibition of Ca (an essential nutrient) uptake by the presence of metals and H{sup +} ions, and (iii) low concentrations of bioavailable nitrogen sources resulting in nitrogen deficiency. This has important implications for the engineering of constructed wetlands in terms of the potential success of plant establishment and vegetation development.

  3. Growth of Phragmites australis (Cav.) Trin ex. Steudel in mine water treatment wetlands: effects of metal and nutrient uptake

    Energy Technology Data Exchange (ETDEWEB)

    Batty, Lesley C.; Younger, Paul L

    2004-11-01

    The abandoned mine of Shilbottle Colliery, Northumberland, UK is an example of acidic spoil heap discharge that contains elevated levels of many metals. Aerobic wetlands planted with the common reed, Phragmites australis, were constructed at the site to treat surface runoff from the spoil heap. The presence of a perched water table within the spoil heap resulted in the lower wetlands receiving acidic metal contaminated water from within the spoil heap while the upper wetland receives alkaline, uncontaminated surface runoff from the revegetated spoil. This unique situation enabled the comparison of metal uptake and growth of plants used in treatment schemes in two cognate wetlands. Results indicated a significant difference in plant growth between the two wetlands in terms of shoot height and seed production. Analyses of metal and nutrient concentrations within plant tissues provided the basis for three hypotheses to explain these differences: (i) the toxic effects of high levels of metals in shoot tissues (ii) the inhibition of Ca (an essential nutrient) uptake by the presence of metals and H{sup +} ions, and (iii) low concentrations of bioavailable nitrogen sources resulting in nitrogen deficiency. This has important implications for the engineering of constructed wetlands in terms of the potential success of plant establishment and vegetation development.

  4. Rhizome age structure of three populations of Phragmites australis (Cav.) TRIN ex STEUD.: Biomass and mineral nutrient concentrations

    Czech Academy of Sciences Publication Activity Database

    Čížková, Hana; Lukavská, Jaroslava

    1999-01-01

    Roč. 34, č. 3 (1999), s. 209-220. ISSN 0015-5551 R&D Projects: GA ČR GA206/96/0589; GA AV ČR KSK2017602 Grant ostatní: EV5V-CT92-0083 (EUREED)(XX) CIPDCT925064 Subject RIV: EF - Botanics Impact factor: 0.393, year: 1999

  5. Auto-stimulatory action of secreted caveolin-1 on the proliferation of Ewing’s sarcoma cells

    OpenAIRE

    Sengupta, Aniruddha; Mateo-Lozano, Silvia; Tirado, Oscar M.; Notario, Vicente

    2011-01-01

    Caveolin-1 (CAV1) is highly expressed in Ewing’s sarcoma (EWS). We previously showed that increased cellular CAV1 is associated with the regulation of the tumorigenicity, drug resistance and metastatic ability of EWS cells. Because several studies reported that melanoma and prostate cancer cells, which express relatively high CAV1 levels, secrete CAV1, and that secreted CAV1 is associated with tumor progression, our study explored the possibility that EWS cells also secreted CAV1 and that sec...

  6. Characterization of mAbs to chicken anemia virus and epitope mapping on its viral protein, VP1.

    Science.gov (United States)

    Trinh, Dai Q; Ogawa, Haruko; Bui, Vuong N; Baatartsogt, Tugsbaatar; Kizito, Mugimba K; Yamaguchi, Shigeo; Imai, Kunitoshi

    2015-05-01

    Three (MoCAV/F2, MoCAV/F8 and MoCAV/F11) of four mouse mAbs established against the A2/76 strain of chicken anemia virus (CAV) showed neutralization activity. Immunoprecipitation showed a band at ~50 kDa in A2/76-infected cell lysates by neutralizing mAbs, corresponding to the 50 kDa capsid protein (VP1) of CAV, and the mAbs reacted with recombinant VP1 proteins expressed in Cos7 cells. MoCAV/F2 and MoCAV/F8 neutralized the 14 CAV strains tested, whereas MoCAV/F11 did not neutralize five of the strains, indicating distinct antigenic variation amongst the strains. In blocking immunofluorescence tests with the A2/76-infected cells, binding of MoCAV/F11 was not inhibited by the other mAbs. MoCAV/F2 inhibited the binding of MoCAV/F8 to the antigens and vice versa, suggesting that the two mAbs recognized the same epitope. However, mutations were found in different parts of VP1 of the escape mutants of each mAb: EsCAV/F2 (deletion of T89+A90), EsCAV/F8 (I261T) and EsCAV/F11 (E144G). Thus, the epitopes recognized by MoCAV/F2 and MoCAV/F8 seemed to be topographically close in the VP1 structure, suggesting that VP1 has at least two different neutralizing epitopes. However, MoCAV/F8 did not react with EsCAV/F2 or EsCAV/F8, suggesting that binding of MoCAV/F8 to the epitope requires coexistence of the epitope recognized by MoCAV/F2. In addition, MoCAV/F2, with a titre of 1 : 12 800 to the parent strain, neutralized EsCAV/F2 and EsCAV/F8 with low titres of 32 and 152, respectively. The similarity of the reactivity of MoCAV/F2 and MoCAV/F8 to VP1 may also suggest the existence of a single epitope recognized by these mAbs. PMID:25568186

  7. The Timothy syndrome mutation differentially affects voltage- and calcium-dependent inactivation of CaV1.2 L-type calcium channels

    OpenAIRE

    Barrett, Curtis F.; Tsien, Richard W.

    2008-01-01

    Calcium entry into excitable cells is an important physiological signal, supported by and highly sensitive to the activity of voltage-gated Ca2+ channels. After membrane depolarization, Ca2+ channels first open but then undergo various forms of negative feedback regulation including voltage- and calcium-dependent inactivation (VDI and CDI, respectively). Inactivation of Ca2+ channel activity is perturbed in a rare yet devastating disorder known as Timothy syndrome (TS), whose features include...

  8. Species biodiversity of microscopic fungi of common reed (Phragmites australis (Cav. Trin. ex Steud. in the rush communities of Lake Glinno

    Directory of Open Access Journals (Sweden)

    Kinga Mazurkiewicz-Zapałowicz

    2012-12-01

    Full Text Available The studies were carried out on common reed (Phragmites australis growing on the shores of Lake Glinno and forming rush communities of the alliance Phragmition. 10 plants with disease symptoms were gathered from each of five sites. The isolation and marking of pathogens were performed twice from fragments of leaf and blade tissues with disease symptoms. First, directly after collecting the plants incubated in sterile humid chambers and microorganism cultures on CDA and PDA medium, and then phytopathogen and saprotroph species occurring on dried green material were identified for 2-4 months. The occurrence of 31 species of microscopic fungi overall was observed on the leaves, blades and inflorescences of P. australis, including 2 mycelia of Mycelia sterilia. The most frequently occurring species, present at all sites of Phragmites australis are: Alternaria alternata, Cladosporium cladosporioides, C. herbarum, Doratomyces stemonitis and Puccinia phragmitis, P. magnussiana and two mycelia of Mycelia sterilia. The most common species occurring on blades and inside them are: Acremoniella atra, Acremonium alternatum and Fusarium sambucinum. Sporadically, Ustilago grandis was also observed inside blades. Tiny necrotic stains on leaves and blades were caused by the presence of three species of the genus Leptoshaeria: L. culmifraga, L. eustoma and L. fuckelli. The occurrence of the sclerote of Claviceps microcephala was found in inflorescences.

  9. Resistencia de campo a la antracnosis de los frutos (Colletotrichum gloeosporioides en tomate de arbol (Cyphomandra (Solanum betacea (betaceum Cav. Sendt.

    Directory of Open Access Journals (Sweden)

    Lobo A. Mario

    2000-12-01

    Full Text Available En el Centro de Investigación "La Selva", de la Corporación Colombiana de Investigación Agropecuaria (CORPOICA, localizado en Rionegro (Antioquia, Colombia,a 2120 msnm, se llevó a cabo una investigación, cuyo objetivo fue evaluar la resistencia de campo a la antracnosis de los frutos de tomate de árbol en la accesión 6975073 de especie C. uniloba, material que en observaciones previas no había presentado incidencia de la enfermedad. En el estudio se incluyeron igualmente 4 poblaciones de la especie cultivada C. betacea de diferentes tipos: rojo común, amarillo, partenocárpico y "mora" o "tamarillo" y una de la especie relacionada C. materna. El trabajo se llevó a cabo durante seis meses, cosechándose quincenalmente, en 5 árboles por accesión, los frutos sanos maduros y los enfermos, clasificados por grado de maduración. Todos los materiales recibieron control quí mico de la enfermedad, con excepción del tomate C. uniloba 6975073. Durante la evaluación se presentaron condiciones favorables para el desarrollo del patógeno, con una precipitación de 1361.1 mm y prevalencia de altas humedades relativas, con un promedio, al respecto, del 80.2%. Durante el período de desarrollo de la evaluación no se presentaron frutos con síntomas de antracnosis en la accesión 6975073, lo cual si ocurrió en los demás materiales, con un porcentaje de frutos enfermos que fluctuó entre el 57.64 y el 98.82%, encontrá ndose que los genotipos con mayor nivel de productividad, a su vez, presentaron la mayor suceptibilidad y pérdidas en rendimiento. Un estimado del valor de los frutos con antracnosis, proyectado a hectárea-año, señaló que las pérdidas de frutos en las diferentes accesiones de C. betacea fluctuaron entre 3006 y 11268 dólares. Complementariamente al estudio, se obtuvieron híbridos interespecí ficos entre el material resistente y una accesión de C. betacea.

  10. Raptor ablation in skeletal muscle decreases Cav1.1 expression and affects the function of the excitation–contraction coupling supramolecular complex

    Science.gov (United States)

    Lopez, Rubén J.; Mosca, Barbara; Treves, Susan; Maj, Marcin; Bergamelli, Leda; Calderon, Juan C.; Bentzinger, C. Florian; Romanino, Klaas; Hall, Michael N.; Rüegg, Markus A.; Delbono, Osvaldo; Caputo, Carlo; Zorzato, Francesco

    2016-01-01

    The protein mammalian target of rapamycin (mTOR) is a serine/threonine kinase regulating a number of biochemical pathways controlling cell growth. mTOR exists in two complexes termed mTORC1 and mTORC2. Regulatory associated protein of mTOR (raptor) is associated with mTORC1 and is essential for its function. Ablation of raptor in skeletal muscle results in several phenotypic changes including decreased life expectancy, increased glycogen deposits and alterations of the twitch kinetics of slow fibres. In the present paper, we show that in muscle-specific raptor knockout (RamKO), the bulk of glycogen phosphorylase (GP) is mainly associated in its cAMP-non-stimulated form with sarcoplasmic reticulum (SR) membranes. In addition, 3[H]–ryanodine and 3[H]–PN200-110 equilibrium binding show a ryanodine to dihydropyridine receptors (DHPRs) ratio of 0.79 and 1.35 for wild-type (WT) and raptor KO skeletal muscle membranes respectively. Peak amplitude and time to peak of the global calcium transients evoked by supramaximal field stimulation were not different between WT and raptor KO. However, the increase in the voltage sensor-uncoupled RyRs leads to an increase of both frequency and mass of elementary calcium release events (ECRE) induced by hyper-osmotic shock in flexor digitorum brevis (FDB) fibres from raptor KO. The present study shows that the protein composition and function of the molecular machinery involved in skeletal muscle excitation–contraction (E–C) coupling is affected by mTORC1 signalling. PMID:25431931

  11. Nueva especie de Condalia Cav. (Rhamnaceae) y notas sobre los géneros de la familia en la flora de Colombia

    OpenAIRE

    1997-01-01

    Se describe e ilustra Condalia thomasiana, una nueva especie de Rhamnaceae, que constituye el primer registro de este genero para Colombia. Se comentan aspectos ecoldgicos y afinidades taxondmicas de la nueva especie. Se presenta una clave para las especies consideradas m8s cercanas y una para los gdneros de ramnáceas conocidos en Colombia.

  12. Accumulation of nutrients and heavy metals in Phragmites australis (Cav.) Trin. ex Steudel and Bolboschoenus maritimus (L.) Palla in a constructed wetland of the Venice lagoon watershed

    Energy Technology Data Exchange (ETDEWEB)

    Bragato, Claudia [Department of Agricultural Biotechnology, University of Padova, Agripolis, 35020 Legnaro (PD) (Italy); Brix, Hans [Department of Biological Sciences, Plant Biology, University of Aarhus, Ole Worms Alle, Building 1135, DK-8000 Arhus C (Denmark); Malagoli, Mario [Department of Agricultural Biotechnology, University of Padova, Agripolis, 35020 Legnaro (PD) (Italy)]. E-mail: mario.malagoli@unipd.it

    2006-12-15

    A recently constructed wetland, located in the Venice lagoon watershed, was monitored to investigate growth dynamics, nutrient and heavy metal shoot accumulation of the two dominating macrophytes: Phragmites australis and Bolboschoenus maritimus. Investigations were conducted over a vegetative season at three locations with different distance to the inlet point to assess effects on vegetation. The distance from the inlet did not affect either shoot biomass or nutrients (N, P, K and Na) and heavy metals (Cr, Ni, Cu and Zn) shoot content. With the exception of Na, nutrient and heavy metal concentrations were higher in shoots of P. australis than in B. maritimus. Heavy metal concentration in the incoming water and in the soil was not correlated to the plant content of either species. Shoot heavy metal concentrations were similar to those reported in the current literature, but accumulation generally increased towards the end of the growing season. - Heavy metal shoot concentration in Phragmites australis and Bolboschoenus maritimus increased significantly at the end of the growing season.

  13. Anatomical localization of Cav3.1 calcium channels and electrophysiological effects of T-type calcium channel blockade in the motor thalamus of MPTP-treated monkeys.

    Science.gov (United States)

    Devergnas, Annaelle; Chen, Erdong; Ma, Yuxian; Hamada, Ikuma; Pittard, Damien; Kammermeier, Stefan; Mullin, Ariana P; Faundez, Victor; Lindsley, Craig W; Jones, Carrie; Smith, Yoland; Wichmann, Thomas

    2016-01-01

    Conventional anti-Parkinsonian dopamine replacement therapy is often complicated by side effects that limit the use of these medications. There is a continuing need to develop nondopaminergic approaches to treat Parkinsonism. One such approach is to use medications that normalize dopamine depletion-related firing abnormalities in the basal ganglia-thalamocortical circuitry. In this study, we assessed the potential of a specific T-type calcium channel blocker (ML218) to eliminate pathologic burst patterns of firing in the basal ganglia-receiving territory of the motor thalamus in Parkinsonian monkeys. We also carried out an anatomical study, demonstrating that the immunoreactivity for T-type calcium channels is strongly expressed in the motor thalamus in normal and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys. At the electron microscopic level, dendrites accounted for >90% of all tissue elements that were immunoreactive for voltage-gated calcium channel, type 3.2-containing T-type calcium channels in normal and Parkinsonian monkeys. Subsequent in vivo electrophysiologic studies in awake MPTP-treated Parkinsonian monkeys demonstrated that intrathalamic microinjections of ML218 (0.5 μl of a 2.5-mM solution, injected at 0.1-0.2 μl/min) partially normalized the thalamic activity by reducing the proportion of rebound bursts and increasing the proportion of spikes in non-rebound bursts. The drug also attenuated oscillatory activity in the 3-13-Hz frequency range and increased gamma frequency oscillations. However, ML218 did not normalize Parkinsonism-related changes in firing rates and oscillatory activity in the beta frequency range. Whereas the described changes are promising, a more complete assessment of the cellular and behavioral effects of ML218 (or similar drugs) is needed for a full appraisal of their anti-Parkinsonian potential. PMID:26538609

  14. Association of the α2δ1 Subunit with Cav3.2 Enhances Membrane Expression and Regulates Mechanically Induced ATP Release in MLO-Y4 Osteocytes

    OpenAIRE

    Thompson, William R.; Majid, Amber S.; Czymmek, Kirk J; Ruff, Albert L.; García, Jesús; Duncan, Randall L.; Farach-Carson, Mary C.

    2011-01-01

    Voltage sensitive calcium channels (VSCCs) mediate signaling events in bone cells in response to mechanical loading. Osteoblasts predominantly express L-type VSCCs composed of the α1 pore-forming subunit and several auxiliary subunits. Osteocytes, in contrast, express T-type VSCCs, but a relatively small amount of L-type α1 subunits. Auxiliary VSCC subunits have several functions including modulating gating kinetics, trafficking of the channel and phosphorylation events. The influence of the ...

  15. Effect of the essential volatile oils isolated from Thymbra capitata (L.) Cav. on olive and sunflower oils

    OpenAIRE

    Pedro, L.; Barroso, J.G.; Martins, D; M.M. Costa; Figueiredo, A. C.; Miguel, M. G.

    2003-01-01

    The chemical composition of the volatile constituents of the oils isolated from different parts of Thymbra capitata collected at different developmental stages were analysed by GC and GC/MS. The antioxidant ability of the oils isolated from T. capitata was evaluated determining the peroxide values, on olive and sunflower oils, stored at 60 ºC. These peroxide values were compared with those obtained when BHT, carvacrol and control (without adding antioxidants) were used and subjecte...

  16. Efecto fitoestrogénico del extracto metanólico de frutos de Prosopis torquata (Cav. ex Lag.) en ratas Wistar hembras púberes

    OpenAIRE

    Carrasco, Mirta; Salinas, Adriana; ANTONIO M MANGIONE; Gil, Esteban R.

    2010-01-01

    Los fitoestrógenos, compuestos de estructura química diversa, actúan sobre la función reproductiva de los mamíferos. Se investigaron los efectos del extracto metanólico (ExM) de frutos de Prosopis torquata sobre las funciones reproductivas de la rata Wistar hembra, inyectadas en el período neonatal. Las crías fueron distribuidas al azar en cuatro grupos. Tres grupos, alimentados con dieta libre de fitoestrógenos, fueron inyectados con ExM, E (estradiol) y vehículo (Clf), respectiv...

  17. Opposite action of beta1- and beta2-adrenergic receptors on Ca(V)1 L-channel current in rat adrenal chromaffin cells.

    Science.gov (United States)

    Cesetti, T; Hernández-Guijo, J M; Baldelli, P; Carabelli, V; Carbone, E

    2003-01-01

    Voltage-gated Ca(2+) channels of chromaffin cells are modulated by locally released neurotransmitters through autoreceptor-activated G-proteins. Clear evidence exists in favor of a Ca(2+) channel gating inhibition mediated by purinergic, opioidergic, and alpha-adrenergic autoreceptors. Few and contradictory data suggest also a role of beta-adrenergic autoreceptors (beta-ARs), the action of which, however, remains obscure. Here, using patch-perforated recordings, we show that rat chromaffin cells respond to the beta-AR agonist isoprenaline (ISO) by either upmodulating or downmodulating the amplitude of Ca(2+) currents through two distinct modulatory pathways. ISO (1 microm) could cause either fast inhibition (approximately 25%) or slow potentiation (approximately 25%), or a combination of the two actions. Both effects were completely prevented by propranolol. Slow potentiation was more evident in cells pretreated with pertussis toxin (PTX) or when beta(1)-ARs were selectively stimulated with ISO + ICI118,551. Potentiation was absent when the beta(2)-AR-selective agonist zinterol (1 microm), the protein kinase A (PKA) inhibitor H89, or nifedipine was applied, suggesting that potentiation is associated with a PKA-mediated phosphorylation of L-channels (approximately 40% L-current increase) through beta(1)-ARs. The ISO-induced inhibition was fast and reversible, preserved in cell treated with H89, and mimicked by zinterol. The action of zinterol was mostly on L-channels (38% inhibition). Zinterol action preserved the channel activation kinetics, the voltage-dependence of the I-V characteristic, and was removed by PTX, suggesting that beta(2)AR-mediated channel inhibition was mainly voltage independent and coupled to G(i)/G(o)-proteins. Sequential application of zinterol and ISO mimicked the dual action (inhibition/potentiation) of ISO alone. The two kinetically and pharmacologically distinct beta-ARs signaling uncover alternative pathways, which may serve the autocrine control of Ca(2+)-dependent exocytosis and other related functions of rat chromaffin cells. PMID:12514203

  18. Structures of Ca(V) Ca**2+/CaM-IQ Domain Complexes Reveal Binding Modes That Underlie Calcium-Dependent Inactivation And Facilitation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, E.Y.; Rumpf, C.H.; Fujiwara, Y.; Cooley, E.S.; Petegem, F.Van; Minor, D.L., Jr.

    2009-05-20

    Calcium influx drives two opposing voltage-activated calcium channel (Ca{sub V}) self-modulatory processes: calcium-dependent inactivation (CDI) and calcium-dependent facilitation (CDF). Specific Ca{sup 2+}/calmodulin (Ca{sup 2+}/CaM) lobes produce CDI and CDF through interactions with the Ca{sub V}{alpha}{sub 1} subunit IQ domain. Curiously, Ca{sup 2+}/CaM lobe modulation polarity appears inverted between Ca{sub V}1s and Ca{sub V}2s. Here, we present crystal structures of Ca{sub V}2.1, Ca{sub V}2.2, and Ca{sub V}2.3 Ca{sup 2+}/CaM-IQ domain complexes. All display binding orientations opposite to Ca{sub V}1.2 with a physical reversal of the CaM lobe positions relative to the IQ {alpha}-helix. Titration calorimetry reveals lobe competition for a high-affinity site common to Ca{sub V}1 and Ca{sub V}2 IQ domains that is occupied by the CDI lobe in the structures. Electrophysiological experiments demonstrate that the N-terminal Ca{sub V}2 Ca{sup 2+}/C-lobe anchors affect CDF. Together, the data unveil the remarkable structural plasticity at the heart of Ca{sub V} feedback modulation and indicate that Ca{sub V}1 and Ca{sub V}2 IQ domains bear a dedicated CDF site that exchanges Ca{sup 2+}/CaM lobe occupants.

  19. Detección de los virus amv, cmv y plrv en cultivos de tomate de árbol (solanum betaceum cav.) en antioquia, colombia.

    OpenAIRE

    Jaramillo Zapata, Margarita; Gutiérrez Sánchez, Pablo Andrés; Cotes Torres, José Miguel; González Jaimes, Elena Paola; Marín Montoya, Mauricio

    2012-01-01

    El tomate de árbol es uno de los principales frutales cultivados en la región andina de Colombia. Durante los últimos años las áreas dedicadas a su plantación se han reducido drásticamente, debido entre otras razones, al efecto detrimental de la enfermedad conocida como Virosis. A pesar de la importancia económica de esta enfermedad, el nivel de conocimiento que se tiene en el país, sobre la identidad taxonómica de los virus asociados a su ocurrencia es precario y por tanto, se carece de buen...

  20. Accumulation of nutrients and heavy metals in Phragmites australis (Cav.) Trin. ex Steudel and Bolboschoenus maritimus (L.) Palla in a constructed wetland of the Venice lagoon watershed

    International Nuclear Information System (INIS)

    A recently constructed wetland, located in the Venice lagoon watershed, was monitored to investigate growth dynamics, nutrient and heavy metal shoot accumulation of the two dominating macrophytes: Phragmites australis and Bolboschoenus maritimus. Investigations were conducted over a vegetative season at three locations with different distance to the inlet point to assess effects on vegetation. The distance from the inlet did not affect either shoot biomass or nutrients (N, P, K and Na) and heavy metals (Cr, Ni, Cu and Zn) shoot content. With the exception of Na, nutrient and heavy metal concentrations were higher in shoots of P. australis than in B. maritimus. Heavy metal concentration in the incoming water and in the soil was not correlated to the plant content of either species. Shoot heavy metal concentrations were similar to those reported in the current literature, but accumulation generally increased towards the end of the growing season. - Heavy metal shoot concentration in Phragmites australis and Bolboschoenus maritimus increased significantly at the end of the growing season

  1. TARIM DIŞI ALANLARDA KAMIŞA (Phragmites australis (Cav.) Trin. Ex Steudel) KARŞI GLYPHOSATE VE IMAZAPYR UYGULAMA ETKİNLİKLERİNİN DEĞERLENDİRİLMESİ (EFFICACY EVALUATION OF GLYPHOSATE AND IMAZAPYR TREATMENTS AGAINST COMMON REED ((Phragmites australis (Cav.) Trin. Ex Steudel) IN NON-CROP LANDS)

    OpenAIRE

    TURABİ, Taha Semih

    2009-01-01

    Farklı glyphosate (500 ml/da, 750 ml/da, 1000 ml/da ve 500+500 ml/da split) ve imazapyr (200 ml/da, 300 ml/da ve 400 ml/da) uygulamalarının kamışa (Phragmites australis) karşı etkinliklerinin değerlendirilmesi amacıyla, Ankara İli’nde belirlenen iki farklı türden tarım dışı alanda denemeler kurularak Nisan-Eylül 2008 aylarında gözlemlenmiştir. Araştırma sonucunda en etkili uygulamanın glyphosate’nin 500+500 ml/da (split)’ dan elde edildiği, glyphosate’nin 1000 ml/da ve imazapyr’in 400 ml/da d...

  2. Comparative analysis of the heat transfer rates in constant (CAV) and variable (VAV) volumes type multi zone acclimation system operating in hot and humid climate; Analise comparativa das taxas transferencia de calor em sistemas de climatizacao do tipo volume de ar constante (CAV) e volume de ar variavel (VAV) multizona operando em clima quente e umido

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Cesar A.G.; Correa, Jorge E. [Para Univ., Belem (Brazil). Centro Tecnologico. Dept. de Engenharia Mecanica]. E-mails: gsantos@ufpa.br; jecorrea@amazon.com.br

    2000-07-01

    This work performs a comparative analysis among the constant and variable air volume multi zones acclimation systems, used for provide the thermal comfort in buildings. The work used the simulation HVAC2KIT computer program. The results of sensible and latent heats transfer rates on the cooling and dehumidification, inflating fan capacity, and heat transfer on the final heating condenser were obtained and analysed for the climate conditions of the Brazilian city of Belem from Para State, presenting hot and humid climate during all the year.

  3. Germination of seeds of the invasive plant Sesbania virgata (cav. pers. under effects of light, temperature, and dormancy overcoming Germinação de sementes da invasora Sesbania virgata (cav. pers. sob efeito de luz, temperatura e superação de dormência

    Directory of Open Access Journals (Sweden)

    Irenice Gomes de Oliveira

    2010-12-01

    Full Text Available The work had as objectives to evaluate the influence of the light, temperature, and chemical scarification on the germination of seeds of S. virgata, species still little studied and that is invasive in several niches of the caatinga ecosystem. The seeds of S. virgata were collected in the municipal district of Natuba-PB in March/2008. Eight treatments were used as follows: intact seeds in the presence and absence of light, at 25ºC; intact seeds in the presence and absence of light, at 30ºC; submerged seeds in sulfuric acid for five minutes in the presence and absence of light, at 25ºC; submerged seeds in sulfuric acid for five minutes in the presence and absence of light, at 30ºC. The germination tests were accomplished in germination cameras under light and continuous darkness at constant temperatures of 25ºC and 30ºC, in a 12 hours photoperiod. The seeds were put to germinate in plastic boxes (gerbox with filter paper. The seeds of S. virgata have tegument dormancy and show low germination tax when intact, which is a strategy for seed banks formation. The acid scarification increased germination percentage and speed germination index. Este trabalho teve como objetivos avaliar a influência da luz, da temperatura e da escarificação química sobre a germinação de sementes de S. virgata, espécie ainda pouco estudada e que se constata invasora em diversos nichos do bioma Caatinga. Sementes de S. virgata foram coletadas no município de Natuba-PB em março de 2008 e submetidas a oito tratamentos, descritos a seguir: sementes intactas, na presença e ausência de luz, a 25ºC; sementes intactas na presença e ausência de luz, a 30ºC; sementes imersas em ácido sulfúrico por cinco minutos na presença e ausência de luz, a 25ºC; sementes imersas em ácido sulfúrico por cinco minutos na presença e ausência de luz, a 30ºC. Para estimar os efeitos dos tratamentos foram realizados testes de germinação em câmaras de germinação sob luz e escuro contínuo a temperaturas constantes de 25ºC e 30ºC, em fotoperíodo de 12 horas. As sementes foram colocadas para germinar em caixas Gerbox forradas com folha de papel de filtro. Constatou-se que as sementes de S. virgata possuem dormência tegumentar e tem baixa porcentagem de germinação quando intactas, o que constitui uma estratégia para formação de bancos de sementes. A escarificação ácida provocou aumento da germinação e da velocidade de germinação das sementes.

  4. Germination of seeds of the invasive plant Sesbania virgata (cav.) pers. under effects of light, temperature, and dormancy overcoming
    Germinação de sementes da invasora Sesbania virgata (cav.) pers. sob efeito de luz, temperatura e superação de dormência

    OpenAIRE

    Irenice Gomes de Oliveira; Leonaldo Alves de Andrade; Pollyanna Freire Montenegro Agra; Vênia Camelo de Souza; Lamartine Soares de Oliveira

    2010-01-01

    The work had as objectives to evaluate the influence of the light, temperature, and chemical scarification on the germination of seeds of S. virgata, species still little studied and that is invasive in several niches of the caatinga ecosystem. The seeds of S. virgata were collected in the municipal district of Natuba-PB in March/2008. Eight treatments were used as follows: intact seeds in the presence and absence of light, at 25ºC; intact seeds in the presence and absence of light, at 30ºC; ...

  5. STABILITY OF ANTHOCYANINS FROM Rubus glaucus AND Solanum betaceum Cav.dark-red strain AS AFFECTED BY TEMPERATURE, STORAGE AND WATER ACTIVITY Efecto de la temperatura, almacenamiento y la actividad de agua sobre la estabilidad de antocianinas de Rubus glaucus y Solanum betaceum Cav.dark-red strain

    Directory of Open Access Journals (Sweden)

    C.M OLAYA

    Full Text Available The stability of sprayed-dried microencapsulated anthocyanins from Andes berry (Rubus glaucus and Tamarillo (Solanum betaceum, as affected by storage time, water activity (Aw and temperature was compared. The fruits were osmotically dehydrated with ethanol and the anthocyanin extract was microencapsulated with maltodextrin DE 20 by spray drying. Half life of the anthocyanins; changes in color, total phenolics, and antioxidant activity of the powders, were analyzed during storage at two different temperatures ( 25 °C and 40 °C and two Aw levels (0.20 and 0.35. A decrease in monomeric anthocyanin was observed in both samples. The half life of the Andes berry pigments ranged between 11 and 32 days while the half life of the tamarillo pigments ranged between 9 and 21 days. A darkening effect occurred in both samples as a result of storage time. The antioxidant activity decreased while the phenolic content increased with time. Antioxidant activity of Andes berry samples was highly correlated with anthocyanin content and total phenolic content while the antioxidant activity of tamarillo samples was highly correlated with total phenolic content. These results would be useful in developing applications for spray-dried anthocyanins as powdered food-grade colorants.Se comparó el efecto del tiempo de almacenamiento, la temperatura y la actividad de agua (Aw sobre la estabilidad de antocianinas microencapsuladas de Mora de Castilla (Rubus glaucus y tamarillo (Solanum betaceum. Las frutas se sometieron a deshidratación osmótica con etanol y el extracto antociánico se microencapsuló con maltodextrina ED 20 por atomización. La vida media de las antocianinas; los cambios en color, fenoles totales y actividad antioxidante se analizaron durante el almacenamiento a dos temperaturas ( 25 °C y 40 °C y dos niveles de Aw (0,20 y 0,35. Se observó una disminución de la antocianina monomérica en las dos muestras. La vida media de los pigmentos de mora de Castilla varió entre 11 y 32 días, mientras que la vida media de los pigmentos del tamarillo varió entre 9 y 21 días. Hubo oscurecimiento de las muestras como resultado del tiempo de almacenamiento. La actividad antioxidante de las dos muestras dismi-nuyó, mientras que el contenido fenólico aumentó con el tiempo. La actividad antioxi-dante de las muestras de mora de Castilla presentó una alta correlación con los con-tenidos de antocianinas y fenoles totales mientras que la actividad antioxidante de las muestras de tamarillo se correlacionó con el contenido de fenoles totales. Estos resultados son útiles en el desarrollo de aplicaciones de antocianinas microencapsu-ladas como colorantes alimenticios.

  6. Identification of a major protein on the cytosolic face of caveolae

    DEFF Research Database (Denmark)

    Vinten, J.; Johnsen, A.H; Roepstorff, P.; Harpøth, J.; Tranum-Jensen, Jørgen

    Rat adipocyte, caveolin, cav-p60, electron microscopy, polymerase and transscript release factor, mass spectrometry, cavin......Rat adipocyte, caveolin, cav-p60, electron microscopy, polymerase and transscript release factor, mass spectrometry, cavin...

  7. Molecular characterization and functional expression of the Apis mellifera voltage-dependent Ca2+ channels.

    Science.gov (United States)

    Cens, Thierry; Rousset, Matthieu; Collet, Claude; Charreton, Mercedes; Garnery, Lionel; Le Conte, Yves; Chahine, Mohamed; Sandoz, Jean-Christophe; Charnet, Pierre

    2015-03-01

    Voltage-gated Ca(2+) channels allow the influx of Ca(2+) ions from the extracellular space upon membrane depolarization and thus serve as a transducer between membrane potential and cellular events initiated by Ca(2+) transients. Most insects are predicted to possess three genes encoding Cavα, the main subunit of Ca(2+) channels, and several genes encoding the two auxiliary subunits, Cavβ and Cavα2δ; however very few of these genes have been cloned so far. Here, we cloned three full-length cDNAs encoding the three Cavα subunits (AmelCav1a, AmelCav2a and AmelCav3a), a cDNA encoding a novel variant of the Cavβ subunit (AmelCavβc), and three full-length cDNAs encoding three Cavα2δ subunits (AmelCavα2δ1 to 3) of the honeybee Apis mellifera. We identified several alternative or mutually exclusive exons in the sequence of the AmelCav2 and AmelCav3 genes. Moreover, we detected a stretch of glutamine residues in the C-terminus of the AmelCav1 subunit that is reminiscent of the motif found in the human Cav2.1 subunit of patients with Spinocerebellar Ataxia type 6. All these subunits contain structural domains that have been identified as functionally important in their mammalian homologues. For the first time, we could express three insect Cavα subunits in Xenopus oocytes and we show that AmelCav1a, 2a and 3a form Ca(2+) channels with distinctive properties. Notably, the co-expression of AmelCav1a or AmelCav2a with AmelCavβc and AmCavα2δ1 produces High Voltage-Activated Ca(2+) channels. On the other hand, expression of AmelCav3a alone leads to Low Voltage-Activated Ca(2+) channels. PMID:25602183

  8. Caveolin-1 plays a critical role in host immunity against Klebsiella pneumoniae by regulating STAT5 and Akt activity

    OpenAIRE

    Guo, Qiang; Shen, Nan; Yuan, Kefei; Li, Jiaxin; Wu, Hong; Zeng, Yong; Fox, John; Bansal, Arvind K.; Singh, Brij B; Gao, Hongwei; Wu, Min

    2012-01-01

    Caveolin-1 (Cav1) is a structural protein of caveolae. Although Cav1 is associated with certain bacterial infections, it is unknown whether Cav1 is involved in host immunity against Klebsiella pneumoniae, the third most commonly isolated microorganism from bacterial sepsis patients. Here, we showed that cav1 knockout mice succumbed to K. pneumoniae infection with markedly decreased survival rates, increased bacterial burdens, intensified tissue injury, hyperactive proinflammatory cytokines, a...

  9. Detection and characterization of chicken anemia virus from commercial broiler breeder chickens

    OpenAIRE

    Omar Abdul; Hailemariam Zerihun; Hair-Bejo Mohd; Giap Tan

    2008-01-01

    Abstract Background Chicken anemia virus (CAV) is the causative agent of chicken infectious anemia (CIA). Study on the type of CAV isolates present and their genetic diversity, transmission to their progeny and level of protection afforded in the breeder farms is lacking in Malaysia. Hence, the present study was aimed to detect CAV from commercial broiler breeder farms and characterize CAV positive samples based on sequence and phylogenetic analysis of partial VP1 gene. Results A total of 12 ...

  10. Cross-talk between Dopachrome Tautomerase and Caveolin-1 Is Melanoma Cell Phenotype-specific and Potentially Involved in Tumor Progression.

    Science.gov (United States)

    Popa, Ioana L; Milac, Adina L; Sima, Livia E; Alexandru, Petruta R; Pastrama, Florin; Munteanu, Cristian V A; Negroiu, Gabriela

    2016-06-10

    l-Dopachrome tautomerase (l-DCT), also called tyrosinase-related protein-2 (TRP-2), is a melanoma antigen overexpressed in most chemo-/radiotherapeutic stress-resistant tumor clones, and caveolin-1 (CAV1) is a main regulator of numerous signaling processes. A structural and functional relationship between DCT and CAV1 is first presented here in two human amelanotic melanoma cell lines, derived from vertical growth phase (MelJuSo) and metastatic (SKMel28) melanomas. DCT co-localizes at the plasma membrane with CAV1 and Cavin-1, another molecular marker for caveolae in both cell phenotypes. Our novel structural model proposed for the DCT-CAV1 complex, in addition to co-immunoprecipitation and mass spectrometry data, indicates a possible direct interaction between DCT and CAV1. The CAV1 control on DCT gene expression, DCT post-translational processing, and subcellular distribution is cell phenotype-dependent. DCT is a modulator of CAV1 stability and supramolecular assembly in both cell phenotypes. During autocrine stimulation, the expressions of DCT and CAV1 are oppositely regulated; DCT increases while CAV1 decreases. Sub-confluent MelJuSo clones DCT(high)/CAV1(low) are proliferating and acquire fibroblast-like morphology, forming massive, confluent clusters as demonstrated by immunofluorescent staining and TissueFAXS quantitative image cytometry analysis. CAV1 down-regulation directly contributes to the expansion of MelJuSo DCT(high) subtype. CAV1 involved in the perpetuation of cell phenotype-overexpressing anti-stress DCT molecule supports the concept that CAV1 functions as a tumor suppressor in early stages of melanoma. DCT is a regulator of the CAV1-associated structures and is possibly a new molecular player in CAV1-mediated processes in melanoma. PMID:27053106

  11. Differential regulation of cell functions by CSD peptide subdomains

    OpenAIRE

    Reese, Charles; Dyer, Shanice; Perry, Beth; Bonner, Michael; Oates, James; Hofbauer, Ann; Sessa, William; Bernatchez, Pascal; Visconti, Richard P; Zhang, Jing; Hatfield, Corey M; Silver, Richard M.; Hoffman, Stanley; Tourkina, Elena

    2013-01-01

    Background In fibrotic lung diseases, expression of caveolin-1 is decreased in fibroblasts and monocytes. The effects of this deficiency are reversed by treating cells or animals with the caveolin-1 scaffolding domain peptide (CSD, amino acids 82–101 of caveolin-1) which compensates for the lack of caveolin-1. Here we compare the function of CSD subdomains (Cav-A, Cav-B, Cav-C, Cav-AB, and Cav-BC) and mutated versions of CSD (F92A and T90A/T91A/F92A). Methods Migration toward the chemokine CX...

  12. Functional expression of T-type Ca2+ channels in spinal motoneurons of the adult turtle.

    Directory of Open Access Journals (Sweden)

    Martha Canto-Bustos

    Full Text Available Voltage-gated Ca2+ (CaV channels are transmembrane proteins comprising three subfamilies named CaV1, CaV2 and CaV3. The CaV3 channel subfamily groups the low-voltage activated Ca2+ channels (LVA or T-type a significant role in regulating neuronal excitability. CaV3 channel activity may lead to the generation of complex patterns of action potential firing such as the postinhibitory rebound (PIR. In the adult spinal cord, these channels have been found in dorsal horn interneurons where they control physiological events near the resting potential and participate in determining excitability. In motoneurons, CaV3 channels have been found during development, but their functional expression has not yet been reported in adult animals. Here, we show evidence for the presence of CaV3 channel-mediated PIR in motoneurons of the adult turtle spinal cord. Our results indicate that Ni2+ and NNC55-0396, two antagonists of CaV3 channel activity, inhibited PIR in the adult turtle spinal cord. Molecular biology and biochemical assays revealed the expression of the CaV3.1 channel isotype and its localization in motoneurons. Together, these results provide evidence for the expression of CaV3.1 channels in the spinal cord of adult animals and show also that these channels may contribute to determine the excitability of motoneurons.

  13. Sequence and phylogenetic analysis of chicken anaemia virus obtained from backyard and commercial chickens in Nigeria : research communication

    Directory of Open Access Journals (Sweden)

    D.O. Oluwayelu

    2008-09-01

    Full Text Available This work reports the first molecular analysis study of chicken anaemia virus (CAV in backyard chickens in Africa using molecular cloning and sequence analysis to characterize CAV strains obtained from commercial chickens and Nigerian backyard chickens. Partial VP1 gene sequences were determined for three CAVs from commercial chickens and for six CAV variants present in samples from a backyard chicken. Multiple alignment analysis revealed that the 6 % and 4 % nucleotide diversity obtained respectively for the commercial and backyard chicken strains translated to only 2 % amino acid diversity for each breed. Overall, the amino acid composition of Nigerian CAVs was found to be highly conserved. Since the partial VP1 gene sequence of two backyard chicken cloned CAV strains (NGR/Cl-8 and NGR/Cl-9 were almost identical and evolutionarily closely related to the commercial chicken strains NGR-1, and NGR-4 and NGR-5, respectively, we concluded that CAV infections had crossed the farm boundary.

  14. Differential effect of T-type voltage-gated calcium channel disruption on renal plasma flow and glomerular filtration rate in vivo

    DEFF Research Database (Denmark)

    Thuesen, Anne D; Andersen, Henrik; Cardel, Majken;

    2014-01-01

    Voltage-gated calcium channels (Cav) play an essential role in regulation of renal blood flow and GFR. Because T-type Cavs are differentially expressed in pre- and postglomerular vessels it was hypothesized that they impact renal blood flow and GFR differentially. The question was addressed by use...... in Cav 3.1 KO compared to Wt with no difference in blood pressure. T-type antagonists significantly inhibited the constriction of human intra-renal arteries in response to a small depolarization. In conclusion, Cav 3.2 channels support dilatation of efferent arterioles and affect GFR while Cav 3.......1 channels in vivo contribute to renal vascular resistance. It is suggested that endothelial and nerve localization of Cav 3.2 and Cav 3.1, respectively, may account for the observed effects....

  15. Caveolin-1 Is Necessary for Hepatic Oxidative Lipid Metabolism: Evidence for Crosstalk between Caveolin-1 and Bile Acid Signaling

    Directory of Open Access Journals (Sweden)

    Manuel A. Fernández-Rojo

    2013-07-01

    Full Text Available Caveolae and caveolin-1 (CAV1 have been linked to several cellular functions. However, a model explaining their roles in mammalian tissues in vivo is lacking. Unbiased expression profiling in several tissues and cell types identified lipid metabolism as the main target affected by CAV1 deficiency. CAV1−/− mice exhibited impaired hepatic peroxisome proliferator-activated receptor α (PPARα-dependent oxidative fatty acid metabolism and ketogenesis. Similar results were recapitulated in CAV1-deficient AML12 hepatocytes, suggesting at least a partial cell-autonomous role of hepatocyte CAV1 in metabolic adaptation to fasting. Finally, our experiments suggest that the hepatic phenotypes observed in CAV1−/− mice involve impaired PPARα ligand signaling and attenuated bile acid and FXRα signaling. These results demonstrate the significance of CAV1 in (1 hepatic lipid homeostasis and (2 nuclear hormone receptor (PPARα, FXRα, and SHP and bile acid signaling.

  16. Évolution de la sensibilité des glumelles du riz à Pyricularia oryzae Cav. et à Drechslera oryzae (Br. de Haan) Sub. et Jain : conséquences pour la transmission des maladies

    OpenAIRE

    Bernaux, P.

    1981-01-01

    Les contaminations artificielles réalisées sur les panicules de riz ont permis de constater la courte durée de la sensibilité des glumelles à la pénétration par Pyricularia oryzae et Drechslera oryzae. Dès l’anthèse, l’évolution anatomique des tissus des glumelles aboutit à une barrière s’opposant à toute pénétration ultérieure. Les contaminations avant l’anthèse ont pour conséquence l’invasion des glumelles par les champignons étudiés et la localisation du mycélium et des conidies entre...

  17. Growth of seedlings of pigeon pea (Cajanus cajan (l. millsp, wand riverhemp (Sesbania virgata (cav. pers., and lead tree (Leucaena leucocephala (lam. de wit in an arsenic-contaminated soil

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo Dias

    2010-06-01

    Full Text Available Phytoremediation strategies utilize plants to decontaminate or immobilize soil pollutants. Among soil pollutants, metalloid As is considered a primary concern as a toxic element to organisms. Arsenic concentrations in the soil result from anthropogenic activities such as: the use of pesticides (herbicides and fungicides; some fertilizers; Au, Pb, Cu and Ni mining; Fe and steel production; coal combustion; and as a bi-product during natural gas extraction. This study evaluated the potential of pigeon pea (Cajanus cajan, wand riverhemp (Sesbania virgata, and lead tree (Leucaena leucocephala as phytoremediators of soils polluted by As. Soil samples were placed in plastic pots, incubated with different As doses (0; 50; 100 and 200 mg dm-3 and then sown with seeds of the three species. Thirty (pigeon pea and 90 days after sowing, the plants were evaluated for height, collar diameter and dry matter of young, intermediate and basal leaves, stems and roots. Arsenic concentration was determined in different aged leaves, stems and roots to establish the translocation index (TI between the plant root system and aerial plant components and the bioconcentration factors (BF. The evaluated species showed distinct characteristics regarding As tolerance, since the lead tree and wand riverhemp were significantly more tolerant than pigeon pea. The high As levels found in wand riverhemp roots suggest the existence of an efficient accumulation and compartmentalization mechanism in order to reduce As translocation to shoot tissues. Pigeon pea is a sensitive species and could serve as a potential bioindicator plant, whereas the other two species have potential for phytoremediation programs in As polluted areas. However, further studies are needed with longer exposure times in actual field conditions to reach definite conclusions on relative phytoremediation potentials.A fitorremediação é uma estratégia que utiliza plantas para descontaminar ou imobilizar poluentes do solo. Entre os poluentes do solo, o As é considerado o principal elemento tóxico aos organismos vivos. A presença de As no solo pode ser decorrente de atividades antrópicas, como: uso de pesticidas (herbicidas e fungicidas; alguns fertilizantes; extração de Au, Pb, Cu e Ni; produção de Fe e aço; combustão de carvão; e coprodução durante a extração de gás natural. Este trabalho avaliou o potencial de feijão-guandu (Cajanus cajan, sesbania (Sesbania virgata e leucena (Leucaena leucocephala como fitorremediadoras de solos contaminados por As. Amostras de solo foram colocadas em potes de plástico, incubadas com diferentes doses de As (0, 50, 100 e 200 mg dm-3 e semeados com sementes das três espécies. Aos 30 (feijão-guandu e aos 90 dias após a semeadura, as plantas foram avaliadas quanto a altura, diâmetro de colo e matéria seca de folhas jovens, intermediárias e basais, ramos e raízes. A concentração de As foi determinada nas diferentes folhas, ramos e raízes, a fim de estabelecer o índice de translocação (TI entre o sistema radicular e a parte aere das plantas e o fator de bioconcentração (BF. As espécies avaliadas apresentaram comportamento distinto em relação à tolerância ao As, uma vez que a sesbania e a leucena foram significativamente mais tolerantes que o feijão guandu. A elevada concentração de As encontrada nas raízes de sesbania sugere a existência de um eficiente mecanismo de acumulação e compartimentalização, no sentido de reduzir a translocação para os tecidos da parte aérea. O feijão-guandu é uma espécie sensível ao As, podendo ser utilizado como planta indicadora, enquanto as outras duas espécies apresentam potencial para serem utilizadas em programas de fitorremediação de áreas contaminadas por As. Entretanto, estudos futuros em condições de campo e de longa duração são necessários para a obtenção de conclusões definitivas em relação ao potencial de fitorremediação dessas espécies.

  18. SU-E-T-448: On the Perturbation Factor P-cav of the Markus Parallel Plate Ion Chambers in Clinical Electron Beams, Monte Carlo Based Reintegration of An Historical Experiment

    Energy Technology Data Exchange (ETDEWEB)

    Voigts-Rhetz, P von; Zink, K [Technische Hochschule Mittelhessen - University of Applied Sciences, Giessen, Hessen (Germany)

    2014-06-01

    Purpose: All present dosimetry protocols recommend well-guarded parallel-plate ion chambers for electron dosimetry. For the guard-less Markus chamber an energy dependent fluence perturbation correction pcav is given. This perturbation correction was experimentally determined by van der Plaetsen by comparison of the read-out of a Markus and a NACP chamber, which was assumed to be “perturbation-free”. Aim of the present study is a Monte Carlo based reiteration of this experiment. Methods: Detailed models of four parallel-plate chambers (Roos, Markus, NACP and Advanced Markus) were designed using the Monte Carlo code EGSnrc and placed in a water phantom. For all chambers the dose to the active volume filled with low density water was calculated for 13 clinical electron spectra (E{sub 0}=6-21 MeV) at the depth of maximum and at the reference depth under reference conditions. In all cases the chamber's reference point was positioned at the depth of measurement. Moreover, the dose to water DW was calculated in a small water voxel positioned at the same depth. Results: The calculated dose ratio D{sub NACP}/D{sub Markus}, which according to van der Plaetsen reflects the fluence perturbation correction of the Markus chamber, deviates less from unity than the values given by van der Plaetsen's but exhibits a similar energy dependence. The same holds for the dose ratios of the other well guarded chambers. But, in comparison to water, the Markus chamber reveals the smallest overall perturbation correction which is nearly energy independent at both investigated depths. Conclusion: The simulations principally confirm the energy dependence of the dose ratio D{sub NACP}/D{sub Markus} as published by van der Plaetsen. But, as shown by our simulations of the ratio D{sub W}/D{sub Markus}, the conclusion drawn in all dosimetry protocols is questionable: in contrast to all well-guarded chambers the guard-less Markus chamber reveals the smallest overall perturbation correction and also the smallest energy dependence.

  19. Características fisicoquímicas y organolépticas de frutos de tomate de árbol (Solanum betaceum Cav a dos temperaturas de almacenamiento y tipos de cera

    Directory of Open Access Journals (Sweden)

    Jorge Contreras

    2011-02-01

    Full Text Available El objetivo de este estudio fue observar y analizar el comportamiento poscosecha de frutos de tomate de árbol, variedad Rojo Común, provenientes de Turmequé (Boyacá; a 2.100 msnm, precipitación anual: 1.600 mm, temperatura media: 19ºC. Los frutos fueron tratados con dos ceras naturales (Ceratec y Cerabrix y almacenados a temperaturas de 6°C y 18°C (temperatura ambiente, con humedades relativas del 90% y 75% respectivamente. Características como firmeza, pH, peso fresco, índice de madurez, acidez titulable y sólidos solubles fueron medidas durante 24 días. Al final de este periodo, se realizó un análisis sensorial, dirigido a los frutos que presentaron las mejores características de consumo (los almacenados a 6ºC. A temperatura ambiente, el encerado no se presentó como una práctica conveniente para la conservación de los frutos de tomate de árbol. Frutos a 6ºC tuvieron un excelente comportamiento poscosecha, pero el encerado no se manifestó como una fuente de variación, ya que no aparecieron diferencias estadísticas significativas entre los tratamientos en frío. La menor pérdida de peso (por transpiración y respiración se obtuvó en frutos a 6ºC, sin cera, seguidos por los encerados con Cerabrix. Características como firmeza, peso y acidez titulable de los frutos mostraron una tendencia a la disminución a lo largo del experimento, Por otra parte, variables como sólidos solubles, pH e índice de madurez aumentaron sus valores iniciales en general, pero se hicieron más notables en los tratamientos a 18ºC. Con respecto al análisis sensorial, se obtuvo una gran aceptación de los frutos expuestos a 6ºC, tanto en apariencia como en sabor.

  20. 二硫苏糖醇提取纯化Cavl.2钙通道片段CT3蛋白%Purification of CT3 Fragment of Cav1.2 with Dithiothreitol

    Institute of Scientific and Technical Information of China (English)

    孙宇; 封瑞; 杨磊; 毛楠; 胡慧媛; 孙雪菲; 郝丽英

    2015-01-01

    目的:探讨二硫苏糖醇(DTT)有助于PreScission蛋白酶切除重组蛋白GST⁃CT3标签GST的方法。方法在大肠埃希菌BL21中转化入pGEX⁃6P⁃3/CT3重组质粒并诱导其表达,用B⁃PER提取纯化重组蛋白GST⁃CT3,在10 mmol/L DTT存在的情况下PreScission蛋白酶切除GST标签得到高浓度的CT3蛋白,进行SDS⁃PAGE电泳鉴定CT3蛋白。结果不含DTT情况下, PreScission蛋白酶很难切除重组蛋白GST⁃CT3的标签蛋白GST,而在10 mmol/L DTT存在的情况下,PreScission蛋白酶能够切除重组蛋白GST⁃CT3的标签蛋白GST,得到高浓度的CT3蛋白。结论10 mmol/L DTT有助于PreScission蛋白酶切除重组蛋白GST⁃CT3的标签蛋白GST,得到高浓度的CT3蛋白。%Objective To explore whether dithiothreitol(DTT)is helpful for PreScission Protease to cut off the GST from GST⁃CT3 protein. Meth⁃ods The pGEX⁃6P⁃3/CT3 recombinant plasmid was transfected into Escherichia coli BL21,and the GST⁃CT3 fusion protein was purified by B⁃PER method. PreScission Protease was applied with 10 mmol/L DTT to cut off the GST,then the SDS⁃PAGE was performed for identification of the CT3 protein. Results Without DTT,it was very difficult for PreScission Protease to cut off the GST from GST⁃CT3 protein. However,in the pres⁃ence of 10 mmol/L DTT,PreScission Protease could cut off the GST easily as identified by SDS⁃PAGE. Conclusion 10 mmol/L DTT can help Pre⁃Scission Protease to cut GST from GST⁃CT3 protein,so as to achieve high concentration of CT3.

  1. Cav1.3 L-type Ca2+ channels mediate long-term adaptation in dopamine D2L-mediated GluA1 trafficking in the dorsal striatum following cocaine exposure

    OpenAIRE

    Schierberl, Kathryn; Giordano, Thomas; Satpute, Shirish; HAO, JIN; Kaur, Gagandeep; Hofmann, Franz; Moosmang, Sven; Striessnig, Joerg; Rajadhyaksha, Anjali

    2012-01-01

    AMPA receptor (AMPAR) plasticity at glutamatergic synapses in the mesostriatal dopaminergic pathway has been implicated in persistent cocaine-induced behavioral responses; however, the precise mechanism underlying these changes remains unknown. Utilizing cocaine psychomotor sensitization in mice we find that repeated cocaine results in a basal reduction of Ser 845 GluA1 and cell surface GluA1 levels in the dorsal striatum (dStr) following a protracted withdrawal period, an adaptation that is ...

  2. Studies on the morphology, anatomy and ecology of ophrys lutea cav. subsp. minor (guss.) o. danesch and e. danesch ex golz and h.r. reinhard (orchidaceae) in turkey

    International Nuclear Information System (INIS)

    In this study, morphological, anatomical and ecological characteristics of Ophrys lutea subsp. minor in Turkey were investigated. The plant material was collected from 30 different populations during 2010-2013. In all 19 morphological and 20 anatomical features were investigated, and habitat characteristics recorded. The soil samples were also collected from the plant sampling sites and subjected to an analysis for 18 characteristics. The minimum plant length was 69 mm and maximum 323 mm, minimum length of underground part was 29 mm, and maximum 49 mm, and the number of leaves varied between 1-10. The ecological characteristics revealed that O. lutea subsp. minor flourishes well from sea level to 800 m, and the most common habitats of O. lutea subsp. minor are rocky limestones, phrygana, macchie, olive gorves, woodland margins. In this study, the morphological and anatomical characteristics of Crocus biflorus subsp. pulchricolor (Iridaceae) were investigated. The subsp. pulchricolor has 4 leaves, 1 mm broad; bracts drying brownish. These properties are characteristics of these plants. In anatomical studies, cross-sections of the root, stem and leaves were examined. These parts photographed and compared with the other Crocus species and Iridaceae family species. (author)

  3. SU-E-T-448: On the Perturbation Factor P-cav of the Markus Parallel Plate Ion Chambers in Clinical Electron Beams, Monte Carlo Based Reintegration of An Historical Experiment

    International Nuclear Information System (INIS)

    Purpose: All present dosimetry protocols recommend well-guarded parallel-plate ion chambers for electron dosimetry. For the guard-less Markus chamber an energy dependent fluence perturbation correction pcav is given. This perturbation correction was experimentally determined by van der Plaetsen by comparison of the read-out of a Markus and a NACP chamber, which was assumed to be “perturbation-free”. Aim of the present study is a Monte Carlo based reiteration of this experiment. Methods: Detailed models of four parallel-plate chambers (Roos, Markus, NACP and Advanced Markus) were designed using the Monte Carlo code EGSnrc and placed in a water phantom. For all chambers the dose to the active volume filled with low density water was calculated for 13 clinical electron spectra (E0=6-21 MeV) at the depth of maximum and at the reference depth under reference conditions. In all cases the chamber's reference point was positioned at the depth of measurement. Moreover, the dose to water DW was calculated in a small water voxel positioned at the same depth. Results: The calculated dose ratio DNACP/DMarkus, which according to van der Plaetsen reflects the fluence perturbation correction of the Markus chamber, deviates less from unity than the values given by van der Plaetsen's but exhibits a similar energy dependence. The same holds for the dose ratios of the other well guarded chambers. But, in comparison to water, the Markus chamber reveals the smallest overall perturbation correction which is nearly energy independent at both investigated depths. Conclusion: The simulations principally confirm the energy dependence of the dose ratio DNACP/DMarkus as published by van der Plaetsen. But, as shown by our simulations of the ratio DW/DMarkus, the conclusion drawn in all dosimetry protocols is questionable: in contrast to all well-guarded chambers the guard-less Markus chamber reveals the smallest overall perturbation correction and also the smallest energy dependence

  4. Ion fluxes through KCa2 (SK) and Cav1 (L-type) channels contribute to chronoselectivity of adenosine A1 receptor-mediated actions in spontaneously beating rat atria

    OpenAIRE

    Paulo eCorreia-De-Sá

    2016-01-01

    Impulse generation in supraventricular tissue is inhibited by adenosine and acetylcholine via the activation of A1 and M2 receptors coupled to inwardly rectifying GIRK/KIR3.1/3.4 channels, respectively. Unlike M2 receptors, bradycardia produced by A1 receptors activation predominates over negative inotropy. Such difference suggests that other ion currents may contribute to adenosine chronoselectivity. In isolated spontaneously beating rat atria, blockade of KCa2/SK channels with apamin and Ca...

  5. Scanning mutagenesis of the I-II loop of the Cav2.2 calcium channel identifies residues Arginine 376 and Valine 416 as molecular determinants of voltage dependent G protein inhibition

    Directory of Open Access Journals (Sweden)

    Tedford Hugo W

    2010-02-01

    Full Text Available Abstract Direct interaction with the β subunit of the heterotrimeric G protein complex causes voltage-dependent inhibition of N-type calcium channels. To further characterize the molecular determinants of this interaction, we performed scanning mutagenesis of residues 372-387 and 410-428 of the N-type channel α1 subunit, in which individual residues were replaced by either alanine or cysteine. We coexpressed wild type Gβ1γ2 subunits with either wild type or point mutant N-type calcium channels, and voltage-dependent, G protein-mediated inhibition of the channels (VDI was assessed using patch clamp recordings. The resulting data indicate that Arg376 and Val416 of the α1 subunit, residues which are surface-exposed in the presence of the calcium channel β subunit, contribute significantly to the functional inhibition by Gβ1. To further characterize the roles of Arg376 and Val416 in this interaction, we performed secondary mutagenesis of these residues, coexpressing the resulting mutants with wild type Gβ1γ2 subunits and with several isoforms of the auxiliary β subunit of the N-type channel, again assessing VDI using patch clamp recordings. The results confirm the importance of Arg376 for G protein-mediated inhibition and show that a single amino acid substitution to phenylalanine drastically alters the abilities of auxiliary calcium channel subunits to regulate G protein inhibition of the channel.

  6. Exposure to cAMP and β-adrenergic stimulation recruits CaV3 T-type channels in rat chromaffin cells through Epac cAMP-receptor proteins

    Science.gov (United States)

    Novara, M; Baldelli, P; Cavallari, D; Carabelli, V; Giancippoli, A; Carbone, E

    2004-01-01

    T-type channels are expressed weakly or not at all in adult rat chromaffin cells (RCCs) and there is contrasting evidence as to whether they play a functional role in catecholamine secretion. Here we show that 3–5 days after application of pCPT-cAMP, most RCCs grown in serum-free medium expressed a high density of low-voltage-activated T-type channels without altering the expression and characteristics of high-voltage-activated channels. The density of cAMP-recruited T-type channels increased with time and displayed the typical biophysical and pharmacological properties of low-voltage-activated Ca2+ channels: (1) steep voltage-dependent activation from −50 mV in 10 mm Ca2+, (2) slow deactivation but fast and complete inactivation, (3) full inactivation following short conditioning prepulses to −30 mV, (4) effective block of Ca2+ influx with 50 μm Ni2+, (5) comparable permeability to Ca2+ and Ba2+, and (6) insensitivity to common Ca2+ channel antagonists. The action of exogenous pCPT-cAMP (200 μm) was prevented by the protein synthesis inhibitor anisomycin and mimicked in most cells by exposure to forskolin and 1-methyl-3-isobutylxanthine (IBMX) or isoprenaline. The protein kinase A (PKA) inhibitor H89 (0.3 μm) and the competitive antagonist of cAMP binding to PKA, Rp-cAMPS, had weak or no effect on the action of pCPT-cAMP. In line with this, the selective Epac agonist 8CPT-2Me-cAMP nicely mimicked the action of pCPT-cAMP and isoprenaline, suggesting the existence of a dominant Epac-dependent recruitment of T-type channels in RCCs that may originate from the activation of β-adrenoceptors. Stimulation of β-adrenoceptors occurs autocrinally in RCCs and thus, the neosynthesis of low-voltage-activated channels may represent a new form of ‘chromaffin cell plasticity’, which contributes, by lowering the threshold of action potential firing, to increasing cell excitability and secretory activity during sustained sympathetic stimulation and/or increased catecholamine circulation. PMID:15133061

  7. Exposure to cAMP and beta-adrenergic stimulation recruits Ca(V)3 T-type channels in rat chromaffin cells through Epac cAMP-receptor proteins.

    Science.gov (United States)

    Novara, M; Baldelli, P; Cavallari, D; Carabelli, V; Giancippoli, A; Carbone, E

    2004-07-15

    T-type channels are expressed weakly or not at all in adult rat chromaffin cells (RCCs) and there is contrasting evidence as to whether they play a functional role in catecholamine secretion. Here we show that 3-5 days after application of pCPT-cAMP, most RCCs grown in serum-free medium expressed a high density of low-voltage-activated T-type channels without altering the expression and characteristics of high-voltage-activated channels. The density of cAMP-recruited T-type channels increased with time and displayed the typical biophysical and pharmacological properties of low-voltage-activated Ca(2+) channels: (1) steep voltage-dependent activation from -50 mV in 10 mm Ca(2+), (2) slow deactivation but fast and complete inactivation, (3) full inactivation following short conditioning prepulses to -30 mV, (4) effective block of Ca(2+) influx with 50 microM Ni(2+), (5) comparable permeability to Ca(2+) and Ba(2+), and (6) insensitivity to common Ca(2+) channel antagonists. The action of exogenous pCPT-cAMP (200 microM) was prevented by the protein synthesis inhibitor anisomycin and mimicked in most cells by exposure to forskolin and 1-methyl-3-isobutylxanthine (IBMX) or isoprenaline. The protein kinase A (PKA) inhibitor H89 (0.3 microM) and the competitive antagonist of cAMP binding to PKA, Rp-cAMPS, had weak or no effect on the action of pCPT-cAMP. In line with this, the selective Epac agonist 8CPT-2Me-cAMP nicely mimicked the action of pCPT-cAMP and isoprenaline, suggesting the existence of a dominant Epac-dependent recruitment of T-type channels in RCCs that may originate from the activation of beta-adrenoceptors. Stimulation of beta-adrenoceptors occurs autocrinally in RCCs and thus, the neosynthesis of low-voltage-activated channels may represent a new form of 'chromaffin cell plasticity', which contributes, by lowering the threshold of action potential firing, to increasing cell excitability and secretory activity during sustained sympathetic stimulation and/or increased catecholamine circulation. PMID:15133061

  8. Production of Mannitol by Fungi from Cotton Dust

    OpenAIRE

    Domelsmith, Linda N.; Klich, Maren A.; Goynes, Wilton R.

    1988-01-01

    Cotton dust associated with high pulmonary function decrements contains relatively high levels of mannitol. In this study, cotton leaf and bract tissue and dust isolated from cotton leaf tissue were examined by optical microscopy, scanning electron microscopy, and capillary gas chromatography. Alternaria alternata, Cladosporium herbarum, Epicoccum purpurascens, and Fusarium pallidoroseum were isolated from cotton leaf dust. The fungal samples, cotton dust, and cotton leaf contained mannitol. ...

  9. Fractions of carbohydrates and of nitrogenous compounds of tropical grasses at different cutting ages Fracionamento de carboidratos e compostos nitrogenados de gramíneas tropicais em diferentes idades de corte

    OpenAIRE

    Patrícia Regina de Souza Siqueira Campos; José Fernando Coelho da Silva; Hernán Maldonado Vásquez; Andréa Vittori; Martinho de Almeida e Silva

    2010-01-01

    It was evaluated by the Cornell System carbohidrates fractions and nitrogenous compounds of the following grasses at the cutting ages of 14, 28, 42, and 56 days: nilo grass (Acroceras macrum), angola grass (Brachiaria purpurascens), aleman grass (Echinochloa polystachya), limpo grass (Hemarthria altíssima), setaria grass (Setaria anceps), tanner grass (Brachiaria arrecta), and tifton-85 grass (Cynodon spp). The experiment was carried out in a complete randomized block design, in a split plot ...

  10. Fungi causing dying out of heather seedlings

    Directory of Open Access Journals (Sweden)

    Maria Kowalik

    2012-12-01

    Full Text Available The aim of the study was to determine the fungi causing dying out of one-yearold heather seedlings. Observations were carried out on: 'Amethyst', 'Annemarie', 'Colette', 'Perestroika' and 'Reini'. The shoots revealing necrotic symptoms were plated on PDA medium. 25 species of fungi were isolated. Among them Pestalotia sydowiana, Alternaria alternata, Cylindrocarpon destructans, Leptosphaeria coniothyrium and Epicoccum purpurascens were dominant, while Mammaria echinobotryoides, Phoma leveillei, Kaissleriella subalpina, Botrytis cinerea and Phytophthora cinnamomi occurred less frequently.

  11. Studies on Total Polyphenols Content and Antioxidant Activity of Methanolic Extracts from Selected Salvia Species

    OpenAIRE

    Ana Viorica Pop Cuceu; Maria Tofană; Sonia A. Socaci; Melinda Nagy; Maria Doiniţa Borş; Liana Salanţă; Romina Vlaic

    2015-01-01

    Salvia is one of the largest genera in the family of Labiatae, comprising about 900 species distributed widely throughout the world. Many species of Salvia are commonly used as herbal tea and for food flavoring, as well as in cosmetics, perfumery and in the pharmaceutical industry. The present study compares the antioxidant properties of four methanolic extracts, obtained by two extraction methods, from Salvia elegans, Salvia officinalis Purpurascens, Salvia officinalis Tricolor and Salvia la...

  12. Binding of nuclear caveolin-1 to promoter elements of growth-associated genes in ovarian carcinoma cells

    International Nuclear Information System (INIS)

    Caveolin-1 (cav-1), a member of a protein family associated mainly with cell membrane microdomains in many cell types, acts as a tumor suppressor in ovarian carcinoma cells. Biochemical analyses demonstrated that cav-1 was also localized in the nuclei of ovarian carcinoma cells, endogenously (SKOV3) or ectopically (IGtC3) expressing cav-1. By confocal analyses, the same cell lines as well as IGROV1 and SKOV3 cells transiently transfected with green fluorescent protein-cav-1 fusion protein showed nuclear punctate speckled pattern. Subnuclear distribution analysis revealed cav-1 mainly associated with the nuclear matrix, but also slightly with chromatin. Cav-1 was found in nuclear high-molecular weight complexes and by confocal analysis was found to co-localized with the inner nuclear membrane protein emerin. Cyclin D1 and folate receptor promoters were modulated by cav-1 in SKOV3 cells as demonstrated by transient transfection with or silencing of cav-1. Chromatin immunoprecipitation and supershift assays indicated that nuclear cav-1 can bind in vitro and in vivo to promoter sequences of both cyclin D1 and folate receptor genes. These data suggest that in ovarian carcinoma cells cav-1, localized in transcriptionally inactive chromatin, exerts a functional activity mediated, at least in part, by directly binding to sequences of genes involved in proliferation

  13. Expression of Caveolin-1 in tongue squamous cell carcinoma by quantum dots

    Directory of Open Access Journals (Sweden)

    J. Xue

    2010-04-01

    Full Text Available Quantum dots (QDs are a new class of fluorescent probes to detect biomarker expression. The role of caveolin-1 (Cav-1 in tongue squamous cell carcinoma (TSCC is still unknown. This study aimed to investigate the expression profile of Cav-1 in carcinogenesis and development of TSCC by QDs immunofluorescence histochemistry (QDs-IHC and discuss the relationship between the Cav-1 expression and the clinicopathological outcomes. QDs-IHC was used to detect Cav-1 expression in tissue microarrays including normal tongue mucosa (NTM; n=10, hyperplastic tongue mucosa (HTM; n=10, tongue pre-cancer lesions (TPL; n=15 and primary tongue squamous cell carcinoma (PTSCC; n=61. Correlations between the Cav-1 expression and clinicopathologic variables were evaluated statistically. Cells positive for Cav-1 were clearly detected and bright images were obtained in a fine, granular pattern at the cell membrane and cytoplasm using QDs-IHC. The rate of Cav-1 immunoreactivity increased progressively from NTM (0%, HTM (0%, TPL (36% to PTSCC (74%. When compared with each other, there was statistical significance among PTSCC, TPL and NTM as well as among PTSCC, TPL and HTM. Moreover, Cav-1 expression level in PTSCC was correlated positively with clinical stage and histologic grade. QDs-IHC could accurately detect protein location in tongue mucosa. An increased expression of Cav-1 in the stepwise carcinogenesis from NTM, HTM, TPL to PTSCC suggested that Cav-1 might be an oncogene in the development of tongue squamous cell carcinoma.

  14. Caveolin-1 Expression Level in Cancer Associated Fibroblasts Predicts Outcome in Gastric Cancer

    Science.gov (United States)

    Gao, Jun; Fan, Lifang; Li, Zonghuan; Yang, Guifang; Chen, Honglei

    2013-01-01

    Aims Altered expression of epithelial or stromal caveolin-1 (Cav-1) is observed in various types of human cancers. However, the clinical significance of Cav-1 expression in gastric cancer (GC) remains largely unknown. The present study aims to explore the clinicopathological significance and prognostic value of both tumor cells and cancer associated fibroblasts (CAFs) Cav-1 in GC. Methods and Results Quantum dots immunofluorescence histochemistry was performed to examine the expression of Cav-1 in 20 cases of gastritis without intestinal metaplasia (IM), 20 cases of gastritis with IM and 286 cases of GC. Positive rates of epithelial Cav-1 in gastritis without IM, gastritis with IM and GC showed a decreasing trend (P = 0.012). Low expression of Cav-1 in CAFs but not in tumor cells was an independent predictor of poor prognosis in GC patients (P = 0.034 and 0.005 respectively in disease free survival and overall survival). Cav-1 level in tumor cells and CAFs showed no significant correlation with classic clinicopathological features. Conclusions Loss of epithelial Cav-1 may promote malignant progression and low CAFs Cav-1 level herald worse outcome of GC patient, suggesting CAFs Cav-1 may be a candidate therapeutic target and a useful prognostic marker of GC. PMID:23527097

  15. Integrin α1β1 Promotes Caveolin-1 Dephosphorylation by Activating T Cell Protein-tyrosine Phosphatase*

    Science.gov (United States)

    Borza, Corina M.; Chen, Xiwu; Mathew, Sijo; Mont, Stacey; Sanders, Charles R.; Zent, Roy; Pozzi, Ambra

    2010-01-01

    Integrin α1β1 is a collagen receptor that down-regulates collagen and reactive oxygen species (ROS) production, and mice lacking this receptor show increased ROS levels and exacerbated glomerular sclerosis following injury. Caveolin-1 (Cav-1) is a multifunctional protein that is tyrosine-phosphorylated in response to injury and has been implicated in ROS-mediated injury. Cav-1 interacts with integrins, and integrin α1β1 binds/activates T cell protein-tyrosine phosphatase (TCPTP), which is homologous to the tyrosine phosphatase PTP1B known to dephosphorylate Cav-1. In this study, we analyzed whether phosphorylated Cav-1 (pCav-1) is a substrate of TCPTP and if integrin α1β1 is essential for promoting TCPTP-mediated Cav-1 dephosphorylation. We found that Cav-1 phosphorylation is significantly higher in cells lacking integrin α1β1 at base line and following oxidative stress. Overexpression of TCPTP leads to reduced pCav-1 levels only in cells expressing integrin α1β1. Using solid phase binding assays, we demonstrated that 1) purified Cav-1 directly interacts with TCPTP and the integrin α1 subunit, 2) pCav-1 is a substrate of TCPTP, and 3) TCPTP-mediated Cav-1 dephosphorylation is highly increased by the addition of purified integrin α1β1 or an integrin α1 cytoplasmic peptide to which TCPTP has been shown to bind. Thus, our results demonstrate that pCav-1 is a new substrate of TCPTP and that integrin α1β1 acts as a negative regulator of Cav-1 phosphorylation by activating TCPTP. This could explain the protective function of integrin α1β1 in oxidative stress-mediated damage and why integrin α1-null mice are more susceptible to fibrosis following injury. PMID:20940300

  16. Integrin {alpha}1{beta}1 promotes caveolin-1 dephosphorylation by activating T cell protein-tyrosine phosphatase.

    Science.gov (United States)

    Borza, Corina M; Chen, Xiwu; Mathew, Sijo; Mont, Stacey; Sanders, Charles R; Zent, Roy; Pozzi, Ambra

    2010-12-17

    Integrin α1β1 is a collagen receptor that down-regulates collagen and reactive oxygen species (ROS) production, and mice lacking this receptor show increased ROS levels and exacerbated glomerular sclerosis following injury. Caveolin-1 (Cav-1) is a multifunctional protein that is tyrosine-phosphorylated in response to injury and has been implicated in ROS-mediated injury. Cav-1 interacts with integrins, and integrin α1β1 binds/activates T cell protein-tyrosine phosphatase (TCPTP), which is homologous to the tyrosine phosphatase PTP1B known to dephosphorylate Cav-1. In this study, we analyzed whether phosphorylated Cav-1 (pCav-1) is a substrate of TCPTP and if integrin α1β1 is essential for promoting TCPTP-mediated Cav-1 dephosphorylation. We found that Cav-1 phosphorylation is significantly higher in cells lacking integrin α1β1 at base line and following oxidative stress. Overexpression of TCPTP leads to reduced pCav-1 levels only in cells expressing integrin α1β1. Using solid phase binding assays, we demonstrated that 1) purified Cav-1 directly interacts with TCPTP and the integrin α1 subunit, 2) pCav-1 is a substrate of TCPTP, and 3) TCPTP-mediated Cav-1 dephosphorylation is highly increased by the addition of purified integrin α1β1 or an integrin α1 cytoplasmic peptide to which TCPTP has been shown to bind. Thus, our results demonstrate that pCav-1 is a new substrate of TCPTP and that integrin α1β1 acts as a negative regulator of Cav-1 phosphorylation by activating TCPTP. This could explain the protective function of integrin α1β1 in oxidative stress-mediated damage and why integrin α1-null mice are more susceptible to fibrosis following injury. PMID:20940300

  17. Interaction of caveolin-1, nitric oxide, and nitric oxide synthases in hypoxic human SK-N-MC neuroblastoma cells.

    Science.gov (United States)

    Shen, Jiangang; Lee, Waisin; Li, Yue; Lau, Chi Fai; Ng, Kwong Man; Fung, Man Lung; Liu, Ke Jian

    2008-10-01

    Neuroblastoma cells are capable of hypoxic adaptation, but the mechanisms involved are not fully understood. We hypothesized that caveolin-1 (cav-1), a plasma membrane signal molecule, might play a role in protecting neuroblastoma cells from oxidative injury by modulating nitric oxide (NO) production. We investigated the alterations of cav-1, cav-2, nitric oxide synthases (NOS), and NO levels in human SK-N-MC neuroblastoma cells exposed to hypoxia with 2% [O2]. The major discoveries include: (i) cav-1 but not cav-2 was up-regulated in the cells exposed to 15 h of hypoxia; (ii) NO donor 1-[N, N-di-(2-aminoethyl) amino] diazen-1-ium-1, 2-diolate up-regulated the expression of cav-1, whereas the non-selective NOS inhibitor N(G)-nitro-L-arginine methyl ester and inducible NOS (iNOS) inhibitor 1400W each abolished the increase in cav-1 expression in the hypoxic SK-N-MC cells. These results suggest that iNOS-induced NO production contributes to the up-regulation of cav-1 in the hypoxic SK-N-MC cells. Furthermore, we studied the roles played by cav-1 in regulating NO, NOS, and apoptotic cell death in the SK-N-MC cells subjected to 15 h of hypoxic treatment. Both cav-1 transfection and cav-1 scaffolding domain peptide abolished the induction of iNOS, reduced the production of NO, and reduced the rates of apoptotic cell death in the hypoxic SK-N-MC cells. These results suggest that increased expression of cav-1 in response to hypoxic stimulation could prevent oxidative injury induced by reactive oxygen species. The interactions of cav-1, NO, and NOS could be an important signal pathway in protecting the neuroblastoma cells from oxidative injury, contributing to the hypoxic tolerance of neuroblastoma cells. PMID:18717816

  18. Presence of two types of flowers with respect to nectar sugar in two gregariously flowering species

    Indian Academy of Sciences (India)

    Chaitali Anand; Chaitrali Umranikar; Pooja Shintre; Anuja Damle; Janhavi Kale; Jahnavi Joshi; Milind Watve

    2007-06-01

    Many species of animal-pollinated flowers are known to vary widely in the nectar content of flowers. Some proportion of flowers in many species is apparently nectarless, and such flowers are believed to be ‘cheaters’. Cheating may explain a part of the variability in nectar content. If cheating exists as a qualitatively different strategy then we expect bimodality in the distribution of nectar content of flowers. It has been shown in a multispecies study that gregarious species have a higher proportion of cheater flowers. We studied the frequency distribution of total nectar sugar in two gregariously flowering species Lantana camara and Utricularia purpurascens, which differed in other floral and ecological characters. At the population level, both the species showed significant bimodality in the total sugar content of flowers. The obvious sources of heterogeneity in the data did not explain bimodality. In Lantana camara, bimodality was observed within flowers of some of the individual plants sampled. In Utricularia purpurascens the proportion of nectarless flowers was more in high-density patches, suggesting that the gregariousness hypothesis may work within a species as well. The results support the hypothesis of cheating as a distinct strategy since two distinct types of flowers were observed in both the species. The effect of density in Utricularia purpurascens also supports the gregariousness hypothesis.

  19. Site specific estimation of cumulative absolute velocity

    International Nuclear Information System (INIS)

    The presented paper shows some recent results for correlation between cumulative absolute velocity (CAV) and the macro-seismic intensity, magnitude and distance (attenuation functions). The analyses are based mainly on European strong motion data. The processing is performed separately for intermediate depth earthquakes (Vrancea seismic region), regional shallow earthquakes and moderate local earthquakes. The results show that CAV correlates with the intensity, magnitude and distance in a similar way as the peak values of strong motion. There is significant difference of expected CAV from local earthquakes and from strong regional seismic excitations. The local earthquakes, although producing high accelerations, are developing small CAV and respectively small damage potential. The analyses show that intermediate depth earthquakes may produce significant CAV on very large distances, i.e. they may affect large territories and produce damage. The attenuation functions developed are used for prediction of CAV on the site of Kozloduy NPP in Bulgaria. (author)

  20. Phylogenetic and molecular characterization of chicken anemia virus in southern China from 2011 to 2012.

    Science.gov (United States)

    Zhang, Xinheng; Liu, Yuanjia; Wu, Boliang; Sun, Baoli; Chen, Feng; Ji, Jun; Ma, Jingyun; Xie, Qingmei

    2013-01-01

    Chicken anemia virus (CAV) is an important pathogen that causes severe immunosuppression in young chickens. We have characterized 13 CAVs isolated from different commercial farms in southern China between 2011 and 2012. We discovered 92 variable residues compared to 37 other CAV complete genome sequences from other parts of the world listed in GenBank; these residues have not been previously observed. All of the Chinese CAV genomes that were characterized in this study had a glutamine at position 394, a hallmark of highly pathogenic CAVs. We also discovered that intra-group genetic recombination plays a role in generating genetic diversity in natural populations of CAV. The GD-J-12 isolate was a possible recombinant between GD-C-12 and GD-M-12 in the genomic region that encompassed both the coding and non-coding regions. PMID:24343380

  1. Deletion of Caveolin-1 Protects against Oxidative Lung Injury via Up-Regulation of Heme Oxygenase-1

    Science.gov (United States)

    Jin, Yang; Kim, Hong Pyo; Chi, Minli; Ifedigbo, Emeka; Ryter, Stefan W.; Choi, Augustine M. K.

    2008-01-01

    Acute lung injury (ALI) is a major cause of morbidity and mortality in critically ill patients. Hyperoxia causes lung injury in animals and humans, and is an established model of ALI. Caveolin-1, a major constituent of caveolae, regulates numerous biological processes, including cell death and proliferation. Here we demonstrate that caveolin-1–null mice (cav-1−/−) were resistant to hyperoxia-induced death and lung injury. Cav-1−/− mice sustained reduced lung injury after hyperoxia as determined by protein levels in bronchoalveolar lavage fluid and histologic analysis. Furthermore, cav-1−/− fibroblasts and endothelial cells and cav-1 knockdown epithelial cells resisted hyperoxia-induced cell death in vitro. Basal and inducible expression of the stress protein heme oxygenase-1 (HO-1) were markedly elevated in lung tissue or fibroblasts from cav-1−/− mice. Hyperoxia induced the physical interaction between cav-1 and HO-1 in fibroblasts assessed by co-immunoprecipitation studies, which resulted in attenuation of HO activity. Inhibition of HO activity with tin protoporphyrin-IX abolished the survival benefits of cav-1−/− cells and cav-1−/− mice exposed to hyperoxia. The cav-1−/− mice displayed elevated phospho-p38 mitogen-activated protein kinase (MAPK) and p38β expression in lung tissue/cells under basal conditions and during hyperoxia. Treatment with SB202190, an inhibitor of p38 MAPK, decreased hyperoxia-inducible HO-1 expression in wild-type and cav-1−/− fibroblasts. Taken together, our data demonstrated that cav-1 deletion protects against hyperoxia-induced lung injury, involving in part the modulation of the HO-1–cav-1 interaction, and the enhanced induction of HO-1 through a p38 MAPK–mediated pathway. These studies identify caveolin-1 as a novel component involved in hyperoxia-induced lung injury. PMID:18323531

  2. Lipid raft/caveolae signaling is required for Cryptococcus neoformans invasion into human brain microvascular endothelial cells

    Directory of Open Access Journals (Sweden)

    Long Min

    2012-02-01

    Full Text Available Abstract Background Cryptococcus neoformans has a predilection for central nervous system infection. C. neoformans traversal of the blood brain barrier, composed of human brain microvascular endothelial cells (HBMEC, is the crucial step in brain infection. However, the molecular mechanism of the interaction between Cryptococcus neoformans and HBMEC, relevant to its brain invasion, is still largely unknown. Methods In this report, we explored several cellular and molecular events involving the membrane lipid rafts and caveolin-1 (Cav1 of HBMEC during C. neoformans infection. Immunofluorescence microscopy was used to examine the roles of Cav1. The knockdown of Cav1 by the siRNA treatment was performed. Phosphorylation of Cav1 relevant to its invasion functions was investigated. Results We found that the host receptor CD44 colocalized with Cav1 on the plasma membrane, and knockdown of Cav1 significantly reduced the fungal ability to invade HBMEC. Although the CD44 molecules were still present, HBMEC membrane organization was distorted by Cav1 knockdown. Concomitantly, knockdown of Cav1 significantly reduced the fungal crossing of the HBMEC monolayer in vitro. Upon C. neoformans engagement, host Cav1 was phosphorylated in a CD44-dependent manner. This phosphorylation was diminished by filipin, a disrupter of lipid raft structure. Furthermore, the phosphorylated Cav1 at the lipid raft migrated inward to the perinuclear localization. Interestingly, the phospho-Cav1 formed a thread-like structure and colocalized with actin filaments but not with the microtubule network. Conclusion These data support that C. neoformans internalization into HBMEC is a lipid raft/caveolae-dependent endocytic process where the actin cytoskeleton is involved, and the Cav1 plays an essential role in C. neoformans traversal of the blood-brain barrier.

  3. The Serological and Virological Investigation of Canine Adenovirus Infection on the Dogs

    OpenAIRE

    Oya Bulut; Orhan Yapici; Oguzhan Avci; Atilla Simsek; Kamil Atli; Irmak Dik; Sibel Yavru; Sibel Hasircioglu; Mehmet Kale; Nuri Mamak

    2013-01-01

    Two types of Canine Adenovirus (CAVs), Canine Adenovirus type 1 (CAV-1), the virus which causes infectious canine hepatitis, and Canine Adenovirus type 2 (CAV-2), which causes canine infectious laryngotracheitis, have been found in dogs. In this study, blood samples taken from 111 dogs, which were admitted to the Internal Medicine Clinic of Selcuk University, Faculty of Veterinary Medicine, with clinical symptoms. Seventy-seven dogs were sampled from Isparta and Burdur dog shelters by random ...

  4. Caveolin-2 Is Required for Apical Lipid Trafficking and Suppresses Basolateral Recycling Defects in the Intestine of Caenorhabditis elegans

    OpenAIRE

    Parker, Scott; Walker, Denise S.; Ly, Sung; Baylis, Howard A.

    2009-01-01

    Caveolins are plasma membrane–associated proteins that colocalize with, and stabilize caveolae. Their functions remain unclear although they are known to be involved in specific events in cell signaling and endocytosis. Caenorhabditis elegans encodes two caveolin genes, cav-1 and cav-2. We show that cav-2 is expressed in the intestine where it is localized to the apical membrane and in intracellular bodies. Using the styryl dye FM4-64 and BODIPY-labeled lactosylceramide, we show that the inte...

  5. Caveolin-1 Protects B6129 Mice against Helicobacter pylori Gastritis

    OpenAIRE

    Hitkova, Ivana; Yuan, Gang; Anderl, Florian; Gerhard, Markus; Kirchner, Thomas; Reu, Simone; Röcken, Christoph; Schäfer, Claus; Schmid, Roland M; Vogelmann, Roger; Ebert, Matthias P A; Burgermeister, Elke

    2013-01-01

    Caveolin-1 (Cav1) is a scaffold protein and pathogen receptor in the mucosa of the gastrointestinal tract. Chronic infection of gastric epithelial cells by Helicobacter pylori (H. pylori) is a major risk factor for human gastric cancer (GC) where Cav1 is frequently down-regulated. However, the function of Cav1 in H. pylori infection and pathogenesis of GC remained unknown. We show here that Cav1-deficient mice, infected for 11 months with the CagA-delivery deficient H. pylori strain SS1, deve...

  6. Atypical properties of a conventional calcium channel β subunit from the platyhelminth Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Schneider Toni

    2008-03-01

    Full Text Available Abstract Background The function of voltage-gated calcium (Cav channels greatly depends on coupling to cytoplasmic accessory β subunits, which not only promote surface expression, but also modulate gating and kinetic properties of the α1 subunit. Schistosomes, parasitic platyhelminths that cause schistosomiasis, express two β subunit subtypes: a structurally conventional β subunit and a variant β subunit with unusual functional properties. We have previously characterized the functional properties of the variant Cavβ subunit. Here, we focus on the modulatory phenotype of the conventional Cavβ subunit (SmCavβ using the human Cav2.3 channel as the substrate for SmCavβ and the whole-cell patch-clamp technique. Results The conventional Schistosoma mansoni Cavβ subunit markedly increases Cav2.3 currents, slows macroscopic inactivation and shifts steady state inactivation in the hyperpolarizing direction. However, currents produced by Cav2.3 in the presence of SmCavβ run-down to approximately 75% of their initial amplitudes within two minutes of establishing the whole-cell configuration. This suppressive effect was independent of Ca2+, but dependent on intracellular Mg2+-ATP. Additional experiments revealed that SmCavβ lends the Cav2.3/SmCavβ complex sensitivity to Na+ ions. A mutant version of the Cavβ subunit lacking the first forty-six amino acids, including a string of twenty-two acidic residues, no longer conferred sensitivity to intracellular Mg2+-ATP and Na+ ions, while continuing to show wild type modulation of current amplitude and inactivation of Cav2.3. Conclusion The data presented in this article provide insights into novel mechanisms employed by platyhelminth Cavβ subunits to modulate voltage-gated Ca2+ currents that indicate interactions between the Ca2+ channel complex and chelated forms of ATP as well as Na+ ions. These results have potentially important implications for understanding previously unknown mechanisms by

  7. Identification of a novel prostate cancer biomarker, caveolin-1: Implications and potential clinical benefit

    International Nuclear Information System (INIS)

    While prostate cancer is a common disease in men, it is uncommonly life-threatening. To better understand this phenomenon, tumor biologists have sought to elucidate the mechanisms that contribute to the development of virulent prostate cancer. The recent discovery that caveolin-1 (Cav-1) functions as an important oncogene involved in prostate cancer progression reflects the success of this effort. Cav-1 is a major structural coat protein of caveolae, specialized plasma membrane invaginations involved in multiple cellular functions, including molecular transport, cell adhesion, and signal transduction. Cav-1 is aberrantly overexpressed in human prostate cancer, with higher levels evident in metastatic versus primary sites. Intracellular Cav-1 promotes cell survival through activation of Akt and enhancement of additional growth factor pro-survival pathways. Cav-1 is also secreted as a biologically active molecule that promotes cell survival and angiogenesis within the tumor microenvironment. Secreted Cav-1 can be reproducibly detected in peripheral blood using a sensitive and specific immunoassay. Cav-1 levels distinguish men with prostate cancer from normal controls, and preoperative Cav-1 levels predict which patients are at highest risk for relapse following radical prostatectomy for localized disease. Thus, secreted Cav-1 is a promising biomarker in identifying clinically significant prostate cancer

  8. Specific T-type calcium channel isoforms are associated with distinct burst phenotypes in deep cerebellar nuclear neurons

    OpenAIRE

    Molineux, Michael L.; McRory, John E.; McKay, Bruce E.; Hamid, Jawed; Mehaffey, W. Hamish; Rehak, Renata; Snutch, Terrance P; Gerald W Zamponi; Turner, Ray W

    2006-01-01

    T-type calcium channels are thought to transform neuronal output to a burst mode by generating low voltage-activated (LVA) calcium currents and rebound burst discharge. In this study we assess the expression pattern of the three different T-type channel isoforms (Cav3.1, Cav3.2, and Cav3.3) in cerebellar neurons and focus on their potential role in generating LVA spikes and rebound discharge in deep cerebellar nuclear (DCN) neurons. We detected expression of one or more Cav3 channel isoforms ...

  9. Insulin resistance in skeletal muscles of caveolin-3-null mice

    OpenAIRE

    Oshikawa, Jin; Otsu, Koji; Toya, Yoshiyuki; Tsunematsu, Takashi; Hankins, Raleigh; Kawabe, Jun-ichi; Minamisawa, Susumu; Umemura, Satoshi; Hagiwara, Yasuko; Ishikawa, Yoshihiro

    2004-01-01

    Type 2 diabetes is preceded by the development of insulin resistance, in which the action of insulin is impaired, largely in skeletal muscles. Caveolin-3 (Cav3) is a muscle-specific subtype of caveolin, an example of a scaffolding protein found within membranes. Cav is also known as growth signal inhibitor, although it was recently demonstrated that the genetic disruption of Cav3 did not augment growth in mice. We found, however, that the lack of Cav3 led to the development of insulin resista...

  10. Caveolin-1 Protects B6129 Mice against Helicobacter pylori Gastritis

    Science.gov (United States)

    Hitkova, Ivana; Yuan, Gang; Anderl, Florian; Gerhard, Markus; Kirchner, Thomas; Reu, Simone; Röcken, Christoph; Schäfer, Claus; Schmid, Roland M.; Vogelmann, Roger; Ebert, Matthias P. A.; Burgermeister, Elke

    2013-01-01

    Caveolin-1 (Cav1) is a scaffold protein and pathogen receptor in the mucosa of the gastrointestinal tract. Chronic infection of gastric epithelial cells by Helicobacter pylori (H. pylori) is a major risk factor for human gastric cancer (GC) where Cav1 is frequently down-regulated. However, the function of Cav1 in H. pylori infection and pathogenesis of GC remained unknown. We show here that Cav1-deficient mice, infected for 11 months with the CagA-delivery deficient H. pylori strain SS1, developed more severe gastritis and tissue damage, including loss of parietal cells and foveolar hyperplasia, and displayed lower colonisation of the gastric mucosa than wild-type B6129 littermates. Cav1-null mice showed enhanced infiltration of macrophages and B-cells and secretion of chemokines (RANTES) but had reduced levels of CD25+ regulatory T-cells. Cav1-deficient human GC cells (AGS), infected with the CagA-delivery proficient H. pylori strain G27, were more sensitive to CagA-related cytoskeletal stress morphologies (“humming bird”) compared to AGS cells stably transfected with Cav1 (AGS/Cav1). Infection of AGS/Cav1 cells triggered the recruitment of p120 RhoGTPase-activating protein/deleted in liver cancer-1 (p120RhoGAP/DLC1) to Cav1 and counteracted CagA-induced cytoskeletal rearrangements. In human GC cell lines (MKN45, N87) and mouse stomach tissue, H. pylori down-regulated endogenous expression of Cav1 independently of CagA. Mechanistically, H. pylori activated sterol-responsive element-binding protein-1 (SREBP1) to repress transcription of the human Cav1 gene from sterol-responsive elements (SREs) in the proximal Cav1 promoter. These data suggested a protective role of Cav1 against H. pylori-induced inflammation and tissue damage. We propose that H. pylori exploits down-regulation of Cav1 to subvert the host's immune response and to promote signalling of its virulence factors in host cells. PMID:23592983

  11. Roles of Caveolin-1 in Angiotensin II-Induced Hypertrophy and Inward Remodeling of Cerebral Pial Arterioles.

    Science.gov (United States)

    Umesalma, Shaikamjad; Houwen, Frederick Keith; Baumbach, Gary L; Chan, Siu-Lung

    2016-03-01

    Angiotensin II (Ang II) is a major determinant of inward remodeling and hypertrophy in pial arterioles that may have an important role in stroke during chronic hypertension. Previously, we found that epidermal growth factor receptor is critical in Ang II-mediated hypertrophy that may involve caveolin-1 (Cav-1). In this study, we examined the effects of Cav-1 and matrix metalloproteinase-9 (MMP9) on Ang II-mediated structural changes in pial arterioles. Cav-1-deficient (Cav-1(-/-)), MMP9-deficient (MMP9(-/-)), and wild-type mice were infused with either Ang II (1000 ng/kg per minute) or saline via osmotic minipumps for 28 days (n=6-8 per group). Systolic arterial pressure was measured by a tail-cuff method. Pressure and diameter of pial arterioles were measured through an open cranial window in anesthetized mice. Cross-sectional area of the wall was determined histologically in pressurized fixed pial arterioles. Expression of Cav-1, MMP9, phosphorylated epidermal growth factor receptor, and Akt was determined by Western blotting and immunohistochemistry. Deficiency of Cav-1 or MMP9 did not affect Ang II-induced hypertension. Ang II increased the expression of Cav-1, phosphorylated epidermal growth factor receptor, and Akt in wild-type mice, which was attenuated in Cav-1(-/-) mice. Ang II-induced hypertrophy, inward remodeling, and increased MMP9 expression in pial arterioles were prevented in Cav-1(-/-) mice. Ang II-mediated increases in MMP9 expression and inward remodeling, but not hypertrophy, were prevented in MMP9(-/-) mice. In conclusion, Cav-1 is essential in Ang II-mediated inward remodeling and hypertrophy in pial arterioles. Cav-1-induced MMP9 is exclusively involved in inward remodeling, not hypertrophy. Further studies are needed to determine the role of Akt in Ang II-mediated hypertrophy. PMID:26831194

  12. Biological characteristics of chicken anemia virus regenerated from clinical specimen by PCR.

    Science.gov (United States)

    van Santen, Vicky L; Toro, Haroldo; Hoerr, Frederic J

    2007-03-01

    Our previous genetic characterization of chicken anemia virus (CAV) in commercial broiler chickens in Alabama revealed a previously undetected polymorphism: a glutamine codon at VP1 position 22, in 7 of the 14 sequences. The novel glutamine codon was always found in association with a VP1 "hypervariable region" identical to CAV field isolates that replicate poorly in culture. The complete genome of CAV73, representative of the sequences with the novel polymorphism, was generated from cloned polymerase chain reaction (PCR) fragments amplified directly from naturally infected tissues. CAV73 had been detected in 31-day-old broilers submitted for examination for reasons unrelated to anemia. After electroporation of the cloned genomes into MDCC-CU147 lymphoblastoid cells, the regenerated CAV caused the culture to fail within 9 days, and the medium contained 5 X 10(6) TCID50 CAV/ml. Use of MDCC-CU147 cells was essential, as identical electroporation of MDCC-MSB1 cells failed to generate CAV able to destroy the culture within 8 wk. Regenerated CAV73 produced anemia and severe lymphocytic depletion of the thymus when inoculated into susceptible 3-day-old chickens and was reisolated from these chickens. Furthermore, it replicated in low- and high-passage MDCC-MSB1 cells similarly to a low-passage CAV field isolate that contains a different VP 1 "hypervariable region." The regeneration of CAV from PCR products directly from naturally infected carcasses, as performed in this study, provides a tool for the evaluation of distinct genetic polymorphisms that may be detected in specimens where infective virions are no longer available. Our results also provide some insight into the differential susceptibility of cell lines for low-passage CAV field isolates. PMID:17461269

  13. Caveolin-1 protects against hepatic ischemia/reperfusion injury through ameliorating peroxynitrite-mediated cell death.

    Science.gov (United States)

    Gao, Lei; Chen, Xingmiao; Peng, Tao; Yang, Dan; Wang, Qi; Lv, Zhiping; Shen, Jiangang

    2016-06-01

    Nitrative stress is considered as an important pathological process of hepatic ischemia and reperfusion injury but its regulating mechanisms are largely unknown. In this study, we tested the hypothesis that caveolin-1 (Cav-1), a plasma membrane scaffolding protein, could be an important cellular signaling against hepatic I/R injury through inhibiting peroxynitrite (ONOO(-))-induced cellular damage. Male wild-type mice and Cav-1 knockout (Cav-1(-/-)) were subjected to 1h hepatic ischemia following 1, 6 and 12h of reperfusion by clipping and releasing portal vessels respectively. Immortalized human hepatocyte cell line (L02) was subjected to 1h hypoxia and 6h reoxygenation and treated with Cav-1 scaffolding domain peptide. The major discoveries included: (1) the expression of Cav-1 in serum and liver tissues of wild-type mice was time-dependently elevated during hepatic ischemia-reperfusion injury. (2) Cav-1 scaffolding domain peptide treatment inhibited cleaved caspase-3 expression in the hypoxia-reoxygenated L02 cells; (3) Cav-1 knockout (Cav-1(-/-)) mice had significantly higher levels of serum transaminases (ALT&AST) and TNF-α, and higher rates of apoptotic cell death in liver tissues than wild-type mice after subjected to 1h hepatic ischemia and 6hour reperfusion; (4) Cav-1(-/-) mice revealed higher expression levels of iNOS, ONOO(-) and 3-nitrotyrosine (3-NT) in the liver than wild-type mice, and Fe-TMPyP, a representative peroxynitrite decomposition catalyst (PDC), remarkably reduced level of ONOO(-) and 3-NT and ameliorated the serum ALT, AST and TNF-α levels in both wild-type and Cav-1(-/-) mice. Taken together, we conclude that Cav-1 could play a critical role in preventing nitrative stress-induced liver damage during hepatic ischemia-reperfusion injury. PMID:27021966

  14. New glimpses of caveolin-1 functions in embryonic development and human diseases

    Institute of Scientific and Technical Information of China (English)

    Saijun MO; Shengli YANG; Zongbin CUI

    2011-01-01

    Caveolin-1 (Cav-1) isoforms,including Cav-1α and Cav-1β,were identified as integral membrane proteins and the major components of caveolae.Cav-1 proteins are highly conserved during evolution from Caenorhabditis elegans to human and are capable of interacting with many signaling molecules through their caveolin scaffolding domains to regulate the activities of multiple signaling pathways.Thus,Cav-1 plays crucial roles in the regulation of cellular proliferation,differentiation and apoptosis in a cell-specific and contextual manner.In addition,Cav-1 is essential for embryonic development of vertebrates owing to its regulation of BMP,Wnt,TGF-β and other key signaling molecules.Moreover,Cav-1 is mainly expressed in terminally differentiated cells and its abnormal expression is often associated with human diseases,such as tumor progression,cardiovascular diseases,fibrosis,lung regeneration,and diseases related to virus.In this review,we will further discuss the potential of Cav-1 as a target for disease therapy and multiple drug resistance.

  15. Temperature effects for high pressure processing of Picornaviruses

    Science.gov (United States)

    Investigation of the effects of pre-pressurization temperature on the high pressure inactivation for single strains of aichivirus (AiV), coxsackievirus A9 (CAV9) and B5 (CBV5) viruses, as well as human parechovirus -1 (HPeV), was performed. For CAV9, an average 1.99 log10 greater inactivation was ...

  16. The administration of a single dose of a multivalent (DHPPiL4R vaccine prevents clinical signs and mortality following virulent challenge with canine distemper virus, canine adenovirus or canine parvovirus

    Directory of Open Access Journals (Sweden)

    Stephen Wilson

    2014-01-01

    In conclusion, we demonstrated that a single administration of a minimum titre, multivalent vaccine to dogs of six weeks of age is efficacious and prevents clinical signs and mortality caused by CAV-1 and CDV; prevents clinical signs and significantly reduces virus shedding caused by CAV-2; and prevents clinical signs, leucopoenia and viral excretion caused by CPV.

  17. Fragile X mental retardation protein controls synaptic vesicle exocytosis by modulating N-type calcium channel density

    Science.gov (United States)

    Ferron, Laurent; Nieto-Rostro, Manuela; Cassidy, John S.; Dolphin, Annette C.

    2014-04-01

    Fragile X syndrome (FXS), the most common heritable form of mental retardation, is characterized by synaptic dysfunction. Synaptic transmission depends critically on presynaptic calcium entry via voltage-gated calcium (CaV) channels. Here we show that the functional expression of neuronal N-type CaV channels (CaV2.2) is regulated by fragile X mental retardation protein (FMRP). We find that FMRP knockdown in dorsal root ganglion neurons increases CaV channel density in somata and in presynaptic terminals. We then show that FMRP controls CaV2.2 surface expression by targeting the channels to the proteasome for degradation. The interaction between FMRP and CaV2.2 occurs between the carboxy-terminal domain of FMRP and domains of CaV2.2 known to interact with the neurotransmitter release machinery. Finally, we show that FMRP controls synaptic exocytosis via CaV2.2 channels. Our data indicate that FMRP is a potent regulator of presynaptic activity, and its loss is likely to contribute to synaptic dysfunction in FXS.

  18. Transplant allograft vasculopathy: Role of multimodality imaging in surveillance and diagnosis.

    Science.gov (United States)

    Payne, Gregory A; Hage, Fadi G; Acharya, Deepak

    2016-08-01

    Cardiac allograft vasculopathy (CAV) is a challenging long-term complication of cardiac transplantation and remains a leading long-term cause of graft failure, re-transplantation, and death. CAV is an inflammatory vasculopathy distinct from traditional atherosclerotic coronary artery disease. Historically, the surveillance and diagnosis of CAV has been dependent on serial invasive coronary angiography with intravascular imaging. Although commonly practiced, angiography is not without significant limitations. Technological advances have provided sophisticated imaging techniques for CAV assessment. It is now possible to assess the vascular lumen, vessel wall characteristics, absolute blood flow, perfusion reserve, myocardial contractile function, and myocardial metabolism and injury in a noninvasive, expeditious manner with little risk. The current article will review key imaging modalities for the surveillance, diagnosis, and prognosis of CAV and discuss coronary physiology of transplanted hearts with emphasis on the clinical implications for provocative and vasodilator stress testing. PMID:26711101

  19. Integrin α1β1 Regulates Epidermal Growth Factor Receptor Activation by Controlling Peroxisome Proliferator-Activated Receptor γ-Dependent Caveolin-1 Expression ▿ # ‖

    Science.gov (United States)

    Chen, Xiwu; Whiting, Carrie; Borza, Corina; Hu, Wen; Mont, Stacey; Bulus, Nada; Zhang, Ming-Zhi; Harris, Raymond C.; Zent, Roy; Pozzi, Ambra

    2010-01-01

    Integrin α1β1 negatively regulates the generation of profibrotic reactive oxygen species (ROS) by inhibiting epidermal growth factor receptor (EGFR) activation; however, the mechanism by which it does this is unknown. In this study, we show that caveolin-1 (Cav-1), a scaffolding protein that binds integrins and controls growth factor receptor signaling, participates in integrin α1β1-mediated EGFR activation. Integrin α1-null mesangial cells (MCs) have reduced Cav-1 levels, and reexpression of the integrin α1 subunit increases Cav-1 levels, decreases EGFR activation, and reduces ROS production. Downregulation of Cav-1 in wild-type MCs increases EGFR phosphorylation and ROS synthesis, while overexpression of Cav-1 in the integrin α1-null MCs decreases EGFR-mediated ROS production. We further show that integrin α1-null MCs have increased levels of activated extracellular signal-regulated kinase (ERK), which leads to reduced activation of peroxisome proliferator-activated receptor γ (PPARγ), a transcription factor that positively regulates Cav-1 expression. Moreover, activation of PPARγ or inhibition of ERK increases Cav-1 levels in the integrin α1-null MCs. Finally, we show that glomeruli of integrin α1-null mice have reduced levels of Cav-1 and activated PPARγ but increased levels of phosphorylated EGFR both at baseline and following injury. Thus, integrin α1β1 negatively regulates EGFR activation by positively controlling Cav-1 levels, and the ERK/PPARγ axis plays a key role in regulating integrin α1β1-dependent Cav-1 expression and consequent EGFR-mediated ROS production. PMID:20368353

  20. Integrin alpha1beta1 regulates epidermal growth factor receptor activation by controlling peroxisome proliferator-activated receptor gamma-dependent caveolin-1 expression.

    Science.gov (United States)

    Chen, Xiwu; Whiting, Carrie; Borza, Corina; Hu, Wen; Mont, Stacey; Bulus, Nada; Zhang, Ming-Zhi; Harris, Raymond C; Zent, Roy; Pozzi, Ambra

    2010-06-01

    Integrin alpha1beta1 negatively regulates the generation of profibrotic reactive oxygen species (ROS) by inhibiting epidermal growth factor receptor (EGFR) activation; however, the mechanism by which it does this is unknown. In this study, we show that caveolin-1 (Cav-1), a scaffolding protein that binds integrins and controls growth factor receptor signaling, participates in integrin alpha1beta1-mediated EGFR activation. Integrin alpha1-null mesangial cells (MCs) have reduced Cav-1 levels, and reexpression of the integrin alpha1 subunit increases Cav-1 levels, decreases EGFR activation, and reduces ROS production. Downregulation of Cav-1 in wild-type MCs increases EGFR phosphorylation and ROS synthesis, while overexpression of Cav-1 in the integrin alpha1-null MCs decreases EGFR-mediated ROS production. We further show that integrin alpha1-null MCs have increased levels of activated extracellular signal-regulated kinase (ERK), which leads to reduced activation of peroxisome proliferator-activated receptor gamma (PPARgamma), a transcription factor that positively regulates Cav-1 expression. Moreover, activation of PPARgamma or inhibition of ERK increases Cav-1 levels in the integrin alpha1-null MCs. Finally, we show that glomeruli of integrin alpha1-null mice have reduced levels of Cav-1 and activated PPARgamma but increased levels of phosphorylated EGFR both at baseline and following injury. Thus, integrin alpha1beta1 negatively regulates EGFR activation by positively controlling Cav-1 levels, and the ERK/PPARgamma axis plays a key role in regulating integrin alpha1beta1-dependent Cav-1 expression and consequent EGFR-mediated ROS production. PMID:20368353

  1. Curcumin attenuates high glucose-induced podocyte apoptosis by regulating functional connections between caveolin-1 phosphorylation and ROS

    Science.gov (United States)

    Sun, Li-na; Liu, Xiang-chun; Chen, Xiang-jun; Guan, Guang-ju; Liu, Gang

    2016-01-01

    Aim: Caveolin-1 (cav-1) is a major multifunctional scaffolding protein of caveolae. Cav-1 is primarily expressed in mesangial cells, renal proximal tubule cells and podocytes in kidneys. Recent evidence shows that the functional connections between cav-1 and ROS play a key role in many diseases. In this study we investigated whether regulating the functional connections between cav-1 and ROS in kidneys contributed to the beneficial effects of curcumin in treating diabetic nephropathy in vitro and in vivo. Methods: Cultured mouse podocytes (mpc5) were incubated in a high glucose (HG, 30 mmol/L) medium for 24, 48 or 72 h. Male rats were injected with STZ (60 mg/kg, ip) to induce diabetes. ROS generation, SOD activity, MDA content and caspase-3 activity in the cultured cells and kidney cortex homogenate were determined. Apoptotic proteins and cav-1 phosphorylation were analyzed using Western blot analyses. Results: Incubation in HG-containing medium time-dependently increased ROS production, oxidative stress, apoptosis, and cav-1 phosphorylation in podocytes. Pretreatment with curcumin (1, 5, and 10 μmol/L) dose-dependently attenuated these abnormalities in HG-treated podocytes. Furthermore, in HG-containing medium, the podocytes transfected with a recombinant plasmid GFP-cav-1 Y14F (mutation at a cav-1 phosphorylation site) exhibited significantly decreased ROS production and apoptosis compared with the cells transfected with empty vector. In diabetic rats, administration of curcumin (100 or 200 mg/kg body weight per day, ig, for 8 weeks) not only significantly improved the renal function, but also suppressed ROS levels, oxidative stress, apoptosis and cav-1 phosphorylation in the kidneys. Conclusion: Curcumin attenuates high glucose-induced podocyte apoptosis in vitro and diabetic nephropathy in vivo partly through regulating the functional connections between cav-1 phosphorylation and ROS. PMID:26838071

  2. Potential role of caveolin-1 in acetaminophen-induced hepatotoxicity

    International Nuclear Information System (INIS)

    Caveolin-1 (Cav-1) is a membrane scaffolding protein, which functions to regulate intracellular compartmentalization of various signaling molecules. In the present studies, transgenic mice with a targeted disruption of the Cav-1 gene (Cav-1-/-) were used to assess the role of Cav-1 in acetaminophen-induced hepatotoxicity. Treatment of wild-type mice with acetaminophen (300 mg/kg) resulted in centrilobular hepatic necrosis and increases in serum transaminases. This was correlated with decreased expression of Cav-1 in the liver. Acetaminophen-induced hepatotoxicity was significantly attenuated in Cav-1-/- mice, an effect that was independent of acetaminophen metabolism. Acetaminophen administration resulted in increased hepatic expression of the oxidative stress marker, lipocalin 24p3, as well as hemeoxygenase-1, but decreased glutathione and superoxide dismutase-1; no differences were noted between the genotypes suggesting that reduced toxicity in Cav-1-/- mice is not due to alterations in antioxidant defense. In wild-type mice, acetaminophen increased mRNA expression of the pro-inflammatory cytokines, interleukin-1β, and monocyte chemoattractant protein-1 (MCP-1), as well as cyclooxygenase-2, while 15-lipoxygenase (15-LOX), which generates anti-inflammatory lipoxins, decreased. Acetaminophen-induced changes in MCP-1 and 15-LOX expression were greater in Cav-1-/- mice. Although expression of tumor necrosis factor-α, a potent hepatocyte mitogen, was up-regulated in the liver of Cav-1-/- mice after acetaminophen, expression of proliferating cell nuclear antigen and survivin, markers of cellular proliferation, were delayed, which may reflect the reduced need for tissue repair. Taken together, these data demonstrate that Cav-1 plays a role in promoting inflammation and toxicity during the pathogenesis of acetaminophen-induced injury.

  3. Fungi in a changing world: growth rates will be elevated, but spore production may decrease in future climates

    Science.gov (United States)

    Damialis, Athanasios; Mohammad, Aqilah B.; Halley, John M.; Gange, Alan C.

    2015-09-01

    Very little is known about the impact of climate change on fungi and especially on spore production. Fungal spores can be allergenic, thus being important for human health. The aim of this study was to investigate how climate change influences the responsive ability of fungi by simulating differing environmental regimes. Fungal species with high spore allergenic potential and atmospheric abundance were grown and experimentally examined under a variety of temperatures and different nutrient availability. Each represented the average decadal air temperature of the 1980s, 1990s and 2000s in the UK, along with an Intergovernmental Panel on Climate Change (IPCC) climate change scenario for 2100. All tests were run on six fungal species: Alternaria alternata, Aspergillus niger, Botrytis cinerea, Cladosporium cladosporioides, Cladosporium oxysporum and Epicoccum purpurascens. Mycelium growth rate and spore production were examined on each single species and competitive capacity among species combinations in pairs. All fungal species grew faster at higher temperatures, and this was more pronounced for the temperature projection in 2100. Most species grew faster when there was lower nutrient availability. Exceptions were the species with the highest growth rate ( E. purpurascens) and with the highest competition capacity ( A. alternata). Most species (except for E. purpurascens) produced more spores in the richer nutrient medium but fewer as temperature increased. C. cladosporioides was an exception, exponentially increasing its spore production in the temperature of the 2100 scenario. Regarding competitive capacity, no species displayed any significant alterations within the environmental range checked. It is suggested that in future climates, fungi will display dramatic growth responses, with faster mycelium growth and lower spore production, with questions risen on relevant allergen potential.

  4. Frações de carboidratos de quatro gramíneas tropicais em diferentes idades de corte e doses de adubação nitrogenada Carbohydrate fractions of tropical grasses at different cutting ages and under nitrogen fertilization

    OpenAIRE

    L.T. Henriques; J.F. Coelho da Silva; E. Detmann; H.M. Vasquez; O.G. Pereira

    2007-01-01

    Avaliaram-se as frações de carboidratos dos capins setária (Setaria anceps Stapf), hemarthria (Hemarthria altissima [Poir] Stapf. & Hubbard), angola (Brachiaria purpurascens [Raddi] Henr.) e acroceres (Acroceras macrum Stapf.) adubados com 0, 100, 200, 300 e 400kg de N/ha na forma de sulfato de amônio e colhidos aos 28, 42, 56 e 70 dias de idade. Os cortes foram realizados a 20cm do solo para o capim-setária e a 10cm para as demais espécies. As frações dos carboidratos das forrageiras foram i...

  5. Occurrence of pathotypes of Ascochyta pisi (Lib. in the Olsztyn district

    Directory of Open Access Journals (Sweden)

    Helena Furgał-Węgrzycka

    2013-12-01

    Full Text Available The causal agents of leaf and pods spot-pot of peas and field peas in the Olsztyn district was the fungus Ascochyta pisi (Lib.. Investigated isolates of A. pisi differed by macroscopic and microscopic features. From diseases leaves and pods of peas and field peas obtained also saprophitic fungi among which Alternaria alternata, Botrytis cinerea, Cladosporium c1adosporioides, Epicoccum purpurascens, Stemphylium botryosum, Sordaria fimicola and mycelia starilia were dominated. In the study of the identification and distribution of pathotypes of A. pisi five pathotypes were identified among which pathotype 2 were dominated. The identified the pathotypes differed by macroscopic and microscopic features.

  6. Aportaciones a la flora de Galicia, IX

    OpenAIRE

    Silva Pando, F.J.; Pino Pérez, R.; Pino Pérez, J.J.; García Martínez, X.R.; Morla Juaristi, C.; Cebolla Lozano, C.; Gómez Vigide, F.; Camaño Portela, J.L.; Rial Pousa, S.; Álvarez Graña, D.; Blanco Dios, J.B.; Paz Rosales, M.

    2009-01-01

    En este trabajo se mencionan 53 plantas de diverso interés para la flora de Galicia o zonas próximas. Se incluyen 3 novedades para España (Sesamoides minor (Lange) O. Kuntze x S. purpurascens (L.) G. López, Hedychium gardnerianum Sheppard ex Ker-Gawler y Oxalis incarnata L.) y otra para Portugal (Bromus hordeaceus L. subsp. ferronii (Mabille) P.M. Smith), 3 novedades regionales (Oxalis incarnata L., Antirrhinum latifolium Mill., Sisyrinchium striatum Mill.), 16 novedades provincia...

  7. Discovery and Characterization of the Short κA-Conotoxins: A Novel Subfamily of Excitatory Conotoxins

    OpenAIRE

    Teichert, Russell W.; Jacobsen, Richard; Terlau, Heinrich; Yoshikami, Doju; Olivera, Baldomero M.

    2006-01-01

    We have characterized the defining members of a novel subfamily of excitatory conotoxins, the short κA-conotoxins (κAS-conotoxins). κA-conotoxins PIVE and PIVF (κA-PIVE and κA-PIVF) were purified from Conus purpurascens venom. Both peptides elicited excitatory activity upon injection into fish. κA-PIVE was synthesized for further characterization. The excitatory effects of κA-PIVE in vivo were dose dependent, causing hyperactivity at low doses and rapid immobilization at high doses, symptomat...

  8. Serum Caveolin-1 as a Novel Biomarker in Idiopathic Pulmonary Artery Hypertension

    Directory of Open Access Journals (Sweden)

    Kuo-Yang Wang

    2015-01-01

    Full Text Available Pulmonary arterial hypertension (PAH is a rare disease but with significant morbidity and high mortality. There is no specific way to diagnose PAH. Thus, an easy used with good sensitivity and specificity biomarker of PAH is highly desirable to aid in the screening, diagnosis, and follow-up. Caveolin-1 (Cav1 is the structural protein of caveolae and is highly expressed in type I pneumocytes. Lungs tissues from idiopathic PAH (IPAH patients showed decreased expression of Cav1 in vascular endothelial cells. Therefore, we developed a direct sandwich immunoassay for the determination of Cav1 in IAPH patient’s serum. The result disclosed serum Cav1 level was significantly lower in IPAH than control groups. Using serum Cav1, 17.17 pg/mL as a cutoff value, the sensitivity was 0.59 and the specificity was 1.0. There were two major findings in our results. First, serum Cav1 might be a novel biomarker in the diagnosis of IPAH with fare sensitivity and good specificity. Second, Cav1 might be used to make differential diagnosis between COPD-PH and IPAH group.

  9. T-type calcium channel expression in cultured human neuroblastoma cells

    Institute of Scientific and Technical Information of China (English)

    Xianjie Wen; Shiyuan Xu; Lingling Wang; Hua Liang; Chengxiang Yang; Hanbing Wang; Hongzhen Liu

    2011-01-01

    Human neuroblastoma cells (SH-SY5Y) have similar structures and functions as neural cells and have been frequently used for cell culture studies of neural cell functions. Previous studies have revealed Land N-type calcium channels in SH-SY5Y cells. However, the distribution of the low -voltage activated calcium channel (namely called T-type calcium channel, including Cav3.1, Cav3.2, and Cav3.3) in SH-SY5Y cells remains poorly understood. The present study detected mRNA and protein expression of the T-type calcium channel (Cav3.1, Cav3.2, and Cav3.3) in cultured SH-SY5Y cells using real-time polymerase chain reaction (PCR) and western blot analysis. Results revealed mRNA and protein expression from all three T-type calcium channel subtypes in SH-SY5Y cells. Moreover,Cav3.1 was the predominant T-type calcium channel subtype in SH-SY5Y cells.

  10. Canine adenovirus type 2 vector generation via I-Sce1-mediated intracellular genome release.

    Directory of Open Access Journals (Sweden)

    Sandy Ibanes

    Full Text Available When canine adenovirus type 2 (CAdV-2, or also commonly referred to as CAV-2 vectors are injected into the brain parenchyma they preferentially transduce neurons, are capable of efficient axonal transport to afferent regions, and allow transgene expression for at last >1 yr. Yet, translating these data into a user-friendly vector platform has been limited because CAV-2 vector generation is challenging. Generation of E1-deleted adenovirus vectors often requires transfection of linear DNA fragments of >30 kb containing the vector genome into an E1-transcomplementing cell line. In contrast to human adenovirus type 5 vector generation, CAV-2 vector generation is less efficient due, in part, to a reduced ability to initiate replication and poor transfectibility of canine cells with large, linear DNA fragments. To improve CAV-2 vector generation, we generated an E1-transcomplementing cell line expressing the estrogen receptor (ER fused to I-SceI, a yeast meganuclease, and plasmids containing the I-SceI recognition sites flanking the CAV-2 vector genome. Using transfection of supercoiled plasmid and intracellular genome release via 4-OH-tamoxifen-induced nuclear translocation of I-SceI, we improved CAV-2 vector titers 1,000 fold, and in turn increased the efficacy of CAV-2 vector generation.

  11. Preparation of monoclonal antibody against celangulin V and immunolocalization of receptor in the oriental armyworm, Mythimna separata walker (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Qi, Zhijun; Xue, Xiaoping; Wu, Wenjun; Zhang, Jiwen; Yang, Runya

    2006-10-01

    The botanical insecticide celangulin V (CA-V) is an insect digestive poison acting on midgut tissue of the target insect larvae. With the aim of localizing the receptor enacted by CA-V, monoclonal antibodies (MAbs) specific to the compound were developed. A hapten was synthesized by introducing a succinoyl into the CA-V structure and conjugated with three carrier proteins. From mice immunized with one conjugate, three MAbs were obtained with a potential capacity of detecting protein-bound residue forms of CA-V in the biological tissues. The oriental armyworm larvae ingested CA-V were examined by the technique of immuno-electron-microscopy (IEM) using the anti-CA-V MAb as the primary antibody and goat anti-mouse/IgG labeled with colloidal gold as the secondary antibody. Electron micrographs of the armyworm midgut tissues showed that the CA-V was associated with the midgut epithelia of the insects. These results demonstrated the existence of a receptor enacted by CA-V on the midgut cells of the oriental armyworm larvae. PMID:17002428

  12. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    Science.gov (United States)

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology. PMID:25952731

  13. Patterns of Exposure of Iberian Wolves (Canis lupus) to Canine Viruses in Human-Dominated Landscapes.

    Science.gov (United States)

    Millán, Javier; López-Bao, José Vicente; García, Emilio J; Oleaga, Álvaro; Llaneza, Luis; Palacios, Vicente; de la Torre, Ana; Rodríguez, Alejandro; Dubovi, Edward J; Esperón, Fernando

    2016-03-01

    Wildlife inhabiting human-dominated landscapes is at risk of pathogen spill-over from domestic species. With the aim of gaining knowledge in the dynamics of viral infections in Iberian wolves (Canis lupus) living in anthropized landscapes of northern Spain, we analysed between 2010 and 2013 the samples of 54 wolves by serology and polymerase chain reaction (PCR) for exposure to four pathogenic canine viruses: canine distemper virus (CDV), canine parvovirus-2 (CPV), canine adenovirus 1 and 2 (CAV-1 and CAV-2) and canine herpesvirus. Overall, 76% of the studied wolves presented evidence of exposure to CPV (96% by HI, 66% by PCR) and 75% to CAV (75% by virus neutralization (VN), 76% by PCR, of which 70% CAV-1 and 6% CAV-2). This represents the first detection of CAV-2 infection in a wild carnivore. CPV/CAV-1 co-infection occurred in 51% of the wolves. The probability of wolf exposure to CPV was positively and significantly correlated with farm density in a buffer zone around the place where the wolf was found, indicating that rural dogs might be the origin of CPV infecting wolves. CPV and CAV-1 appear to be enzootic in the Iberian wolf population, which is supported by the absence of seasonal and inter-annual variations in the proportion of positive samples detected. However, while CPV may depend on periodical introductions by dogs, CAV-1 may be maintained within the wolf population. All wolves were negative for exposure to CDV (by VN and PCR) and CHV (by PCR). The absence of acquired immunity against CDV in this population may predispose it to an elevated rate of mortality in the event of a distemper spill-over via dogs. PMID:26589403

  14. Sex-dependent expression of caveolin 1 in response to sex steroid hormones is closely associated with development of obesity in rats.

    Directory of Open Access Journals (Sweden)

    Rajib Mukherjee

    Full Text Available Caveolin-1 (CAV1 is a conserved group of structural membrane proteins that form special cholesterol and sphingolipid-rich compartments, especially in adipocytes. Recently, it has been reported that CAV1 is an important target protein in sex hormone-dependent regulation of various metabolic pathways, particularly in cancer and diabetes. To clarify distinct roles of CAV1 in sex-dependent obesity development, we investigated the effects of high fat diet (HFD and sex steroid hormones on CAV1 expression in adipose tissues of male and female rats. Results of animal experiments revealed that estrogen (17-β-estradiol, E2 and androgen (dihydrotestosterone, DHT had opposite effects on body weight gain as well as on the regulation of CAV1, hormone sensitive lipase (HSL and uncoupling protein 1 (UCP1 in adipose tissues. Furthermore, sex hormone receptors and aromatase were differentially expressed in a sex-dependent manner in response to E2 and DHT treatments. In vivo data were confirmed using 3T3-L1 and HIB1B cell lines, where Cav1 knock down stimulated lipogenesis but suppressed sex hormone receptor signaling proteins. Most importantly, co-immunoprecipitation enabled the identification of previously unrecognized CAV1-interacting mitochondrial or lipid oxidative pathway proteins in adipose tissues. Taken together, current data showed that CAV1 may play important preventive role in the development of obesity, with more prominent effects in females, and proved to be an important target protein for the hormonal regulation of adipose tissue metabolism by manipulating sex hormone receptors and mitochondrial oxidative pathways. Therefore, we can report, for the first time, the molecular mechanism underlying the effects of sex steroid hormones in the sex-dimorphic regulation of CAV1.

  15. Functional role of voltage gated Ca2+ channels in heart automaticity

    Directory of Open Access Journals (Sweden)

    Pietro eMesirca

    2015-02-01

    Full Text Available Pacemaker activity of automatic cardiac myocytes controls the heartbeat in everyday life. Cardiac automaticity is under the control of several neurotransmitters and hormones and is constantly regulated by the autonomic nervous system to match the physiological needs of the organism. Several classes of ion channels and proteins involved in intracellular Ca2+ dynamics contribute to pacemaker activity. The functional role of voltage-gated calcium channels (VGCCs in heart automaticity and impulse conduction has been matter of debate for 30 years. However, growing evidence shows that VGCCs are important regulators of the pacemaker mechanisms and play also a major role in atrio-ventricular impulse conduction. Incidentally, studies performed in genetically modified mice lacking L-type Cav1.3 (Cav1.3-/- or T-type Cav3.1 (Cav3.1-/- channels show that genetic inactivation of these channels strongly impacts pacemaking. In cardiac pacemaker cells, VGCCs activate at negative voltages at the beginning of the diastolic depolarization and importantly contribute to this phase by supplying inward current. Loss-of-function of these channels also impairs atrio-ventricular conduction. Furthermore, inactivation of Cav1.3 channels promotes also atrial fibrillation and flutter in knockout mice suggesting that these channels can play a role in stabilizing atrial rhythm. Genomic analysis demonstrated that Cav1.3 and Cav3.1 channels are widely expressed in pacemaker tissue of mice, rabbits and humans. Importantly, human diseases of pacemaker activity such as congenital bradycardia and heart block have been attributed to loss-of-function of Cav1.3 and Cav3.1 channels. In this article, we will review the current knowledge on the role of VGCCs in the generation and regulation of heart rate and rhythm. We will discuss also how loss of Ca2+ entry through VGCCs could influence intracellular Ca2+ handling and promote atrial arrhythmias.

  16. Signal processing by T-type calcium channel interactions in the cerebellum

    Science.gov (United States)

    Engbers, Jordan D. T.; Anderson, Dustin; Zamponi, Gerald W.; Turner, Ray W.

    2013-01-01

    T-type calcium channels of the Cav3 family are unique among voltage-gated calcium channels due to their low activation voltage, rapid inactivation, and small single channel conductance. These special properties allow Cav3 calcium channels to regulate neuronal processing in the subthreshold voltage range. Here, we review two different subthreshold ion channel interactions involving Cav3 channels and explore the ability of these interactions to expand the functional roles of Cav3 channels. In cerebellar Purkinje cells, Cav3 and intermediate conductance calcium-activated potassium (IKCa) channels form a novel complex which creates a low voltage-activated, transient outward current capable of suppressing temporal summation of excitatory postsynaptic potentials (EPSPs). In large diameter neurons of the deep cerebellar nuclei, Cav3-mediated calcium current (IT) and hyperpolarization-activated cation current (IH) are activated during trains of inhibitory postsynaptic potentials. These currents have distinct, and yet synergistic, roles in the subthreshold domain with IT generating a rebound burst and IH controlling first spike latency and rebound spike precision. However, by shortening the membrane time constant the membrane returns towards resting value at a faster rate, allowing IH to increase the efficacy of IT and increase the range of burst frequencies that can be generated. The net effect of Cav3 channels thus depends on the channels with which they are paired. When expressed in a complex with a KCa channel, Cav3 channels reduce excitability when processing excitatory inputs. If functionally coupled with an HCN channel, the depolarizing effect of Cav3 channels is accentuated, allowing for efficient inversion of inhibitory inputs to generate a rebound burst output. Therefore, signal processing relies not only on the activity of individual subtypes of channels but also on complex interactions between ion channels whether based on a physical complex or by indirect

  17. Quantum Dots-Based Immunofluorescent Imaging of Stromal Fibroblasts Caveolin-1 and Light Chain 3B Expression and Identification of Their Clinical Significance in Human Gastric Cancer

    Science.gov (United States)

    He, Yuyu; Zhao, Xianda; Gao, Jun; Fan, Lifang; Yang, Guifang; Cho, William Chi-shing; Chen, Honglei

    2012-01-01

    Caveolin-1 (Cav-1) expression deficiency and autophagy in tumor stromal fibroblasts (hereafter fibroblasts) are involved in tumor proliferation and progression, particularly in breast and prostate cancer. The aim of this study was to detect the expression of fibroblastic Cav-1 and LC3B, markers of autophagy, in gastric cancer (GC) and to analyze their clinical significances. Furthermore, because Epstein-Barr virus (EBV)-associated GC (EBVaGC) is a unique subtype of GC; we compared the differential expression of fibroblastic Cav-1 and LC3B in EBVaGC and non-EBVaGC. Quantum dots (QDs)-based immunofluorescence histochemistry was used to examine the expression of fibroblastic Cav-1 and LC3B in 118 cases of GC with adequate stroma. QDs-based double immunofluorescence labeling was performed to detect the coexpression of Cav-1 and LC3B proteins. EBV-encoded small RNA was detected by QDs-based fluorescence in situ hybridization to identify EBVaGC. Multivariate analysis indicated that low fibroblastic Cav-1 level was an independent prognosticator (p = 0.029) that predicted poorer survival of GC patients. Positive fibroblastic LC3B was correlated with lower invasion (p = 0.032) and was positively associated with Cav-1 expression (r = 0.432, p < 0.001). EBV infection did not affect fibroblastic Cav-1 and LC3B expression. In conclusion, positive fibroblastic LC3B correlates with lower invasion, and low expression of fibroblastic Cav-1 is a novel predictor of poor GC prognosis. PMID:23203033

  18. Blockade of CD26-mediated T cell costimulation with soluble caveolin-1-Ig fusion protein induces anergy in CD4+T cells

    International Nuclear Information System (INIS)

    CD26 binds to caveolin-1 in antigen-presenting cells (APC), and that ligation of CD26 by caveolin-1 induces T cell proliferation in a TCR/CD3-dependent manner. We report herein the effects of CD26-caveolin-1 costimulatory blockade by fusion protein caveolin-1-Ig (Cav-Ig). Soluble Cav-Ig inhibits T cell proliferation and cytokine production in response to recall antigen, or allogeneic APC. Our data hence suggest that blocking of CD26-associated signaling by soluble Cav-Ig may be an effective approach as immunosuppressive therapy.

  19. Caveolin-1 Deficiency Induces Spontaneous Endothelial-to-Mesenchymal Transition in Murine Pulmonary Endothelial Cells in Vitro

    OpenAIRE

    Li, Zhaodong; Wermuth, Peter J.; Benn, Bryan S.; Lisanti, Michael P.; Jimenez, Sergio A.

    2013-01-01

    It was previously demonstrated that transforming growth factor β (TGF-β) induces endothelial-to-mesenchymal transition (EndoMT) in murine lung endothelial cells (ECs) in vitro. Owing to the important role of caveolin-1 (CAV1) in TGF-β receptor internalization and TGF-β signaling, the participation of CAV1 in the induction of EndoMT in murine lung ECs was investigated. Pulmonary ECs were isolated from wild-type and Cav1 knockout mice using immunomagnetic methods with sequential anti-CD31 and a...

  20. Two-pore channels provide insight into the evolution of voltage-gated Ca2+ and Na+ channels

    OpenAIRE

    Rahman, Taufiq; Cai, Xinjiang; Brailoiu, G. Cristina; Abood, Mary E; Brailoiu, Eugen; Patel, Sandip

    2014-01-01

    Four-domain voltage-gated Ca2+ and Na+ channels (CaV, NaV) underpin nervous system function and likely emerged upon intragenic duplication of a primordial two-domain precursor. To investigate if two-pore channels (TPCs) may represent an intermediate in this evolutionary transition, we performed molecular docking simulations with a homology model of TPC1, which suggested that the pore region could bind antagonists of CaV or NaV. CaV or NaV antagonists blocked NAADP-evoked Ca2+ signals in sea u...

  1. Determinantes de la fuerza específica de contracción en el músculo esquelético del ratón geronte : Participación de Cav1.1, Troponina T, calpaínas y la interacción músculo-tejido adiposo

    OpenAIRE

    Pereyra, Andrea

    2015-01-01

    El músculo esquelético (ME) constituye el tejido más abundante del organismo y representa aproximadamente el 40% del peso corporal total. La función clásica del tejido muscular es la de producir movimiento y generar fuerza siendo también de vital importancia su rol como regulador de la homeostasis energética general. Bajo condiciones fisiológicas, su masa se mantiene por un delicado balance entre la fusión de células precursoras, la hipertrofia de las miofibras y el catabolismo proteico. A pa...

  2. NMR Studies of the Vanadium Spin Dynamics and Spin Structure in LiV2O4, CaV2O4, and (LixV1-x)3BO5 (x is almost equal to 0.33, 0.40)

    Energy Technology Data Exchange (ETDEWEB)

    Xiaopeng Zong

    2007-12-01

    Strong electron correlation is believed to be an essential and unifying factor in diverse properties of condensed matter systems. Ground states that can arise due to electron correlation effects include Mott insulators, heavy fermion, ferromagnetism and antiferromagnetism, spin glasses, and high-temperature superconductivity. The electronic systems in transition metal oxide compounds are often highly correlated. In this thesis, the author presents experimental studies on three strongly correlated vanadium oxide compounds: LiV{sub 2}O{sub 4}, (Li{sub x}V{sub 1-x}){sub 3}BO{sub 5}, and CaV{sub 2}O{sub 4}, which have completely different ground states.

  3. Contenido de compuestos antioxidantes en tres estados de maduración de tomate de árbol (Solanum betaceum Cav. cultivado a diferentes alturas (m.s.n.m. Antioxidant compounds content in red-purple tree tomato grown at different altitudes (m.a.s.l.

    Directory of Open Access Journals (Sweden)

    Lorena Cuesta

    2013-06-01

    Full Text Available (Recibido: 2013/03/07 - Aceptado: 2013/06/17El objetivo de esta investigación fue estudiar el contenido de compuestos antioxidantes entomate de árbol morado cultivado a diferentes alturas (m.s.n.m.. Se emplearon frutosprovenientes de Pelileo (2660 m.s.n.m. y Chiquicha (2440 m.s.n.m. en tres estados demaduración (inmaduros, maduros y sobremaduros. Se realizaron análisis fisicoquímicos (colorsuperficial e interno, pH, acidez total titulable -ATT- y sólidos solubles totales -SST- ybioquímicos por espectrofotometría UV-Vis (fenoles totales -FT-, antocianinas totales -AT-,carotenos totales -CT-, ácido L-ascórbico -AA- y capacidad antioxidante -CA- en el endocarpioy mesocarpio de los frutos. En el color se obtuvieron mayores valores de L* (epicarpio y h*(epicarpio y mesocarpio en frutos inmaduros, maduros y sobremaduros de Chiquicha; mientrasque el valor de C* del mesocarpio y endocarpio fue mayor en aquellos de Pelileo. Lasdiferencias de color fueron evidentes con la maduración, pero no se encontró una relacióndirecta entre la altura de cultivo y los cambios en los parámetros de color analizados. Se obtuvouna diferencia significativamente mayor en el pH de tomates (inmaduros y maduros deChiquicha. Asimismo, estos frutos presentaron mayor ATT (maduros y sobremaduros y SST(sobremaduros que los de Pelileo. El mesocarpio presentó mayor contenido de FT y CT en losfrutos (tres estados de maduración de Chiquicha, mientras que mayor concentración de estoscompuestos presentó el endocarpio de frutos sobremaduros de Pelileo. El contenido de AT yAA fue mayor en frutos maduros de Chiquicha. En general, los frutos de Chiquicha mostraronmayor concentración de compuestos antioxidantes; posiblemente porque dicha plantación,constituida por al menos dos genotipos, se sitúa en una zona montañosa con buenascaracterísticas para el cultivo de este frutal.(Received: 2013/03/07 - Accepted: 2013/06/17The aim of this research was to study the antioxidant compounds content in Red-purple treetomato grown at different altitudes (m.a.s.l.. Fruits from Pelileo (2660 m.a.s.l. and Chiquicha(2440 m.a.s.l. in three maturity stages (unripe, ripe and overripe were used. Physicochemical(surface and internal color, pH, titratable total acidity -TTA- and total soluble solids -SST- andbiochemical analyses using UV-Vis spectrophotometry (total phenol -TF-, total anthocyanins -TA-, total carotenes -CT-, L-ascorbic acid -AA- and antioxidant capacity -AC- in the mesocarpand endocarp of fruits were performed. In the color, higher values of L* (epicarp and h*(epicarp and mesocarp in unripe, ripe and overripe fruits from Chiquicha were obtained, whileC* values of mesocarp and endocarp was higher in those from Pelileo. The color differenceswere evident during fruit ripening, but a direct relationship between crop altitude and changes in color parameters analyzed wasn’t found. There was a significantly higher difference in the pH oftomatoes (unripe and ripe from Chiquicha. Also, these fruits had higher TTA (ripe and overripeand TSS (overripe than those from Pelileo. The mesocarp showed higher content of TF and TCin the fruits (three maturity stages from Chiquicha, while higher concentration of thesecompounds showed the endocarp from Pelileo’s fruit. The TA and AA content was higher in ripefruits from Chiquicha. In general, the fruits from Chiquicha showed higher concentration ofantioxidant compounds, possibly because the plantation, consisting of at least two genotypes, islocated in a mountainous area with good characteristics for this fruit cultivation.

  4. Demographic population structure and fungal associations of plants colonizing High Arctic glacier forelands, Petuniabukta, Svalbard

    Directory of Open Access Journals (Sweden)

    Jakub Těšitel

    2014-04-01

    Full Text Available The development of vegetation in Arctic glacier forelands has been described as unidirectional, non-replacement succession characterized by the gradual establishment of species typical for mature tundra with no species turnover. Our study focused on two early colonizers of High Arctic glacier forelands: Saxifraga oppositifolia (Saxifragaceae and Braya purpurascens (Brassicaceae. While the first species is a common generalist also found in mature old growth tundra communities, the second specializes on disturbed substrate. The demographic population structures of the two study species were investigated along four glacier forelands in Petuniabukta, north Billefjorden, in central Spitsbergen, Svalbard. Young plants of both species occurred exclusively on young substrate, implying that soil conditions are favourable for establishment only before soil crusts develop. We show that while S. oppositifolia persists from pioneer successional stages and is characterized by increased size and flowering, B. purpurascens specializes on disturbed young substrate and does not follow the typical unidirectional, non-replacement succession pattern. Plants at two of the forelands were examined for the presence of root-associated fungi. Fungal genus Olpidium (Fungus incertae sedis was found along a whole successional gradient in one of the forelands.

  5. Differential expression of T- and L-type voltage-dependent calcium channels in renal resistance vessels

    DEFF Research Database (Denmark)

    Hansen, Pernille B. Lærkegaard; Jensen, Boye L.; Andreasen, D;

    2001-01-01

    The distribution of voltage-dependent calcium channels in kidney pre- and postglomerular resistance vessels was determined at the molecular and functional levels. Reverse transcription-polymerase chain reaction analysis of microdissected rat preglomerular vessels and cultured smooth muscle cells...... showed coexpression of mRNAs for T-type subunits (Ca(V)3.1, Ca(V)3.2) and for an L-type subunit (Ca(V)1.2). The same expression pattern was observed in juxtamedullary efferent arterioles and outer medullary vasa recta. No calcium channel messages were detected in cortical efferent arterioles. Ca(V)1.......2 protein was demonstrated by immunochemical labeling of rat preglomerular vasculature and juxtamedullary efferent arterioles and vasa recta. Cortical efferent arterioles were not immunopositive. Recordings of intracellular calcium concentration with digital fluorescence imaging microscopy showed a...

  6. Influence of maternally-derived antibodies in 6-week old dogs for the efficacy of a new vaccine to protect dogs against virulent challenge with canine distemper virus, adenovirus or parvovirus

    Directory of Open Access Journals (Sweden)

    Stephen Wilson

    2014-01-01

    In conclusion, two doses of the DHPPi/L4R vaccine administered to dogs from six weeks of age in the presence of maternal antibodies aided in the protection against virulent challenge with CDV, CAV-1 or CPV.

  7. 不同发育年龄大鼠脑内小窝蛋白的表达与神经可塑性%Expression of Caveolin-1 protein in different aged rat brain and its synaptic plasticity

    Institute of Scientific and Technical Information of China (English)

    吕国枫; 王红霞; 邹伟

    2009-01-01

    目的:证实小窝蛋白(Cav-1)在中枢神经系统内的表达及其与脑发育和神经可塑性的关系.方法:以突触素(Syn)的表达为参照,利用免疫印迹法对Cav-1蛋白在大鼠大脑不同部位表达的年龄特征以及青年大鼠脑内不同脑区的分布特征进行了研究.结果:Cav-1的表达与不同年龄和不同脑区有关.结论:Cav-1蛋白的动态变化可能参与脑发育和神经可塑性活动.

  8. 5,6-EET potently inhibits T-type calcium channels

    DEFF Research Database (Denmark)

    Cazade, M.; Bidaud, I.; Hansen, Pernille B. Lærkegaard;

    2014-01-01

    T-type calcium channels (T-channels) are important actors in neuronal pacemaking, in heart rhythm, and in the control of the vascular tone. T-channels are regulated by several endogenous lipids including the primary eicosanoid arachidonic acid (AA), which display an important role in vasodilation...... the three human recombinant T-channels (Ca(v)3.1, Ca(v)3.2, and Ca(v)3.3), as compared to those of AA. We identified the P450 epoxygenase product, 5,6-EET, as a potent physiological inhibitor of Ca(v)3 currents. The effects of 5,6-EET were observed at sub-micromolar concentrations (IC50 = 0.54 mu M...

  9. Role of T-type calcium channels in myogenic tone of skeletal muscle resistance arteries

    DEFF Research Database (Denmark)

    VanBavel, Ed; Sorop, Oana; Andreasen, Ditte;

    2002-01-01

    T-type calcium channels may be involved in the maintenance of myogenic tone. We tested their role in isolated rat cremaster arterioles obtained after CO(2) anesthesia and decapitation. Total RNA was analyzed by RT-PCR and Southern blotting for calcium channel expression. We observed expression of...... voltage-operated calcium (Ca(V)) channels Ca(V)3.1 (T-type), Ca(V)3.2 (T-type), and Ca(V)1.2 (L-type) in cremaster arterioles (n = 3 rats). Amplification products were observed only in the presence of reverse transcriptase and cDNA. Concentration-response curves of the relatively specific L-type blocker......); K(+) -5.4 +/- 0.3 (n = 4); all log(IC(50)) P <0.05, basal vs. K(+)]. These data suggest that T-type calcium channels are expressed and involved in maintenance of myogenic tone in rat cremaster muscle arterioles....

  10. Genetics Home Reference: isolated hyperCKemia

    Science.gov (United States)

    ... the elevated blood levels of creatine kinase that characterize this condition. In addition to isolated hyperCKemia , CAV3 ... cause the disorder. In most cases, an affected person has one parent with isolated hyperCKemia or another ...

  11. Chronic deficit in nitric oxide elicits oxidative stress and augments T-type calcium-channel contribution to vascular tone of rodent arteries and arterioles

    DEFF Research Database (Denmark)

    Howitt, Lauren; Kuo, Ivana Y; Ellis, Anthie;

    2013-01-01

    /L) significantly increased the T-type, but not the L-type, channel contribution to vascular tone in vitro and in vivo, and altered the smooth muscle expression of the Cav3.1 and Cav3.2 T-type channels. In pressurized mesenteric arteries of Cav3.1ko and Cav3.2ko mice, acutely treated with l-NAME, the contribution...... of T-type channels relative to L-type channels was significantly reduced, compared with arteries from wild-type mice.Chronic l-NAME treatment (40 mg/kg/day; 14-18 days) increased blood pressure, vascular superoxide, and the contribution of T-type channels to vascular tone in vivo. The latter was......, by regulating the bioavailability of reactive oxygen species produced by NADPH oxidase. Our data provide evidence for a novel causal link between nitric oxide deficit, oxidative stress, and T-type calcium channel function....

  12. Quantitative variation of flavonoids and related compounds in Cosmos bipinnatus

    OpenAIRE

    Koho Saito

    2015-01-01

    Quantitative variation of nine flavonoid compounds and two related phenolic acids in several parts of three garden varieties of Cosmos bipinnatus Cav. were examined by; means of paper chromatography followed by a spectrophotometric procedure.

  13. Modeling the Energy Use of a Connected and Automated Transportation System (Poster)

    Energy Technology Data Exchange (ETDEWEB)

    Gonder, J.; Brown, A.

    2014-07-01

    Early research points to large potential impacts of connected and automated vehicles (CAVs) on transportation energy use - dramatic savings, increased use, or anything in between. Due to a lack of suitable data and integrated modeling tools to explore these complex future systems, analyses to date have relied on simple combinations of isolated effects. This poster proposes a framework for modeling the potential energy implications from increasing penetration of CAV technologies and for assessing technology and policy options to steer them toward favorable energy outcomes. Current CAV modeling challenges include estimating behavior change, understanding potential vehicle-to-vehicle interactions, and assessing traffic flow and vehicle use under different automation scenarios. To bridge these gaps and develop a picture of potential future automated systems, NREL is integrating existing modeling capabilities with additional tools and data inputs to create a more fully integrated CAV assessment toolkit.

  14. Interplay between hepatic mitochondria-associated membranes, lipid metabolism and caveolin-1 in mice

    Science.gov (United States)

    Sala-Vila, Aleix; Navarro-Lérida, Inmaculada; Sánchez-Alvarez, Miguel; Bosch, Marta; Calvo, Carlos; López, Juan Antonio; Calvo, Enrique; Ferguson, Charles; Giacomello, Marta; Serafini, Annalisa; Scorrano, Luca; Enriquez, José Antonio; Balsinde, Jesús; Parton, Robert G.; Vázquez, Jesús; Pol, Albert; Del Pozo, Miguel A.

    2016-01-01

    The mitochondria-associated membrane (MAM) is a specialized subdomain of the endoplasmic reticulum (ER) which acts as an intracellular signaling hub. MAM dysfunction has been related to liver disease. We report a high-throughput mass spectrometry-based proteomics characterization of MAMs from mouse liver, which portrays them as an extremely complex compartment involved in different metabolic processes, including steroid metabolism. Interestingly, we identified caveolin-1 (CAV1) as an integral component of hepatic MAMs, which determine the relative cholesterol content of these ER subdomains. Finally, a detailed comparative proteomics analysis between MAMs from wild type and CAV1-deficient mice suggests that functional CAV1 contributes to the recruitment and regulation of intracellular steroid and lipoprotein metabolism-related processes accrued at MAMs. The potential impact of these novel aspects of CAV1 biology on global cell homeostasis and disease is discussed. PMID:27272971

  15. Lectotypification of three Iberian endemic species belonging to monotypic genera described by Cosson

    Directory of Open Access Journals (Sweden)

    Buira, Antoni

    2015-12-01

    Full Text Available Three lectotypes are here designated for Euzomodendron bourgaeanum Coss., Guiraoa arvensis Coss. and Laserpitium scabrum Cav. (Guillonea scabra (Cav. Coss., whose genera are monospecific and endemic to the Iberian Peninsula. The selected types of the two former species are kept at P and belong to Cosson’s personal herbarium, whilst the last one is kept at MA and belongs to the historical herbarium of Cavanilles.Se designan los lectótipos de Euzomodendron bourgaeanum Coss., Guiraoa arvensis Coss. y Laserpitium scabrum Cav. (Guillonea scabra (Cav. Coss., cuyos géneros son monoespecíficos y endémicos de la Península Ibérica. Los tipos seleccionados para las dos primeras especies se encuentran en P y pertenecen al herbario personal de Cosson, mientras que el de la última se encuentra en MA y pertenece al herbario histórico de Cavanilles.

  16. Revision of the genus Dombeya (Sterculiaceae in southern Africa

    Directory of Open Access Journals (Sweden)

    I. C. Verdoon

    1986-12-01

    Full Text Available The species of Dombeya Cav. occurring in southern Africa are revised and a key is provided to the eight species present in the region. A new record for southern Africa, D. quinqueseta (Del. Exell, is included.

  17. Small Interfering RNA-mediated Caveolin-1 Knockout on Plasminogen Activator Inhibitor-1 Expression in Insulin-stimulated Human Vascular Endothelial Cells

    Institute of Scientific and Technical Information of China (English)

    Huiling YANG; Gebo WEN; Weixin HU; Shuya HE; Zhihua QUAN; Weixia PENG; Bin YAN; Jianghua LIU; Fang WEN; Renxian CAO; Yangyan XU

    2007-01-01

    Using human vascular endothelial cells (ECV304) as the target,we studied the effect of caveolin(CAV)-1 in the course of insulin-stimulated expression of plasminogen activator inhibitor(PAI)-1.The appropriate single-stranded oligonucleotides representing the RNAi CAV-1 gene were analyzed by Ambion software.After annealing to generate double-stranded oligonucleotides (ds oligo),it was cloned into the pENTR/U6 entry vector containing RNA polymerase Ⅲ expression element by T4 DNA ligase.The short hairpin (shRNA) sequences transferred from the pENTR/U6 entry were cloned into the pLenti6/BLOCK-iTDEST vector with an LR recombination reaction.After identification by sequencing,we successfully constructed the CAV-1 RNAi lentiviral expression system using Gateway technology.Silencing efficiency was assayed by real-time reverse transcription-polymerase chain reaction,immunofluorescence staining and Western blotting.ECV304 cells were cultured in the medium containing different concentrations of insulin(1×10-9 to 1×10-7M)with the CAV-1 gene silenced or not.The expression level and subcellular localization of PAI-1 and CAV-1 were compared using reverse transcription-polymerase chain reaction,immunofluorescence staining and Western blot assay.The results showed that the potent inhibition of CAV-1 expression could reach 85%,and it was specific to the CAV-1-derived shRNA,not the S100A13-derived shRNA.There was no dramatic difference in PAI-1 expression between the RNAi+ and RNAi-ECV304 cells incubated with physiological insulin,but PAI-1 protein did accumulate under the cell membrane.As the concentration of insulin increased,the expression of PAI-1 was up-regulated,whereas the expression of CAV-1 attenuated.Furthermore,PAl-1 clearly augmented after CAV-1 knockdown.These results indicated that hyperinsulinism could promote PAI-1 expression by inhibiting CAV-1,and stabilizing or up-regulating CAV-1 expression in endothelial cells might reduce complications of the great vessels

  18. H{sub 2}S does not regulate proliferation via T-type Ca{sup 2+} channels

    Energy Technology Data Exchange (ETDEWEB)

    Elies, Jacobo; Johnson, Emily; Boyle, John P.; Scragg, Jason L.; Peers, Chris, E-mail: c.s.peers@leeds.ac.uk

    2015-06-12

    T-type Ca{sup 2+} channels (Cav3.1, 3.2 and 3.3) strongly influence proliferation of various cell types, including vascular smooth muscle cells (VSMCs) and certain cancers. We have recently shown that the gasotransmitter carbon monoxide (CO) inhibits T-type Ca{sup 2+} channels and, in so doing, attenuates proliferation of VSMC. We have also shown that the T-type Ca{sup 2+} channel Cav3.2 is selectively inhibited by hydrogen sulfide (H{sub 2}S) whilst the other channel isoforms (Cav3.1 and Cav3.3) are unaffected. Here, we explored whether inhibition of Cav3.2 by H{sub 2}S could account for the anti-proliferative effects of this gasotransmitter. H{sub 2}S suppressed proliferation in HEK293 cells expressing Cav3.2, as predicted by our previous observations. However, H{sub 2}S was similarly effective in suppressing proliferation in wild type (non-transfected) HEK293 cells and those expressing the H{sub 2}S insensitive channel, Cav3.1. Further studies demonstrated that T-type Ca{sup 2+} channels in the smooth muscle cell line A7r5 and in human coronary VSMCs strongly influenced proliferation. In both cell types, H{sub 2}S caused a concentration-dependent inhibition of proliferation, yet by far the dominant T-type Ca{sup 2+} channel isoform was the H{sub 2}S-insensitive channel, Cav3.1. Our data indicate that inhibition of T-type Ca{sup 2+} channel-mediated proliferation by H{sub 2}S is independent of the channels’ sensitivity to H{sub 2}S. - Highlights: • T-type Ca{sup 2+} channels regulate proliferation and are sensitive to the gasotransmitters CO and H{sub 2}S. • H{sub 2}S reduced proliferation in HEK293 cells expressing the H{sub 2}S sensitive Cav3.2 channel. • H{sub 2}S also inhibited proliferation in non-transfected cells and HEK293 cells expressing Cav3.1. • Native smooth muscle cells primarily express Cav3.1. Their proliferation was also inhibited by H{sub 2}S. • Unlike CO, H{sub 2}S does not regulate smooth muscle proliferation via T-type Ca{sup 2

  19. Detection and characterization of chicken anemia virus from commercial broiler breeder chickens

    Directory of Open Access Journals (Sweden)

    Omar Abdul

    2008-10-01

    Full Text Available Abstract Background Chicken anemia virus (CAV is the causative agent of chicken infectious anemia (CIA. Study on the type of CAV isolates present and their genetic diversity, transmission to their progeny and level of protection afforded in the breeder farms is lacking in Malaysia. Hence, the present study was aimed to detect CAV from commercial broiler breeder farms and characterize CAV positive samples based on sequence and phylogenetic analysis of partial VP1 gene. Results A total of 12 CAV isolates from different commercial broiler breeder farms were isolated and characterized. Detection of CAV positive embryos by the PCR assay in the range of 40 to 100% for different farms indicated high level of occurrence of vertical transmission of viral DNA to the progeny. CAV antigen was detected in the thymus and in the bone marrow but not in spleen, liver, duodenum, ovary and oviduct by indirect immunoperoxidase staining. The 12 CAV isolates were characterized based on partial sequences of VP1 gene. Six isolates (MF1A, MF3C, M3B5, NF4A, P12B and P24A were found to have maximum homology with previously characterized Malaysian isolate SMSC-1, four isolates (M1B1, NF3A, PYT4 and PPW4 with isolate BL-5 and the remaining two (NF1D and NF2C have maximum homology both with isolates 3-1 and BL-5. Meanwhile, seven of the isolates with amino acid profile of 75-I, 97-L, 139-Q and 144-Q were clustered together in cluster I together with other isolates from different geographical places. The remaining five isolates with amino acid profile of 75-V, 97-M, 139-K and 144-E were grouped under cluster II. All the CAV isolates demonstrated omega values (Ka/Ks of less than one (the values ranging from 0.07 to 0.5 suggesting the occurrence of purifying (negative selection in all the studied isolates. Conclusion The present study showed that CAV is widespread in the studied commercial broiler breeder farms. The result also indicated the occurrence of genetic variability in

  20. Phorbol Ester Modulation of Ca2+ Channels Mediates Nociceptive Transmission in Dorsal Horn Neurones

    Directory of Open Access Journals (Sweden)

    Gary J. Stephens

    2013-05-01

    Full Text Available Phorbol esters are analogues of diacylglycerol which activate C1 domain proteins, such as protein kinase C (PKC. Phorbol ester/PKC pathways have been proposed as potential therapeutic targets for chronic pain states, potentially by phosphorylating proteins involved in nociception, such as voltage-dependent Ca2+ channels (VDCCs. In this brief report, we investigate the potential involvement of CaV2 VDCC subtypes in functional effects of the phorbol ester, phorbol 12-myristate 13-acetate (PMA on nociceptive transmission in the spinal cord. Effects of PMA and of selective pharmacological blockers of CaV2 VDCC subtypes on nociceptive transmission at laminae II dorsal horn neurones were examined in mouse spinal cord slices. Experiments were extended to CaV2.3(−/− mice to complement pharmacological studies. PMA increased the mean frequency of spontaneous postsynaptic currents (sPSCs in dorsal horn neurones, without an effect on event amplitude or half-width. sPSC frequency was reduced by selective VDCC blockers, w-agatoxin-IVA (AgTX; CaV2.1, w-conotoxin-GVIA (CTX; CaV2.2 or SNX-482 (CaV2.3. PMA effects were attenuated in the presence of each VDCC blocker and, also, in CaV2.3(−/− mice. These initial data demonstrate that PMA increases nociceptive transmission at dorsal horn neurones via actions on different CaV2 subtypes suggesting potential anti-nociceptive targets in this system.

  1. Production of canine adenovirus type 2 in serum-free suspension cultures of MDCK cells.

    Science.gov (United States)

    Castro, R; Fernandes, P; Laske, T; Sousa, M F Q; Genzel, Y; Scharfenberg, K; Alves, P M; Coroadinha, A S

    2015-09-01

    The potential of adherent Madin Darby Canine Kidney (MDCK) cells for the production of influenza viruses and canine adenovirus type 2 (CAV-2) for vaccines or gene therapy approaches has been shown. Recently, a new MDCK cell line (MDCK.SUS2) that was able to grow in suspension in a fully defined system was established. In this work, we investigated whether the new MDCK.SUS2 suspension cell line is suitable for the amplification of CAV-2 under serum-free culture conditions. Cell growth performance and CAV-2 production were evaluated in three serum-free media: AEM, SMIF8, and EXCELL MDCK. CAV-2 production in shake flasks was maximal when AEM medium was used, resulting in an amplification ratio of infectious particles (IP) of 142 IP out/IP in and volumetric and cell-specific productivities of 2.1 × 10(8) IP/mL and 482 IP/cell, respectively. CAV-2 production was further improved when cells were cultivated in a 0.5-L stirred tank bioreactor. To monitor infection and virus production, cells were analyzed by flow cytometry. A correlation between the side scatter measurement and CAV-2 productivity was found, which represents a key feature to determine the best harvesting time during process development of gene therapy vectors that do not express reporter genes. This work demonstrates that MDCK.SUS2 is a suitable cell substrate for CAV-2 production, constituting a step forward in developing a production process transferable to industrial scales. This could allow for the production of high CAV-2 titers either for vaccination or for gene therapy purposes. PMID:25994255

  2. Production and purification of non replicative canine adenovirus type 2 derived vectors

    OpenAIRE

    Szelechowski, Marion; Bergeron, Corinne; Gonzalez Dunia, Daniel; Klonjkowski, Bernard

    2013-01-01

    Adenovirus (Ad) derived vectors have been widely used for short or long-term gene transfer, both for gene therapy and vaccine applications. Because of the frequent pre-existing immunity against the classically used human adenovirus type 5, canine adenovirus type 2 (CAV2) has been proposed as an alternative vector for human gene transfer. The well-characterized biology of CAV2, together with its ease of genetic manipulation, offer major advantages, notably for gene transfer into the central ne...

  3. Temporal evolution in caveolin 1 methylation levels during human esophageal carcinogenesis

    International Nuclear Information System (INIS)

    Esophageal cancer ranks eighth among frequent cancers worldwide. Our aim was to investigate whether and at which neoplastic stage promoter hypermethylation of CAV1 is involved in human esophageal carcinogenesis. Using real-time quantitative methylation-specific PCR (qMSP), we examined CAV1 promoter hypermethylation in 260 human esophageal tissue specimens. Real-time RT-PCR and qMSP were also performed on OE33 esophageal cancer cells before and after treatment with the demethylating agent, 5-aza-2’-deoxycytidine (5-Aza-dC). CAV1 hypermethylation showed highly discriminative ROC curve profiles, clearly distinguishing esophageal adenocarcinomas (EAC) and esophageal squamous cell carcinomas (ESCC) from normal esophagus (NE) (EAC vs. NE, AUROC = 0.839 and p < 0.0001; ESCC vs. NE, AUROC = 0.920 and p < 0.0001). Both CAV1 methylation frequency and normalized methylation value (NMV) were significantly higher in Barrett’s metaplasia (BE), low-grade and high-grade dysplasia occurring in BE (D), EAC, and ESCC than in NE (all p < 0.01, respectively). Meanwhile, among 41 cases with matched NE and EAC or ESCC, CAV1 NMVs in EAC and ESCC (mean = 0.273) were significantly higher than in corresponding NE (mean = 0.146; p < 0.01, Student’s paired t-test). Treatment of OE33 EAC cells with 5-Aza-dC reduced CAV1 methylation and increased CAV1 mRNA expression. CAV1 promoter hypermethylation is a frequent event in human esophageal carcinomas and is associated with early neoplastic progression in Barrett’s esophagus

  4. Decreased caveolin-1 levels contribute to fibrosis and deposition of extracellular IGFBP-5

    OpenAIRE

    Yamaguchi, Yukie; Yasuoka, Hidekata; Stolz, Donna B.; Feghali-Bostwick, Carol A.

    2010-01-01

    Abstract Our previous studies have demonstrated increased expression of insulin-like growth factor binding protein-5 (IGFBP-5) in fibrotic tissues and IGFBP-5 induction of extracellular matrix (ECM) components. The mechanism resulting in increased IGFBP-5 in the extracellular milieu of fibrotic fibroblasts is unknown. Since Caveolin-1 (Cav-1) has been implicated to play a role in membrane trafficking and signal transduction in tissue fibrosis, we examined the effect of Cav-1 on IGFBP-5 intern...

  5. Regulación de la expresión y activación de la caveolina-1 durante la adipogénesis, y su relación con la señalización insulínica. Efecto de TNF-a y glucosa

    OpenAIRE

    Palacios-Ortega, S. (Sara); Miguel, C.

    2015-01-01

    Knowledge of the regulatory mechanisms involved in adipocyte differentiation and hypertrophy is essential to understand the nexus between obesity and insulin resistance and will help to design new therapeutic approaches for obesity and diabetes. Previous studies published by our group pointed to caveolin-1 (Cav-1) as one of the genes upregulated in rat visceral white adipose tissue in response to a high fat diet. Cav-1 is an essential constituent of adipocyte caveolae which binds the beta sub...

  6. The response to climate factors for the different specificity in M2 physiology of 60Co γ-ray irradiated flowers

    International Nuclear Information System (INIS)

    Physiology and climate factor of different M2 flower (Begonia tuberhybrida Voss and Dahlia pinnata Cav root) after 60Co γ-ray irradiated were determined. The result showed that transpiration rate, stoma conductance, intercellular CO2 concentration of M2 of Dahlia pinnata Cav are affected obviously with climate factors, however, effect on the photosynthetic speed was not found, and M2 of Begonia tuberhybrida Voss was not significant for physiology to climate

  7. Dual-Axis Rotational Angiography is Safe and Feasible to Detect Coronary Allograft Vasculopathy in Pediatric Heart Transplant Patients: A Single-Center Experience.

    Science.gov (United States)

    Rios, Rodrigo; Loomba, Rohit S; Foerster, Susan R; Pelech, Andrew N; Gudausky, Todd M

    2016-04-01

    Coronary allograft vasculopathy (CAV) is the leading cause of graft failure in pediatric heart transplant recipients, also adding to mortality in this patient population. Coronary angiography is routinely performed to screen for CAV, with conventional single-plane or bi-plane angiography being utilized. Dual-axis rotational coronary angiography (RA) has been described, mostly in the adult population, and may offer reduction in radiation dose and contrast volume. Experience with this in the pediatric population is limited. This study describes a single-institution experience with RA for screening for CAV in pediatric patients. The catheterization database at our institution was used to identify pediatric heart transplant recipients having undergone RA to screen for CAV. Procedural data including radiation dose, fluoroscopy time, contrast volume, and procedure time were collected for each catheterization. The number of instances in which RA was not successful, ECG changes were present, and CAV was detected were also collected for each catheterization. A total of 97 patients underwent 345 catheterizations utilizing RA. Median radiation dose-area product per kilogram was found to be 341.7 (mGy cm(2)/kg), total air kerma was 126.8 (mGy), procedure time was 69 min, fluoroscopy time was 9.9 min, and contrast volume was 13 ml. A total of 17 (2 %) coronary artery injections out of 690 could not be successfully imaged using RA. A total of 14 patients had CAV noted at any point, 10 of whom had progressive CAV. Electrocardiographic changes were documented in a total of 10 (3 %) RA catheterizations. Procedural characteristics did not differ between serial catheterizations. RA is safe and feasible for CAV screening in pediatric heart transplant recipients while offering coronary imaging in multiple planes compared to conventional angiography. PMID:26846123

  8. Molecular basis of toxicity of N-type calcium channel inhibitor MVIIA.

    Science.gov (United States)

    Wang, Fei; Yan, Zhenzhen; Liu, Zhuguo; Wang, Sheng; Wu, Qiaoling; Yu, Shuo; Ding, Jiuping; Dai, Qiuyun

    2016-02-01

    MVIIA (ziconotide) is a specific inhibitor of N-type calcium channel, Cav2.2. It is derived from Cone snail and currently used for the treatment of severe chronic pains in patients unresponsive to opioid therapy. However, MVIIA produces severe side-effects, including dizziness, nystagmus, somnolence, abnormal gait, and ataxia, that limit its wider application. We previously identified a novel inhibitor of Cav2.2, ω-conopeptide SO-3, which possesses similar structure and analgesic activity to MVIIA's. To investigate the key residues for MVIIA toxicity, MVIIA/SO-3 hybrids and MVIIA variants carrying mutations in its loop 2 were synthesized. The substitution of MVIIA's loop 1 with the loop 1 of SO-3 resulted in significantly reduced Cav2.2 binding activity in vitro; the replacement of MVIIA loop 2 by the loop 2 of SO-3 not only enhanced the peptide/Cav2.2 binding but also decreased its toxicity on goldfish, attenuated mouse tremor symptom, spontaneous locomotor activity, and coordinated locomotion function. Further mutation analysis and molecular calculation revealed that the toxicity of MVIIA mainly arose from Met(12) in the loop 2, and this residue inserts into a hydrophobic hole (Ile(300), Phe(302) and Leu(305)) located between repeats II and III of Cav2.2. The combinative mutations of the loop 2 of MVIIA or other ω-conopeptides may be used for future development of more effective Cav2.2 inhibitors with lower side effects. PMID:26344359

  9. Caveolin-1 overexpression in benign and malignant salivary gland tumors.

    Science.gov (United States)

    Jaafari-Ashkavandi, Zohreh; Ashraf, Mohammad Javad; Nazhvani, Ali Dehghani; Azizi, Zahra

    2016-02-01

    Caveolin-1, a tyrosine-phosphorylated protein, is supposed to have different regulatory roles as promoter or suppressor in many human cancers. However, no published study concerned its expression in benign and malignant salivary gland tumors. The aim of this study was to evaluate and compare the expression of Cav-1 in the most common benign and malignant salivary gland tumors and evaluate its correlation with proliferation activity. In this cross-sectional retrospective study, immunohistochemical expression of caveolin-1 and Ki67 were evaluated in 49 samples, including 11 normal salivary glands, 15 cases of pleomorphic adenoma (PA), 13 adenoid cystic carcinomas (AdCC), and 10 mucoepidermoid carcinomas (MEC). The expression of Cav-1 was seen in 18 % of normal salivary glands and 85 % of tumors. The immunoreaction in the tumors was significantly higher than normal tissues (P = 0.001), but the difference between benign and malignant tumors was not significant (P = 0.07). Expression of Cav-1 was correlated with Ki67 labeling index in PAs, but not in malignant tumors. Cav-1 expression was not in association with tumor size and stage. Overexpression of Cav-1 was found in salivary gland tumors in comparison with normal tissues, but no significant difference was observed between benign and malignant tumors. Cav-1 was inversely correlated with proliferation in PA. Therefore, this marker may participate in tumorigenesis of salivary gland tumors and may be a potential biomarker for cancer treatments. PMID:26323261

  10. Clinical and serological examination of a parental flock latently infected with chicken anemia virus

    Directory of Open Access Journals (Sweden)

    Kapetanov Miloš

    2003-01-01

    Full Text Available At the beginning of etiopathogenic research on chicken anaemia virus (CAV it was thought that groups of chickens at risk for CAV infection were those coming from a parental flock infected before laying. Therefore it is important to know the level and persistence of transfered maternal antibodies (MaAt and to measure specific antibody development during rearing. The goal of this research was to assess the necessity for prophylactic measures by determining the humoral immune response to CAV and any clinical changes in breeder chickens. Chickens from the parental Hybro flock were examined from the first day until the end of production. Maternal antibodies for CAV, which were present initially, were not detected at 4 weeks old. At 6 weeks old specific antibodies for CAV were found in 45% of the serum samples. These antibodies increased until the 18th week when the experiment was terminated. The state of health of the parental flock in the period when MaAt antibodies could not be detected and until specific antibodies appeared did not differ significantly. The results of these investigations are the first evidence of CAV infection in Yugoslavia, based on serological examination.

  11. [Construction and experimental immunity of recombinant replication-competent canine adenovirus type 2 expressing hemagglutinin gene of H5N1 subtype tiger influenza virus].

    Science.gov (United States)

    Gao, Yu-Wei; Xia, Xian-Zhu; Wang, Li-Gang; Liu, Dan; Huang, Geng

    2006-04-01

    H5N1 highly pathogenic avian influenza virus was highly pathogenic and sometimes even fatal for tigers and cats. To develop a new type of vaccine for Felidae influenza prevention, recombinant replication-competent canine adenovirus Type 2 expressing hemagglutinin gene of H5N1 subtype tiger influenza virus was constructed. A/tiger/Harbin/01/2003 (HSN1) HA gene was cloned into PVAX1. The HA expression cassette which included CMV and HA and PolyA was ligated into the E3 deletion region of pVAXdeltaE. The recombinant plasmid was named pdeltaEHA. The pdelta EHA and the pPoly2-CAV2 were digested with Nru I /Sal I, respectively. The purified Nru I/Sal I DNA fragment containing the HA expression cassette was cloned into pPoly2-CAV2 to generate the recombinant plasmid pCAV-2/HA. The recombinant genome was released from pCAV-2/HA, and was transfected into MDCK cells by Lipofectamine. The recombinant virus named CAV2/HA was gained. Anti-H5N1 influenza virus HI antibody (1:8 - 1:16) was detected in the cat immunized with CAV-2/HA. PMID:16736595

  12. L-Canavanine potentiates the cytotoxicity of doxorubicin and cisplatin in arginine deprived human cancer cells

    Science.gov (United States)

    Wink, Michael

    2016-01-01

    The non-protein amino acid L-canavanine (L-CAV), an antimetabolite of L-arginine (L-ARG), can alter the 3D conformation of proteins when incorporated into a protein instead of L-ARG. L-CAV inhibits the proliferation of some tumour cells. The deprivation of L-ARG in the culture medium enhances the response of cells to L-CAV. This study aimed to investigate the interaction of L-CAV in combination with the chemotherapeutic drugs, doxorubicin (DOX) or cisplatin (CIS), in cancer cells, especially in the absence of L-ARG. A combination method based on the median-effect principle and mass-action law was used. The following cancer cells were employed: HeLa and Caco-2 cells, overexpressing argininosuccinate synthase (ASS), pancreatic cells (MIA PaCa-2 and BxPC-3) and hepatocellular carcinoma cells (Hep G2 and SK-HEP-1), with down-regulated ASS. When constant and non-constant ratios of L-CAV were combined with DOX and CIS, a synergistic potentiation of cytotoxicity was recorded. Cells expressing high levels of ASS were more sensitive to the treatment as compared to the cells with reduced ASS levels. Overall, this study may provide a new approach to targeting some cancer cells with L-CAV in combination with DNA-targeting drugs such as DOX and CIS, especially those cells which overexpress ASS, such as human cervical and colorectal carcinoma cells. PMID:26839743

  13. Estimation of Cumulative Absolute Velocity using Empirical Green's Function Method

    International Nuclear Information System (INIS)

    In recognition of the needs to develop a new criterion for determining when the OBE (Operating Basis Earthquake) has been exceeded at nuclear power plants, Cumulative Absolute Velocity (CAV) was introduced by EPRI. The concept of CAV is the area accumulation with the values more than 0.025g occurred during every one second. The equation of the CAV is as follows. CAV = ∫0max |a(t)|dt (1) tmax = duration of record, a(t) = acceleration (>0.025g) Currently, the OBE exceedance criteria in Korea is Peak Ground Acceleration (PGA, PGA>0.1g). When Odesan earthquake (ML=4.8, January 20th, 2007) and Gyeongju earthquake (ML=3.4, June 2nd, 1999) were occurred, we have had already experiences of PGA greater than 0.1g that did not even cause any damage to the poorly-designed structures nearby. This moderate earthquake has motivated Korea to begin the use of the CAV for OBE exceedance criteria for NPPs. Because the present OBE level has proved itself to be a poor indicator for small-to-moderate earthquakes, for which the low OBE level can cause an inappropriate shut down the plant. A more serious possibility is that this scenario will become a reality at a very high level. Empirical Green's Function method was a simulation technique which can estimate the CAV value and it is hereby introduced

  14. The serological and virological investigation of canine adenovirus infection on the dogs.

    Science.gov (United States)

    Bulut, Oya; Yapici, Orhan; Avci, Oguzhan; Simsek, Atilla; Atli, Kamil; Dik, Irmak; Yavru, Sibel; Hasircioglu, Sibel; Kale, Mehmet; Mamak, Nuri

    2013-01-01

    Two types of Canine Adenovirus (CAVs), Canine Adenovirus type 1 (CAV-1), the virus which causes infectious canine hepatitis, and Canine Adenovirus type 2 (CAV-2), which causes canine infectious laryngotracheitis, have been found in dogs. In this study, blood samples taken from 111 dogs, which were admitted to the Internal Medicine Clinic of Selcuk University, Faculty of Veterinary Medicine, with clinical symptoms. Seventy-seven dogs were sampled from Isparta and Burdur dog shelters by random sampling, regardless of the clinical findings. Dogs showed a systemic disease, characterized by fever, diarrhea, vomiting, oculonasal discharge, conjunctivitis, severe moist cough, signs of pulmonary disease and dehydration. Two dogs had corneal opacity and photophobia. In serological studies, 188 serum samples were investigated on the presence of CAV antibodies by ELISA. Total 103 (103/188-54.7%) blood samples were detected to be positive for CAV antibodies by ELISA. However, 85 (85/188-45.2%) blood samples were negative. Blood leukocyte samples from dogs were processed and inoculated onto confluent monolayers of MDCK cells using standard virological techniques. After third passage, cells were examined by direct immunoflourescence test for virus isolation. But positive result was not detected. In conclusion, this study clearly demonstrates the high prevalence of CAV infection in dogs. PMID:24223508

  15. Transplant coronary heart disease: challenges and solutions

    Directory of Open Access Journals (Sweden)

    Jentzer JC

    2014-12-01

    Full Text Available Jacob C Jentzer,1 Gavin W Hickey,1 Sameer J Khandhar2,3 1Heart and Vascular Institute, University of Pittsburgh Medical Center, Pittsburgh, PA, USA; 2Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; 3Heart and Vascular Institute, Penn-Presbyterian Medical Center, Philadelphia, PA, USAAbstract: Cardiac allograft vasculopathy (CAV remains one of the leading causes of death and graft failure after heart transplantation. A variety of causes, including donor heart characteristics, recipient risk factors, and immune-mediated influences, are associated with developing CAV. In this review, we will focus on the pathophysiology of developing CAV and various methods to screen for this condition. The pathogenesis of CAV likely involves repeated injuries to the endothelium from a variety of factors such as cellular-mediated rejection, and alloimmune factors, including antibody-mediated injury, ischemia-reperfusion injury at time of transplant, cytomegalovirus infections, immunosuppression medications, systemic inflammation, and traditional atherosclerosis risk factors. Patients with significant CAV are often asymptomatic, and therefore early detection by routine screening prior to graft dysfunction is crucial. There are a variety of invasive, noninvasive, and blood tests that have been studied as screening methods, and we will discuss the role of each of these in this review article. Although some treatment regimens have been established for CAV, this is an area where further studies and research are necessary.Keywords: cardiac allograft vasculopathy, orthotopic heart transplantation, intra-vascular imaging

  16. The IQ Motif is Crucial for Ca v 1.1 Function

    Directory of Open Access Journals (Sweden)

    Katarina Stroffekova

    2011-01-01

    Full Text Available Ca2+-dependent modulation via calmodulin, with consensus CaM-binding IQ motif playing a key role, has been documented for most high-voltage-activated Ca2+ channels. The skeletal muscle Cav1.1 also exhibits Ca2+-/CaM-dependent modulation. Here, whole-cell Ca2+ current, Ca2+ transient, and maximal, immobilization-resistant charge movement (Qmax recordings were obtained from cultured mouse myotubes, to test a role of IQ motif in function of Cav1.1. The effect of introducing mutation (IQ to AA of IQ motif into Cav1.1 was examined. In dysgenic myotubes expressing YFP-Cav1.1AA, neither Ca2+ currents nor evoked Ca2+ transients were detectable. The loss of Ca2+ current and excitation-contraction coupling did not appear to be a consequence of defective trafficking to the sarcolemma. The Qmax in dysgenic myotubes expressing YFP-Cav1.1AA was similar to that of normal myotubes. These findings suggest that the IQ motif of the Cav1.1 may be an unrecognized site of structural and functional coupling between DHPR and RyR.

  17. Caveolin-1 Regulates the P2Y2 Receptor Signaling in Human 1321N1 Astrocytoma Cells.

    Science.gov (United States)

    Martinez, Namyr A; Ayala, Alondra M; Martinez, Magdiel; Martinez-Rivera, Freddyson J; Miranda, Jorge D; Silva, Walter I

    2016-06-01

    Damage to the CNS can cause a differential spatio-temporal release of multiple factors, such as nucleotides, ATP and UTP. The latter interact with neuronal and glial nucleotide receptors. The P2Y2 nucleotide receptor (P2Y2R) has gained prominence as a modulator of gliotic responses after CNS injury. Still, the molecular mechanisms underlying these responses in glia are not fully understood. Membrane-raft microdomains, such as caveolae, and their constituent caveolins, modulate receptor signaling in astrocytes; yet, their role in P2Y2R signaling has not been adequately explored. Hence, this study evaluated the role of caveolin-1 (Cav-1) in modulating P2Y2R subcellular distribution and signaling in human 1321N1 astrocytoma cells. Recombinant hP2Y2R expressed in 1321N1 cells and Cav-1 were found to co-fractionate in light-density membrane-raft fractions, co-localize via confocal microscopy, and co-immunoprecipitate. Raft localization was dependent on ATP stimulation and Cav-1 expression. This hP2Y2R/Cav-1 distribution and interaction was confirmed with various cell model systems differing in the expression of both P2Y2R and Cav-1, and shRNA knockdown of Cav-1 expression. Furthermore, shRNA knockdown of Cav-1 expression decreased nucleotide-induced increases in the intracellular Ca(2+) concentration in 1321N1 and C6 glioma cells without altering TRAP-6 and carbachol Ca(2+) responses. In addition, Cav-1 shRNA knockdown also decreased AKT phosphorylation and altered the kinetics of ERK1/2 activation in 1321N1 cells. Our findings strongly suggest that P2Y2R interaction with Cav-1 in membrane-raft caveolae of 1321N1 cells modulates receptor coupling to its downstream signaling machinery. Thus, P2Y2R/Cav-1 interactions represent a novel target for controlling P2Y2R function after CNS injury. PMID:27129210

  18. Lipid rafts are required for signal transduction by angiotensin II receptor type 1 in neonatal glomerular mesangial cells

    International Nuclear Information System (INIS)

    Angiotensin II (ANG-II) receptors (AGTRs) contribute to renal physiology and pathophysiology, but the underlying mechanisms that regulate AGTR function in glomerular mesangium are poorly understood. Here, we show that AGTR1 is the functional AGTR subtype expressed in neonatal pig glomerular mesangial cells (GMCs). Cyclodextrin (CDX)-mediated cholesterol depletion attenuated cell surface AGTR1 protein expression and ANG-II-induced intracellular Ca2+ ([Ca2+]i) elevation in the cells. The COOH-terminus of porcine AGTR1 contains a caveolin (CAV)-binding motif. However, neonatal GMCs express CAV-1, but not CAV-2 and CAV-3. Colocalization and in situ proximity ligation assay detected an association between endogenous AGTR1 and CAV-1 in the cells. A synthetic peptide corresponding to the CAV-1 scaffolding domain (CSD) sequence also reduced ANG-II-induced [Ca2+]i elevation in the cells. Real-time imaging of cell growth revealed that ANG-II stimulates neonatal GMC proliferation. ANG-II-induced GMC growth was attenuated by EMD 66684, an AGTR1 antagonist; BAPTA, a [Ca2+]i chelator; KN-93, a Ca2+/calmodulin-dependent protein kinase II inhibitor; CDX; and a CSD peptide, but not PD 123319, a selective AGTR2 antagonist. Collectively, our data demonstrate [Ca2+]i-dependent proliferative effect of ANG-II and highlight a critical role for lipid raft microdomains in AGTR1-mediated signal transduction in neonatal GMCs. - Highlights: • AGTR1 is the functional AGTR subtype expressed in neonatal mesangial cells. • Endogenous AGTR1 associates with CAV-1 in neonatal mesangial cells. • Lipid raft disruption attenuates cell surface AGTR1 protein expression. • Lipid raft disruption reduces ANG-II-induced [Ca2+]i elevation in neonatal mesangial cells. • Lipid raft disruption inhibits ANG-II-induced neonatal mesangial cell growth

  19. Lipid rafts are required for signal transduction by angiotensin II receptor type 1 in neonatal glomerular mesangial cells

    Energy Technology Data Exchange (ETDEWEB)

    Adebiyi, Adebowale, E-mail: aadebiyi@uthsc.edu; Soni, Hitesh; John, Theresa A.; Yang, Fen

    2014-05-15

    Angiotensin II (ANG-II) receptors (AGTRs) contribute to renal physiology and pathophysiology, but the underlying mechanisms that regulate AGTR function in glomerular mesangium are poorly understood. Here, we show that AGTR1 is the functional AGTR subtype expressed in neonatal pig glomerular mesangial cells (GMCs). Cyclodextrin (CDX)-mediated cholesterol depletion attenuated cell surface AGTR1 protein expression and ANG-II-induced intracellular Ca{sup 2+} ([Ca{sup 2+}]{sub i}) elevation in the cells. The COOH-terminus of porcine AGTR1 contains a caveolin (CAV)-binding motif. However, neonatal GMCs express CAV-1, but not CAV-2 and CAV-3. Colocalization and in situ proximity ligation assay detected an association between endogenous AGTR1 and CAV-1 in the cells. A synthetic peptide corresponding to the CAV-1 scaffolding domain (CSD) sequence also reduced ANG-II-induced [Ca{sup 2+}]{sub i} elevation in the cells. Real-time imaging of cell growth revealed that ANG-II stimulates neonatal GMC proliferation. ANG-II-induced GMC growth was attenuated by EMD 66684, an AGTR1 antagonist; BAPTA, a [Ca{sup 2+}]{sub i} chelator; KN-93, a Ca{sup 2+}/calmodulin-dependent protein kinase II inhibitor; CDX; and a CSD peptide, but not PD 123319, a selective AGTR2 antagonist. Collectively, our data demonstrate [Ca{sup 2+}]{sub i}-dependent proliferative effect of ANG-II and highlight a critical role for lipid raft microdomains in AGTR1-mediated signal transduction in neonatal GMCs. - Highlights: • AGTR1 is the functional AGTR subtype expressed in neonatal mesangial cells. • Endogenous AGTR1 associates with CAV-1 in neonatal mesangial cells. • Lipid raft disruption attenuates cell surface AGTR1 protein expression. • Lipid raft disruption reduces ANG-II-induced [Ca{sup 2+}]{sub i} elevation in neonatal mesangial cells. • Lipid raft disruption inhibits ANG-II-induced neonatal mesangial cell growth.

  20. Caveolin-1, Caveolin-2 and Cavin-1 are strong predictors of adipogenic differentiation in human tumors and cell lines of liposarcoma.

    Science.gov (United States)

    Codenotti, Silvia; Vezzoli, Marika; Poliani, Pietro Luigi; Cominelli, Manuela; Bono, Federica; Kabbout, Hadi; Faggi, Fiorella; Chiarelli, Nicola; Colombi, Marina; Zanella, Isabella; Biasiotto, Giorgio; Montanelli, Alessandro; Caimi, Luigi; Monti, Eugenio; Fanzani, Alessandro

    2016-08-01

    Caveolins (Cav-1, -2 and -3) and Cavins (Cavin-1, -2, -3 and -4) are two protein families controlling the biogenesis and function of caveolae, plasma membrane omega-like invaginations representing the primary site of important cellular processes like endocytosis, cholesterol homeostasis and signal transduction. Caveolae are especially abundant in fat tissue, playing a consistent role in a number of processes, such as the insulin-dependent glucose uptake and transmembrane transport of lipids underlying differentiation, maintenance and adaptive hypertrophy of adipocytes. Based on this premise, in this work we have investigated the expression of caveolar protein components in liposarcoma (LPS), an adipocytic soft tissue sarcoma affecting adults categorized in well-differentiated, dedifferentiated, myxoid and pleomorphic histotypes. By performing an extensive microarray data analysis followed by immunohistochemistry on human LPS tumors, we demonstrated that Cav-1, Cav-2 and Cavin-1 always cluster in all the histotypes, reaching the highest expression in well-differentiated LPS, the least aggressive of the malignant forms composed by tumor cells with a morphology resembling mature adipocytes. In vitro experiments carried out using two human LPS cell lines showed that the expression levels of Cav-1, Cav-2 and Cavin-1 proteins were faintly detectable during cell growth, becoming consistently increased during the accumulation of intracellular lipid droplets characterizing the adipogenic differentiation. Moreover, in differentiated LPS cells the three proteins were also found to co-localize and form molecular aggregates at the plasma membrane, as shown via immunofluorescence and immunoprecipitation analysis. Overall, these data indicate that Cav-1, Cav-2 and Cavin-1 may be considered as reliable markers for identification of LPS tumors characterized by consistent adipogenic differentiation. PMID:27168348

  1. Caveolae-dependent and -independent uptake of albumin in cultured rodent pulmonary endothelial cells.

    Directory of Open Access Journals (Sweden)

    Hui-Hua Li

    Full Text Available Although a critical role for caveolae-mediated albumin transcytosis in pulmonary endothelium is well established, considerably less is known about caveolae-independent pathways. In this current study, we confirmed that cultured rat pulmonary microvascular (RPMEC and pulmonary artery (RPAEC endothelium endocytosed Alexa488-labeled albumin in a saturable, temperature-sensitive mode and internalization resulted in co-localization by fluorescence microscopy with cholera B toxin and caveolin-1. Although siRNA to caveolin-1 (cav-1 in RPAEC significantly inhibited albumin uptake, a remnant portion of albumin uptake was cav-1-independent, suggesting alternative pathways for albumin uptake. Thus, we isolated and cultured mouse lung endothelial cells (MLEC from wild type and cav-1(-/- mice and noted that ~ 65% of albumin uptake, as determined by confocal imaging or live cell total internal reflectance fluorescence microscopy (TIRF, persisted in total absence of cav-1. Uptake of colloidal gold labeled albumin was evaluated by electron microscopy and demonstrated that albumin uptake in MLEC from cav-1(-/- mice was through caveolae-independent pathway(s including clathrin-coated pits that resulted in endosomal accumulation of albumin. Finally, we noted that albumin uptake in RPMEC was in part sensitive to pharmacological agents (amiloride [sodium transport inhibitor], Gö6976 [protein kinase C inhibitor], and cytochalasin D [inhibitor of actin polymerization] consistent with a macropinocytosis-like process. The amiloride sensitivity accounting for macropinocytosis also exists in albumin uptake by both wild type and cav-1(-/- MLEC. We conclude from these studies that in addition to the well described caveolar-dependent pulmonary endothelial cell endocytosis of albumin, a portion of overall uptake in pulmonary endothelial cells is cav-1 insensitive and appears to involve clathrin-mediated endocytosis and macropinocytosis-like process.

  2. Helleborus purpurascens—Amino Acid and Peptide Analysis Linked to the Chemical and Antiproliferative Properties of the Extracted Compounds

    Directory of Open Access Journals (Sweden)

    Adina-Elena Segneanu

    2015-12-01

    Full Text Available There is a strong drive worldwide to discover and exploit the therapeutic potential of a large variety of plants. In this work, an alcoholic extract of Helleborus purpurascens (family Ranunculaceae was investigated for the identification of amino acids and peptides with putative antiproliferative effects. In our work, a separation strategy was developed using solvents of different polarity in order to obtain active compounds. Biochemical components were characterized through spectroscopic (mass spectroscopy and chromatographic techniques (RP-HPLC and GC-MS. The biological activity of the obtained fractions was investigated in terms of their antiproliferative effects on HeLa cells. Through this study, we report an efficient separation of bioactive compounds (amino acids and peptides from a plant extract dependent on solvent polarity, affording fractions with unaffected antiproliferative activities. Moreover, the two biologically tested fractions exerted a major antiproliferative effect, thereby suggesting potential anticancer therapeutic activity.

  3. Ectomycorrhizal inoculation of willows on cut-over bored peatland

    Energy Technology Data Exchange (ETDEWEB)

    Loree, M.; Lumme, I.; Niemi, M.; Toermaelae, T.

    1988-05-01

    Ectomycorrhizal fungal inoculation was explored as a means to improve productivity of experimental short-rotation plantations of the willows Salix viminalis L. and Salix dasyclados Wimm. for biomass production on surface-mined peatlands in northern Finland. Both willow species formed ectomycorrhizas with Amanita spp., Cortinarius purpurascens, Entoloma nidorosum and other Entoloma spp., Hebeloma crustuliniforme, H. pusillum, Laccaria bicolor, and Paxillus involutus in greenhouse experiments. Field trials on a mined peatland site revealed (after one growing season) statistically significant growth stimultion due to inoculation, but these results were sometimes attributable to non-symbiotic effects of inoculation. Inoculation with some Entoloma isolates resulted in additional field growth stimulation due to symbiotic effects both willow species: plants inoculated with Entoloma were sometimes twice as large as control plants. however, such effects were observed only in plots receiving normal phosphate fertilization 40-80 kg as opposed to low 10-40 kg application and were not consistent from season to season.

  4. [Diversity and microstructure of quitons (Mollusca: Polyplacophora) from the Caribbean of Costa Rica].

    Science.gov (United States)

    García-Ríos, Cedar I; Alvarez-Ruiz, Migdalia

    2011-03-01

    Diversity and microstructure of quitons (Mollusca: Polyplacophora) from the Caribbean of Costa Rica. The polyplacophorans of the coral reef on the Caribbean coast of Costa Rica have been insufficiently studied. The examination of coral rubble accumulated in the shallow sublitoral waters on four collection stations in Provincia Limón revealed a higher diversity of chitons than was documented. From the country eight species were previously known: Ischnochiton erythronotus (C.B. Adams 1845); Ischnoplax pectinata (Sowerby 1840); Stenoplax boogii (Haddon 1886); S. purpurascens (C.B. Adams 1845); Acanthopleura granulata (Gmelin 1791); Chiton marmoratus Gmelin 1791; C. tuberculatus Linnaeus 1758 and Acanthochitona rhodea (Pilsbry 1893). This study added five more species that are reported here for the first time: Callistochiton portobelensis Ferreira 1976; Ischnochiton kaasi Ferreira 1987; I. pseudovirgatus Kaas 1972; Acanthochitona balesae Abbott 1954 and Cryptoconchus floridanus (Dall 1889). PMID:21516641

  5. Dicty_cDB: Contig-U15144-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available :none) Streptomyces purpurascens rhodomycin ... 54 2e-06 AM269991_16( AM269991 |pid:none) Aspergillus niger contig...000285_1812( CP000285 |pid:none) Chromohalobacter salexigens DSM... 86 7e-16 CP001615_229( CP001615 |pid:none) Exiguobacter... 1e-15 AE017355_3732( AE017355 |pid:none) Bacillus thuringiensis serovar ... 85 1e-15 CP000903_3758( CP000903 |pid:none) Bacillus wei...r CPRD12 prot... 54 2e-06 BT076081_1( BT076081 |pid:none) Caligus rogercresseyi clone crog-e... 54 2e-06 AK1...AAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAA Gap no gap Contig length 558 Chromosome number (1..6, M) 6 Chromosome length 3595308 Start

  6. Plant-Lore with Reference to Muslim Folksong in Association with Human Perception of Plants in Agricultural and Horticultural Practices

    Directory of Open Access Journals (Sweden)

    M. M. AHMED

    2008-08-01

    Full Text Available The Muslims in the Manipur valley are a Manipuri speaking community. The study of folksong texts of Muslims in Manipur is a parameter for exploring native plant resources which certainly help to take conservation steps by examining the way of the human perception of plants. Man-plant interaction in agricultural as well as horticultural practices unfolds human dependence on plants in their various functions leading to a comfortable living. Some plants are used directly by men without any process, while some are selectively avoided. Tapping off heritage of ethnobotanical lore has been practiced by Muslims, conserving Pogostemon purpurascens Dalz in many graveyard for its religious significance. It has been proved that some plants are prevalent in the human language (proverbs, folksong, similies etc. to make sense of their expression.

  7. Cooperative Role of Mineralocorticoid Receptor and Caveolin-1 in Regulating the Vascular Response to Low Nitric Oxide-High Angiotensin II-Induced Cardiovascular Injury.

    Science.gov (United States)

    Pojoga, Luminita H; Yao, Tham M; Opsasnick, Lauren A; Siddiqui, Waleed T; Reslan, Ossama M; Adler, Gail K; Williams, Gordon H; Khalil, Raouf A

    2015-10-01

    Aldosterone interacts with mineralocorticoid receptor (MR) to stimulate sodium reabsorption in renal tubules and may also affect the vasculature. Caveolin-1 (cav-1), an anchoring protein in plasmalemmal caveolae, binds steroid receptors and also endothelial nitric oxide synthase, thus limiting its translocation and activation. To test for potential MR/cav-1 interaction in the vasculature, we investigated if MR blockade in cav-1-replete or -deficient states would alter vascular function in a mouse model of low nitric oxide (NO)-high angiotensin II (AngII)-induced cardiovascular injury. Wild-type (WT) and cav-1 knockout mice (cav-1(-/-)) consuming a high salt diet (4% NaCl) received Nω-nitro-l-arginine methyl ester (L-NAME) (0.1-0.2 mg/ml in drinking water at days 1-11) plus AngII (0.7-2.8 mg/kg per day via an osmotic minipump at days 8-11) ± MR antagonist eplerenone (EPL) 100 mg/kg per day in food. In both genotypes, blood pressure increased with L-NAME + AngII. EPL minimally changed blood pressure, although its dose was sufficient to block MR and reverse cardiac expression of the injury markers cluster of differentiation 68 and plasminogen activator inhibitor-1 in L-NAME+AngII treated mice. In aortic rings, phenylephrine and KCl contraction was enhanced with EPL in L-NAME+AngII treated WT mice, but not cav-1(-/-) mice. AngII-induced contraction was not different, and angiotensin type 1 receptor expression was reduced in L-NAME + AngII treated WT and cav-1(-/-) mice. In WT mice, acetylcholine-induced relaxation was enhanced with L-NAME + AngII treatment and reversed with EPL. Acetylcholine relaxation in cav-1(-/-) mice was greater than in WT mice, not modified by L-NAME + AngII or EPL, and blocked by ex vivo L-NAME, 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one (ODQ), or endothelium removal, suggesting the role of NO-cGMP. Cardiac endothelial NO synthase was increased in cav-1(-/-) versus WT mice, further increased with L-NAME + AngII, and not affected by EPL

  8. Quis custodiet ipsos custodies: who watches the watchmen?

    Energy Technology Data Exchange (ETDEWEB)

    Ghajar, Cyrus M.; Meier, Roland; Bissell, Mina J.

    2009-06-03

    prognostic for patient survival and predictive of response to chemotherapeutic treatment provide proof of this concept. In the current issue of The American Journal of Pathology, two independent studies identify a novel stromal marker, caveolin (Cav)-1, which predicts clinical outcome of breast cancer patients irrespective of its expression in tumor epithelium. Cav-1 is a scaffolding protein essential to the structure of caveolae, 'little caves' or invaginations in cellular plasma membranes. Cav-1 recruits and arranges lipids and proteins to these membrane sites to function in endocytosis and signal transduction. The observation that Cav-1 expression is attenuated in oncogenically transformed cells led to exploration of whether Cav-1 loss in mammary epithelium was causative. Although mechanistic data suggested that Cav-1 null mice exhibited aberrant epithelial growth, and that forcing Cav-1 expression in breast cancer cell lines inhibited growth and metastases in xenograft models, a clinical link proved elusive. However, MMTV-PyMT tumors transplanted into the fat pads of Cav-1 knockout mice displayed significantly enhanced growth (vs. wild-type mice), motivating investigation of whether stromal Cav-1 expression correlates with human breast cancer patient survival. This is precisely what Sloan et al. and Witkiewicz, Dasgupta, et al. demonstrate in this issue of AJP. Using tissue microarray data in conjunction with breast tumor sections and extensive patient survival data, Sloan et al. demonstrate that strong stromal Cav-1 expression is associated with smaller breast tumor size and grade, and is highly predictive of increased survival (Fig. 1). Patients with positive expression of stromal Cav-1 had a 91% ten year survival rate, vs. a 43% survival rate for patients lacking stromal Cav-1 expression. Importantly, there was no correlation between Cav-1 expression in the tumor epithelium and clinical outcome in either tissue arrays or tumor sections.

  9. Caveolae在高糖诱导的大鼠肾小球系膜细胞细胞外基质合成中的作用%Role of caveolae in high glucose-induced extracellular matrix production in rat mesangial cells

    Institute of Scientific and Technical Information of China (English)

    卢圣霞; 刘元涛; 孔峰; 杜月娟; 刘晔; 傅余芹

    2013-01-01

    目的:探讨胞膜小凹(caveolae)在高糖(HG)诱导大鼠肾小球系膜细胞(MCs)细胞外基质(extra-cellular matrix,ECM)表达过程中的作用.方法:传代培养的大鼠MCs同步化后分为:(1)正常糖组;(2)HG组;(3)HG+甲基-β-环糊精(β-MCD)组;(4) HG+ β-MCD+胆固醇组.用Western blotting检测小凹蛋白1(Cav-1)、磷酸化小凹蛋白1 (p-Cav-1-Y14)和Ⅰ型胶原(Col Ⅰ)蛋白的表达水平,用实时定量PCR检测细胞中Cav-1 mRNA表达变化,用ELISA方法检测上清中纤维连接蛋白(FN)的蛋白含量.结果:(1)高糖状态下,系膜细胞FN和Col Ⅰ蛋白表达水平均显著增加(均P<0.05).(2)高糖培养显著增加p-Cav-1-Y14水平(P<0.01),而对Cav-1 mRNA及蛋白表达无明显影响(P>0.05).(3) β-MCD预处理抑制了高糖诱导的p-Cav-1-Y14升高(P<0.01)及FN表达(P<0.05),但对高糖诱导的Col Ⅰ表达无影响;β-MCD的作用可被胆固醇抑制.结论:高糖可显著增加系膜细胞1型胶原及纤维连接蛋白的合成.高糖诱导的纤维连接蛋白合成与C av-1磷酸化水平增加有关.%AIM: To investigate the role of caveolae in high glucose ( HG ) -induced extracellular matrix (ECM) production in rat mesangial cells (MCs). METHODS: Synchronized rat MCs were divided into normal glucose group, HG group, HG + methyl-g-eyelodextrin (3-MCD) group and HG + β-MCD + cholesterol (Choi) group. Western blotting was used to detect the protein expression of caveolin-1 (Cav-1 ) , phosphorylated caveolin-1 (p-Cav-l-Y14) and collagen type 1 (Col I). The mRNA expression of Cav-1 was determined by real-time PCR. ELISA was used to measure the level of fibronectin (FN) in the supernatant. RESULTS; High glucose significantly increased the expression of FN and Col I. In HG 12, 24 and 48 h groups, the mRNA and protein levels of Cav-1 were not significantly different from those in HG 0 h group, whereas the level of p-Cav-l-Y14 was significantly increased. β-MCD significantly attenuated HG

  10. Early inflammatory markers are independent predictors of cardiac allograft vasculopathy in heart-transplant recipients.

    Directory of Open Access Journals (Sweden)

    Carlos A Labarrere

    Full Text Available BACKGROUND: Identification of risk is essential to prevent cardiac allograft vasculopathy (CAV and graft failure due to CAV (GFDCAV in heart transplant patients, which account for 30% of all deaths. Early CAV detection involves invasive, risky, and expensive monitoring approaches. We determined whether prediction of CAV and GFDCAV improves by adding inflammatory markers to a previously validated atherothrombotic (AT model. METHODS AND FINDINGS: AT and inflammatory markers interleukin-6 (IL-6 and C-reactive protein (CRP were measured in heart biopsies and sera of 172 patients followed prospectively for 8.9±5.0 years. Models were estimated for 5- and 10-year risk using (1 the first post-transplant biopsy only, or (2 all biopsies obtained within 3 months. Multivariate models were adjusted for other covariates and cross-validated by bootstrapping. After adding IL-6 and CRP to the AT models, we evaluated the significance of odds ratios (ORs associated with the additional inflammatory variables and the degree of improvement in the area under the receiver operating characteristic curve (AUROC. When inflammatory markers were tested alone in prediction models, CRP (not IL-6 was a significant predictor of CAV and GFDCAV at 5 (CAV: p<0.0001; GFDCAV: p = 0.005 and 10 years (CAV: p<0.0001; GFDCAV: p = 0.003. Adding CRP (not IL-6 to the best AT models improved discriminatory power to identify patients destined to develop CAV (using 1st biopsy: p<0.001 and p = 0.001; using all 3-month biopsies: p<0.04 and p = 0.008 at 5- and 10-years, respectively and GFDCAV (using 1st biopsy: 0.92 vs. 0.95 and 0.86 vs. 0.89; using all 3-month biopsies: 0.94 vs. 0.96 and 0.88 vs. 0.89 at 5- and 10-years, respectively, as indicated by an increase in AUROC. CONCLUSIONS: Early inflammatory status, measured by a patient's CRP level (a non-invasive, safe and inexpensive test, independently predicts CAV and GFDCAV. Adding CRP to a previously established AT model

  11. The demonstration, embryology and association with a neurological disorder of the brain midline structures cavum septi pellucidi and cavum vergae by neuroradiological methods from the perinatal period until childhood

    International Nuclear Information System (INIS)

    Cavum septi pellucidi (CSP) and Cavum vergae (CaV) are primitive midline structures of the premature brain. Retrospectively we looked up the CSP and the CaV at birth, 1st week, 1st, 2nd, 3rd and 4th month of the done transcranial sonographies. Then we compared preterms with terms and standardized these data to one population. The results were that the CaV begins to close in the 30th week of gestational age (ga), increases in the 31st and 36th week and shows the highest rate of reduction between the 33rd to 35th week of ga. The CSP starts closing in the 35th week of ga. We observed an increase of relative rates in the 36th and 39th week of ga and the period of the greatest reduction between the 2nd and 4th month. In the perinatal period there are unknown mechanisms in the leaves of the Septum pellucidum. By a documentation system we found out in all MRI (n=3743) of the last five years which were done in children between 0 and 15 years. There was a relative rate of Cusps 1,31 %, according to the age 0 to 1 there was a rate of 2,67 %. The rate of CaV was 0,85 % and 2,23 %. The highest period of reduction is after the 5th year for the CSP and after the first year for CaV. In the CCT we observed a relative rate of 0,62 % (CSP) and of 0,25 % (CaV). The association between Cava and neurological disorder showed that 20 % are combined with seizures (MRI) and 20 % with hydrocephalus (CT). More boys than girls show CSP and CaV. In the last examination we compared the rates of CSP/CaV in all MRI (n=3743) with the CSP/CaV associated with the neurological disorder hydrocephalus. In the population hydrocephalus there is a division in obstructive and communicating (ratio 80:20). From birth to the 15th year the relative rates (CSP) with hydrocephalus keeps constant (0,73 %), as the rates in the normal population decreases continuously; these of CaV show an increase between the 2nd and 5th year and are above the normal CaV rate. (author)

  12. A combined method for rescue of modified enteroviruses by mutagenic primers, long PCR and T7 RNA polymerase-driven in vivo transcription.

    Science.gov (United States)

    Heikkilä, Outi; Kainulainen, Markus; Susi, Petri

    2011-01-01

    The current methods for manipulation of enteroviral RNA genomes and production of modified virus particles include stepwise subcloning procedures and in vitro transcription and RNA transfection steps that are both time-consuming and inefficient. Several enteroviral cDNA clones with 5'-terminal T7 promoter and coxsackievirus A9 (CAV9) PCR product with the T7 promoter were transfected successfully into target cells expressing T7 RNA polymerase for the rescue of virus particles. This demonstrated the overall feasibility of the in vivo transcription method. Furthermore, a rapid method using high-fidelity DNA polymerase, Phusion™, for amplification and mutagenesis of CAV9 cDNA was generated. A long PCR method was employed together with mutagenic primers for direct introduction of a unique restriction enzyme site into the VP1-2A junction of the CAV9 cDNA clone during the PCR amplification process. Enhanced green fluorescent protein was subcloned to that site, and CAV9-eGFP cDNA was transfected to the target cells for in vivo transcription and successful rescue of CAV9-eGFP particles. The method allowed a straightforward mutagenesis and in vivo production of infectious enteroviral particles, and may be applicable routinely for rapid production of the modified picornaviruses over the use of the traditional subcloning protocols. PMID:20974179

  13. Targeting voltage-gated calcium channels: developments in peptide and small-molecule inhibitors for the treatment of neuropathic pain.

    Science.gov (United States)

    Vink, S; Alewood, P F

    2012-11-01

    Chronic pain affects approximately 20% of people worldwide and places a large economic and social burden on society. Despite the availability of a range of analgesics, this condition is inadequately treated, with complete alleviation of symptoms rarely occurring. In the past 30 years, the voltage-gated calcium channels (VGCCs) have been recognized as potential targets for analgesic development. Although the majority of the research has been focused on Ca(v) 2.2 in particular, other VGCC subtypes such as Ca(v) 3.2 have recently come to the forefront of analgesic research. Venom peptides from marine cone snails have been proven to be a valuable tool in neuroscience, playing a major role in the identification and characterization of VGCC subtypes and producing the first conotoxin-based drug on the market, the ω-conotoxin, ziconotide. This peptide potently and selectively inhibits Ca(v) 2.2, resulting in analgesia in chronic pain states. However, this drug is only available via intrathecal administration, and adverse effects and a narrow therapeutic window have limited its use in the clinic. Other Ca(v) 2.2 inhibitors are currently in development and offer the promise of an improved route of administration and safety profile. This review assesses the potential of targeting VGCCs for analgesic development, with a main focus on conotoxins that block Ca(v) 2.2 and the developments made to transform them into therapeutics. PMID:22725651

  14. Molecular characterization and expression analysis of caveolin-1 in pig tissues

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Members of the caveolin family played important roles during fundamental cellular processes,such as regulation of cell morphology,migration,and gene expression in muscle cells.In this study,caveolin-1 (Cav-1),one of the caveolins,was identified from longissimus dorsi muscle of Large Yorkshire pig and Chinese indigenous Lantang pig based on the results of mRNA differential display analysis.The deduced amino acids sequence of the porcine Cav-1 contained a caveolin domain,and was very conservative among different species.The Cav-1 mRNA was widely expressed in the eight tissues in this study,including heart,liver,kidney,encephalon,spleen,lung,longissimus dorsi muscle,and back fat, and the highest expression quantity was found in back fat of the two pig breeds.The expression quantity of porcine Cav-1 in back fat and longissimus dorsi muscle of Lantang pig was significantly higher than that of Large Yorkshire(P<0.01,and P<0.05,respectively).These results suggested that the Cav-1 might be a candidate gene for carcass traits,and might provide valuable information for understanding the mechanism of caveolae signaling in fat deposition by using the animal model of pig.

  15. Production and purification of non replicative canine adenovirus type 2 derived vectors.

    Science.gov (United States)

    Szelechowski, Marion; Bergeron, Corinne; Gonzalez-Dunia, Daniel; Klonjkowski, Bernard

    2013-01-01

    Adenovirus (Ad) derived vectors have been widely used for short or long-term gene transfer, both for gene therapy and vaccine applications. Because of the frequent pre-existing immunity against the classically used human adenovirus type 5, canine adenovirus type 2 (CAV2) has been proposed as an alternative vector for human gene transfer. The well-characterized biology of CAV2, together with its ease of genetic manipulation, offer major advantages, notably for gene transfer into the central nervous system, or for inducing a wide range of protective immune responses, from humoral to cellular immunity. Nowadays, CAV2 represents one of the most appealing nonhuman adenovirus for use as a vaccine vector. This protocol describes a simple method to construct, produce and titer recombinant CAV2 vectors. After cloning the expression cassette of the gene of interest into a shuttle plasmid, the recombinant genomic plasmid is obtained by homologous recombination in the E. coli BJ5183 bacterial strain. The resulting genomic plasmid is then transfected into canine kidney cells expressing the complementing CAV2-E1 genes (DK-E1). A viral amplification enables the production of a large viral stock, which is purified by ultracentrifugation through cesium chloride gradients and desalted by dialysis. The resulting viral suspension routinely has a titer of over 10(10) infectious particles per ml and can be directly administrated in vivo. PMID:24326926

  16. Calcium channel-dependent molecular maturation of photoreceptor synapses.

    Directory of Open Access Journals (Sweden)

    Nawal Zabouri

    Full Text Available Several studies have shown the importance of calcium channels in the development and/or maturation of synapses. The Ca(V1.4(α(1F knockout mouse is a unique model to study the role of calcium channels in photoreceptor synapse formation. It features abnormal ribbon synapses and aberrant cone morphology. We investigated the expression and targeting of several key elements of ribbon synapses and analyzed the cone morphology in the Ca(V1.4(α(1F knockout retina. Our data demonstrate that most abnormalities occur after eye opening. Indeed, scaffolding proteins such as Bassoon and RIM2 are properly targeted at first, but their expression and localization are not maintained in adulthood. This indicates that either calcium or the Ca(V1.4 channel, or both are necessary for the maintenance of their normal expression and distribution in photoreceptors. Other proteins, such as Veli3 and PSD-95, also display abnormal expression in rods prior to eye opening. Conversely, vesicle related proteins appear normal. Our data demonstrate that the Ca(V1.4 channel is important for maintaining scaffolding proteins in the ribbon synapse but less vital for proteins related to vesicular release. This study also confirms that in adult retinae, cones show developmental features such as sprouting and synaptogenesis. Overall we present evidence that in the absence of the Ca(V1.4 channel, photoreceptor synapses remain immature and are unable to stabilize.

  17. New Conotoxin SO-3 Targeting N-type Voltage-Sensitive Calcium Channels

    Directory of Open Access Journals (Sweden)

    Lei Wen

    2006-04-01

    Full Text Available Selective blockers of the N-type voltage-sensitive calcium (CaV channels are useful in the management of severe chronic pain. Here, the structure and function characteristics of a novel N-type CaV channel blocker, SO-3, are reviewed. SO-3 is a 25-amino acid conopeptide originally derived from the venom of Conus striatus, and contains the same 4-loop, 6-cysteine framework (C-C-CC-C-C as O-superfamily conotoxins. The synthetic SO-3 has high analgesic activity similar to ω-conotoxin MVIIA (MVIIA, a selective N-type CaV channel blocker approved in the USA and Europe for the alleviation of persistent pain states. In electrophysiological studies, SO-3 shows more selectivity towards the N-type CaV channels than MVIIA. The dissimilarity between SO-3 and MVIIA in the primary and tertiary structures is further discussed in an attempt to illustrate the difference in selectivity of SO-3 and MVIIA towards N-type CaV channels.

  18. Epidemiology of chicken anemia virus in Central African Republic and Cameroon

    Directory of Open Access Journals (Sweden)

    Snoeck Chantal J

    2012-09-01

    Full Text Available Abstract Background Although chicken anemia virus (CAV has been detected on all continents, little is known about this virus in sub-Saharan Africa. This study aimed to detect and characterize CAV for the first time in Central African Republic and in Cameroon. Results An overall flock seroprevalence of 36.7% was found in Central African Republic during the 2008–2010 period. Virus prevalences were 34.2% (2008, 14.3% (2009 and 10.4% (2010 in Central African Republic and 39% (2007 and 34.9% (2009 in Cameroon. CAV DNA was found in cloacal swabs of 76.9% of seropositive chickens, suggesting that these animals excreted the virus despite antibodies. On the basis of VP1 sequences, most of the strains in Central African Republic and Cameroon belonged to 9 distinct phylogenetic clusters at the nucleotide level and were not intermixed with strains from other continent. Several cases of mixed infections in flocks and individual chickens were identified. Conclusions Our results suggest multiple introductions of CAV in each country that later spread and diverged locally. Mixed genotype infections together with the observation of CAV DNA in cloacal samples despite antibodies suggest a suboptimal protection by antibodies or virus persistence.

  19. Molecular Interactions between Tarantula Toxins and Low-Voltage-Activated Calcium Channels.

    Science.gov (United States)

    Salari, Autoosa; Vega, Benjamin S; Milescu, Lorin S; Milescu, Mirela

    2016-01-01

    Few gating-modifier toxins have been reported to target low-voltage-activated (LVA) calcium channels, and the structural basis of toxin sensitivity remains incompletely understood. Studies of voltage-gated potassium (Kv) channels have identified the S3b-S4 "paddle motif," which moves at the protein-lipid interface to drive channel opening, as the target for these amphipathic neurotoxins. Voltage-gated calcium (Cav) channels contain four homologous voltage sensor domains, suggesting multiple toxin binding sites. We show here that the S3-S4 segments within Cav3.1 can be transplanted into Kv2.1 to examine their individual contributions to voltage sensing and pharmacology. With these results, we now have a more complete picture of the conserved nature of the paddle motif in all three major voltage-gated ion channel types (Kv, Nav, and Cav). When screened with tarantula toxins, the four paddle sequences display distinct toxin binding properties, demonstrating that gating-modifier toxins can bind to Cav channels in a domain specific fashion. Domain III was the most commonly and strongly targeted, and mutagenesis revealed an acidic residue that is important for toxin binding. We also measured the lipid partitioning strength of all toxins tested and observed a positive correlation with their inhibition of Cav3.1, suggesting a key role for membrane partitioning. PMID:27045173

  20. Significance of caveolin-1 and matrix metalloproteinase 14 gene expression in canine mammary tumours.

    Science.gov (United States)

    Ebisawa, M; Iwano, H; Nishikawa, M; Tochigi, Y; Komatsu, T; Endou, Y; Hirayama, K; Taniyama, H; Kadosawa, T; Yokota, H

    2015-11-01

    Canine mammary tumours (CMTs) are the most common neoplasms affecting female dogs. There is an urgent need for molecular biomarkers that can detect early stages of the disease in order to improve accuracy of CMT diagnosis. The aim of this study was to examine whether caveolin-1 (Cav-1) and matrix metalloproteinase 14 (MMP14) are associated with CMT histological malignancy and invasion. Sixty-five benign and malignant CMT samples and six normal canine mammary glands were analysed using quantitative reverse transcription-polymerase chain reaction. Cav-1 and MMP14 genes were highly expressed in CMT tissues compared to normal tissues. Cav-1 especially was overexpressed in malignant and invasive CMT tissues. When a CMT cell line was cultured on fluorescent gelatin-coated coverslips, localisation of Cav-1 was observed at invadopodia-mediated degradation sites of the gelatin matrix. These findings suggest that Cav-1 may be involved in CMT invasion and that the markers may be useful for estimating CMT malignancy. PMID:26364240

  1. Cardiac allograft vasculopathy: optical coherence guided innovative treatment options with the bioresorbable vascular scaffold: proof of concept.

    Science.gov (United States)

    Édes, István F; Hajas, Ágota; Sax, Balázs; Bartykowszki, Andrea; Becker, Dávid; Merkely, Béla

    2016-08-01

    The aim of our work was to assess a novel interventional therapy option in cardiac allograft vasculopathy (CAV), a complex form of coronary disease presenting only in heart transplant (HTx) recipients. It is typically a rapidly progressing phenomenon, affecting the entire coronary circulation causing diffuse, severe coronary lesions and has no one unique cause. Treatment options are limited, but where eligible, palliation via percutaneous revascularization (PCI) mainly using new generation drug eluting stents (DES) is recommended. Our working group sought to assess outcomes of CAV PCI using an Absorb (Abbott Vascular, Santa Clara, CA, USA) fully bioresorbable, everolimus eluting vascular scaffold (BVS), under optical coherence tomography (OCT) guidance. Our initial, proof-of-concept case showed a late CAV, macrophage and foam-cell rich lesion, with typical asymmetric intimal hyperplasia and contralateral thin-cap fibroatheroma formation. Post-PCI OCT showed underexpansion, requiring aggressive postdilatation. Ninety-day follow-up CT angiogram identified the scaffold and displayed a patent lumen of the device. BVS use thus seems eligible in CAV, yet needs proper, meticulous implantation. Use may also delay CAV progression as lesion healing is promoted, with restoration of vasomotion and a natural increase in vascular lumen. Furthermore, the chronically present vascular irritation surrounding stent/scaffold struts may subside, as no permanent metal is present as an increased substrate for inflammation. To assess full efficacy, further studies will be needed. PMID:27152623

  2. A single immunization with a recombinant canine adenovirus expressing the rabies virus G protein confers protective immunity against rabies in mice

    International Nuclear Information System (INIS)

    Rabies vaccines based on live attenuated rabies viruses or recombinant pox viruses expressing the rabies virus (RV) glycoprotein (G) hold the greatest promise of safety and efficacy, particularly for oral immunization of wildlife. However, while these vaccines induce protective immunity in foxes, they are less effective in other animals, and safety concerns have been raised for some of these vaccines. Because canine adenovirus 2 (CAV2) is licensed for use as a live vaccine for dogs and has an excellent efficacy and safety record, we used this virus as an expression vector for the RVG. The recombinant CAV2-RV G produces virus titers similar to those produced by wild-type CAV2, indicating that the RVG gene does not affect virus replication. Comparison of RVG expressed by CAV2-RV G with that of vaccinia-RV G recombinant virus (V-RG) revealed similar amounts of RV G on the cell surface. A single intramuscular or intranasal immunization of mice with CAV2-RVG induced protective immunity in a dose-dependent manner, with no clinical signs or discomfort from the virus infection regardless of the route of administration or the amount of virus

  3. Na+-K+ pump location and translocation during muscle contraction  in rat skeletal muscle

    DEFF Research Database (Denmark)

    Kristensen, Michael; Rasmussen, Martin Krøyer; Juel, Carsten

    2008-01-01

    Muscle contraction may up-regulate the number of Na+-K+ pumps in the plasma membrane by translocation of subunits. Since there is still controversy about where this translocation takes place from and if it takes place at all, the present study used different techniques to characterize the...... translocation. Electrical stimulation and biotin labeling of rat muscle revealed a 40% and 18% increase in the amounts of the Na+-K+ pump a2 subunit and caveolin-3 (Cav-3), respectively, in the sarcolemma. Exercise induced a 36% and 19% increase in the relative amounts of the a2 subunit and Cav-3, respectively......% of the a1 subunits were associated with Cav-3 in soleus, extensor digitorum longus, and mixed muscles, respectively. For the a2, the corresponding values were 17%, 5% and 16%. In conclusion; muscle contraction induces translocation of the a subunits, which is suggested to be caused partly by...

  4. The Role of L- and T-Type Calcium Channels in Local and Remote Calcium Responses in Rat Mesenteric Terminal Arterioles

    DEFF Research Database (Denmark)

    Braunstein, Thomas Hartig; Inoue, Ryuji; Cribbs, Leanne;

    2009-01-01

    Background/Aims: The roles of intercellular communication and T-type versus L-type voltage-dependent Ca(2+) channels (VDCCs) in conducted vasoconstriction to local KCl-induced depolarization were investigated in mesenteric arterioles. Methods: Ratiometric Ca(2+) imaging (R) using Fura-PE3 with...... local (DeltaR = 0.54) and remote (DeltaR = 0.17 at 500 mum) increases in intracellular Ca(2+). Remote Ca(2+) responses were inhibited by the gap junction uncouplers carbenoxolone and palmitoleic acid. Ca(V)1.2, Ca(V)3.1 and Ca(V)3.2 channels were immunolocalized in vascular smooth muscle cells and Ca...... arterioles (at 200-300 mum) using micro-application of VDCC blockers. Conclusion: Both L- and T-type channels mediate Ca(2+) entry during conducted vasoconstriction to local KCl in mesenteric arterioles. However, these channels do not participate in the conduction process per se....

  5. Tx1, from Phoneutria nigriventer spider venom, interacts with dihydropyridine sensitive-calcium channels in GH3 cells

    International Nuclear Information System (INIS)

    The aim of this work was to use the binding assay of tritiated-dihydropyridine and radioiodinated Tx1, isolated from the Phoneutria nigriventer venom, in order to show the presence of Cav1 calcium channels on pituitary tumour cell (GH3). We showed that GH3 cells have specific sites for 125I-Tx1, which are sensitive to nifedipine (∼20%). Reverse competition assay with 3H-PN200-110 (40% inhibition) and electrophysiological data (50% inhibition) suggest that Cav1 calcium channels are target sites for this toxin. To summarize, Tx1 binds to specific sites on GH3 cells and this interaction results in Cav1 calcium channel blockade. 3H-PN200-110 and 125I-Tx1 binding assays proved to be useful tools to show the presence of calcium channels on GH3 cells. (author)

  6. 蛇床子素对L-和N-型钙通道的影响%Effect of Osthole on L- and N- type Calcium Channels

    Institute of Scientific and Technical Information of China (English)

    袁春华; 李勃兴; 孙丽荣; 朱心红

    2011-01-01

    Objective: To determine the effects of osthole on voltage-dependent calcium channel subtypes. Methods: tsA-201 cells were transfected with the calcium channel subtypes of Cavl.2, Cavl.3, Cav2.2e [37a],and Cav2.2e [37b]. The calcium currents were recorded in tsA-201 cells using whole cell patch clamp technique. The effects of osthole on calcium currents were investigated. Results: Osthole could inhibit Cavl.2 and Cavl.3 currents in a concentration - dependent manner. The half-effective concentration of the inhibition was 162.1 μmol -L-1, and 56.2 μmo\\ -L-1, respectively. In addition, Osthole reduced the Cav2.2e [37a],and Cav2.2e[37b] currents by 38 %, and 61 %, respectively, at the concentration of 300 μmol-L -1. The inhibition was fast and reversible. Osthole decreased the current amplitudes at all tested potentials, but showed no significant influence on the activation kinetics of the above calcium channels subtypes. Conclusions: Osthole concentration - dependency inhibited calcium currents and displayed different affinity to the calcium channel subtypes.%比较蛇床子素对不同钙通道亚型的作用差异.方法:首先在tsA201细胞上瞬时转染Cavl.2,Cav1.3,Cav2.2e[37a],和Cav2.2e[37b]通道,然后采用全细胞膜片钳技术,记录tsA201细胞上的钙电流,并观察蛇床子素对各种钙通道亚型的影响.结果:蛇床子素可以浓度依赖性抑制Cav1.2和Cav1.3电流,抑制的半有效浓度分别为162.1 μmol·L1和56.2 μmol·L-1.此外,蛇床子素对Cav2.2通道也有一定的抑制作用,在300μmol·L-1的浓度下,抑制38%的Cav2.2e[37a]电流和61%的Cav2.2e[37b]电流.蛇床子素对钙电流的抑制是快速可逆的.蛇床子素在各个测试电位水平均能抑制上述四种钙通道电流,但不改变电流的激活阈值和最大峰值电流的激活电压.结论:蛇床子素以浓度依赖的方式抑制多种钙通道亚型并表现出不同的亲和力.

  7. Functional importance of T-type voltage-gated calcium channels in the cardiovascular and renal system

    DEFF Research Database (Denmark)

    Hansen, Pernille B L

    2015-01-01

    lack of highly specific blockers cast doubt on the conclusions. As new T-type channel antagonists are being designed, the roles of T-type channels in cardiovascular and renal pathology need to be elucidated before T-type blockers can be clinically useful. Two types of T-type channels, Cav3.1 and Cav3...... several cardiovascular pathologies, but the use of T-type blockers should be specifically directed to the disease and to the channel subtype.......Over the years, it has been discussed whether T-type calcium channels Cav3 play a role in the cardiovascular and renal system. T-type channels have been reported to play an important role in renal hemodynamics, contractility of resistance vessels, and pacemaker activity in the heart. However, the...

  8. Molecular characterization of chicken infectious anemia virus circulating in Argentina during 2007.

    Science.gov (United States)

    Craig, M I; Rimondi, A; Delamer, M; Sansalone, P; König, G; Vagnozzi, A; Pereda, A

    2009-09-01

    Chicken infectious anemia virus (CAV) is a worldwide-distributed infectious agent that affects commercial poultry. Although this agent was first detected in Argentina in 1994, no further studies on CAV in this country were reported after that. The recent increased occurrence of clinical cases of immunosuppression that could be caused by CAV has prompted this study. Our results confirmed that CAV is still circulating in commercial flocks in Argentina. Phylogenetic analysis focusing on the VP1 nucleotide sequence showed that all Argentinean isolates grouped together in a cluster, sharing a high similarity (> 97%) with genotype B reference strains. However, Argentinean isolates were distantly related to other strains commonly used for vaccination in this country, such as Del-Ros and Cux-1. Sequence analysis of predicted VP1 peptides showed that most of the Argentinean isolates have a glutamine residue at positions 139 and 144, suggesting that these isolates might have a reduced spread in cell culture compared with Cux-1. In addition, a particular amino acid substitution at position 290 is present in all studied Argentinean isolates, as well as in several VP1 sequences from Malaysia, Australia, and Japan isolates. Our results indicate that it is possible to typify CAV strains by comparison of VPI nucleotide sequences alone because the same tree topology was obtained when using the whole genome sequence. The molecular analysis of native strains sheds light into the epidemiology of CAV in Argentinean flocks. In addition, this analysis could be considered in future control strategies focused not only on breeders but on broilers and layer flocks. PMID:19848068

  9. L-type calcium channels in adrenal chromaffin cells: role in pace-making and secretion.

    Science.gov (United States)

    Marcantoni, A; Baldelli, P; Hernandez-Guijo, J M; Comunanza, V; Carabelli, V; Carbone, E

    2007-01-01

    Voltage-gated L-type (Cav1.2 and Cav1.3) channels are widely expressed in cardiovascular tissues and represent the critical drug-target for the treatment of several cardiovascular diseases. The two isoforms are also abundantly expressed in neuronal and neuroendocrine tissues. In the brain, Cav1.2 and Cav1.3 channels control synaptic plasticity, somatic activity, neuronal differentiation and brain aging. In neuroendocrine cells, they are involved in the genesis of action potential generation, bursting activity and hormone secretion. Recent studies have shown that Cav1.2 and Cav1.3 are also expressed in chromaffin cells but their functional role has not yet been identified despite that L-type channels possess interesting characteristics, which confer them an important role in the control of catecholamine secretion during action potentials stimulation. In intact rat adrenal glands L-type channels are responsible for adrenaline and noradrenaline release following splanchnic nerve stimulation or nicotinic receptor activation. L-type channels can be either up- or down-modulated by membrane autoreceptors following distinct second messenger pathways. L-type channels are tightly coupled to BK channels and activate at relatively low-voltages. In this way they contribute to the action potential hyperpolarization and to the pace-maker current controlling action potential firings. L-type channels are shown also to regulate the fast secretion of the immediate readily releasable pool of vesicles with the same Ca(2+)-efficiency of other voltage-gated Ca(2+) channels. In mouse adrenal slices, repeated action potential-like stimulations drive L-type channels to a state of enhanced stimulus-secretion efficiency regulated by beta-adrenergic receptors. Here we will review all these novel findings and discuss the possible implication for a specific role of L-type channels in the control of chromaffin cells activity. PMID:17561252

  10. Depolarization-Evoked Secretion Requires Two Vicinal Transmembrane Cysteines of Syntaxin 1A

    OpenAIRE

    Cohen, Roy; Marom, Merav; Atlas, Daphne

    2007-01-01

    Background The interactions of the voltage-gated Ca2+ channel (VGCC) with syntaxin 1A (Sx 1A), Synaptosome-associated protein of 25 kD (SNAP-25), and synaptotagmin, couple electrical excitation to evoked secretion. Two vicinal Cys residues, Cys 271 and Cys 272 in the Sx 1A transmembrane domain, are highly conserved and participate in modulating channel kinetics. Each of the Sx1A Cys mutants, differently modify the kinetics of Cav1.2, and neuronal Cav2.2 calcium channel. Methodology/Principle ...

  11. A reverse Warburg metabolism in oral squamous cell carcinoma is not dependent upon myofibroblasts

    DEFF Research Database (Denmark)

    Jensen, David Hebbelstrup; Therkildsen, Marianne Hamilton; Dabelsteen, Erik

    2015-01-01

    expression of MCT-4 have been shown to have prognostic importance, primarily in patients with breast cancer. However, this phenomenon has only scarcely been described in oral squamous cell carcinoma (OSCC). Given the prognostic importance of myofibroblasts in OSCC, we also examined a potential relationship...... cancer observed as a lack of stromal caveolin-1 (CAV-1) and an increased expression of monocarboxylate transporter 4 (MCT-4) in the tumour stroma, with a concomitant increase in the expression of monocarboxylate transporter 1 (MCT-1) in the epithelial, tumour compartment. The lack of CAV-1 and increased...... between the expression of MCT-4 and the presence of myofibroblasts....

  12. Spinocerebellar Ataxia Type 6 Protein Aggregates Cause Deficits in Motor Learning and Cerebellar Plasticity

    OpenAIRE

    Mark, Melanie D.; Krause, Martin; Boele, Henk-Jan; Kruse, Wolfgang; Pollok, Stefan; Kuner, Thomas; Dalkara, Deniz; Koekkoek, Sebastiaan; De Zeeuw, Chris I.; Herlitze, Stefan

    2015-01-01

    Spinocerebellar ataxia type 6 (SCA6) is linked to poly-glutamine (polyQ) within the C terminus (CT) of the pore-forming subunits of P/Q-type Ca(2+) channels (Cav2.1) and is characterized by CT protein aggregates found in cerebellar Purkinje cells (PCs). One hypothesis regarding SCA6 disease is that a CT fragment of the Cav2.1 channel, which is detected specifically in cytosolic and nuclear fractions in SCA6 patients, is associated with the SCA6 pathogenesis. To test this hypothesis, we expres...

  13. Therapeutic potential of RQ-00311651, a novel T-type Ca2+ channel blocker, in distinct rodent models for neuropathic and visceral pain.

    Science.gov (United States)

    Sekiguchi, Fumiko; Kawara, Yuma; Tsubota, Maho; Kawakami, Eri; Ozaki, Tomoka; Kawaishi, Yudai; Tomita, Shiori; Kanaoka, Daiki; Yoshida, Shigeru; Ohkubo, Tsuyako; Kawabata, Atsufumi

    2016-08-01

    T-type Ca channels (T channels), particularly Cav3.2 among the 3 isoforms, play a role in neuropathic and visceral pain. We thus characterized the effects of RQ-00311651 (RQ), a novel T-channel blocker, in HEK293 cells transfected with human Cav3.1 or Cav3.2 by electrophysiological and fluorescent Ca signaling assays, and also evaluated the antiallodynic/antihyperalgesic activity of RQ in somatic, visceral, and neuropathic pain models in rodents. RQ-00311651 strongly suppressed T currents when tested at holding potentials of -65 ∼ -60 mV, but not -80 mV, in the Cav3.1- or Cav3.2-expressing cells. RQ-00311651 also inhibited high K-induced Ca signaling in those cells. In mice, RQ, administered intraperitoneally (i.p.) at 5 to 20 mg/kg or orally at 20 to 40 mg/kg, significantly suppressed the somatic hyperalgesia and visceral pain-like nociceptive behavior/referred hyperalgesia caused by intraplantar and intracolonic administration of NaHS or Na2S, H2S donors, respectively, which involve the enhanced activity of Cav3.2 channels. RQ-00311651, given i.p. at 5 to 20 mg/kg, exhibited antiallodynic or antihyperalgesic activity in rats with spinal nerve injury-induced neuropathy or in rats and mice with paclitaxel-induced neuropathy. Oral and i.p. RQ at 10 to 20 mg/kg also suppressed the visceral nociceptive behavior and/or referred hyperalgesia accompanying cerulein-induced acute pancreatitis and cyclophosphamide-induced cystitis in mice. The analgesic and antihyperalgesic/antiallodynic doses of oral and i.p. RQ did not significantly affect the locomotor activity and motor coordination. Together, RQ is considered a state-dependent blocker of Cav3.1/Cav3.2 T channels and may serve as an orally available analgesic for treatment of neuropathic and inflammatory pain including distinct visceral pain with minimum central side effects. PMID:27023424

  14. Produção de vectores de adenovírus caninos:optimização do processo na perspectiva de aumento de escala

    OpenAIRE

    Fernandes, Paulo David Machado dos Santos

    2009-01-01

    Introdução: Os vectores derivados de adenovírus caninos tipo 2 (CAV-2) possuem umgrande potencial em terapia génica uma vez que circunscrevem os problemas encontrados na utilização dos vectores virais mais comuns adenovírus humanos nomeadamente a memória imunitária o que implica o aumento de doses a administrar por pessoa. Até à data os vectores de CAV-2 são produzidos em frascos de cultura utilizando células DK como produtoras. Por isso é necessário desenvolver um sistema de cultura que ...

  15. Gabapentin suppresses cortical spreading depression susceptibility

    OpenAIRE

    Hoffmann, Ulrike; Dileköz, Ergin; Kudo, Chiho; Ayata, Cenk

    2010-01-01

    Cortical spreading depression (CSD) is an intense depolarization wave implicated in the pathophysiology of brain injury states and migraine aura. As Cav2.1 channels modulate CSD susceptibility, we tested gabapentin, which inhibits Cav2.1 through high-affinity binding to its α2δ subunit, on CSD susceptibility in anesthetized rats. Gabapentin, 100 or 200 mg/kg, elevated the electrical threshold for CSD and diminished recurrent CSDs evoked by topical KCl, when administered 1 hour before testing....

  16. Caveolin-1 is important for nitric oxide-mediated angiogenesis in fibrin gels with human umbilical vein endothelial cells

    Institute of Scientific and Technical Information of China (English)

    Yi-ming PAN; Yong-zhong YAO; Zhang-hua ZHU; Xi-tai SUN; Yu-dong QIU; Yi-tao DING

    2006-01-01

    Aim: The role of caveolin-l (Cav-1) in angiogenesis remains poorly understood. The endothelial nitric oxide (NO) synthase (eNOS), a caveolin-interacting protein, was demonstrated to play a predominant role in vascular endothelial growth factor (VEGF) -induced angiogenesis. The purpose of our study was to examine the role of Cav-1 and the eNOS complex in NO-mediated angiogenesis. Methods: Human umbilical vein endothelial cells (HUVEC) were isolated and cultured in 3-D fibrin gels to form capillary-like tubules by VEGF stimulation. The expression of Cav-1 and eNOS was detected by semiquantitative RT-PCR. The HUVEC were treated with antisense oligonucleotides to downregulate Cav-1 expression. Both transduced and non-infected HUVEC were cultured in fibrin gels in the presence or absence of VEGF (20 ng/mL) and NG-nitro-L-arginine methyl ester (L-NAME; 5 mmol/L). NO was measured using a NO assay kit and capillary-like tubules were quantified by tubule formation index using the Image J program. Results: RT-PCR analysis revealed that Cav-1 levels steadily increased in a time-dependent manner and reached their maximum after 5 d of incubation, but there were no obvious changes in eNOS mRNA expression in response to VEGF in the fibrin gel model. VEGF (20 ng/mL) can promote NO production and the formation of capillary-like tubules, and this promoting effect of VEGF was blocked by the addition of L-NAME (5 mmol/L). When transduced HUVEC with the antisense Cav-1 oligonucleotides were plated in the fibrin gels, the capillary-like tubules were significantly fewer than those of the non-infected cells. The capillary-like tubules formation and NO production of transduced HUVEC with the antisense Cav-1 oligonucleotides cultured in fibrin gels showed no responses to the addition of VEGF (20 ng/mL) and L-NAME (5.0 mmol/L). Conclusion: NO was a critical angiogenic mediator in this model. Cav-1 was essential for NO-mediated angiogenesis and may be an important target of anti

  17. Smooth muscle NOS, colocalized with caveolin-1, modulates contraction in mouse small intestine

    OpenAIRE

    El-Yazbi, Ahmed F.; Cho, Woo Jung; Cena, Jonathan; Schulz, Richard; Daniel, Edwin E

    2008-01-01

    Neuronal nitric oxide synthase (nNOS) in myenteric neurons is activated during peristalsis to produce nitric oxide which relaxes intestinal smooth muscle. A putative nNOS is also found in the membrane of intestinal smooth muscle cells in mouse and dog. In this study we studied the possible functions of this nNOS expressed in mouse small intestinal smooth muscle colocalized with caveolin-1(Cav-1). Cav-1 knockout mice lacked nNOS in smooth muscle and provided control tissues. 60 mM KCl was used...

  18. PCB 126 toxicity is modulated by cross-talk between caveolae and Nrf2 signaling

    Energy Technology Data Exchange (ETDEWEB)

    Petriello, Michael C. [Graduate Center for Toxicology, College of Medicine, University of Kentucky, Lexington, KY 40536 (United States); University of Kentucky Superfund Research Center, Lexington, KY 40536 (United States); Han, Sung Gu [University of Kentucky Superfund Research Center, Lexington, KY 40536 (United States); Department of Food Science and Biotechnology of Animal Resources, College of Animal Bioscience and Technology, Konkuk University, Seoul 143-701 (Korea, Republic of); Newsome, Bradley J. [University of Kentucky Superfund Research Center, Lexington, KY 40536 (United States); Department of Chemistry, College of Arts and Sciences, University of Kentucky, Lexington, KY 40506 (United States); Hennig, Bernhard [Graduate Center for Toxicology, College of Medicine, University of Kentucky, Lexington, KY 40536 (United States); University of Kentucky Superfund Research Center, Lexington, KY 40536 (United States); Department of Animal and Food Sciences, College of Agriculture, Food and Environment, University of Kentucky, KY 40506 (United States)

    2014-06-01

    Environmental toxicants such as polychlorinated biphenyls (PCBs) have been implicated in the promotion of multiple inflammatory disorders including cardiovascular disease, but information regarding mechanisms of toxicity and cross-talk between relevant cell signaling pathways is lacking. To examine the hypothesis that cross-talk between membrane domains called caveolae and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathways alters PCB-induced inflammation, caveolin-1 was silenced in vascular endothelial cells, resulting in a decreased PCB-induced inflammatory response. Cav-1 silencing (siRNA treatment) also increased levels of Nrf2-ARE transcriptional binding, resulting in higher mRNA levels of the antioxidant genes glutathione s-transferase and NADPH dehydrogenase quinone-1 in both vehicle and PCB-treated systems. Along with this upregulated antioxidant response, Cav-1 siRNA treated cells exhibited decreased mRNA levels of the Nrf2 inhibitory protein Keap1 in both vehicle and PCB-treated samples. Silencing Cav-1 also decreased protein levels of Nrf2 inhibitory proteins Keap1 and Fyn kinase, especially in PCB-treated cells. Further, endothelial cells from wildtype and Cav-1 −/− mice were isolated and treated with PCB to better elucidate the role of functional caveolae in PCB-induced endothelial inflammation. Cav-1 −/− endothelial cells were protected from PCB-induced cellular dysfunction as evidenced by decreased vascular cell adhesion molecule (VCAM-1) protein induction. Compared to wildtype cells, Cav-1 −/− endothelial cells also allowed for a more effective antioxidant response, as observed by higher levels of the antioxidant genes. These data demonstrate novel cross-talk mechanisms between Cav-1 and Nrf2 and implicate the reduction of Cav-1 as a protective mechanism for PCB-induced cellular dysfunction and inflammation. - Highlights: • Reduction of caveolin-1 protein protects against polychlorinated biphenyl toxicity. • Decreasing

  19. PCB 126 toxicity is modulated by cross-talk between caveolae and Nrf2 signaling

    International Nuclear Information System (INIS)

    Environmental toxicants such as polychlorinated biphenyls (PCBs) have been implicated in the promotion of multiple inflammatory disorders including cardiovascular disease, but information regarding mechanisms of toxicity and cross-talk between relevant cell signaling pathways is lacking. To examine the hypothesis that cross-talk between membrane domains called caveolae and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathways alters PCB-induced inflammation, caveolin-1 was silenced in vascular endothelial cells, resulting in a decreased PCB-induced inflammatory response. Cav-1 silencing (siRNA treatment) also increased levels of Nrf2-ARE transcriptional binding, resulting in higher mRNA levels of the antioxidant genes glutathione s-transferase and NADPH dehydrogenase quinone-1 in both vehicle and PCB-treated systems. Along with this upregulated antioxidant response, Cav-1 siRNA treated cells exhibited decreased mRNA levels of the Nrf2 inhibitory protein Keap1 in both vehicle and PCB-treated samples. Silencing Cav-1 also decreased protein levels of Nrf2 inhibitory proteins Keap1 and Fyn kinase, especially in PCB-treated cells. Further, endothelial cells from wildtype and Cav-1 −/− mice were isolated and treated with PCB to better elucidate the role of functional caveolae in PCB-induced endothelial inflammation. Cav-1 −/− endothelial cells were protected from PCB-induced cellular dysfunction as evidenced by decreased vascular cell adhesion molecule (VCAM-1) protein induction. Compared to wildtype cells, Cav-1 −/− endothelial cells also allowed for a more effective antioxidant response, as observed by higher levels of the antioxidant genes. These data demonstrate novel cross-talk mechanisms between Cav-1 and Nrf2 and implicate the reduction of Cav-1 as a protective mechanism for PCB-induced cellular dysfunction and inflammation. - Highlights: • Reduction of caveolin-1 protein protects against polychlorinated biphenyl toxicity. • Decreasing

  20. Leaky channels make weak muscles

    OpenAIRE

    George, Alfred L.

    2012-01-01

    Mutations in the skeletal muscle voltage-gated calcium channel (CaV1.1) have been associated with hypokalemic periodic paralysis, but how the pathogenesis of this disorder relates to the functional consequences of mutations was unclear. In this issue of the JCI, Wu and colleagues recapitulate the disease by generating a novel knock-in CaV1.1 mutant mouse and use this model to investigate the cellular and molecular features of pathogenesis. They demonstrated an aberrant muscle cell current con...

  1. Conditional intrinsic voltage oscillations in mature vertebrate neurons undergo specific changes in culture

    DEFF Research Database (Denmark)

    Guertin, Pierre A; Hounsgaard, Jørn

    2006-01-01

    cord of adult turtles maintainable for several weeks in culture conditions. N-methyl-D-aspartate (NMDA)-induced-tetrodotoxin (TTX)-resistant voltage oscillations in motoneurons were approximately 10 times faster in culture than in acute preparations. Oscillations in culture were abolished by NMDA...... receptor antagonists or by high extracellular Mg2+ concentrations. However, in contrast with results from motoneurons in the acute slice, NMDA-induced oscillations in culture did not depend on CaV1.3 channel activation as they still remained after nifedipine application. Other CaV1.3 channel...

  2. Deletion of Caveolin-1 Protects against Oxidative Lung Injury via Up-Regulation of Heme Oxygenase-1

    OpenAIRE

    Jin, Yang; Kim, Hong Pyo; Chi, Minli; Ifedigbo, Emeka; Stefan W. Ryter; Choi, Augustine M. K.

    2008-01-01

    Acute lung injury (ALI) is a major cause of morbidity and mortality in critically ill patients. Hyperoxia causes lung injury in animals and humans, and is an established model of ALI. Caveolin-1, a major constituent of caveolae, regulates numerous biological processes, including cell death and proliferation. Here we demonstrate that caveolin-1–null mice (cav-1−/−) were resistant to hyperoxia-induced death and lung injury. Cav-1−/− mice sustained reduced lung injury after hyperoxia as determin...

  3. Cell Volume Regulation and Signaling in 3T3-L1 Pre-adipocytes and Adipocytes

    DEFF Research Database (Denmark)

    Eduardsen, Kathrine; Larsen, Susanne; Novak, Ivana;

    2011-01-01

    osmotic cell shrinkage (RVI), accompanied by increased swelling-activated taurine efflux. However, caveolin-1 distribution was not detectably altered after osmotic swelling or shrinkage, and caveolae integrity, as studied by cholesterol depletion or expression of dominant negative Cav-1, was not required...... had no detectable effect on InsR, yet increased ERK1/2 phosphorylation. In conclusion, differentiated 3T3-L1 adipocytes exhibit greatly accelerated RVD and RVI responses and increased swelling-activated taurine efflux compared to pre-adipocytes. Furthermore, in pre-adipocytes, Cav-1/caveolae integrity...

  4. Virtual histology assessment of cardiac allograft vasculopathy following introduction of everolimus--results of a multicenter trial

    DEFF Research Database (Denmark)

    Arora, Stina Jørgensen; Erikstad, I; Ueland, T;

    2012-01-01

    In this 12-month multicenter Scandinavian study, 78 maintenance heart transplant (HTx) recipients randomized to everolimus with reduced calcineurin inhibitor (CNI) exposure or continued standard CNI-therapy underwent matched virtual histology (VH) examination to evaluate morphological progression...... of cardiac allograft vasculopathy (CAV). Parallel measurement of a range of inflammatory markers was also performed. A similar rate of quantitative CAV progression was observed in the everolimus (n = 30) and standard CNI group (n = 48) (plaque index 1.9 ± 3.8% and 1.6 ± 3.9%, respectively; p = 0...

  5. Calcineurin Controls Voltage-Dependent-Inactivation (VDI) of the Normal and Timothy Cardiac Channels

    OpenAIRE

    Cohen-Kutner, Moshe; Yahalom, Yfat; Trus, Michael; Atlas, Daphne

    2012-01-01

    Ca2+-entry in the heart is tightly controlled by Cav1.2 inactivation, which involves Ca2+-dependent inactivation (CDI) and voltage-dependent inactivation (VDI) components. Timothy syndrome, a subtype-form of congenital long-QT syndrome, results from a nearly complete elimination of VDI by the G406R mutation in the α11.2 subunit of Cav1.2. Here, we show that a single (A1929P) or a double mutation (H1926A-H1927A) within the CaN-binding site at the human C-terminal tail of α11.2, accelerate the ...

  6. G Protein Regulation of Neuronal Calcium Channels: Back to the Future

    Czech Academy of Sciences Publication Activity Database

    Proft, Juliane; Weiss, Norbert

    2015-01-01

    Roč. 87, č. 6 (2015), s. 890-906. ISSN 0026-895X R&D Projects: GA ČR GA15-13556S Institutional support: RVO:61388963 Keywords : voltage gated calcium channels Cav * G proteins * GPCR Subject RIV: CE - Biochemistry Impact factor: 4.128, year: 2014

  7. T-type channels: release a brake, engage a gear

    Czech Academy of Sciences Publication Activity Database

    Weiss, Norbert; Laconova, L.

    2016-01-01

    Roč. 10, č. 2 (2016), s. 78-80. ISSN 1933-6950 Institutional support: RVO:61388963 Keywords : gating brake * pore opening * Ca(V)3.3 Subject RIV: CE - Biochemistry Impact factor: 2.410, year: 2014 http://www.tandfonline.com/doi/pdf/10.1080/19336950.2015.1106655

  8. Pharmacological Inhibition of Voltage-gated Ca(2+) Channels for Chronic Pain Relief.

    Science.gov (United States)

    Lee, Seungkyu

    2013-12-01

    Chronic pain is a major therapeutic problem as the current treatment options are unsatisfactory with low efficacy and deleterious side effects. Voltage-gated Ca2+ channels (VGCCs), which are multi-complex proteins consisting of α1, β, γ, and α2δ subunits, play an important role in pain signaling. These channels are involved in neurogenic inflammation, excitability, and neurotransmitter release in nociceptors. It has been previously shown that N-type VGCCs (Cav2.2) are a major pain target. U.S. FDA approval of three Cav2.2 antagonists, gabapentin, pregabalin, and ziconotide, for chronic pain underlies the importance of this channel subtype. Also, there has been increasing evidence that L-type (Cav1.2) or T-type (Cav3.2) VGCCs may be involved in pain signaling and chronic pain. In order to develop novel pain therapeutics and to understand the role of VGCC subtypes, discovering subtype selective VGCC inhibitors or methods that selectively target the inhibitor into nociceptors would be essential. This review describes the various VGCC subtype inhibitors and the potential of utilizing VGCC subtypes as targets of chronic pain. Development of VGCC subtype inhibitors and targeting them into nociceptors will contribute to a better understanding of the roles of VGCC subtypes in pain at a spinal level as well as development of a novel class of analgesics for chronic pain. PMID:24396337

  9. siRNA-induced in vivo downregulation of L-type calcium channels in rat small mesenteric arteries

    DEFF Research Database (Denmark)

    Matchkov, Vladimir; Larsen, Per; Kold-Petersen, Henrik Houmann;

    2008-01-01

    Ca2+ entry via L-type voltage-gated Ca2+ channels (Cav1.2) is a key factor in regulation of excitation-contraction coupling in smooth muscle cells (SMCs). Previous gene deletion studies have provided insight into the critical role of the pore-forming α1C subunit in regulation of blood pressure...

  10. P/Q-type and T-type voltage-gated calcium channels are involved in the contraction of mammary and brain blood vessels from hypertensive patients

    DEFF Research Database (Denmark)

    Thuesen, A D; Lyngsø, K S; Rasmussen, L;

    2016-01-01

    AIM: Calcium channel blockers are widely used in cardiovascular diseases. Besides L-type channels, T- and P/Q-type calcium channels are involved in the contraction of human renal blood vessels. It was hypothesized that T- and P/Q-type channels are involved in the contraction of human brain and...... P/Q-type antagonist ω-agatoxin IVA (10(-8) mol L(-1) ) and the T-type calcium blocker mibefradil (10(-7) mol L(-1) ) inhibited KCl depolarization-induced contraction in mammary arteries from hypertensive patients with no effect on blood vessels from normotensive patients. ω-Agatoxin IVA decreased...... contraction in cerebral arterioles from hypertensive patients. L-type blocker nifedipine abolished the contraction in mammary arteries. PCR analysis showed expression of P/Q-type (Cav 2.1), T-type (Cav 3.1 and Cav 3.2) and L-type (Cav 1.2) calcium channels in mammary and cerebral arteries. Immunohistochemical...

  11. Premières données sur la pharmacopée traditionnelle des Maya K'iche (Guatemala)

    OpenAIRE

    Nicolas, J. P.; Saravia, A.; Delelis, A.

    1996-01-01

    First data of traditional pharmacopoeias from the Maya K'iché (Guatemala). Practice of traditional medicine/ethnopharmacological investigation on #Tagetes lucida$ Cav. Ic., #Psidium guajava$ L., #Plantago major$ L., #Chenopodium ambrosioides$ L. and #Aloe vera$ L. Mains indications about the traditional pharmacopoeias : the diarrhoeas, acute breathing difficulties, parasites transmitted diseases, dermatological aliments. (Résumé d'auteur)

  12. Shoot demographics for melaleuca and impacts of simulated herbivory on vegetative development

    Science.gov (United States)

    We monitored branches of the invasive tree Melaleuca quinquenervia (Cav.) Blake (melaleuca) at two sites in southern Florida between January 2002 and May 2003 to elucidate patterns in vegetative shoot maturation. Six shoot stages characterize development from dormancy to fully expanded and lignified...

  13. Influence of Temperature, Humidity, and Plant Terpenoid Profiles on Life History Characteristics of Boreioglycaspis melaleucae (Hemiptera: Psyllidae), a Biological Control Agent of the Invasive Tree Melaleuca quinquenervia

    Science.gov (United States)

    Although the introduced weed biological control agent Boreioglycaspis melaleucae Moore is widely established among stands of its host Melaleuca quinquenervia (Cav.) Blake (Myrtaceae) in south Florida, it’s population densities decline markedly during summer months. We investigated the hypothesis tha...

  14. The effects of litter on growth and plasticity of Phragmites australis clones originating from infertile, fertile or eutrophicated habitats

    NARCIS (Netherlands)

    Clevering, O.A.

    1999-01-01

    In many European countries a strong decline of Phragmites australis (Cav.) Trin. Ex Steudel (common reed) has been observed. In some instances this decline has been related to the accumulation of litter. A greenhouse experiment was conducted with P. australis cuttings from different stable and die-b

  15. Cardiac voltage-gated calcium channel macromolecular complexes.

    Science.gov (United States)

    Rougier, Jean-Sébastien; Abriel, Hugues

    2016-07-01

    Over the past 20years, a new field of research, called channelopathies, investigating diseases caused by ion channel dysfunction has emerged. Cardiac ion channels play an essential role in the generation of the cardiac action potential. Investigators have largely determined the physiological roles of different cardiac ion channels, but little is known about the molecular determinants of their regulation. The voltage-gated calcium channel Cav1.2 shapes the plateau phase of the cardiac action potential and allows the influx of calcium leading to cardiomyocyte contraction. Studies suggest that the regulation of Cav1.2 channels is not uniform in working cardiomyocytes. The notion of micro-domains containing Cav1.2 channels and different calcium channel interacting proteins, called macro-molecular complex, has been proposed to explain these observations. The objective of this review is to summarize the currently known information on the Cav1.2 macromolecular complexes in the cardiac cell and discuss their implication in cardiac function and disorder. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel. PMID:26707467

  16. Sodium current and potassium transient outward current genes in Brugada syndrome

    DEFF Research Database (Denmark)

    Holst, Anders G; Saber, Siamak; Houshmand, Massoud;

    2012-01-01

    -frequency genetic variants and true monogenetic disease-causing variants. We aimed to screen the genes SCN1B through SCN4B, MOG1, CAV3, and KCND3 for variations in a population of SCN5A negative Danish and Iranian BrS patients, as well as research prior associations using newly released exome data....

  17. he land manager’s handbook on integrated pest management of Melaleuca quinquenervia

    Science.gov (United States)

    The adventive Australian tree Melaleuca quinquenervia (Cav.) S.T. Blake is an invasive pest plant in the greater Everglades region of Florida. Public agencies and organizations responsible for natural areas management have developed effective chemical and mechanical strategies for treating infestati...

  18. Fluid composition analysis device and method

    DEFF Research Database (Denmark)

    2014-01-01

    The invention relates in particular to a device for determining one or more fluid properties of a fluid. The device preferably comprises a flow distributor comprising a cavity, an inlet to the cavity, one or more outlets from the cavity, and one or more flow deflecting elements present in the cav...

  19. Management of Solanum elaeagnifolium in the Mediterranean Basin

    Science.gov (United States)

    Solanum elaeagnifolium Cav. (silverleaf nightshade, SOLEL) is a prominent invasive alien weed in many countries of the Mediterranean Basin since its introduction in the mid-20th century, originating from the southwestern United States and northern Mexico. It reproduces vegetatively and by seeds that...

  20. siRNA-induced in vivo downregulation of L-type calcium channels in rat small mesenteric arteries

    DEFF Research Database (Denmark)

    Møller, Kate; Aalkjær, Christian; Matchkov, Vladimir

    2009-01-01

    Previous gene deletion studies have provided insight into the critical role of L-type voltage-gated Ca2+ channels (Cav1.2) in regulation of blood pressure. Homozygous knockout is, however, lethal but this limitation can be overcome by a small RNA interference (siRNA). A specific downregulation of...

  1. Genetics Home Reference: Timothy syndrome

    Science.gov (United States)

    ... and the regulation of certain genes. CaV1.2 calcium channels are particularly important for the normal function of ... Severe arrhythmia disorder caused by cardiac L-type calcium channel mutations. Proc Natl Acad Sci U S A. 2005 ...

  2. Stac gets the skeletal L-type calcium channel unstuck

    Czech Academy of Sciences Publication Activity Database

    Weiss, Norbert

    2015-01-01

    Roč. 34, č. 2 (2015), s. 101-103. ISSN 0231-5882 Institutional support: RVO:61388963 Keywords : calcium channel * L-type calcium channel * Ca(v)1.1 channel * Stac adaptor protein * excitation-contraction coupling * trafficking Subject RIV: CE - Biochemistry Impact factor: 1.173, year: 2014

  3. Overzicht der in Nederland gevonden inheemse en adventieve Erodium-soorten

    NARCIS (Netherlands)

    Bruggeman, Catharina J.M.

    1964-01-01

    An enumeration of the species of Erodium, found in the Netherlands. E. glutinosum Dum. and E. cicutarium (L.) L’Hérit. are native; E. malacoides (L.) L’Hérit., E. crinitum Carolin, E. botrys (Cav.) Bertol., E. gruinum (L.) L’Herit., E. ciconium (L.) L’Herit., E. stephanianum Willd., E. aethiopicum (

  4. Saving energy for ventilation using decentralised duct fans

    DEFF Research Database (Denmark)

    Gunner, Amalie; Bergsøe, Niels Christian; Afshari, Alireza

    2014-01-01

    pressure drops which in turn will increase energy use. This paper presents a novel procedure for balancing CAV-systems using decentralised duct fans instead of flat plate dampers. The procedure will result in lower pressure drops and consequently in a reduction of energy use for ventilation when compared...... mechanical ventilation systems with low energy use for new and renovation of buildings....

  5. Molecular characterization and expression analysis of caveolin-1 in pig tissues

    Institute of Scientific and Technical Information of China (English)

    WANG Chong; MEI YingJie; LI Li; MO DeLin; LI JiaQi; ZHANG Hao; TIAN XingGuo; CHEN YaoSheng

    2008-01-01

    Members of the caveolin family played important roles during fundamental cellular processes, such as regulation of cell morphology, migration, and gene expression in muscle cells. In this study, caveolin-1 (Cav-1), one of the caveolins, was identified from Iongissimus dorsi muscle of Large Yorkshire pig and Chinese indigenous Lantang pig based on the results of mRNA differential display analysis. The de-duced amino acids sequence of the porcine Cav-1 contained a caveolin domain, and was very conser-vative among different species. The Cav-1 mRNA was widely expressed in the eight tissues in this study, including heart, liver, kidney, encephalon, spleen, lung, Iongissimus dorsi muscle, and back fat, and the highest expression quantity was found in back fat of the two pig breeds. The expression quan-tity of porcine Car-1 in back fat and Iongissimus dorsi muscle of Lantang pig was significantly higher than that of Large Yorkshire (P<0.01, and P<0.05, respectively). These results suggested that the Cav-1 might be a candidate gene for carcass traits, and might provide valuable information for understanding the mechanism of caveolae signaling in fat deposition by using the animal model of pig.

  6. Tyrosine-phosphorylated caveolin-1 blocks bacterial uptake by inducing Vav2-RhoA-mediated cytoskeletal rearrangements.

    Directory of Open Access Journals (Sweden)

    Jan Peter Boettcher

    Full Text Available Certain bacterial adhesins appear to promote a pathogen's extracellular lifestyle rather than its entry into host cells. However, little is known about the stimuli elicited upon such pathogen host-cell interactions. Here, we report that type IV pili (Tfp-producing Neisseria gonorrhoeae (P(+GC induces an immediate recruitment of caveolin-1 (Cav1 in the host cell, which subsequently prevents bacterial internalization by triggering cytoskeletal rearrangements via downstream phosphotyrosine signaling. A broad and unbiased analysis of potential interaction partners for tyrosine-phosphorylated Cav1 revealed a direct interaction with the Rho-family guanine nucleotide exchange factor Vav2. Both Vav2 and its substrate, the small GTPase RhoA, were found to play a direct role in the Cav1-mediated prevention of bacterial uptake. Our findings, which have been extended to enteropathogenic Escherichia coli, highlight how Tfp-producing bacteria avoid host cell uptake. Further, our data establish a mechanistic link between Cav1 phosphorylation and pathogen-induced cytoskeleton reorganization and advance our understanding of caveolin function.

  7. Exercise-Induced Changes in Caveolin-1, Depletion of Mitochondrial Cholesterol, and the Inhibition of Mitochondrial Swelling in Rat Skeletal Muscle but Not in the Liver

    Directory of Open Access Journals (Sweden)

    Damian Jozef Flis

    2016-01-01

    Full Text Available The reduction in cholesterol in mitochondria, observed after exercise, is related to the inhibition of mitochondrial swelling. Caveolin-1 (Cav-1 plays an essential role in the regulation of cellular cholesterol metabolism and is required by various signalling pathways. Therefore, the aim of this study was to investigate the effect of prolonged swimming on the mitochondrial Cav-1 concentration; additionally, we identified the results of these changes as they relate to the induction of changes in the mitochondrial swelling and cholesterol in rat skeletal muscle and liver. Male Wistar rats were divided into a sedentary control group and an exercise group. The exercised rats swam for 3 hours and were burdened with an additional 3% of their body weight. After the cessation of exercise, their quadriceps femoris muscles and livers were immediately removed for experimentation. The exercise protocol caused an increase in the Cav-1 concentration in crude muscle mitochondria; this was related to a reduction in the cholesterol level and an inhibition of mitochondrial swelling. There were no changes in rat livers, with the exception of increased markers of oxidative stress in mitochondria. These data indicate the possible role of Cav-1 in the adaptive change in the rat muscle mitochondria following exercise.

  8. Blockade of T-type calcium channels prevents tonic-clonic seizures in a maximal electroshock seizure model.

    Science.gov (United States)

    Sakkaki, Sophie; Gangarossa, Giuseppe; Lerat, Benoit; Françon, Dominique; Forichon, Luc; Chemin, Jean; Valjent, Emmanuel; Lerner-Natoli, Mireille; Lory, Philippe

    2016-02-01

    T-type (Cav3) calcium channels play important roles in neuronal excitability, both in normal and pathological activities of the brain. In particular, they contribute to hyper-excitability disorders such as epilepsy. Here we have characterized the anticonvulsant properties of TTA-A2, a selective T-type channel blocker, in mouse. Using the maximal electroshock seizure (MES) as a model of tonic-clonic generalized seizures, we report that mice treated with TTA-A2 (0.3 mg/kg and higher doses) were significantly protected against tonic seizures. Although no major change in Local Field Potential (LFP) pattern was observed during the MES seizure, analysis of the late post-ictal period revealed a significant increase in the delta frequency power in animals treated with TTA-A2. Similar results were obtained for Cav3.1-/- mice, which were less prone to develop tonic seizures in the MES test, but not for Cav3.2-/- mice. Analysis of extracellular signal-regulated kinase 1/2 (ERK) phosphorylation and c-Fos expression revealed a rapid and elevated neuronal activation in the hippocampus following MES clonic seizures, which was unchanged in TTA-A2 treated animals. Overall, our data indicate that TTA-A2 is a potent anticonvulsant and that the Cav3.1 isoform plays a prominent role in mediating TTA-A2 tonic seizure protection. PMID:26456350

  9. Identification of a major protein on the cytosolic face of caveolae

    DEFF Research Database (Denmark)

    Vinten, J; Johnsen, A H; Roepstorff, Peter;

    2005-01-01

    Cav-p60, a specific and ubiquitous caveolar protein, was immunoprecipitated from solubilized rat adipocyte plasma membranes and identified as similar to a GeneBank entry annotated mouse polymerase transcript release factor (PTRF) by MALDI-TOF and MS-MS of major fragments. Cloning and virtual tran...

  10. Diseases of the nervous system associated with calcium channelopathies

    NARCIS (Netherlands)

    Todorov, Boyan Bogdanov

    2010-01-01

    The aim of the studies described in this thesis was to investigate how abnormal CaV2.1 channel function can cause disease, in particular motor coordination dysfunction. The chapters illustrate how various neuronal cell types in the periphery (peripheral nervous system) and the central nervous system

  11. Inhibition of Voltage-Gated Calcium Channels by RGK Proteins.

    Science.gov (United States)

    Buraei, Zafir; Yang, Jian

    2015-01-01

    Due to their essential biological roles, voltage-gated calcium channels (VGCCs) are regulated by a myriad of molecules and mechanisms. Fifteen years ago, RGK proteins were discovered to bind the VGCC β subunit (Cavβ) and potently inhibit high-voltage activated Ca(2+) channels. RGKs (Rad, Rem, Rem2 and Gem/Kir) are a family of monomeric small GTPases belonging to the superfamily of Ras GTPases. They exert dual inhibitory effects on VGCCs, decreasing surface expression and suppressing surface channels through immobilization of the voltage sensor or reduction of channel open probability. While Cavβ is required for all forms of RGK inhibition, not all inhibition is mediated by the RGK-Cavβ interaction. Some RGK proteins also interact directly with the pore-forming α1 subunit of some types of VGCCs (Cavα1). Importantly, RGK proteins tonically inhibit VGCCs in native cells, regulating cardiac and neural functions. This minireview summarizes the mechanisms, molecular determinants, and physiological impact of RGK inhibition of VGCCs. PMID:25966691

  12. Uma nova espécie de Centris Fabricius (Hymenoptera, Apoidea, Anthophoridae do Nordeste do Brasil A new species of Centris Fabricius (Hymenoptera, Apoidea, Anthophoridae from Northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Jesus Santiago Moure

    2001-07-01

    Full Text Available A new species of Centris Fabricius, 1804 is described as C. xanthomelaena (body size 15 mm, forewing length 10.33 mm, head width 5.25 mm, second abdominal tergum width 5.75 mm. The specimens were captured when visiting the yellow flowers of Chamaecrista amiciela (I. & B. I. & B., Caesalpiniaceae and of Stimaphyllom auriculatum (Cav. Adr. Juss, Malpighiaceae.

  13. MURC deficiency in smooth muscle attenuates pulmonary hypertension

    Science.gov (United States)

    Nakanishi, Naohiko; Ogata, Takehiro; Naito, Daisuke; Miyagawa, Kotaro; Taniguchi, Takuya; Hamaoka, Tetsuro; Maruyama, Naoki; Kasahara, Takeru; Nishi, Masahiro; Matoba, Satoaki; Ueyama, Tomomi

    2016-01-01

    Emerging evidence suggests that caveolin-1 (Cav1) is associated with pulmonary arterial hypertension. MURC (also called Cavin-4) is a member of the cavin family, which regulates caveolar formation and functions together with caveolins. Here, we show that hypoxia increased Murc mRNA expression in the mouse lung, and that Murc-null mice exhibited attenuation of hypoxia-induced pulmonary hypertension (PH) accompanied by reduced ROCK activity in the lung. Conditional knockout mice lacking Murc in smooth muscle also resist hypoxia-induced PH. MURC regulates the proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) through Rho/ROCK signalling. Cav1 suppresses RhoA activity in PASMCs, which is reversed by MURC. MURC binds to Cav1 and inhibits the association of Cav1 with the active form of Gα13, resulting in the facilitated association of the active form of Gα13 with p115RhoGEF. These results reveal that MURC has a function in the development of PH through modulating Rho/ROCK signalling. PMID:27546070

  14. Cellular Prion Protein and Caveolin-1 Interaction in a Neuronal Cell Line Precedes Fyn/Erk 1/2 Signal Transduction

    Directory of Open Access Journals (Sweden)

    Mattia Toni

    2006-01-01

    Full Text Available It has been reported that cellular prion protein (PrPc is enriched in caveolae or caveolae-like domains with caveolin-1 (Cav-1 participating to signal transduction events by Fyn kinase recruitment. By using the Glutathione-S-transferase (GST-fusion proteins assay, we observed that PrPc strongly interacts in vitro with Cav-1. Thus, we ascertained the PrPc caveolar localization in a hypothalamic neuronal cell line (GN11, by confocal microscopy analysis, flotation on density gradient, and coimmunoprecipitation experiments. Following the anti-PrPc antibody-mediated stimulation of live GN11 cells, we observed that PrPc clustered on plasma membrane domains rich in Cav-1 in which Fyn kinase converged to be activated. After these events, a signaling cascade through p42/44 MAP kinase (Erk 1/2 was triggered, suggesting that following translocations from rafts to caveolae or caveolae-like domains PrPc could interact with Cav-1 and induce signal transduction events.

  15. Cytoprotective effects of chalcones from Zuccagnia punctata and melatonin on the gastroduodenal tract in rats.

    Science.gov (United States)

    de la Rocha, N; María, A O M; Gianello, J C; Pelzer, L

    2003-07-01

    The effects of 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone from Zuccagnia punctata Cav. (Fabaceae) and melatonin administration on ethanol-induced gastroduodenal injury were investigated in rats. Both chalcones showed significant preventive effects in treatment with melatonin previous to the necrotising agent. These effects could be due, in part, to the radical scavenging activity of the melatonin. PMID:12770521

  16. Biophysical Methods to Analyze Direct G-Protein Regulation of Neuronal Voltage-Gated Calcium Channels

    Czech Academy of Sciences Publication Activity Database

    Weiss, Norbert; De Waard, M.

    New York: Humana Press, 2016 - (Luján, R.; Ciruela, F.), s. 357-368. (Neuromethods. 110). ISBN 978-1-4939-3063-0 Institutional support: RVO:61388963 Keywords : calciumchannel * Ca(v)2 channel * G-protein-coupled receptor * G-proteins * G beta gamma-dimer Subject RIV: CE - Biochemistry

  17. Modelling collapse aggresiveness of cavitation bubbles in hydromachinery

    Czech Academy of Sciences Publication Activity Database

    Zima, Patrik; Sedlář, M.; Müller, M.

    Ann Arbor: University of Michigan, 2009. s. 21-21. [CAV2009 International Symposium on Cavitation /7./. 16.08.2009-20.08.2009, Ann Arbor] R&D Projects: GA ČR(CZ) GA101/07/1612 Institutional research plan: CEZ:AV0Z20760514 Keywords : cavitation * erosion * pumps Subject RIV: BK - Fluid Dynamics

  18. AN RNA EXTRACTION PROTOCOL FOR SHELLFISH-BORNE VIRUSES

    Science.gov (United States)

    The GPTT virus RNA extraction method, originally developed for extraction of human norovirus and hepatitis A virus RNAs from contaminated shellfish, was evaluated for extraction of RNA from Aichi virus strain A846/88 (AiV), coxsackievirus strains A9 (CAV9) and B5 (CBV5), murine norovirus (strain MNV...

  19. Soroprevalência das infecções por parvovírus, adenovírus, coronavírus canino e pelo vírus da cinomose em cães de Santa Maria, Rio Grande do Sul, Brasil Seroprevalence of parvovirus, adenovirus, coronavirus and canine distemper virus infections in dogs of Santa Maria, Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Renata Dezengrini

    2007-02-01

    Full Text Available As infecções pelo vírus da cinomose (CDV, por parvovírus (CPV, adenovírus (CAV e coronavírus (CCoV são importantes causas de morbidade e de mortalidade em cães de todo o mundo, porém pouco se sabe sobre a sua incidência e prevalência no Brasil. Para determinar-se a prevalência dessas infecções na população canina de Santa Maria, RS, Brasil, amostras de sangue foram coletadas de 817 cães não-vacinados de 14 bairros do município e testadas para a presença de anticorpos específicos. Anticorpos contra o CDV foram detectados em 27,3% (223/817 das amostras, contra o CPV em 68,7% (561/817, contra o CAV em 43% (353/817 e contra o CCoV em 50,4% (412/817 dos cães. Observou-se um aumento gradativo da prevalência de anticorpos de acordo com a idade para o CDV, o CAV e o CCoV. Os índices de positividade para o CPV, o CAV e o CCoV foram um pouco superiores entre machos, e semelhantes entre os sexos para o CDV. Os animais que convivem com outros cães em casa ou na rua apresentaram prevalência maior de anticorpos para o CDV e o CCoV do que cães sem contato ou convívio, enquanto que, para o CPV e o CAV, não houve diferença. Esses resultados demonstram que esses vírus estão difundidos na população canina dos bairros da cidade. Por outro lado, demonstram também que uma parte considerável da população é soronegativa e, portanto, está desprotegida frente a esses agentes, indicando a necessidade de se ampliar a cobertura vacinal.Canine distemper virus (CDV, parvovirus (CPV, adenovirus (CAV and coronavirus (CCoV infections have been associated with significant morbidity and mortality among dogs worldwide yet very little is known about these infections in Brazil. As to determine the prevalence of these infections in the canine population of Santa Maria, RS, Brazil, 817 blood samples were collected from non-vaccinated dogs of 14 neighborhoods and tested for specific antibodies. Antibodies to CDV were detected in 27.3% (223/817 of

  20. Epithelial membrane protein 2 regulates sphingosylphosphorylcholine-induced keratin 8 phosphorylation and reorganization: Changes of PP2A expression by interaction with alpha4 and caveolin-1 in lung cancer cells.

    Science.gov (United States)

    Lee, Eun Ji; Park, Mi Kyung; Kim, Hyun Ji; Kim, Eun Ji; Kang, Gyeoung-Jin; Byun, Hyun Jung; Lee, Chang Hoon

    2016-06-01

    Sphingosylphosphorylcholine (SPC) is found at increased in the malignant ascites of tumor patients and induces perinuclear reorganization of keratin 8 (K8) filaments that contribute to the viscoelasticity of metastatic cancer cells. However, the detailed mechanism of SPC-induced K8 phosphorylation and reorganization is not clear. We observed that SPC dose-dependently reduced the expression of epithelial membrane protein 2 (EMP2) in lung cancer cells. Then, we examined the role of EMP2 in SPC-induced phosphorylation and reorganization of K8 in lung cancer cells. We found that SPC concentration-dependently reduced EMP2 in A549, H1299, and other lung cancer cells. This was verified at the mRNA level by RT-PCR and real-time PCR (qPCR), and intracellular variation through confocal microscopy. EMP2 gene silencing and stable lung cancer cell lines established using EMP2 lentiviral shRNA induced K8 phosphorylation and reorganization. EMP2 overexpression reduced K8 phosphorylation and reorganization. We also observed that SPC-induced loss of EMP2 induces phosphorylation of JNK and ERK via reduced expression of protein phosphatase 2A (PP2A). Loss of EMP2 induces ubiquitination of protein phosphatase 2A (PP2A). SPC induced caveolin-1 (cav-1) expression and EEA1 endosome marker protein but not cav-2. SPC treatment enhanced the binding of cav-1 and PP2A and lowered binding of PP2A and alpha4. Gene silencing of EMP2 increased and gene silencing of cav-1 reduced migration of A549 lung cancer cells. Overall, these results suggest that SPC induces EMP2 down-regulation which reduces the PP2A via ubiquitination induced by cav-1, which sequestered alpha4, leading to the activation of ERK and JNK. PMID:26876307

  1. Stimulation of ICa by basal PKA activity is facilitated by caveolin-3 in cardiac ventricular myocytes.

    Science.gov (United States)

    Bryant, Simon; Kimura, Tomomi E; Kong, Cherrie H T; Watson, Judy J; Chase, Anabelle; Suleiman, M Saadeh; James, Andrew F; Orchard, Clive H

    2014-03-01

    L-type Ca channels (LTCC), which play a key role in cardiac excitation-contraction coupling, are located predominantly at the transverse (t-) tubules in ventricular myocytes. Caveolae and the protein caveolin-3 (Cav-3) are also present at the t-tubules and have been implicated in localizing a number of signaling molecules, including protein kinase A (PKA) and β2-adrenoceptors. The present study investigated whether disruption of Cav-3 binding to its endogenous binding partners influenced LTCC activity. Ventricular myocytes were isolated from male Wistar rats and LTCC current (ICa) recorded using the whole-cell patch-clamp technique. Incubation of myocytes with a membrane-permeable peptide representing the scaffolding domain of Cav-3 (C3SD) reduced basal ICa amplitude in intact, but not detubulated, myocytes, and attenuated the stimulatory effects of the β2-adrenergic agonist zinterol on ICa. The PKA inhibitor H-89 also reduced basal ICa; however, the inhibitory effects of C3SD and H-89 on basal ICa amplitude were not summative. Under control conditions, myocytes stained with antibody against phosphorylated LTCC (pLTCC) displayed a striated pattern, presumably reflecting localization at the t-tubules. Both C3SD and H-89 reduced pLTCC staining at the z-lines but did not affect staining of total LTCC or Cav-3. These data are consistent with the idea that the effects of C3SD and H-89 share a common pathway, which involves PKA and is maximally inhibited by H-89, and suggest that Cav-3 plays an important role in mediating stimulation of ICa at the t-tubules via PKA-induced phosphorylation under basal conditions, and in response to β2-adrenoceptor stimulation. PMID:24412535

  2. L-type voltage-operated calcium channels contribute to astrocyte activation In vitro.

    Science.gov (United States)

    Cheli, Veronica T; Santiago González, Diara A; Smith, Jessica; Spreuer, Vilma; Murphy, Geoffrey G; Paez, Pablo M

    2016-08-01

    We have found a significant upregulation of L-type voltage-operated Ca(++) channels (VOCCs) in reactive astrocytes. To test if VOCCs are centrally involved in triggering astrocyte reactivity, we used in vitro models of astrocyte activation in combination with pharmacological inhibitors, siRNAs and the Cre/lox system to reduce the activity of L-type VOCCs in primary cortical astrocytes. The endotoxin lipopolysaccharide (LPS) as well as high extracellular K(+) , glutamate, and ATP promote astrogliosis in vitro. L-type VOCC inhibitors drastically reduce the number of reactive cells, astrocyte hypertrophy, and cell proliferation after these treatments. Astrocytes transfected with siRNAs for the Cav1.2 subunit that conducts L-type Ca(++) currents as well as Cav1.2 knockout astrocytes showed reduce Ca(++) influx by ∼80% after plasma membrane depolarization. Importantly, Cav1.2 knock-down/out prevents astrocyte activation and proliferation induced by LPS. Similar results were found using the scratch wound assay. After injuring the astrocyte monolayer, cells extend processes toward the cell-free scratch region and subsequently migrate and populate the scratch. We found a significant increase in the activity of L-type VOCCs in reactive astrocytes located in the growing line in comparison to quiescent astrocytes situated away from the scratch. Moreover, the migration of astrocytes from the scratching line as well as the number of proliferating astrocytes was reduced in Cav1.2 knock-down/out cultures. In summary, our results suggest that Cav1.2 L-type VOCCs play a fundamental role in the induction and/or proliferation of reactive astrocytes, and indicate that the inhibition of these Ca(++) channels may be an effective way to prevent astrocyte activation. GLIA 2016. GLIA 2016;64:1396-1415. PMID:27247164

  3. Methamphetamine blunts Ca(2+) currents and excitatory synaptic transmission through D1/5 receptor-mediated mechanisms in the mouse medial prefrontal cortex.

    Science.gov (United States)

    González, Betina; Rivero-Echeto, Celeste; Muñiz, Javier A; Cadet, Jean Lud; García-Rill, Edgar; Urbano, Francisco J; Bisagno, Verónica

    2016-05-01

    Psychostimulant addiction is associated with dysfunctions in frontal cortex. Previous data demonstrated that repeated exposure to methamphetamine (METH) can alter prefrontal cortex (PFC)-dependent functions. Here, we show that withdrawal from repetitive non-contingent METH administration (7 days, 1 mg/kg) depressed voltage-dependent calcium currents (ICa ) and increased hyperpolarization-activated cation current (IH ) amplitude and the paired-pulse ratio of evoked excitatory postsynaptic currents (EPSCs) in deep-layer pyramidal mPFC neurons. Most of these effects were blocked by systemic co-administration of the D1/D5 receptor antagonist SCH23390 (0.5 and 0.05 mg/kg). In vitro METH (i.e. bath-applied to slices from naïve-treated animals) was able to emulate its systemic effects on ICa and evoked EPSCs paired-pulse ratio. We also provide evidence of altered mRNA expression of (1) voltage-gated calcium channels P/Q-type Cacna1a (Cav 2.1), N-type Cacna1b (Cav 2.2), T-type Cav 3.1 Cacna1g, Cav 3.2 Cacna1h, Cav 3.3 Cacna1i and the auxiliary subunit Cacna2d1 (α2δ1); (2) hyperpolarization-activated cyclic nucleotide-gated channels Hcn1 and Hcn2; and (3) glutamate receptors subunits AMPA-type Gria1, NMDA-type Grin1 and metabotropic Grm1 in the mouse mPFC after repeated METH treatment. Moreover, we show that some of these changes in mRNA expression were sensitive D1/5 receptor blockade. Altogether, these altered mechanisms affecting synaptic physiology and transcriptional regulation may underlie PFC functional alterations that could lead to PFC impairments observed in METH-addicted individuals. PMID:25871318

  4. The Physiology, Pathology, and Pharmacology of Voltage-Gated Calcium Channels and Their Future Therapeutic Potential.

    Science.gov (United States)

    Zamponi, Gerald W; Striessnig, Joerg; Koschak, Alexandra; Dolphin, Annette C

    2015-10-01

    Voltage-gated calcium channels are required for many key functions in the body. In this review, the different subtypes of voltage-gated calcium channels are described and their physiologic roles and pharmacology are outlined. We describe the current uses of drugs interacting with the different calcium channel subtypes and subunits, as well as specific areas in which there is strong potential for future drug development. Current therapeutic agents include drugs targeting L-type Ca(V)1.2 calcium channels, particularly 1,4-dihydropyridines, which are widely used in the treatment of hypertension. T-type (Ca(V)3) channels are a target of ethosuximide, widely used in absence epilepsy. The auxiliary subunit α2δ-1 is the therapeutic target of the gabapentinoid drugs, which are of value in certain epilepsies and chronic neuropathic pain. The limited use of intrathecal ziconotide, a peptide blocker of N-type (Ca(V)2.2) calcium channels, as a treatment of intractable pain, gives an indication that these channels represent excellent drug targets for various pain conditions. We describe how selectivity for different subtypes of calcium channels (e.g., Ca(V)1.2 and Ca(V)1.3 L-type channels) may be achieved in the future by exploiting differences between channel isoforms in terms of sequence and biophysical properties, variation in splicing in different target tissues, and differences in the properties of the target tissues themselves in terms of membrane potential or firing frequency. Thus, use-dependent blockers of the different isoforms could selectively block calcium channels in particular pathologies, such as nociceptive neurons in pain states or in epileptic brain circuits. Of important future potential are selective Ca(V)1.3 blockers for neuropsychiatric diseases, neuroprotection in Parkinson's disease, and resistant hypertension. In addition, selective or nonselective T-type channel blockers are considered potential therapeutic targets in epilepsy, pain, obesity, sleep

  5. Aspectos imunoistoquímicos da hepatite infecciosa canina Immunohistochemical aspects of canine infectious hepatitis

    Directory of Open Access Journals (Sweden)

    Maria Andréia Inkelmann

    2008-12-01

    Full Text Available Tecidos de 27 cães que morreram de hepatite infecciosa canina (HIC foram avaliados por imunoistoquímica (IHQ para detecção de antígeno de adenovírus canino tipo 1 (CAV-1, por meio de um anticorpo monoclonal. Os tecidos examinados incluíram fígado, rim, baço, linfonodos, tonsilas, pulmão, intestino delgado, encéfalo e medula óssea. Para cada órgão foram atribuídos graus crescentes (de leve a acentuada de intensidade de imunomarcação. O antígeno de CAV-1 estava presente na maioria dos órgãos examinados, principalmente em células endoteliais. Relacionando o tempo de evolução clínica da HIC com a intensidade da imunomarcação, o maior número de casos com evolução clínica hiperaguda ou aguda coincidiu com a maior intensidade de marcação do antígeno viral. A IHQ demonstrou ser um teste adequado para a detecção do antígeno de CAV-1 e pode ser usado para o estudo da patogênese da doença.Tissues from 27 dogs died from infectious canine hepatitis (ICH were evaluated by immunohistochemestry (IHC for detection of canine adenovirus type 1 (CAV-1 antigen using a monoclonal antibody. Evaluated tissues included liver, kidney, spleen, lymph nodes, tonsils, lungs, small intestine, brain, and spinal cord. For each organ increasing degrees (from mild to marked of intensity of immune staining were ascribed. CAV-1 was detected in most of the evaluated organs, mainly in the endothelial cells. Considering the clinical course of ICH and the intensity of IHC staining, cases with acute and peracute courses had more marked staining. IHC proved to be a reliable test for CAV-1 antigen detection and can be used for pathogenesis studies on ICH either in the spontaneous and experimental disease.

  6. Performance comparison between broilers positive and negative for antibodies against the chicken anemia virus Comparação de desempenho entre frangos positivos e negativos para anticorpos contra o vírus da anemia das galinhas

    Directory of Open Access Journals (Sweden)

    Lauricio Librelotto Rubin

    2003-11-01

    Full Text Available The chicken anemia virus (CAV is present in virtually every country investigated, Brazil including. The aim of this study was to determine what the difference in performance is between positive (progeny of breeders vaccinated or with natural infection and negative broilers to the presence of antibodies against the CAV in currently intensive raising systems. As a result, it was observed that negative broilers were significantly heavier than positive broilers. Negative males had a final weight 5.43% higher than positive males. There was no significant difference among different treatments in relation to parameters as mortality and feeding conversion. These study indicated that the presence of antibodies against CAV in broilers - may it be through vaccination or natural infection of breeders - did not generate progeny with superior performance under the tested raising conditions.O vírus da anemia das galinhas (CAV - "chicken anemia virus" está presente em praticamente todos os países investigados, inclusive no Brasil. O objetivo deste trabalho foi determinar qual a diferença de desempenho, comparando frangos positivos (progênie de matrizes vacinadas ou com infecção natural com frangos negativos para a presença de anticorpos contra o CAV, no sistema atual de criação intensiva. Como resultado, foi observado que os frangos negativos foram significativamente mais pesados que os frangos positivos. Os machos negativos tiveram um peso final 5,43% superior ao dos machos positivos. Não houve diferença significativa entre os tratamentos em relação aos parâmetros de mortalidade e conversão alimentar. Este estudo indicou que a presença de anticorpos contra o CAV em frangos de corte, seja através da vacinação ou infecção natural das matrizes, não gerou uma progênie com melhor desempenho nas condições de criação testadas.

  7. Fungi inhabiting healthy grapevine canes (Vitis spp. in some nurseries

    Directory of Open Access Journals (Sweden)

    Ewa Król

    2012-12-01

    Full Text Available The purpose of this study, conducted in the years 2000 - 2002, was to identify fungi species colonizing apparently healthy canes and to investigate whether canes storage modify the quantitative and qualitative composition of these fungi. The plant material was collected from 5 commercial plantations growing in various regions of Poland, taking into consideration 8 cultivars which were the most frequently cultivated. From each plantation and cultivar 20 apparently healthy canes were randomly sampled in two terms: before storage - November/December (term I and 3-4 months after storage - February/March (term II. The results showed that from asymptomatic canes 2746 isolates of fungi belonging to 23 species were obtained, but the majority of them origined from canes analysed after storage. It was found that P. viticola is able to live latently within grapevine tissue in Polish conditions because isolates of this fungus from visually healty canes the all studied plantations and terms were obtained. Among the other fungi species inhabiting grapevine canes Alternaria alternata and Fusarium spp. dominated. Moreover, both in term I and term II Botrytis cinerea, Phoma spp., Epicoccum purpurascens and Cladosporium cladosporioides were frequently isolated, whereas fungi from the genus Acremonium only in the term I. Each time isolates of Trichoderma spp. and Gliocladium spp. were also obtained. Inhabitation of grapevine canes by various fungi species shown in the present experiment indicate the danger of pathogens spread with propagation material on the new plantations.

  8. Occurrence of fungal diseases on angelica (Archangelica officinalis Hoffm. in the region of Małopolska

    Directory of Open Access Journals (Sweden)

    Stanisław Mazur

    2012-12-01

    Full Text Available The field experiment on angelica (Archangelica officinalis Hoffm. was carried out in the years 2001-2002 at the plantations situated in Małopolska (Mydlniki near Kraków and Królówka near Bochnia. The results of two years' investigations pointed out, that angelica plants during vegetation were infested by many pathogens. The most often noticed on the plants were angelica rust (Puccinia anglicae, powdery mildew (Erysiphe umbelliferarum and Cercospora leaf spot (Cercospora sp.. Moreover, many various types of disease symptoms, which were described and made a diagnosis. There were mainly spots on leaves and steams, from which there were isolated most often fungi from Alternaria genus. The investigations on the salubrity of the seed material (angelica fruits coming from different sources showed that fungi from genus Alternaria were likely to transfer by seeds and caused disease symptoms on plants during vegetation. Fungi from genus Alternaria were most often isolated from angelica fruits. Saprotrophic fungi (Epicoccum purpurascens, Penicillium spp., Mucor hiemalis were also often isolated from angelica fruits. Disinfection on fruits reduced number of isolated fungi and bacteria.

  9. Variability in automated assignment of NOESY spectra and three-dimensional structure determination: A test case on three small disulfide-bonded proteins

    Energy Technology Data Exchange (ETDEWEB)

    Savarin, Philippe; Zinn-Justin, Sophie; Gilquin, Bernard [CEA-Saclay, Departement d' Ingenierie et d' Etudes des Proteines (Bat. 152) (France)

    2001-01-15

    Three independent runs of automatic assignment and structure calculations were performed on three small proteins, calcicludine from the venom of the green mamba Dendroaspis angusticeps, {kappa}-conotoxin PVIIA from the purple cone Conus purpurascens and HsTX1, a short scorpion toxin from the venom of Heterometrus spinnifer. At the end of all the runs, the number of cross peaks which remained unassigned (0.6%, 1.4% and 2% for calcicludine, {kappa}-conotoxin and HsTX1, respectively), as well as the number of constraints which were rejected as producing systematic violations (2.7%, 1.0%, and 1.4% for calcicludine, {kappa}-conotoxin and HsTX1, respectively) were low. The conformation of the initial model used in the procedure (linear model or constructed by homology) has no influence on the final structures. Mainly two parameters control the procedure: the chemical shift tolerance and the cut-off distance. Independent runs of structure calculations, using the same parameters, yield structures for which the rmsd between averaged structures and the rmsd around each averaged structure were of the same order of magnitude. A different cut-off distance and a different chemical shift tolerance yield rmsd values on final average structures which did not differ more than 0.5 A compared to the rmsd obtained around the averaged structure for each calculation. These results show that the procedure is robust when applied to such a small disulfide-bonded protein.

  10. Solution structure of α-conotoxin PIA, a novel antagonist of α6 subunit containing nicotinic acetylcholine receptors

    International Nuclear Information System (INIS)

    α-Conotoxin PIA is a novel nicotinic acetylcholine receptor (nAChR) antagonist isolated from Conus purpurascens that targets nAChR subtypes containing α6 and α3 subunits. α-conotoxin PIA displays 75-fold higher affinity for rat α6/α3β2β3 nAChRs than for rat α3β2 nAChRs. We have determined the three-dimensional structure of α-conotoxin PIA by nuclear magnetic resonance spectroscopy. The α-conotoxin PIA has an 'ω-shaped' overall topology as other α4/7 subfamily conotoxins. Yet, unlike other neuronally targeted α4/7-conotoxins, its N-terminal tail Arg1-Asp2-Pro3 protrudes out of its main molecular body because Asp2-Pro3-Cys4-Cys5 forms a stable type I β-turn. In addition, a kink introduced by Pro15 in the second loop of this toxin provides a distinct steric and electrostatic environment from those in α-conotoxins MII and GIC. By comparing the structure of α-conotoxin PIA with other functionally related α-conotoxins we suggest structural features in α-conotoxin PIA that may be associated with its unique receptor recognition profile

  11. A novel D-leucine-containing Conus peptide: diverse conformational dynamics in the contryphan family.

    Science.gov (United States)

    Jacobsen, R B; Jimenez, E C; De la Cruz, R G; Gray, W R; Cruz, L J; Olivera, B M

    1999-08-01

    A Conus peptide family (the contryphans) is noteworthy because of the presence of a post-translationally modified D-amino acid in all members of the family. A new contryphan peptide, Leu-contryphan-P, was isolated from the venom of Conus purpurascens; the sequence of this peptide is: Gly-Cys-Val-D-Leu-Leu-Pro-Trp-Cys-OH. This is the first known occurrence of D-leucine in a Conus peptide. The discovery of Leu-contryphan-P suggests that there may be branches of the contryphan peptide family that diverge much more in sequence than previously anticipated. Several natural contryphans provide dramatic examples of interconversion between multiple conformational states in small constrained peptides. The contryphans that have 4-trans-hydroxyproline and D-tryptophan in positions 3 and 4, respectively, exhibit two peaks under reverse-phase HPLC conditions, indicating interconversion between two discrete conformations. However, [L-Trp4]contryphan-Sm (with L-Trp substituted for D-Trp) exhibits a single, broad peak that elutes later than the natural peptide, suggesting that D-Trp stabilizes a conformation in which hydrophobic residues are buried. Leucontryphan-P which has valine and D-leucine instead of 4-trans-hydroxyproline and D-tryptophan shows only a single peak that elutes much later than the other contryphans. PMID:10461743

  12. Studies on Total Polyphenols Content and Antioxidant Activity of Methanolic Extracts from Selected Salvia Species

    Directory of Open Access Journals (Sweden)

    Ana Viorica Pop Cuceu

    2015-05-01

    Full Text Available Salvia is one of the largest genera in the family of Labiatae, comprising about 900 species distributed widely throughout the world. Many species of Salvia are commonly used as herbal tea and for food flavoring, as well as in cosmetics, perfumery and in the pharmaceutical industry. The present study compares the antioxidant properties of four methanolic extracts, obtained by two extraction methods, from Salvia elegans, Salvia officinalis Purpurascens, Salvia officinalis Tricolor and Salvia lavandulifolia. The amount of total phenolics was quantified using the Folin-Ciocalteu method, while the antioxidant activity of selected herbs was determined with 1,1-diphenyl-2-picrylhydrazyl (DPPH reagent. The total amount of phenolics was between 1122.50 and 3672.16 mg GAE/100g dry plant for the first methanolic extract, while for the second extract was between 767.66 and 2725.04mg GAE/100g dry plant. A positive linear correlation was observed between total phenolics content and antioxidant activity of the extracts. The results suggested that the extracts of Salvia species, notably Salvia officinalis Tricolor with the highest antioxidant activity, can be used as natural antioxidants in the food industry.

  13. Historical ecology meets conservation and evolutionary genetics: a secondary contact zone between Carabus violaceus (Coleoptera, Carabidae populations inhabiting ancient and recent woodlands in north-western Germany

    Directory of Open Access Journals (Sweden)

    Andrea Matern

    2011-05-01

    Full Text Available Only very few cases have documented that an increase in connectivity after a period of fragmentation in ecological time has had an effect on the distribution, genetic structure and morphology of stenotopic species. In this study we present an example of clinal variability in a woodland ground beetle as a result of changes in the connectivity of a landscape during the last two centuries. The study area hosts both the nominate form C. violaceus s. str. and the subspecies C. v. purpurascens, which is ranked as a distinct species by some authors. We studied 12 Carabus violaceus populations from a 30 km transect of ancient and recent forests in north-western Germany. We analyzed three polymorphic enzyme loci, classified the elytron sculpture and measured the shape of the aedeagus tip of the specimens. C. violaceus showed secondary gradients both in allozyme markers and morphometric characters in our study area. A genetic differentiation of 16% between the populations is high but lies within the range of intraspecific variability in habitat specialists of the genus Carabus. Populations had no significant deficit of heterozygotes. We found many hybrid populations in terms of morphological properties. This study highlights the conservation value of ancient woodland and the consequences of landscape connectivity and defragmentation on the genetic setting of a ground beetle. Moreover, it shows that differences in the external shape of male genitalia do not prevent gene flow within the genus Carabus. Thus, the establishment of species status should not exclusively be based on this property.

  14. Variability in automated assignment of NOESY spectra and three-dimensional structure determination: A test case on three small disulfide-bonded proteins

    International Nuclear Information System (INIS)

    Three independent runs of automatic assignment and structure calculations were performed on three small proteins, calcicludine from the venom of the green mamba Dendroaspis angusticeps, κ-conotoxin PVIIA from the purple cone Conus purpurascens and HsTX1, a short scorpion toxin from the venom of Heterometrus spinnifer. At the end of all the runs, the number of cross peaks which remained unassigned (0.6%, 1.4% and 2% for calcicludine, κ-conotoxin and HsTX1, respectively), as well as the number of constraints which were rejected as producing systematic violations (2.7%, 1.0%, and 1.4% for calcicludine, κ-conotoxin and HsTX1, respectively) were low. The conformation of the initial model used in the procedure (linear model or constructed by homology) has no influence on the final structures. Mainly two parameters control the procedure: the chemical shift tolerance and the cut-off distance. Independent runs of structure calculations, using the same parameters, yield structures for which the rmsd between averaged structures and the rmsd around each averaged structure were of the same order of magnitude. A different cut-off distance and a different chemical shift tolerance yield rmsd values on final average structures which did not differ more than 0.5 A compared to the rmsd obtained around the averaged structure for each calculation. These results show that the procedure is robust when applied to such a small disulfide-bonded protein

  15. Comparative anatomical and ecological investigations on some centaurea (asteracae) taxa from turkey and their taxonomic significance

    International Nuclear Information System (INIS)

    The anatomical and ecological characteristics of 7 Centaurea L. s. l. taxa (C. urvillei DC. ssp. stepposa Wagenitz, C. pseudoreflexa Hayek, C. simplicicaulis Boiss. and Huet, C. pecho Albow, C. hypoleuca DC., C. cheiranthifolia Willd.var. purpurascens (DC.) Wagenitz, C. woronowii Bornm.), which three of them are endemic (C. pseudoreflexa, C. pecho, C. woronowii) to Turkey has been carried out. Cross sections of stem, leaf blade and midrib and peripheral sections of the leaves, and pH, total N, P, K and organic matter contents of soil samples have been investigated. Variance analysis and Tukey's Honest Significant Difference test were performed on ecological and anatomical data. Ecological characteristics obtained in this investigation were found mostly similar. On the other hand, anatomically important differences were determined in diversity of anticlinal cell walls, length and width of abaxial epidermis, number of bundles in the midrib and number of stomata per mm/sup 2/ both on the epidermis of the leaf, and the shape of transverse section of stem. According to Tukey's Honest Significant Difference test, especially length and width of abaxial epidermis, number of stomata both on the adaxial and abaxial epidermises have displayed three distinct groups. These results are in accordance with sectional delimitation in the Flora of Turkey and also support upgrading the sum of species of the genus Centaurea. (author)

  16. Demand controlled ventilation in a bathroom

    DEFF Research Database (Denmark)

    Mortensen, Dorthe Kragsig; Nielsen, Toke Rammer; Topp, Claus

    2008-01-01

    climate and energy consumption of a Constant Air Volume (CAV) system and a Demand Controlled Ventilation (DCV) system for two different bathroom designs. The air change rate of the CAV system corresponded to 0.5h-1. The ventilation rate of the DCV system was controlled by occupancy and by the relative...... humidity in the bathroom. The two designs differed by the construction of the shower cubicle which in one case was sealed and in the other case unsealed. The construction influenced the relative humidity within the bathroom during a shower, i.e. the pollution source the ventilation rate was controlled by....... The indoor climate and the energy consumption were estimated based on a simplified calculation of the variation of the water content within the bathroom during a day. The results showed that the DCV system controlled by occupancy and relative humidity had an improved energy performance and an improved...

  17. Effect of plastic spacer handling on salbutamol lung deposition in asthmatic children

    DEFF Research Database (Denmark)

    Lipworth, Brian J; Lee, Daniel K C; Anhøj, Jacob;

    2002-01-01

    AIMS: To study the effects of electrostatics in a plastic spacer on the lung deposition of salbutamol in asthmatic children. METHODS: Twenty-five children (5-12 years) with mild asthma were given salbutamol hydrofluoroalkane pressurized metered dose inhaler 400 micro g via a 750 ml plastic spacer...... repeated daily use (Used ), a spacer rinsed after repeated use (UsedRinsed ) and a spacer primed with benzalkonium chloride to avoid electrostatics (Primed1). In addition, spacers were evaluated using a 15 s inhalation delay following actuation with primed (PrimedDelay) and rinsed (RinsedDelay) spacers....... Data were log transformed and expressed as geometric mean fold difference for the average plasma salbutamol concentration (Cav) over 20 min. RESULTS: There were significant differences (P < 0.05) in Cav (as geometric mean fold difference and 95% CI) between Primed1 vs NewRinsed 1.92 fold (95% CI 1...

  18. Functional Importance of L- and P/Q-Type Voltage-Gated Calcium Channels in Human Renal Vasculature

    DEFF Research Database (Denmark)

    Hansen, Pernille B; Poulsen, Christian B; Walter, Steen;

    2011-01-01

    revealed signals for Ca(v) 2.1 and Ca(v) 3.1 associated with smooth muscle cells of preglomerular and postglomerular vessels. In human intrarenal arteries, depolarization with potassium induced a contraction inhibited by the L-type antagonist nifedipine, EC(50) 1.2×10(-8) mol/L. The T-type antagonist...... L- and P/Q-type channels are of functional importance for the depolarization-induced vasoconstriction. The contribution of P/Q-type channels to contraction in the human vasculature is a novel mechanism for the regulation of renal blood flow and suggests that clinical treatment with calcium blockers......Calcium channel blockers are widely used for treatment of hypertension, because they decrease peripheral vascular resistance through inhibition of voltage-gated calcium channels. Animal studies of renal vasculature have shown expression of several types of calcium channels that are involved in...

  19. Clinical features, exercise hemodynamics, and determinants of left ventricular elevated filling pressure in heart transplanted patients

    DEFF Research Database (Denmark)

    Clemmensen, Tor Skibsted; Eiskjaer, Hans; Løgstrup, Brian Bridal; Mellemkjaer, Søren; Andersen, Mads Jønsson; Tolbod, Lars Poulsen; Harms, Hendrik J; Poulsen, Steen Hvitfeldt

    2015-01-01

    PURPOSE: This study aimed to assess clinical, functional, and hemodynamic characteristics of heart-transplanted (HTX) patients during exercise. METHODS: We performed comprehensive echocardiographic graft function assessment during invasive hemodynamic semi-supine exercise test in 57 HTX patients......-one patients (54%) had normal LV-FP and 26 patients (46%) had elevated LV-FP. The latter had higher cumulative rejection burden (p<0.01), were more symptomatic (NYHA class >1) (p<0.05), and CAV was more prevalent (p<0.05). With exercise, the changes in both left and right sided filling pressures were...... in approximately 50% of HTX patients. Patients with elevated LV-FP have impaired myocardial deformation capacity, higher prevalence of CAV, higher rejection burden, and were more symptomatic. Exercise test with assessment of longitudinal myocardial deformation should be considered in routine...

  20. Algebraic Reconstruction Algorithm of Vapor Tomography

    Directory of Open Access Journals (Sweden)

    HE Lin

    2015-01-01

    Full Text Available While applying algebraic reconstruction algorithm in vapor tomography, problems have to be solved with respect to constructing the constraint condition, selecting the initial value, calculating optimal relaxation factor and deciding the iteration termination condition. Golden section search method and NCP termination rule are given to solve the latter two problems, respectively. Eight algebraic reconstruction algorithms, including Kaczmarz, Randkaczmarz, Symkaczmarz, SART, Landweber, Cimmino, CAV and DROP algorithm, are comparatively analyzed and tested by the data from SatRef station in Hong Kong. The results show that all the eight algorithms can satisfy the requirements of vapor tomography and the iteration termination condition is more important than the relaxation condition. While the golden section method and NCP method are used, the CAV algorithm performs best, and then the Cimmino algorithm.

  1. Optimal Control and Coordination of Connected and Automated Vehicles at Urban Traffic Intersections

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yue J. [Boston University; Malikopoulos, Andreas [ORNL; Cassandras, Christos G. [Boston University

    2016-01-01

    We address the problem of coordinating online a continuous flow of connected and automated vehicles (CAVs) crossing two adjacent intersections in an urban area. We present a decentralized optimal control framework whose solution yields for each vehicle the optimal acceleration/deceleration at any time in the sense of minimizing fuel consumption. The solu- tion, when it exists, allows the vehicles to cross the intersections without the use of traffic lights, without creating congestion on the connecting road, and under the hard safety constraint of collision avoidance. The effectiveness of the proposed solution is validated through simulation considering two intersections located in downtown Boston, and it is shown that coordination of CAVs can reduce significantly both fuel consumption and travel time.

  2. The prostaglandin E2/EP4 receptor/cyclic AMP/T-type Ca(2+) channel pathway mediates neuritogenesis in sensory neuron-like ND7/23 cells.

    Science.gov (United States)

    Mitani, Kenji; Sekiguchi, Fumiko; Maeda, Takashi; Tanaka, Yukari; Yoshida, Shigeru; Kawabata, Atsufumi

    2016-03-01

    We investigated mechanisms for the neuritogenesis caused by prostaglandin E2 (PGE2) or intracellular cyclic AMP (cAMP) in sensory neuron-like ND7/23 cells. PGE2 caused neuritogenesis, an effect abolished by an EP4 receptor antagonist or inhibitors of adenylyl cyclase (AC) or protein kinase A (PKA) and mimicked by the AC activator forskolin, dibutyryl cAMP (db-cAMP), and selective activators of PKA or Epac. ND7/23 cells expressed both Cav3.1 and Cav3.2 T-type Ca(2+) channels (T-channels). The neuritogenesis induced by db-cAMP or PGE2 was abolished by T-channel blockers. T-channels were functionally upregulated by db-cAMP. The PGE2/EP4/cAMP/T-channel pathway thus appears to mediate neuritogenesis in sensory neurons. PMID:27032908

  3. Role of T-type channels in vasomotor function

    DEFF Research Database (Denmark)

    Kuo, Ivana Y-T; Howitt, Lauren; Sandow, Shaun L;

    2014-01-01

    Low-voltage-activated T-type calcium channels play an important role in regulating cellular excitability and are implicated in conditions, such as epilepsy and neuropathic pain. T-type channels, especially Cav3.1 and Cav3.2, are also expressed in the vasculature, although patch clamp studies of...... make a small contribution to vascular tone at low intraluminal pressures, although the relevance of this contribution is unclear. By contrast, these channels play a larger role in vascular tone of small arterioles, which would be expected to function at lower intra-vascular pressures. Upregulation of T......-type channel function following decrease in nitric oxide bioavailability and increase in oxidative stress, which occurs during cardiovascular disease, suggests that a more important role could be played by these channels in pathophysiological situations. The ability of T-type channels to be rapidly recruited...

  4. Electroconvulsive stimulations prevent chronic stress-induced increases in L-type calcium channel mRNAs in the hippocampus and basolateral amygdala

    DEFF Research Database (Denmark)

    Maigaard, Katrine; Pedersen, Ida Hageman; Jørgensen, Anders;

    2012-01-01

    chronic restraint stress model of affective disorder (6 h restraint per day for 21 days) in combination with electroconvulsive stimulations to examine the effects of stress and an effective antidepressive treatment modality on L-type voltage gated calcium channel subunit mRNA expression patterns in the......, while stress only upregulated Ca(v)1.3 channel expression significantly in the dentate gyrus. ECS effects on Ca(v)1.2 channel expression were generally specific to stressed animals. Our findings are consistent with and extent previous studies on the involvement of intracellular calcium ion dysfunction...... in affective disorders. Selective modulation of neuronal L-type voltage gated calcium channels appears to be a promising target for the development of novel antidepressive treatment modalities....

  5. Risk factors of cardiac allograft vasculopathy

    Science.gov (United States)

    Szczurek, Wioletta; Gąsior, Mariusz; Zembala, Marian

    2015-01-01

    Despite advances in prevention and treatment of heart transplant rejection, development of cardiac allograft vasculopathy (CAV) remains the leading factor limiting long-term survival of the graft. Cardiac allograft vasculopathy etiopathogenesis is not fully understood, but a significant role is attributed to endothelial cell damage, caused by immunological and non-immunological mechanisms. Immunological factors include the differences between the recipient's and the donor's HLA systems, the presence of alloreactive antibodies and episodes of acute rejection. Among the non-immunological factors the most important are the age of the donor, ischemia-reperfusion injury and cytomegalovirus infection. The classical cardiovascular risk factors (diabetes, hypertension, obesity and hyperlipidemia) are also important. This study presents an up-to-date overview of current knowledge on the vasculopathy etiopathogenesis and the role played by endothelium and inflammatory processes in CAV, and it also investigates the factors which may serve as risk markers of cardiac allograft vasculopathy. PMID:26855649

  6. Aspectos imunoistoquímicos da hepatite infecciosa canina Immunohistochemical aspects of canine infectious hepatitis

    OpenAIRE

    Maria Andréia Inkelmann; Bruno Leite dos Anjos; Glaucia Denise Kommers; Rafael Almeida Fighera; Claudio Severo Lombardo de Barros

    2008-01-01

    Tecidos de 27 cães que morreram de hepatite infecciosa canina (HIC) foram avaliados por imunoistoquímica (IHQ) para detecção de antígeno de adenovírus canino tipo 1 (CAV-1), por meio de um anticorpo monoclonal. Os tecidos examinados incluíram fígado, rim, baço, linfonodos, tonsilas, pulmão, intestino delgado, encéfalo e medula óssea. Para cada órgão foram atribuídos graus crescentes (de leve a acentuada) de intensidade de imunomarcação. O antígeno de CAV-1 estava presente na maioria dos órgão...

  7. Duplex PCR assay for the detection of avian adeno virus and chicken anemia virus prevalent in Pakistan

    Directory of Open Access Journals (Sweden)

    Iqbal Aqib

    2011-09-01

    Full Text Available Abstract Avian Adeno viruses and Chicken Anemia Viruses cause serious economic losses to the poultry industry of Pakistan each year. Timely and efficient diagnosis of the viruses is needed in order to practice prevention and control strategies. In the first part of this study, we investigated broilers, breeder and Layer stocks for morbidity and mortality rates due to AAV and CAV infections and any co-infections by examining signs and symptoms typical of their infestation or post mortem examination. In the second part of the study, we developed a duplex PCR assay for the detection of AAV and CAV which is capable to simultaneously detect both the viral types prevalent in Pakistan with high sensitivity and 100% specificity.

  8. Selective modulation of cellular voltage dependent calcium channels by hyperbaric pressure - a suggested HPNS partial mechanism

    Directory of Open Access Journals (Sweden)

    Ben eAviner

    2014-05-01

    Full Text Available Professional deep sea divers experience motor and cognitive impairment, known as High Pressure Neurological Syndrome (HPNS, when exposed to pressures of 100 msw (1.1MPa and above, considered to be the result of synaptic transmission alteration. Previous studies have indicated modulation of presynaptic Ca2+ currents at high pressure. We directly measured for the first time pressure effects on the currents of voltage dependent Ca2+ channels (VDCCs expressed in Xenopus oocytes. Pressure selectivity augmented the current in CaV1.2 and depressed it in CaV3.2 channels. Pressure application also affected the channels' kinetics, such as ƮRise, ƮDecay. Pressure modulation of VDCCs seems to play an important role in generation of HPNS signs and symptoms.

  9. Evaluating the performance of vehicular platoon control under different network topologies of initial states

    Science.gov (United States)

    Li, Yongfu; Li, Kezhi; Zheng, Taixiong; Hu, Xiangdong; Feng, Huizong; Li, Yinguo

    2016-05-01

    This study proposes a feedback-based platoon control protocol for connected autonomous vehicles (CAVs) under different network topologies of initial states. In particularly, algebraic graph theory is used to describe the network topology. Then, the leader-follower approach is used to model the interactions between CAVs. In addition, feedback-based protocol is designed to control the platoon considering the longitudinal and lateral gaps simultaneously as well as different network topologies. The stability and consensus of the vehicular platoon is analyzed using the Lyapunov technique. Effects of different network topologies of initial states on convergence time and robustness of platoon control are investigated. Results from numerical experiments demonstrate the effectiveness of the proposed protocol with respect to the position and velocity consensus in terms of the convergence time and robustness. Also, the findings of this study illustrate the convergence time of the control protocol is associated with the initial states, while the robustness is not affected by the initial states significantly.

  10. Serologic evidence for selected infectious diseases in Marsican brown bears (Ursus arctos marsicanus) in Italy (2004-09).

    Science.gov (United States)

    Di Francesco, Cristina Esmeralda; Gentile, Leonardo; Di Pirro, Vincenza; Ladiana, Lara; Tagliabue, Silvia; Marsilio, Fulvio

    2015-01-01

    We tested 30 serum samples collected during 2004-09 from 22 free-ranging Marsican brown bears (Ursus arctos marsicanus) in the National Park of Abruzzo, Lazio, and Molise, Italy, for antibodies against canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and eight Leptospira interrogans sensu lato serovars. Antibody to CDV was detected in 11 samples (37%); only two bears (10%) had detectable CAV-2 and Brucella spp. antibodies; three bears were positive for L. interrogans serovar Bratislava; and one sample had antibody against L. interrogans serovar Copenhageni. All samples were positive for CPV-2 antibody. The CPV-2 antibody titers varied from 1∶640 to 1∶10,240, suggesting that transmission was still active. Fifty percent of bears were positive for antibody to two or more pathogens. Our results highlight the need to consider infectious diseases as a potential risk for Marsican brown bear conservation. PMID:25375945

  11. Subsolidus phase relations in the systems M2+-M2+O-V2O5 (M+-Li, Na, K, Rb, Cs; M2+-Mg, Ca)

    International Nuclear Information System (INIS)

    One studied phase composition of M2O(M2CO3)-MgO-V2O5 and M2O(M2CO3)-CaO-V2O5 systems, where M-Li, Na, K, Rb, Cs under the subsolidus temperatures. One synthesized 20 binary vanadates 5 ones of which (Rb2CaV2O7, Cs2CaV2O7, LiMg4(VO4)3, RbCaVO4 and CsCaVO4) were prepared for the first time. Paper presents summary table of structural characteristics of all binary vanadates. For 14 ones of them one presents structure and parameters of lattice (for 6 vanadates data were obtained for the first time). One shows formation of terminal solid solution based on calcium orthovanadate. One studied phase diagrams of 6 systems (M+-Li, Na, K, Rb, Cs; M2+-Mg, Ca)

  12. T-type Ca(2+) channels facilitate NO-formation, vasodilatation and NO-mediated modulation of blood pressure

    DEFF Research Database (Denmark)

    Svenningsen, Per; Andersen, Kenneth; Thuesen, Anne D;

    2014-01-01

    Voltage-gated calcium channels are involved in the vascular excitation-contraction mechanism and regulation of arterial blood pressure. It was hypothesized that T-type channels promote formation of nitric oxide from the endothelium. The present experiments determine the involvement of T-type chan......Voltage-gated calcium channels are involved in the vascular excitation-contraction mechanism and regulation of arterial blood pressure. It was hypothesized that T-type channels promote formation of nitric oxide from the endothelium. The present experiments determine the involvement of T......-type channels in depolarization-dependent dilatation of mesenteric arteries and blood pressure regulation in Cav3.1 knock-out mice. Nitric oxide-dependent vasodilatation following depolarization-mediated vasoconstriction was reduced significantly in mesenteric arteries from Cav3.1(-/-) compared to wild type...

  13. Strategies for Investigating G-Protein Modulation of Voltage-Gated Ca2+ Channels.

    Science.gov (United States)

    Lu, Van B; Ikeda, Stephen R

    2016-01-01

    G-protein-coupled receptor modulation of voltage-gated ion channels is a common means of fine-tuning the response of channels to changes in membrane potential. Such modulation impacts physiological processes such as synaptic transmission, and hence therapeutic strategies often directly or indirectly target these pathways. As an exemplar of channel modulation, we examine strategies for investigating G-protein modulation of CaV2.2 or N-type voltage-gated Ca(2+) channels. We focus on biochemical and genetic tools for defining the molecular mechanisms underlying the various forms of CaV2.2 channel modulation initiated following ligand binding to G-protein-coupled receptors. PMID:27140924

  14. Distinct Roles of Endothelial and Adipocyte Caveolin-1 in Macrophage Infiltration and Adipose Tissue Metabolic Activity

    OpenAIRE

    Briand, N.; Le Lay, S.; Sessa, W. C.; Ferre, P.; Dugail, I.

    2011-01-01

    OBJECTIVE Defective caveolin-1 expression is now recognized as a cause of lipoatrophic diabetes in patients, due to primary caveolin gene mutations or secondary caveolin deficiency caused by PTRF/cavin gene defects. The goal of this study was to establish the relative contribution of endothelial cells and adipocytes, both highly expressing caveolin-1 to the lipoatrophic phenotype of mice with global caveolin-1 gene invalidation (Cav1-KO). RESEARCH DESIGN AND METHODS We compared adipose tissue...

  15. RICE BLAST CONTROL WITH RELEASE OF RESISTANT VARIETIES Борьба с пирикуляриозом риса путем создания устойчивых сортов

    OpenAIRE

    Zelenskiy G. L.

    2013-01-01

    Among fungus diseases of rice, blast is the most harm-ful. The disease is caused by Pyricularia oryzae Cav. Rice is sensitive to blast at all fazes of vegetation. In Russia, the purposeful breeding of rice varieties re-sistant to this disease began in 1982. Over the past period, the rice varieties which are genetically protect-ed from blast and not requiring crop protection have been created

  16. A homology model of the pore domain of a voltage-gated calcium channel is consistent with available SCAM data

    OpenAIRE

    Bruhova, Iva; Zhorov, Boris S.

    2010-01-01

    In the absence of x-ray structures of calcium channels, their homology models are used to rationalize experimental data and design new experiments. The modeling relies on sequence alignments between calcium and potassium channels. Zhen et al. (2005. J. Gen. Physiol. doi:10.1085/jgp.200509292) used the substituted cysteine accessibility method (SCAM) to identify pore-lining residues in the Cav2.1 channel and concluded that their data are inconsistent with the symmetric architecture of the pore...

  17. High-resolution digital 3D models of Algar do Penico Chamber: limitations, challenges, and potential

    OpenAIRE

    Silvestre, Ivo; Rodrigues, J. I.; Figueiredo, Mauro; C. Veiga-Pires

    2015-01-01

    The study of karst and its geomorphological structures is important for understanding the relationships between hydrology and climate over geological time. In that context, we conducted a terrestrial laser-scan survey to map geomorphological structures in the karst cave of Algar do Penico in southern Portugal. The point cloud data set obtained was used to generate 3D meshes with different levels of detail, allowing the limitations of mapping capabilities to be explored. In addition to cav...

  18. Relationship with spatial memory in diabetic rats and protein kinase Cγ,caveolin-1 in the hippocampus and neuroprotective effect of catalpol

    Institute of Scientific and Technical Information of China (English)

    Zhou Haicheng; Liu Jing; Ren Liyuan; Liu Wei; Xing Qian; Men Lili; Song Guirong

    2014-01-01

    Background The mechanisms underlying diabetic encephalopathy are largely unknown,and no effective treatments are available.Catalpol has received much attention due to its numerous biological effects,especially in neuroprotective studies.The aim of this study was to investigate the effects of catalpol on cognitive functions in diabetic rats and the underlying mechanisms.Methods A rat model of diabetes was established by streptozotocin injection,followed by intraperitoneal infusion of catalpol after 10 weeks.Two weeks later,the Morris water maze was used to test the spatial learning performance.Nissl staining was performed to evaluate the morphological changes in the hippocampus.Expression of protein kinase Cy (PKCy) and caveolin-1 (Cav-1) in the hippocampus were assessed by reverse transcription PCR and Western blotting.Activities of anti-oxidative enzymes such as glutathione (GSH),superoxide dismutase (SOD) and catalase (CAT) and levels of malonaldehyde (MDA) were measured using commercial kits.Results Significant hippocampal neuronal injury was observed in rats with streptozotocin-induced diabetes.Moreover,cognitive dysfunction was associated with markedly increased oxidative stress in the brain.Catalpol treatment significantly attenuated cognitive deficits,neuronal damage,and oxidative stress in the brain of diabetic rats.Biochemical analyses showed that catalpol reversed the down-regulation of PKCγ and Cav-1 expression in the diabetic rats.Conclusions Spatial memory in diabetic rats is associated with the expression of PKCy and Cav-1.Catalpol treatment markedly attenuated oxidative stress,reversed the alteration of PKCy,Cav-1 and spatial memory deficits.

  19. Targeting voltage-gated calcium channels: developments in peptide and small-molecule inhibitors for the treatment of neuropathic pain

    OpenAIRE

    Vink, S.; Alewood, PF

    2012-01-01

    Chronic pain affects approximately 20% of people worldwide and places a large economic and social burden on society. Despite the availability of a range of analgesics, this condition is inadequately treated, with complete alleviation of symptoms rarely occurring. In the past 30 years, the voltage-gated calcium channels (VGCCs) have been recognized as potential targets for analgesic development. Although the majority of the research has been focused on Cav2.2 in particular, other VGCC subtypes...

  20. Assessment of cadmium and lead mobility in the rhizosphere using voltammetry and electrospray ionization mass spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Šestáková, Ivana; Jaklová Dytrtová, Jana; Jakl, M.; Navrátil, Tomáš

    2011-01-01

    Roč. 5, č. 3 (2011), s. 347-355. ISSN 1109-9577 R&D Projects: GA AV ČR IAA400400806 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z40550506 Keywords : cadmium * electrochemical impedance spectroscopy (EIS) * oxalic acid Subject RIV: CG - Electrochemistry http://www.library.sk/i2/i2.entry.cls?ictx=cav&language=2&op=esearch

  1. Actividad antimicrobiana de Verbesina encelioides Antimicrobial activity of Verbesina encelioides

    OpenAIRE

    M.S Toribio; D.S Oriani; Fernández, J G; Skliar, M.I.

    2005-01-01

    Verbesina encelioides (Cav.) Benth. & Hook. (Compuestas) es una especie ampliamente distribuida en la Argentina, que se conoce vulgarmente con el nombre de «girasolcito» o «mirasolcito del campo», debido a que presenta flores amarillas en capítulos terminales pedunculados. El objetivo del presente trabajo es evaluar la actividad antimicrobiana de esta especie frente a bacterias Gram positivas, Gram negativas y Candida albicans. Se utilizaron concentraciones crecientes del extracto vegetal par...

  2. Retrograde optogenetic characterization of the pontospinal module of the locus coeruleus with a canine adenoviral vector.

    Science.gov (United States)

    Li, Yong; Hickey, Louise; Perrins, Ray; Werlen, Emilie; Patel, Amisha A; Hirschberg, Stefan; Jones, Matt W; Salinas, Sara; Kremer, Eric J; Pickering, Anthony E

    2016-06-15

    Noradrenergic neurons of the brainstem extend projections throughout the neuraxis to modulate a wide range of processes including attention, arousal, autonomic control and sensory processing. A spinal projection from the locus coeruleus (LC) is thought to regulate nociceptive processing. To characterize and selectively manipulate the pontospinal noradrenergic neurons in rats, we implemented a retrograde targeting strategy using a canine adenoviral vector to express channelrhodopsin2 (CAV2-PRS-ChR2-mCherry). LC microinjection of CAV2-PRS-ChR2-mCherry produced selective, stable, transduction of noradrenergic neurons allowing reliable opto-activation in vitro. The ChR2-transduced LC neurons were opto-identifiable in vivo and functional control was demonstrated for >6 months by evoked sleep-wake transitions. Spinal injection of CAV2-PRS-ChR2-mCherry retrogradely transduced pontine noradrenergic neurons, predominantly in the LC but also in A5 and A7. A pontospinal LC (ps:LC) module was identifiable, with somata located more ventrally within the nucleus and with a discrete subset of projection targets. These ps:LC neurons had distinct electrophysiological properties with shorter action potentials and smaller afterhyperpolarizations compared to neurons located in the core of the LC. In vivo recordings of ps:LC neurons showed a lower spontaneous firing frequency than those in the core and they were all excited by noxious stimuli. Using this CAV2-based approach we have demonstrated the ability to retrogradely target, characterise and optogenetically manipulate a central noradrenergic circuit and show that the ps:LC module forms a discrete unit. This article is part of a Special Issue entitled SI: Noradrenergic System. PMID:26903420

  3. A criterion for determining exceedance of the operating basis earthquake

    International Nuclear Information System (INIS)

    A criterion is recommended for determining whether the operating basis earthquake (OBE) has been exceeded after the occurrence of a seismic event at a nuclear power plant. The technical procedure provides a fast and effective means for assessing the damage potential of any felt earthquake in the first few hours after occurrence. For OBE exceedance to occur, the recommended criterion requires exceedance of both a response spectrum parameter and a second damage parameter, referred to as the cumulative absolute velocity (CAV)

  4. Actin Depolymerization Disrupts Tight Junctions via Caveolae-mediated EndocytosisV⃞

    OpenAIRE

    Shen, Le; Turner, Jerrold R.

    2005-01-01

    The tight junction (TJ) determines epithelial barrier function. Actin depolymerization disrupts TJ structure and barrier function, but the mechanisms of this effect remain poorly understood. The goal of this study was to define these mechanisms. Madin-Darby canine kidney (MDCK) cells expressing enhanced green fluorescent protein-, enhanced yellow fluorescent protein-, or monomeric red fluorescent protein 1-fusion proteins of β-actin, occludin, claudin-1, ZO-1, clathrin light chain A1, and cav...

  5. Matrix metalloproteinase-2, caveolins, focal adhesion kinase and c-Kit in cells of the mouse myocardium

    OpenAIRE

    Cho, Woo Jung; Chow, Ava K; Schulz, Richard; Daniel, Edwin E

    2007-01-01

    Abstract Matrix metalloproteinase-2 (MMP-2) may play roles at intracellular and extracellular sites of the heart in ischaemia/reperfusion injury. Caveolins (Cav-1, -2 and -3) are lipid raft proteins which play roles in cell sig-nalling. This study examined, using immunohistochemistry and two photon confocal microscopy, if MMP-2 and caveolins co-localize at the plasma membrane of cardiac cells: cardiomyocytes (CM), fibroblasts (FB) and capillary endothelial cells (CEC) in the left ventricle (L...

  6. Simulation-Based Optimization of Cure Cycle of Large Area Compression Molding for LED Silicone Lens

    OpenAIRE

    Min-Jae Song; Kwon-Hee Kim; Seok-Kwan Hong; Jeong-Won Lee; Jeong-Yeon Park; Gil-Sang Yoon; Heung-Kyu Kim

    2015-01-01

    Three-dimensional heat transfer-curing simulation was performed for the curing process by introducing a large area compression molding for simultaneous forming and mass production for the lens and encapsulants in the LED molding process. A dynamic cure kinetics model for the silicone resin was adopted and cure model and analysis result were validated and compared through a temperature measurement experiment for cylinder geometry with cure model. The temperature deviation between each lens cav...

  7. CHARACTERIZATION QUALITATIVE OF SOEL FOR A VALORIZATION BEST OF THE BIOMASS PRODUCED

    OpenAIRE

    Y. M’Sadak; Saad, I.

    2015-01-01

    The main objective of this study was to qualitatively characterize biomass of Solanum elaeagnifolium Cav. (or SOEL) to highlight the essential alternatives its valorization. At first, we studied the potential of composting biomass of this plant. The physico-chemical characterization of biomass silverleaf nightshade has detected a wealth in organic carbon, which highlighted the potential value of this plant in the Co-composting with other biodegradable waste. In a second step, we assessed the ...

  8. Leukocytic acetylcholine in chronic rejection of renal allografts

    OpenAIRE

    Wilczynska, Joanna

    2011-01-01

    Leukocytes, which accumulate in graft blood vessels during fatal acute rejection of experimental renal allografts, synthesise and release acetylcholine (ACh). In this study, I tested the hypothesis that ACh produced by leukocytes accumulating in graft blood vessels contributes to the pathogenesis of chronic renal allograft vasculopathy (CAV). Kidneys were transplanted in the allogeneic Fischer 344 to Lewis rat strain combination. Isogeneic transplantations were performed in Lew...

  9. Mechanism of PKA-dependent and lipid-raft independent stimulation of Connexin43 expression by oxytoxin in mouse embryonic stem cells.

    Science.gov (United States)

    Yun, Seung Pil; Park, Su Shin; Ryu, Jung Min; Park, Jae Hong; Kim, Mi Ok; Lee, Jang-Hern; Han, Ho Jae

    2012-07-01

    Previous studies shows that connexins appear very early during murine embryo development, the gap junctional intercellular communication found in the inner cell mass of early embryo is also maintained in embryonic stem cells (ESC), and expression of oxytocin receptor (OTR) is developmentally regulated at early embryonic development. However, effect of oxytocin (OT) on the regulation of the connexin43 (Cx43) and maintenance of undifferentiation is not fully understood in stem cells. Therefore, we investigated the effect of OT on Cx43 expression and related signaling cascades in mouse ESC. OT increased Cx43 expression that was inhibited by the OTR inhibitor atosiban. In experiments to examine whether the effect of OT depends on lipid rafts, caveolin-1 (cav-1), cav-2, and flotillin-2, but not OTR, were detected in lipid raft fractions. Also, colocalization of OTR, cav-1, and cav-2 was not detected. Moreover, the lipid raft disruptor methyl-β-cyclodextrin did not attenuate OT-induced Cx43 expression. In experiments to examine related signaling pathways, OT activated cAMP/protein kinase A (PKA) which was inhibited by adenylyl cyclase inhibitor SQ 22536 and PKA inhibitor PKI. OT increased nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) phosphorylation which was inhibited by PKI. OT also increased cAMP response element-binding (CREB)/CREB-binding protein (CBP) expression in the nucleus and induced the formation of CREB1/NF-κB/CBP complexes, which was blocked by the NF-κB-specific small interfering RNA, NF-κB inhibitors, SN50, and bay11-7082. Complex disruption by NF-κB inhibitors decreased OT-induced Cx43 expression. In conclusion, OT stimulates Cx43 expression through the NF-κB/CREB/CBP complex via the lipid raft-independent OTR-mediated cAMP/PKA in mouse ESC. PMID:22564436

  10. Ablation of triadin causes loss of cardiac Ca2+ release units, impaired excitation-contraction coupling, and cardiac arrhythmias.

    Science.gov (United States)

    Chopra, Nagesh; Yang, Tao; Asghari, Parisa; Moore, Edwin D; Huke, Sabine; Akin, Brandy; Cattolica, Robert A; Perez, Claudio F; Hlaing, Thinn; Knollmann-Ritschel, Barbara E C; Jones, Larry R; Pessah, Isaac N; Allen, Paul D; Franzini-Armstrong, Clara; Knollmann, Björn C

    2009-05-01

    Heart muscle excitation-contraction (E-C) coupling is governed by Ca(2+) release units (CRUs) whereby Ca(2+) influx via L-type Ca(2+) channels (Cav1.2) triggers Ca(2+) release from juxtaposed Ca(2+) release channels (RyR2) located in junctional sarcoplasmic reticulum (jSR). Although studies suggest that the jSR protein triadin anchors cardiac calsequestrin (Casq2) to RyR2, its contribution to E-C coupling remains unclear. Here, we identify the role of triadin using mice with ablation of the Trdn gene (Trdn(-/-)). The structure and protein composition of the cardiac CRU is significantly altered in Trdn(-/-) hearts. jSR proteins (RyR2, Casq2, junctin, and junctophilin 1 and 2) are significantly reduced in Trdn(-/-) hearts, whereas Cav1.2 and SERCA2a remain unchanged. Electron microscopy shows fragmentation and an overall 50% reduction in the contacts between jSR and T-tubules. Immunolabeling experiments show reduced colocalization of Cav1.2 with RyR2 and substantial Casq2 labeling outside of the jSR in Trdn(-/-) myocytes. CRU function is impaired in Trdn(-/-) myocytes, with reduced SR Ca(2+) release and impaired negative feedback of SR Ca(2+) release on Cav1.2 Ca(2+) currents (I(Ca)). Uninhibited Ca(2+) influx via I(Ca) likely contributes to Ca(2+) overload and results in spontaneous SR Ca(2+) releases upon beta-adrenergic receptor stimulation with isoproterenol in Trdn(-/-) myocytes, and ventricular arrhythmias in Trdn(-/-) mice. We conclude that triadin is critically important for maintaining the structural and functional integrity of the cardiac CRU; triadin loss and the resulting alterations in CRU structure and protein composition impairs E-C coupling and renders hearts susceptible to ventricular arrhythmias. PMID:19383796

  11. Inhibition of Recombinant Human T-type Calcium Channels by Δ9-Tetrahydrocannabinol and Cannabidiol*

    OpenAIRE

    Ross, Hamish Redmond; Napier, Ian; Connor, Mark

    2008-01-01

    Δ9-Tetrahydrocannabinol (THC) and cannabidiol (CBD) are the most prevalent biologically active constituents of Cannabis sativa. THC is the prototypic cannabinoid CB1 receptor agonist and is psychoactive and analgesic. CBD is also analgesic, but it is not a CB1 receptor agonist. Low voltage-activated T-type calcium channels, encoded by the CaV3 gene family, regulate the excitability of many cells, including neurons involved in nociceptive processing. We examined the eff...

  12. Nutritional status and specific leaf area of mahogany and tonka bean under two light environments Estado nutricional e área foliar específica de mogno e cumaru sob dois ambientes de luz

    OpenAIRE

    José Francisco de C. Gonçalves; Gil Vieira; Ricardo A. Marenco; João Baptista S. Ferraz; Ulysses Moreira dos Santos Junior; Francisco Cleber F. Barros

    2005-01-01

    Studies on nutritional status and leaf traits were carried out in two tropical tree species Swietenia macrophylla King (mahogany) and Dipetryx odorata Aubl. Willd. (tonka bean) planted under contrasting light environments in Presidente Figueiredo-AM, Brazil. Leaves of S. macrophylla and D. odorata were collected in three year-old trees grown under full sunlight (about 2000 µmol m-2 s-1) and natural shade under a closed canopy of Balsa-wood plantation (Ochroma pyramidale Cav. Ex. Lam.Urb) abou...

  13. Physiochemical analysis and centesimal composition of Pleurotus ostreatus mushroom grown in residues from the Amazon

    OpenAIRE

    Ceci Sales-Campos; Lidia Medina Araujo; Marli Teixeira de Almeida Minhoni; Meire Cristina Nogueira de Andrade

    2011-01-01

    The objective of the present work was to evaluate the nutritional composition of mushrooms produced in alternative substrates in agricultural and agro-industrial residues from the Amazon. C, N, pH, moisture, soluble solids, protein, lipids, total fibers, ashes, carbohydrates and energy were determined. Substrates were formulated from Simarouba amara Aubl. and Ochroma piramidale Cav. ex. Lam. Sawdust and from Bactris gasipaes Kunth and Saccharum officinarum stipe. Results showed that the nutri...

  14. Differentiated neuroprogenitor cells incubated with human or canine adenovirus, or lentiviral vectors have distinct transcriptome profiles.

    Directory of Open Access Journals (Sweden)

    Stefania Piersanti

    Full Text Available Several studies have demonstrated the potential for vector-mediated gene transfer to the brain. Helper-dependent (HD human (HAd and canine (CAV-2 adenovirus, and VSV-G-pseudotyped self-inactivating HIV-1 vectors (LV effectively transduce human brain cells and their toxicity has been partly analysed. However, their effect on the brain homeostasis is far from fully defined, especially because of the complexity of the central nervous system (CNS. With the goal of dissecting the toxicogenomic signatures of the three vectors for human neurons, we transduced a bona fide human neuronal system with HD-HAd, HD-CAV-2 and LV. We analysed the transcriptional response of more than 47,000 transcripts using gene chips. Chip data showed that HD-CAV-2 and LV vectors activated the innate arm of the immune response, including Toll-like receptors and hyaluronan circuits. LV vector also induced an IFN response. Moreover, HD-CAV-2 and LV vectors affected DNA damage pathways--but in opposite directions--suggesting a differential response of the p53 and ATM pathways to the vector genomes. As a general response to the vectors, human neurons activated pro-survival genes and neuron morphogenesis, presumably with the goal of re-establishing homeostasis. These data are complementary to in vivo studies on brain vector toxicity and allow a better understanding of the impact of viral vectors on human neurons, and mechanistic approaches to improve the therapeutic impact of brain-directed gene transfer.

  15. Demand controlled ventilation; Behovsstyrt ventilasjon

    Energy Technology Data Exchange (ETDEWEB)

    Soerensen, Henning Holm

    2006-07-01

    The terms CAV and VAV have been known terms for many years in the ventilation business. The terms are also included in building regulations, but the time is now right to focus on demand controlled ventilation (DCV). The new building regulations and the accompanying energy framework underline the need for a more nuanced thinking when it comes to controlling ventilation systems. Descriptions and further details of the ventilation systems are provided (ml)

  16. Mechanisms of Intracellular Calcium Homeostasis in MC3T3-E1 Cells and Bone Tissues of Sprague-Dawley Rats Exposed to Fluoride.

    Science.gov (United States)

    Duan, Xiao-Qin; Li, Yan-Hui; Zhang, Xiu-Yun; Zhao, Zhi-Tao; Wang, Ying; Wang, Huan; Li, Guang-Sheng; Jing, Ling

    2016-04-01

    Calcium homeostasis of osteoblasts (OBs) has an important role in the physiology and pathology of bone tissue. In order to study the mechanisms of intracellular calcium homeostasis, MC3T3-E1 cells and Sprague-Dawley rats were treated with different concentrations of fluoride. Then, we examined intracellular-free calcium ion ([Ca(2+)]i) in MC3T3-E1 cells as well as mRNA and protein levels of Cav1.2, the main subunit of L-type voltage-dependent calcium channels (VDCCs), Na(+)/Ca(2+) exchange carriers (NCS), and plasma membrane Ca(2+)-ATPase (PMCA), inositol 1,4,5-trisphosphate receptor (IP3R) channels, sarco/endoplasmic reticulum calcium ATPase 2b (SERCA2b)/ATP2A2 in vitro, and rat bone tissues in vivo. Our results showed that [Ca(2+)]i of fluoride-treated OBs increased in a concentration-dependent manner with an increase in the concentration of fluoride. We also found that the low dose of fluoride led to high expression levels of Cav1.2, NCS-1, and PMCA and low expression levels of IP3R and SERCA2b/ATP2A2, while the high dose of fluoride induced an increase in SERCA2b/ATP2A2 levels and decrease in Cav1.2, PMCA, NCS-1, and IP3R levels. These results demonstrate that calcium channels and calcium pumps of plasma and endoplasmic reticulum (ER) membranes keep intracellular calcium homeostasis by regulating Cav1.2, NCS-1, PMCA, IP3R, and SERCA2b/ATP2A2 expression. PMID:26276564

  17. (S)-lacosamide inhibition of CRMP2 phosphorylation reduces postoperative and neuropathic pain behaviors through distinct classes of sensory neurons identified by constellation pharmacology.

    Science.gov (United States)

    Moutal, Aubin; Chew, Lindsey A; Yang, Xiaofang; Wang, Yue; Yeon, Seul Ki; Telemi, Edwin; Meroueh, Seeneen; Park, Ki Duk; Shrinivasan, Raghuraman; Gilbraith, Kerry B; Qu, Chaoling; Xie, Jennifer Y; Patwardhan, Amol; Vanderah, Todd W; Khanna, May; Porreca, Frank; Khanna, Rajesh

    2016-07-01

    Chronic pain affects the life of millions of people. Current treatments have deleterious side effects. We have advanced a strategy for targeting protein interactions which regulate the N-type voltage-gated calcium (CaV2.2) channel as an alternative to direct channel block. Peptides uncoupling CaV2.2 interactions with the axonal collapsin response mediator protein 2 (CRMP2) were antinociceptive without effects on memory, depression, and reward/addiction. A search for small molecules that could recapitulate uncoupling of the CaV2.2-CRMP2 interaction identified (S)-lacosamide [(S)-LCM], the inactive enantiomer of the Food and Drug Administration-approved antiepileptic drug (R)-lacosamide [(R)-LCM, Vimpat]. We show that (S)-LCM, but not (R)-LCM, inhibits CRMP2 phosphorylation by cyclin dependent kinase 5, a step necessary for driving CaV2.2 activity, in sensory neurons. (S)-lacosamide inhibited depolarization-induced Ca influx with a low micromolar IC50. Voltage-clamp electrophysiology experiments demonstrated a commensurate reduction in Ca currents in sensory neurons after an acute application of (S)-LCM. Using constellation pharmacology, a recently described high content phenotypic screening platform for functional fingerprinting of neurons that uses subtype-selective pharmacological agents to elucidate cell-specific combinations (constellations) of key signaling proteins that define specific cell types, we investigated if (S)-LCM preferentially acts on certain types of neurons. (S)-lacosamide decreased the dorsal root ganglion neurons responding to mustard oil, and increased the number of cells responding to menthol. Finally, (S)-LCM reversed thermal hypersensitivity and mechanical allodynia in a model of postoperative pain, and 2 models of neuropathic pain. Thus, using (S)-LCM to inhibit CRMP2 phosphorylation is a novel and efficient strategy to treat pain, which works by targeting specific sensory neuron populations. PMID:26967696

  18. Quantitative analytical technique applied to histopathology of birds infected experimentally by the virus of chicken anemia virus

    OpenAIRE

    García, Luz; Bermudez, Victor; Brett, Mariela; Peroza, Luzmila; Landa, Juan de; Borregales, Franklin

    2008-01-01

    This research was conducted on ten glass slides selected from the histopathology evaluation chickens. Five slides of control's chickens healthy and five slides of chickens infected experimentally with chicken anemia virus (CAV slide) between one and twenty-one days post infection (PI), they were analyzed in magnifications of 200× and 400×. Histopathology showed severe bone marrow hypoplasia to complete aplasia, fully depletion of the erythrocytic and granulocytic series, both accompanied by s...

  19. Quantitative analytical technique applied to histopathology of birds infected experimentally by the virus of chicken anemia virus

    OpenAIRE

    Landa Juan; Peroza Luzmila; Brett Mariela; Bermudez Victor; García Luz; Borregales Franklin

    2008-01-01

    Abstract This research was conducted on ten glass slides selected from the histopathology evaluation chickens. Five slides of control's chickens healthy and five slides of chickens infected experimentally with chicken anemia virus (CAV slide) between one and twenty-one days post infection (PI), they were analyzed in magnifications of 200× and 400×. Histopathology showed severe bone marrow hypoplasia to complete aplasia, fully depletion of the erythrocytic and granulocytic series, both accompa...

  20. Heme oxygenase-1 regulates cell proliferation via carbon monoxide-mediated inhibition of T-type Ca2+ channels

    OpenAIRE

    Duckles, Hayley; Boycott, Hannah E.; Al-Owais, Moza M.; Elies, Jacobo; Johnson, Emily; Dallas, Mark L.; Porter, Karen E.; Giuntini, Francesca; Boyle, John P.; Scragg, Jason L.; Peers, Chris

    2014-01-01

    Induction of the antioxidant enzyme heme oxygenase-1 (HO-1) affords cellular protection and suppresses proliferation of vascular smooth muscle cells (VSMCs) associated with a variety of pathological cardiovascular conditions including myocardial infarction and vascular injury. However, the underlying mechanisms are not fully understood. Over-expression of Cav3.2 T-type Ca2+ channels in HEK293 cells raised basal [Ca2+]i and increased proliferation as compared with non-transfected cells. Prolif...