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Sample records for boldoa purpurascens cav

  1. Evaluación fotoquímica y farmacológica del extracto acuoso de las hojas de la especie Boldoa purpurascens, Cav

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    González Mosquera, Dulce María

    2007-01-01

    Boldoa purpurascens,Cav es una especie silvestre que crece en Latinoamérica y el Caribe; es utilizada en varios países como diurético y para combatir las afecciones del tracto urinario, existen escasos informes sobre dichos usos y no hay estudios desde el punto de vista químico que validen su uso tradicional. Se establecieron los parámetros de calidad de la droga. Se informa por primera vez que el extracto acuoso no presenta toxicidad ni es genotóxico. Se comprobó, que el mayor porcentaje de ...

  2. 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.

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    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.

  3. Antihyperglycemic activity of extracts from Boldoa purpurascens leaves in alloxan-induced diabetic rats.

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    González Mosquera, D M; Hernandez Ortega, Y; By, B; Vicet Muro, L; Saucedo Hernandez, Y; Grau Ábalos, R; Dehaen, W; Pieters, L; Apers, S

    2013-05-01

    In order to investigate the potential use of Boldoa purpurascens against diabetes, the antihyperglycemic effect of an ethanol extract obtained from its leaves was evaluated at doses of 50, 100 and 200 mg/kg in rats after induction of hyperglycemia by alloxan. Insulin 5 IU/kg was used as positive control and NaCl 0.9% as negative control. A similar experiment was performed with the aqueous extract used at doses of 50 and 100 mg/kg using metformin at a dose of 50mg/kg as positive control. Statistical analysis was carried using the Kruskal-Wallis test with an interval of trust of 99%. The ethanolic and aqueous extract of B. purpurascens showed a significant decrease of blood glucose levels 72 h after administration. Phytochemical analysis of the ethanol extract showed the presence of D-pinitol, a compound known for its hypoglycemic properties. In conclusion, ethanolic as well as aqueous extracts of B. purpurascens leaves show antihyperglycemic activity, possibly due to the presence of D-pinitol and flavonoids.

  4. Eluma purpurascens, nieuw voor de Nederlandse fauna (Crustacea: Isopoda: Oniscoidea)

    NARCIS (Netherlands)

    Tempelman, D.; Berg, M.P.

    1997-01-01

    Eluma purpurascens, new for the Dutch fauna (Crustacea: Isopoda: Oniscoidea). The first Dutch specimen of Eluma purpurascens was found on 29 April 1994 on the Kaloot, east of Vlissingen, in the south-western province of Zeeland. One year later the species proved to be locally common along the southe

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

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    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. Typification of Statice dichotoma Cav. (Plumbaginaceae

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    Ferrer-Gallego, P. Pablo

    2013-12-01

    Full Text Available A lectotype and an epitype are here designated for Statice dichotoma Cav. (Plumbaginaceae, an endemic taxon from the central part of the Iberian Peninsula, which is currently named Limonium dichotomum (Cav. KuntzeSe designan un lectótipo y un epítipo para Statice dichotoma Cav. (Plumbaginaceae, un taxon endémico del centro de la península Ibérica, actualmente denominado Limonium dichotomum (Cav. Kuntze.

  7. Vegetative and reproductive phenology of Butia purpurascens Glassman (Arecaceae under the effects of leaf harvesting

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    FAG Guilherme

    Full Text Available Butia purpurascens is an endemic and threatened palm tree species that occurs in open areas of the Brazilian Cerrado, predominantly in southwestern Goiás. The leaves of this palm tree are harvested by local people to fabricate brooms. This study evaluated changes in vegetative and reproductive phenology in two different natural populations of this palm tree: one population with leaf harvesting and another non-harvested population. Twenty plants were monitored in each area for 23 months. The phenophases were related to the temperature and precipitation averages for a 30-year period. Leaf sprouting occurred throughout the year, with a slight reduction in periods of low temperatures and low rainfall. The first spathes emerged in March and flowering began during the dry season (June, continuing until January of the following year, concurrent with the period of most intense fruiting. Flowering and fruiting appear to be triggered by periods of drought, which are commonly observed in the Cerrado. The harvested sites produced significantly fewer leaves, spathes, inflorescences and infructescences than the non-harvested sites. Thus, the supply of resources to the local fauna is possibly reduced in sites under leaf exploitation, which in the long term can represent damage to the palm tree population’s structure and dynamics. Other socioeconomic and ecological studies about the effects of leaf harvesting in B. purpurascens are necessary to enable strategies for sustainable use, devise management alternatives and conserve this threatened palm species.

  8. Vegetative and reproductive phenology of Butia purpurascens Glassman (Arecaceae) under the effects of leaf harvesting.

    Science.gov (United States)

    Guilherme, F A G; Vasconcelos, E I; Coelho, C P; Ressel, K; Batista, N T F; Souza, L F

    2015-01-01

    Butia purpurascens is an endemic and threatened palm tree species that occurs in open areas of the Brazilian Cerrado, predominantly in southwestern Goiás. The leaves of this palm tree are harvested by local people to fabricate brooms. This study evaluated changes in vegetative and reproductive phenology in two different natural populations of this palm tree: one population with leaf harvesting and another non-harvested population. Twenty plants were monitored in each area for 23 months. The phenophases were related to the temperature and precipitation averages for a 30-year period. Leaf sprouting occurred throughout the year, with a slight reduction in periods of low temperatures and low rainfall. The first spathes emerged in March and flowering began during the dry season (June), continuing until January of the following year, concurrent with the period of most intense fruiting. Flowering and fruiting appear to be triggered by periods of drought, which are commonly observed in the Cerrado. The harvested sites produced significantly fewer leaves, spathes, inflorescences and infructescences than the non-harvested sites. Thus, the supply of resources to the local fauna is possibly reduced in sites under leaf exploitation, which in the long term can represent damage to the palm tree population's structure and dynamics. Other socioeconomic and ecological studies about the effects of leaf harvesting in B. purpurascens are necessary to enable strategies for sustainable use, devise management alternatives and conserve this threatened palm species.

  9. Studies in the genus Riccia (Marchantiales from southern Africa. 21. R. stricta, R. purpurascens and R. fluitans, subgenus Ricciella

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    S. M. Perold

    1990-12-01

    Full Text Available Lindenberg (1836 regarded and published Riccia stricta as a variety of R. fluitans L. Subsequently, Nees (1838 and Gottsche et al. (1846 also treated it as a variety. Trevisan (1877 raised its rank and published the epithet,  Ricciella stricta Trevis.Ricciella is, however, regarded as a subgenus; Ricciella stricta is, therefore, transferred to Riccia stricta (Lindenb. Perold.It is described in detail and illustrated.  R. purpurascens Lehm. & Lindenb., a related endemic species, is also more fully described than before and illustrated.  R. fluitans L. apparently does not occur naturally in southern Africa. As far as is known, a single local specimen of it was introduced.

  10. The chemopreventive potential of Curcuma purpurascens rhizome in reducing azoxymethane-induced aberrant crypt foci in rats.

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    Rouhollahi, Elham; Moghadamtousi, Soheil Zorofchian; Al-Henhena, Nawal; Kunasegaran, Thubasni; Hasanpourghadi, Mohadeseh; Looi, Chung Yeng; Abd Malek, Sri Nurestri; Awang, Khalijah; Abdulla, Mahmood Ameen; Mohamed, Zahurin

    2015-01-01

    Curcuma purpurascens BI. rhizome, a member of the Zingiberaceae family, is a popular spice in Indonesia that is traditionally used in assorted remedies. Dichloromethane extract of C. purpurascens BI. rhizome (DECPR) has previously been shown to have an apoptosis-inducing effect on colon cancer cells. In the present study, we examined the potential of DECPR to prevent colon cancer development in rats treated with azoxymethane (AOM) (15 mg/kg) by determining the percentage inhibition in incidence of aberrant crypt foci (ACF). Starting from the day immediately after AOM treatment, three groups of rats were orally administered once a day for 2 months either 10% Tween 20 (5 mL/kg, cancer control), DECPR (250 mg/kg, low dose), or DECPR (500 mg/kg, high dose). Meanwhile, the control group was intraperitoneally injected with 5-fluorouracil (35 mg/kg) for 5 consecutive days. After euthanizing the rats, the number of ACF was enumerated in colon tissues. Bax, Bcl-2, and proliferating cell nuclear antigen (PCNA) protein expressions were examined using immunohistochemical and Western blot analyses. Antioxidant enzymatic activity was measured in colon tissue homogenates and associated with malondialdehyde level. The percentage inhibition of ACF was 56.04% and 68.68% in the low- and high-dose DECPR-treated groups, respectively. The ACF inhibition in the treatment control group was 74.17%. Results revealed that DECPR exposure at both doses significantly decreased AOM-induced ACF formation, which was accompanied by reduced expression of PCNA. Upregulation of Bax and downregulation of Bcl-2 suggested the involvement of apoptosis in the chemopreventive effect of DECPR. In addition, the oxidative stress resulting from AOM treatment was significantly attenuated after administration of DECPR, which was shown by the elevated antioxidant enzymatic activity and reduced malondialdehyde level. Taken together, the present data clearly indicate that DECPR significantly inhibits ACF formation

  11. Differential zinc permeation and blockade of L-type Ca2+ channel isoforms Cav1.2 and Cav1.3.

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    Park, So-Jung; Min, Se-Hong; Kang, Ho-Won; Lee, Jung-Ha

    2015-10-01

    Certain voltage-activated Ca2+ channels have been reported to act as potential zinc entry routes. However, it remains to be determined whether zinc can permeate individual Ca2+ channel isoforms. We expressed recombinant Ca2+ channel isoforms in Xenopus oocytes and attempted to record zinc currents from them using a two-electrode voltage clamp method. We found that, in an extracellular zinc solution, inward currents arising from zinc permeation could be recorded from Xenopus oocytes expressing L-type Cav1.2 or Cav1.3 isoforms, but not from oocytes expressing Cav2.2, Cav2.3, Cav3.1, or Cav3.2. Zinc currents through Cav1.2 and Cav1.3 were blocked by nimodipine, but enhanced by (±)Bay K8644, supporting the finding that zinc can permeate both L-type Cav1.2 and Cav1.3 channel isoforms. We also examined the blocking effects of low concentrations of zinc on Ca2+ currents through the L-type channel isoforms. Low micro-molar zinc potently blocked Ca2+ currents through Cav1.2 and Cav1.3 with different sensitivities (IC50 for Cav1.2 and Cav1.3=18.4 and 34.1 μM) and de-accelerated the activation and inactivation kinetics in a concentration-dependent manner. Notably, mild acidifications of the external zinc solution increased zinc currents through Cav1.2 and Cav1.3, with the increment level for Cav1.3 being greater than that for Cav1.2. In overall, we provide evidence that Cav1.2 and Cav1.3 isoforms are capable of potentially functioning as zinc permeation routes, through which zinc entry can be differentially augmented by mild acidifications.

  12. The chemopreventive potential of Curcuma purpurascens rhizome in reducing azoxymethane-induced aberrant crypt foci in rats

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    Rouhollahi E

    2015-07-01

    Full Text Available Elham Rouhollahi,1 Soheil Zorofchian Moghadamtousi,2 Nawal Al-Henhena,3 Thubasni Kunasegaran,1 Mohadeseh Hasanpourghadi,4 Chung Yeng Looi,4 Sri Nurestri Abd Malek,2 Khalijah Awang,5 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, 4Cell Biology and Drug Discovery Laboratory, Department of Pharmacology, Faculty of Medicine, 5Department of Chemistry, University of Malaya, Kuala Lumpur, Malaysia Abstract: Curcuma purpurascens BI. rhizome, a member of the Zingiberaceae family, is a popular spice in Indonesia that is traditionally used in assorted remedies. Dichloromethane extract of C. purpurascens BI. rhizome (DECPR has previously been shown to have an apoptosis-inducing effect on colon cancer cells. In the present study, we examined the potential of DECPR to prevent colon cancer development in rats treated with azoxymethane (AOM (15 mg/kg by determining the percentage inhibition in incidence of aberrant crypt foci (ACF. Starting from the day immediately after AOM treatment, three groups of rats were orally administered once a day for 2 months either 10% Tween 20 (5 mL/kg, cancer control, DECPR (250 mg/kg, low dose, or DECPR (500 mg/kg, high dose. Meanwhile, the control group was intraperitoneally injected with 5-fluorouracil (35 mg/kg for 5 consecutive days. After euthanizing the rats, the number of ACF was enumerated in colon tissues. Bax, Bcl-2, and proliferating cell nuclear antigen (PCNA protein expressions were examined using immunohistochemical and Western blot analyses. Antioxidant enzymatic activity was measured in colon tissue homogenates and associated with malondialdehyde level. The percentage inhibition of ACF was 56.04% and 68.68% in the low- and high-dose DECPR-treated groups, respectively. The ACF inhibition in the treatment control group was 74.17%. Results revealed that

  13. Cav1.2, but not Cav1.3, L-type calcium channel subtype mediates nicotine-induced conditioned place preference in miceo.

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    Liu, Yudan; Harding, Meghan; Dore, Jules; Chen, Xihua

    2017-04-03

    Nicotine use is one of the most common forms of drug addiction. Although L-type calcium channels (LTCCs) are involved in nicotine addiction, the contribution of the two primary LTCC subtypes (Cav1.2 and 1.3) is unknown. This study aims to determine the contribution of these two LTCC subtypes to nicotine-induced conditioned place preference (CPP) responses by using transgenic mouse models that do not express Cav1.3 (Cav1.3(-/-)) or contain a mutation in the dihydropyridine (DHP) site of the Cav1.2 (Cav1.2DHP(-/-)). We found a hyperbolic dose dependent nicotine (0.1-1mg/kg; 0.5mg/kg optimum) effect on place preference in wild type (WT) mice, that could be prevented by the DHP LTCC blocker nifedipine pretreatment. Similarly, Cav1.3(-/-) mice showed nicotine-induced place preference which was antagonized by nifedipine. In contrast, nifedipine pretreatment of Cav1.2DHP(-/-) mice had no effect on nicotine-induced CPP responses, suggesting an involvement of Cav1.2 subtype in the nicotine-induced CPP response. Nifedipine alone failed to produce either conditioned place aversion or CPP in WT mice. These results collectively indicate Cav1.2, but not Cav1.3 LTCC subtype regulates, at least in part, the reinforcing effects of nicotine use.

  14. Ca(V)1 and Ca(V)2 channels engage distinct modes of Ca(2+) signaling to control CREB-dependent gene expression.

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    Wheeler, Damian G; Groth, Rachel D; Ma, Huan; Barrett, Curtis F; Owen, Scott F; Safa, Parsa; Tsien, Richard W

    2012-05-25

    Activity-dependent gene expression triggered by Ca(2+) entry into neurons is critical for learning and memory, but whether specific sources of Ca(2+) act distinctly or merely supply Ca(2+) to a common pool remains uncertain. Here, we report that both signaling modes coexist and pertain to Ca(V)1 and Ca(V)2 channels, respectively, coupling membrane depolarization to CREB phosphorylation and gene expression. Ca(V)1 channels are advantaged in their voltage-dependent gating and use nanodomain Ca(2+) to drive local CaMKII aggregation and trigger communication with the nucleus. In contrast, Ca(V)2 channels must elevate [Ca(2+)](i) microns away and promote CaMKII aggregation at Ca(V)1 channels. Consequently, Ca(V)2 channels are ~10-fold less effective in signaling to the nucleus than are Ca(V)1 channels for the same bulk [Ca(2+)](i) increase. Furthermore, Ca(V)2-mediated Ca(2+) rises are preferentially curbed by uptake into the endoplasmic reticulum and mitochondria. This source-biased buffering limits the spatial spread of Ca(2+), further attenuating Ca(V)2-mediated gene expression.

  15. CAV1 promotes HCC cell progression and metastasis through Wnt/β-catenin pathway.

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    Hongxiu Yu

    Full Text Available Caveolin-1 (CAV1 has significant roles in many primary tumors and metastasis, despite the fact that malignant cells from different cancer types have different profiles of CAV1 expression. There is little information concerning CAV1 expression and role in hepatocellular carcinoma (HCC progresion and metastasis. The role of CAV1 in HCC progression was explored in this study. We reported that CAV1 was overexpressed in highly invasive HCC cell lines compared with poorly invasive ones. The immunohistochemical staining was obviously stronger in metastatic HCC samples than in the non-metastatic specimens via tissue microarrays. Furthermore, CAV1 overexpression enhanced HCC cell invasiveness in vitro, and promoted tumorigenicity and lung metastasis in vivo. By contrast, CAV1 stable knockdown markedly reduced these malignant behaviors. Importantly, we found that CAV1 could induce EMT process through Wnt/β-catenin pathway to promote HCC metastasis. We also identify MMP-7 as a novel downstream target of CAV1. We have determined that CAV1 acts as a mediator between hyperactive ERK1/2 signaling and regulation of MMP-7 transcription. Together, these studies mechanistically show a previously unrecognized interplay between CAV1, EMT, ERK1/2 and MMP-7 that is likely significant in the progression of HCC toward metastasis.

  16. 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!

  17. Expression and pharmacology of endogenous Cav channels in SH-SY5Y human neuroblastoma cells.

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    Silmara R Sousa

    Full Text Available SH-SY5Y human neuroblastoma cells provide a useful in vitro model to study the mechanisms underlying neurotransmission and nociception. These cells are derived from human sympathetic neuronal tissue and thus, express a number of the Cav channel subtypes essential for regulation of important physiological functions, such as heart contraction and nociception, including the clinically validated pain target Cav2.2. We have detected mRNA transcripts for a range of endogenous expressed subtypes Cav1.3, Cav2.2 (including two Cav1.3, and three Cav2.2 splice variant isoforms and Cav3.1 in SH-SY5Y cells; as well as Cav auxiliary subunits α2δ1-3, β1, β3, β4, γ1, γ4-5, and γ7. Both high- and low-voltage activated Cav channels generated calcium signals in SH-SY5Y cells. Pharmacological characterisation using ω-conotoxins CVID and MVIIA revealed significantly (∼ 10-fold higher affinity at human versus rat Cav2.2, while GVIA, which interacts with Cav2.2 through a distinct pharmacophore had similar affinity for both species. CVID, GVIA and MVIIA affinity was higher for SH-SY5Y membranes vs whole cells in the binding assays and functional assays, suggesting auxiliary subunits expressed endogenously in native systems can strongly influence Cav2.2 channels pharmacology. These results may have implications for strategies used to identify therapeutic leads at Cav2.2 channels.

  18. A description of the larva of Coomaniella (Coomaniella) purpurascens Baudon, 1966 and a discussion on the tribal placement of Coomaniellini Bílý, 1974 (Coleoptera: Buprestidae).

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    Bílý, Svatopluk; Volkovitsh, Mark G

    2015-02-02

    A detailed description of the mature larva of Coomaniella (Coomaniella) purpurascens Baudon, 1966 and a discussion of the taxonomic position of the tribe Coomaniellini Bílý, 1974 are presented. Larva is fully illustrated including details of the mouth parts and inner structures of the proventriculus. The larval characters support the tribal status of Coomaniellini with close relations to the tribe Dicercini Gistel, 1884 in contrast with the former placement of the tribe near Anthaxiini Gory & Laporte, 1839. 

  19. 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.

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    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.

  20. A genetic polymorphism in the CAV1 gene associates with the development of bronchiolitis obliterans syndrome after lung transplantation

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    Kastelijn Elisabeth A

    2011-11-01

    Full Text Available Abstract Background Caveolin 1 (Cav-1 is the primary structural component of cell membrane invaginations called 'caveolae'. Expression of Cav-1 is implicated in the pathogenesis of pulmonary fibrosis. Genetic polymorphisms in the CAV1 gene influence the function of Cav-1 in malignancies and associate with renal allograft fibrosis. Chronic allograft rejection after lung transplantation, called 'bronchiolitis obliterans syndrome' (BOS, is also characterised by the development of fibrosis. In this study, we investigated whether CAV1 genotypes associate with BOS and whether Cav-1 serum levels are influenced by the CAV1 genotype and can be used as a biomarker to predict the development of BOS. Methods Twenty lung transplant recipients with BOS (BOSpos, ninety without BOS (BOSneg and four hundred twenty-two healthy individuals donated DNA samples. Four SNPs in CAV1 were genotyped. Serial Cav-1 serum levels were measured in a matched cohort of 10 BOSpos patients and 10 BOSneg patients. Furthermore, single-time point Cav-1 serum levels were measured in 33 unmatched BOSneg patients and 60 healthy controls. Results Homozygosity of the minor allele of rs3807989 was associated with an increased risk for BOS (odds ratio: 6.13; P = 0.0013. The median Cav-1 serum level was significantly higher in the BOSpos patients than in the matched BOSneg patients (P = 0.026. Longitudinal analysis did not show changes in Cav-1 serum levels over time in both groups. The median Cav-1 serum level in the group of 43 BOSneg patients was lower than that in the healthy control group (P = 0.046. In lung transplant recipients, homozygosity of the minor allele of rs3807989 and rs3807994 was associated with increased Cav-1 serum levels. Conclusion In lung transplant recipients, the CAV1 SNP rs3807989 was associated with the development of BOS and Cav-1 serum levels were influenced by the CAV1 genotype.

  1. Essential oil content of the rhizome of Curcuma purpurascens Bl. (Temu Tis) and its antiproliferative effect on selected human carcinoma cell lines.

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

    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, followed by germacrone, ar-turmerone, germacrene-B, and curlone. The rhizome oil of temu tis exhibited strong cytotoxicity against HT29 cells (IC50 value of 4.9 ± 0.4 μg/mL), weak cytotoxicity against A549, Ca Ski, and HCT116 cells (with IC50 values of 46.3 ± 0.7, 32.5 ± 1.1, and 35.0 ± 0.3 μg/mL, resp.), and no inhibitory effect against MCF7 cells. It exhibited mild cytotoxicity against a noncancerous human lung fibroblast cell line (MRC5), with an IC50 value of 25.2 ± 2.7 μg/mL. This is the first report on the chemical composition of this rhizome's oil and its selective antiproliferative effect on HT29. The obtained data provided a basis for further investigation of the mode of cell death.

  2. Schizophrenia risk gene CAV1 is both pro-psychotic and required for atypical antipsychotic drug actions in vivo

    OpenAIRE

    Allen, J A; Yadav, P N; Setola, V.; Farrell, M.; Roth, B L

    2011-01-01

    Caveolin-1 (Cav-1) is a scaffolding protein important for regulating receptor signaling cascades by partitioning signaling molecules into membrane microdomains. Disruption of the CAV1 gene has recently been identified as a rare structural variant associated with schizophrenia. Although Cav-1 knockout (KO) mice displayed no baseline behavioral disruptions, Cav-1 KO mice, similar to schizophrenic individuals, exhibited increased sensitivity to the psychotomimetic N-methyl--aspartate receptor an...

  3. CaV1.2 calcium channels: just cut out to be regulated?

    Science.gov (United States)

    Groth, Rachel D; Tirko, Natasha N; Tsien, Richard W

    2014-06-04

    Tight regulation of calcium entry through the L-type calcium channel CaV1.2 ensures optimal excitation-response coupling. In this issue of Neuron, Michailidis et al. (2014) demonstrate that CaV1.2 activity triggers negative feedback regulation through proteolytic cleavage of the channel within the core of the pore-forming subunit.

  4. Intrathecal administration of Cav3.2 and Cav3.3 antisense oligonucleotide reverses tactile allodynia and thermal hyperalgesia in rats following chronic compression of dorsal root of ganglion

    Institute of Scientific and Technical Information of China (English)

    Xian-jie WEN; Zhang-jun LI; Zhi-xin CHEN; Zhi-yuan FANG; Chen-xiang YANG; Heng LI; Yin-ming ZENG

    2006-01-01

    Aim: The present study aimed to elucidate the role of T-subtype calcium channels (Cav3.1, Cav3.2, and Cav3.3) in the pathogenesis of neuropathic pain at spinal level. Methods: The chronic compression of the dorsal root ganglion (CCD) rat model was adopted. The antisense oligonucleotide of Cav3.1, Cav3.2, and Cav3.3 or normal saline (NS) were intrathecally administered twice per day from the first day to the fourth day after operation. Paw mechanical withdrawal threshold and paw thermal withdrawal latency were measured to evaluate the tactile allodynia and thermal hyperalgesia, respectively. Results: CCD rats developed reliable tactile allodynia and thermal hyperalgesia after operation. Intrathecal administration of antisense oligonucleotide of Cav3.2 and Cav3.3 significantly relieved tactile allodynia and thermal hyperalgesia in CCD rats, but not Cav3.1. Conclusion: Cav2 and Cav3.3 subtype calcium channels in the spinal cord may play an important role in the pathogenesis of neuropathic pain, which may contribute to the management of the neuropathic pain.

  5. T Cell Receptor Mediated Calcium Entry Requires Alternatively Spliced Cav1.1 Channels.

    Directory of Open Access Journals (Sweden)

    Didi Matza

    Full Text Available The process of calcium entry in T cells is a multichannel and multi-step process. We have studied the requirement for L-type calcium channels (Cav1.1 α1S subunits during calcium entry after TCR stimulation. High expression levels of Cav1.1 channels were detected in activated T cells. Sequencing and cloning of Cav1.1 channel cDNA from T cells revealed that a single splice variant is expressed. This variant lacks exon 29, which encodes the linker region adjacent to the voltage sensor, but contains five new N-terminal exons that substitute for exons 1 and 2, which are found in the Cav1.1 muscle counterpart. Overexpression studies using cloned T cell Cav1.1 in 293HEK cells (that lack TCR suggest that the gating of these channels was altered. Knockdown of Cav1.1 channels in T cells abrogated calcium entry after TCR stimulation, suggesting that Cav1.1 channels are controlled by TCR signaling.

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

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    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.

  7. The Brugada syndrome mutation A39V does not affect surface expression of neuronal rat Cav1.2 channels

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    Simms Brett A

    2012-03-01

    Full Text Available Abstract Background A loss of function of the L-type calcium channel, Cav1.2, results in a cardiac specific disease known as Brugada syndrome. Although many Brugada syndrome channelopathies reduce channel function, one point mutation in the N-terminus of Cav1.2 (A39V has been shown to elicit disease a phenotype because of a loss of surface trafficking of the channel. This lack of cell membrane expression could not be rescued by the trafficking chaperone Cavβ. Findings We report that despite the striking loss of trafficking described previously in the cardiac Cav1.2 channel, the A39V mutation while in the background of the brain isoform traffics and functions normally. We detected no differences in biophysical properties between wild type Cav1.2 and A39V-Cav1.2 in the presence of either a cardiac (Cavβ2b, or a neuronal beta subunit (Cavβ1b. In addition, the A39V-Cav1.2 mutant showed a normal Cavβ2b mediated increase in surface expression in tsA-201 cells. Conclusions The Brugada syndrome mutation A39V when introduced into rat brain Cav1.2 does not trigger the loss-of-trafficking phenotype seen in a previous study on the human heart isoform of the channel.

  8. Environmental perspectives of Phragmites australis (Cav.) Trin. Ex. Steudel

    Science.gov (United States)

    Srivastava, Jatin; Kalra, Swinder J. S.; Naraian, Ram

    2014-09-01

    Extensive research is being conducted worldwide to find alternative and efficient systems to lessen the impacts of climate change and reduce environmental pollution. The genus Phragmites has proven ability to mitigate the environmental pollution of its surroundings. Common reed ( Phragmites australis (Cav.) Trin. Ex. Steudel), a graminaceous plant of cosmopolitan nature, has been extensively studied especially for the mitigation of environmental contamination. The capability of common reed to grow well at extreme environmental conditions such as elevated CO2 and high temperature is conferred by several factors such as change of carbon trapping mechanism (from C3 to C4 and vice versa), microbial association and biochemical adaptations. P. australis has been a most preferred unique plant system, especially in ecological engineering for improving the quality of wastewater. This paper reviews the current state of knowledge regarding the suitability of Phragmites australis for environmental remediation and summarizes recent advancements in our understanding of this grass.

  9. DEMONSTRATION BULLETIN: CAV-OX ULTRAVIOLET OXIDATION PROCESS MAGNUM WATER TECHNOLOGY

    Science.gov (United States)

    The CAV-OX® technology (see Fig- ure 1) destroys organic contaminants, including chlorinated hy- drocarbons, in water. The process uses hydrogen peroxide, hy- drodynamic cavitation, and ultraviolet (UV) radiation to photolyze and oxidize organic compounds present in water at ...

  10. Variability of Phragmites australis (Cav.) Trin. ex Streudel in relation to the environment

    NARCIS (Netherlands)

    Toorn, Jaap van der

    1972-01-01

    Een onderzoek werd uitgevoerd naar de variabiliteit van Phragmites australis (Cav.) Trin ex Steudel, het Riet, in Nederland, waarbij werd nagegaan in hoeverre deze in verband staat met het milieu. ... Zie: Samenvatting

  11. CaV3.2 Channels and the Induction of Negative Feedback in Cerebral Arteries

    Science.gov (United States)

    Harraz, Osama F.; Abd El-Rahman, Rasha R.; Bigdely-Shamloo, Kamran; Wilson, Sean M.; Brett, Suzanne E.; Romero, Monica; Gonzales, Albert L.; Earley, Scott; Vigmond, Edward J.; Nygren, Anders; Menon, Bijoy K.; Mufti, Rania E.; Watson, Tim; Starreveld, Yves; Furstenhaupt, Tobias; Muellerleile, Philip R.; Kurjiaka, David T.; Kyle, Barry D.; Braun, Andrew P.; Welsh, Donald G.

    2015-01-01

    Rationale T-type (CaV3.1/CaV3.2) Ca2+ channels are expressed in rat cerebral arterial smooth muscle. Although present, their functional significance remains uncertain with findings pointing to a variety of roles. Objective This study tested whether CaV3.2 channels mediate a negative feedback response by triggering Ca2+ sparks, discrete events that initiate arterial hyperpolarization by activating large-conductance Ca2+-activated K+ channels. Methods and Results Micromolar Ni2+, an agent that selectively blocks CaV3.2 but not CaV1.2/CaV3.1, was first shown to depolarize/constrict pressurized rat cerebral arteries; no effect was observed in CaV3.2−/− arteries. Structural analysis using 3-dimensional tomography, immunolabeling, and a proximity ligation assay next revealed the existence of microdomains in cerebral arterial smooth muscle which comprised sarcoplasmic reticulum and caveolae. Within these discrete structures, CaV3.2 and ryanodine receptor resided in close apposition to one another. Computational modeling revealed that Ca2+ influx through CaV3.2 could repetitively activate ryanodine receptor, inducing discrete Ca2+-induced Ca2+ release events in a voltage-dependent manner. In keeping with theoretical observations, rapid Ca2+ imaging and perforated patch clamp electrophysiology demonstrated that Ni2+ suppressed Ca2+ sparks and consequently spontaneous transient outward K+ currents, large-conductance Ca2+-activated K+ channel mediated events. Additional functional work on pressurized arteries noted that paxilline, a large-conductance Ca2+-activated K+ channel inhibitor, elicited arterial constriction equivalent, and not additive, to Ni2+. Key experiments on human cerebral arteries indicate that CaV3.2 is present and drives a comparable response to moderate constriction. Conclusions These findings indicate for the first time that CaV3.2 channels localize to discrete microdomains and drive ryanodine receptor–mediated Ca2+ sparks, enabling large

  12. 黑附球菌抗紫外线保护剂的筛选%Screening of ultraviolet protectants for Epicoccum purpurascens

    Institute of Scientific and Technical Information of China (English)

    夏丽琴; 王亚南; 周岱超; 王树桐; 张瑜; 胡同乐; 曹克强

    2012-01-01

    In order to reduce the impact of UV on conidia of Epicoccum purpurascens strain XF1 and improve the biological activity of conidia in the field,the protective efficiency of ultraviolet protectant (UVP) combinations of 6 UV-absorbers and 4 light stabilizers against UV for conidia of E. purpurascens strain XF1 was tested in laboratory. The results showed that the protective efficiency of different UVP combinations was different, combinations of benzotriazole and hexamethyl phosphorus triamide (a3bl), and combinations of oxybenzone and hexamethyl phosphorus triamide ( a4bl and a5bl) ,were confirmed to be more effective than the other UV-protectants, with protective efficiency over 90% after 40 min exposure to UV light (at λ253.7N,) ,while that of combinations of salicylic acid benzyl ester and hexamethyl phosphorus triamide (albl) was only about 37. 12% . UVP combinations without protective efficiency included combinations of salicylic acid benzyl esters and light stabilizer 770,benzotriazole and light stabilizer 770,benzophenones and light stabilizer 770 (alb2,a3b2,a4b2, a5b2). The appropriate solvent combination was V( solvent n-hexane): V( peanut oil) =1:1 for (a3bl, a4bl,a5bl). For the three UVP complexes selected, the effective concentration of a4bl and a5bl should be higher than 7.5 mg/mL,while that of a3bl should be higher than 10 mg/mL. After 90 min and 180 min exposure of conidia to UV light (at λ253.7N,) , the protective efficiency of the three selective UVP combinations were over 88% and 63% Respectively.%为减少紫外线对黑附球菌Epicoccum purpurascens分生孢子的影响,提高分生孢子在田间的生物活性,在室内检测了6种紫外线吸收剂和4种光稳定剂之间的不同组合(复合型抗紫外线保护剂)对黑附球菌XF1菌株分生孢子抗紫外线的保护作用.结果表明:不同复合型抗紫外线保护剂对XF1分生孢子的保护效率差异较大,其中苯并三唑类与六甲基磷酰三胺的组合(a3b1)及苯

  13. Composition and Antidiarrheal Activity of Bidens odorata Cav.

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    Daniel Zavala-Mendoza

    2013-01-01

    Full Text Available The antidiarrheal effects of chloroform, methanol, and aqueous extracts of Bidens odorata Cav. were investigated at doses of 200 mg/kg on castor-oil-induced diarrhea. The chloroform extract of B. odorata (CBO reduced diarrhea by 72.72%. The effect of CBO was evaluated on mice with diarrhea induced by castor oil, MgSO4, arachidonic acid, or prostaglandin E2. CBO inhibited the contraction induced by carbachol chloride on ileum (100 µg/mL and intestinal transit (200 mg/kg in Wistar rats. The active fraction of CBO (F4 at doses of 100 mg/kg inhibited the diarrhea induced by castor oil (90.1% or arachidonic acid (72.9% but did not inhibit the diarrhea induced by PGE2. The active fraction of F4 (FR5 only was tested on diarrhea induced with castor oil and inhibited this diarrhea by 92.1%. The compositions of F4 and FR5 were determined by GC-MS, and oleic, palmitic, linoleic, and stearic acids were found. F4 and a mixture of the four fatty acids inhibited diarrhea at doses of 100 mg/kg (90.1% and 70.6%, resp.. The results of this study show that B. odorata has antidiarrheal effects, as is claimed by folk medicine, and could possibly be used for the production of a phytomedicine.

  14. Composition and Antidiarrheal Activity of Bidens odorata Cav.

    Science.gov (United States)

    Zavala-Mendoza, Daniel; Alarcon-Aguilar, Francisco J.; Pérez-Gutierrez, Salud; Escobar-Villanueva, M. Carmen; Zavala-Sánchez, Miguel A.

    2013-01-01

    The antidiarrheal effects of chloroform, methanol, and aqueous extracts of Bidens odorata Cav. were investigated at doses of 200 mg/kg on castor-oil-induced diarrhea. The chloroform extract of B. odorata (CBO) reduced diarrhea by 72.72%. The effect of CBO was evaluated on mice with diarrhea induced by castor oil, MgSO4, arachidonic acid, or prostaglandin E2. CBO inhibited the contraction induced by carbachol chloride on ileum (100 µg/mL) and intestinal transit (200 mg/kg) in Wistar rats. The active fraction of CBO (F4) at doses of 100 mg/kg inhibited the diarrhea induced by castor oil (90.1%) or arachidonic acid (72.9%) but did not inhibit the diarrhea induced by PGE2. The active fraction of F4 (FR5) only was tested on diarrhea induced with castor oil and inhibited this diarrhea by 92.1%. The compositions of F4 and FR5 were determined by GC-MS, and oleic, palmitic, linoleic, and stearic acids were found. F4 and a mixture of the four fatty acids inhibited diarrhea at doses of 100 mg/kg (90.1% and 70.6%, resp.). The results of this study show that B. odorata has antidiarrheal effects, as is claimed by folk medicine, and could possibly be used for the production of a phytomedicine. PMID:24282432

  15. Levels of CaV1.2 L-Type Ca2+ Channels Peak in the First Two Weeks in Rat Hippocampus Whereas CaV1.3 Channels Steadily Increase through Development

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    Audra A. Kramer

    2012-01-01

    Full Text Available Influx of calcium through voltage-dependent channels regulates processes throughout the nervous system. Specifically, influx through L-type channels plays a variety of roles in early neuronal development and is commonly modulated by G-protein-coupled receptors such as GABAB receptors. Of the four isoforms of L-type channels, only CaV1.2 and CaV1.3 are predominately expressed in the nervous system. Both isoforms are inhibited by the same pharmacological agents, so it has been difficult to determine the role of specific isoforms in physiological processes. In the present study, Western blot analysis and confocal microscopy were utilized to study developmental expression levels and patterns of CaV1.2 and CaV1.3 in the CA1 region of rat hippocampus. Steady-state expression of CaV1.2 predominated during the early neonatal period decreasing by day 12. Steady-state expression of CaV1.3 was low at birth and gradually rose to adult levels by postnatal day 15. In immunohistochemical studies, antibodies against CaV1.2 and CaV1.3 demonstrated the highest intensity of labeling in the proximal dendrites at all ages studied (P1–72. Immunohistochemical studies on one-week-old hippocampi demonstrated significantly more colocalization of GABAB receptors with CaV1.2 than with CaV1.3, suggesting that modulation of L-type calcium current in early development is mediated through CaV1.2 channels.

  16. Exploring the dominant role of Cav1 channels in signalling to the nucleus.

    Science.gov (United States)

    Ma, Huan; Cohen, Samuel; Li, Boxing; Tsien, Richard W

    2012-12-20

    Calcium is important in controlling nuclear gene expression through the activation of multiple signal-transduction pathways in neurons. Compared with other voltage-gated calcium channels, Ca(V)1 channels demonstrate a considerable advantage in signalling to the nucleus. In this review, we summarize the recent progress in elucidating the mechanisms involved. Ca(V)1 channels, already advantaged in their responsiveness to depolarization, trigger communication with the nucleus by attracting colocalized clusters of activated CaMKII (Ca(2+)/calmodulin-dependent protein kinase II). Ca(V)2 channels lack this ability, but must work at a distance of >1 μm from the Ca(V)1-CaMKII co-clusters, which hampers their relative efficiency for a given rise in bulk [Ca(2+)](i) (intracellular [Ca(2+)]). Moreover, Ca(2+) influx from Ca(V)2 channels is preferentially buffered by the ER (endoplasmic reticulum) and mitochondria, further attenuating their effectiveness in signalling to the nucleus.

  17. Exploring the dominant role of Cav1 channels in signalling to the nucleus

    Science.gov (United States)

    Ma, Huan; Cohen, Samuel; Li, Boxing; Tsien, Richard W.

    2012-01-01

    Calcium is important in controlling nuclear gene expression through the activation of multiple signal-transduction pathways in neurons. Compared with other voltage-gated calcium channels, CaV1 channels demonstrate a considerable advantage in signalling to the nucleus. In this review, we summarize the recent progress in elucidating the mechanisms involved. CaV1 channels, already advantaged in their responsiveness to depolarization, trigger communication with the nucleus by attracting colocalized clusters of activated CaMKII (Ca2+/calmodulin-dependent protein kinase II). CaV2 channels lack this ability, but must work at a distance of >1 μm from the CaV1-CaMKII co-clusters, which hampers their relative efficiency for a given rise in bulk [Ca2+]i (intracellular [Ca2+]). Moreover, Ca2+ influx from CaV2 channels is preferentially buffered by the ER (endoplasmic reticulum) and mitochondria, further attenuating their effectiveness in signalling to the nucleus. PMID:23088728

  18. 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.

  19. Prevalence of antibodies against chicken anaemia virus (CAV in broiler breeders in Southern Brazil Prevalência de anticorpos contra o vírus da anemia das galinhas (CAV em matrizes de corte no Sul do Brasil

    Directory of Open Access Journals (Sweden)

    Cláudio W. Canal

    2004-06-01

    Full Text Available Chicks infected during the first two weeks of life with chicken anaemia virus (CAV manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA. All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.A doença clínica causada pelo vírus da anemia das galinhas (CAV ocorre quando os pintos são infectados durante as primeiras duas semanas de vida e pode ser prevenida se as matrizes transferirem anticorpos suficientes para a sua progênie. Em vista disso, este estudo foi realizado visando determinar a prevalência de anticorpos contra o CAV em alguns lotes de matrizes pesadas no Brasil. Buscou-se ainda verificar em que fase da vida as reprodutoras seriam infectadas e quais seriam os títulos de anticorpos nessas aves. Um total de 1709 amostras de soro de 64 lotes de reprodutoras não vacinadas e 12 lotes de reprodutoras vacinadas contra o CAV foram analisados por ELISA. Todos os lotes de aves não vacinadas apresentaram anticorpos. Dentre esses, 89% dos indivíduos foram positivos, 52% com títulos de anticorpos capazes de conferir proteção a sua progênie contra o CAV. Igualmente, todos os lotes de matrizes vacinadas

  20. The CAV program for numerical evaluation of laminar natural convection heat transfer in vertical rectangular cavities

    Science.gov (United States)

    Novak, Milos H.; Nowak, Edwin S.

    1993-12-01

    To analyze the laminar natural convection heat transfer and fluid flow distribution in vertical rectangular cavities with or without inner partitions, the personal computer finite difference program entitled CAV is used. The CAV program was tested successfully for slender cavities with aspect ratios as high as R = H/ L = 90 and for the Grashof numbers, based on the cavity height, up to GrH = 3 x10 9. To make the CAV program useful for a number of applications, various types of boundary conditions can also be imposed on the program calculations. Presented are program applications dealing with the 2-D numerical analysis of natural convection heat transfer in very slender window cavities with and without small inner partitions and recommendations are made for window design.

  1. β-Adrenergic stimulation increases Cav3.1 activity in cardiac myocytes through protein kinase A.

    Directory of Open Access Journals (Sweden)

    Yingxin Li

    Full Text Available The T-type Ca(2+ channel (TTCC plays important roles in cellular excitability and Ca(2+ regulation. In the heart, TTCC is found in the sinoatrial nodal (SAN and conduction cells. Cav3.1 encodes one of the three types of TTCCs. To date, there is no report regarding the regulation of Cav3.1 by β-adrenergic agonists, which is the topic of this study. Ventricular myocytes (VMs from Cav3.1 double transgenic (TG mice and SAN cells from wild type, Cav3.1 knockout, or Cav3.2 knockout mice were used to study β-adrenergic regulation of overexpressed or native Cav3.1-mediated T-type Ca(2+ current (I(Ca-T(3.1. I(Ca-T(3.1 was not found in control VMs but was robust in all examined TG-VMs. A β-adrenergic agonist (isoproterenol, ISO and a cyclic AMP analog (dibutyryl-cAMP significantly increased I(Ca-T(3.1 as well as I(Ca-L in TG-VMs at both physiological and room temperatures. The ISO effect on I(Ca-L and I(Ca-T in TG myocytes was blocked by H89, a PKA inhibitor. I(Ca-T was detected in control wildtype SAN cells but not in Cav3.1 knockout SAN cells, indicating the identity of I(Ca-T in normal SAN cells is mediated by Cav3.1. Real-time PCR confirmed the presence of Cav3.1 mRNA but not mRNAs of Cav3.2 and Cav3.3 in the SAN. I(Ca-T in SAN cells from wild type or Cav3.2 knockout mice was significantly increased by ISO, suggesting native Cav3.1 channels can be upregulated by the β-adrenergic (β-AR system. In conclusion, β-adrenergic stimulation increases I(Ca-T(3.1 in cardiomyocytes(, which is mediated by the cAMP/PKA pathway. The upregulation of I(Ca-T(3.1 by the β-adrenergic system could play important roles in cellular functions involving Cav3.1.

  2. Purkinje cell-specific ablation of Cav2.1 channels is sufficient to cause cerebellar ataxia in mice.

    Science.gov (United States)

    Todorov, Boyan; Kros, Lieke; Shyti, Reinald; Plak, Petra; Haasdijk, Elize D; Raike, Robert S; Frants, Rune R; Hess, Ellen J; Hoebeek, Freek E; De Zeeuw, Chris I; van den Maagdenberg, Arn M J M

    2012-03-01

    The Cacna1a gene encodes the α(1A) subunit of voltage-gated Ca(V)2.1 Ca(2+) channels that are involved in neurotransmission at central synapses. Ca(V)2.1-α(1)-knockout (α1KO) mice, which lack Ca(V)2.1 channels in all neurons, have a very severe phenotype of cerebellar ataxia and dystonia, and usually die around postnatal day 20. This early lethality, combined with the wide expression of Ca(V)2.1 channels throughout the cerebellar cortex and nuclei, prohibited determination of the contribution of particular cerebellar cell types to the development of the severe neurobiological phenotype in Cacna1a mutant mice. Here, we crossed conditional Cacna1a mice with transgenic mice expressing Cre recombinase, driven by the Purkinje cell-specific Pcp2 promoter, to specifically ablate the Ca(V)2.1-α(1A) subunit and thereby Ca(V)2.1 channels in Purkinje cells. Purkinje cell Ca(V)2.1-α(1A)-knockout (PCα1KO) mice aged without difficulties, rescuing the lethal phenotype seen in α1KO mice. PCα1KO mice exhibited cerebellar ataxia starting around P12, much earlier than the first signs of progressive Purkinje cell loss, which appears in these mice between P30 and P45. Secondary cell loss was observed in the granular and molecular layers of the cerebellum and the volume of all individual cerebellar nuclei was reduced. In this mouse model with a cell type-specific ablation of Ca(V)2.1 channels, we show that ablation of Ca(V)2.1 channels restricted to Purkinje cells is sufficient to cause cerebellar ataxia. We demonstrate that spatial ablation of Ca(V)2.1 channels may help in unraveling mechanisms of human disease.

  3. Splice variants of the CaV1.3 L-type calcium channel regulate dendritic spine morphology

    Science.gov (United States)

    Stanika, Ruslan; Campiglio, Marta; Pinggera, Alexandra; Lee, Amy; Striessnig, Jörg; Flucher, Bernhard E.; Obermair, Gerald J.

    2016-01-01

    Dendritic spines are the postsynaptic compartments of glutamatergic synapses in the brain. Their number and shape are subject to change in synaptic plasticity and neurological disorders including autism spectrum disorders and Parkinson’s disease. The L-type calcium channel CaV1.3 constitutes an important calcium entry pathway implicated in the regulation of spine morphology. Here we investigated the importance of full-length CaV1.3L and two C-terminally truncated splice variants (CaV1.342A and CaV1.343S) and their modulation by densin-180 and shank1b for the morphology of dendritic spines of cultured hippocampal neurons. Live-cell immunofluorescence and super-resolution microscopy of epitope-tagged CaV1.3L revealed its localization at the base-, neck-, and head-region of dendritic spines. Expression of the short splice variants or deletion of the C-terminal PDZ-binding motif in CaV1.3L induced aberrant dendritic spine elongation. Similar morphological alterations were induced by co-expression of densin-180 or shank1b with CaV1.3L and correlated with increased CaV1.3 currents and dendritic calcium signals in transfected neurons. Together, our findings suggest a key role of CaV1.3 in regulating dendritic spine structure. Under physiological conditions it may contribute to the structural plasticity of glutamatergic synapses. Conversely, altered regulation of CaV1.3 channels may provide an important mechanism in the development of postsynaptic aberrations associated with neurodegenerative disorders. PMID:27708393

  4. Schizophrenia risk gene CAV1 is both pro-psychotic and required for atypical antipsychotic drug actions in vivo.

    Science.gov (United States)

    Allen, J A; Yadav, P N; Setola, V; Farrell, M; Roth, B L

    2011-08-16

    Caveolin-1 (Cav-1) is a scaffolding protein important for regulating receptor signaling cascades by partitioning signaling molecules into membrane microdomains. Disruption of the CAV1 gene has recently been identified as a rare structural variant associated with schizophrenia. Although Cav-1 knockout (KO) mice displayed no baseline behavioral disruptions, Cav-1 KO mice, similar to schizophrenic individuals, exhibited increased sensitivity to the psychotomimetic N-methyl-D-aspartate receptor antagonist phencyclidine (PCP). Thus, PCP disruption of prepulse inhibition (PPI) and PCP-induced mouse locomotor activity were both enhanced by genetic deletion of Cav-1. Interestingly, genetic deletion of Cav-1 rendered the atypical antipsychotics clozapine and olanzapine and the 5-HT(2A)-selective antagonist M100907 ineffective at normalizing PCP-induced disruption of PPI. We also discovered that genetic deletion of Cav-1 attenuated 5-HT(2A)-induced c-Fos and egr-1 expression in mouse frontal cortex and also reduced 5-HT(2A)-mediated Ca(2+) mobilization in primary cortical neuronal cultures. The behavioral effects of the 5-HT(2A) agonist (2,5-dimethoxy-4-iodoamphetamine) including head twitch responses and disruption of PPI were also attenuated by genetic deletion of Cav-1, indicating that Cav-1 is required for both inverse agonist (that is, atypical antipsychotic drug) and agonist actions at 5-HT(2A) receptors. This study demonstrates that disruption of the CAV1 gene--a rare structural variant associated with schizophrenia--is not only pro-psychotic but also attenuates atypical antipsychotic drug actions.

  5. [Effect of overexpression of CAV1 mediated by lentivirus on proliferation and apoptosis of HL-60 cells].

    Science.gov (United States)

    Ma, Wei; Wang, Di-Di; Wang, Zhao; Zhu, Gui-Ming; Zhang, Peng-Xia

    2013-08-01

    This study was purposed to explore the effect of lentivirus-mediated CAV1 overexpression on proliferation and apoptosis in HL-60 cells. Recombinant lentiviral expression vector pcDNA-EF1-CAV1 was constructed, and cotransfected the 293TN cells with a mixture of pPACK packaging plasmids. Then collecting virus suspension infects the HL-60 cells, which make CAV1 gene stable transfection and high expression in the cells. The CAV1 protein expression status in HL-60 cells transfected was evaluated through Western blot method. Proliferative activity and apoptosis of HL-60 cells before and after transfection were detected by CCK-8 method and flow cytometry, respectively. The results showed that the PCR-positive clone screening and results of nucleotide sequencing confirmed that the CAV1 gene inserted into the expression vector pcDNA-EF1-GFP correctly, recombinant lentiviral particles Lv-CAV1 transfected HL-60 cells successfully and with transfection rate up to 90%. The result of Western blot showed that CAV1 protein expression in HL-60 cells significantly increased at 48 hours after transfection. CCK-8 result indicated that cell proliferation activity increased at 48 h after transfection (P HL-60 cells obviously decreased after transfection (P HL-60 cells can inhibit cell proliferation activity and promote cell apoptosis.

  6. Caldendrin, a neuron-specific modulator of Cav/1.2 (L-type) Ca2+ channels.

    Science.gov (United States)

    Tippens, Alyssa L; Lee, Amy

    2007-03-16

    EF-hand Ca2+-binding proteins such as calmodulin and CaBP1 have emerged as important regulatory subunits of voltage-gated Ca2+ channels. Here, we show that caldendrin, a variant of CaBP1 enriched in the brain, interacts with and distinctly modulates Cav1.2 (L-type) voltage-gated Ca2+ channels relative to other Ca2+-binding proteins. Caldendrin binds to the C-terminal IQ-domain of the pore-forming alpha1-subunit of Cav1.2 (alpha(1)1.2) and competitively displaces calmodulin and CaBP1 from this site. Compared with CaBP1, caldendrin causes a more modest suppression of Ca2+-dependent inactivation of Cav1.2 through a different subset of molecular determinants. Caldendrin does not bind to the N-terminal domain of alpha11.2, a site that is critical for functional interactions of the channel with CaBP1. Deletion of the N-terminal domain inhibits CaBP1, but spares caldendrin modulation of Cav1.2 inactivation. In contrast, mutations of the IQ-domain abolish physical and functional interactions of caldendrin and Cav1.2, but do not prevent channel modulation by CaBP1. Using antibodies specific for caldendrin and Cav1.2, we show that caldendrin coimmunoprecipitates with Cav1.2 from the brain and colocalizes with Cav1.2 in somatodendritic puncta of cortical neurons in culture. Our findings reveal functional diversity within related Ca2+-binding proteins, which may enhance the specificity of Ca2+ signaling by Cav1.2 channels in different cellular contexts.

  7. 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

  8. Interaction between Snapin and Cardiac L-type Calcium Channel Cav1.3%Snapin蛋白与心肌L型钙离子通道Cav1.3的相互作用

    Institute of Scientific and Technical Information of China (English)

    袁秀杰; 钱辉; 刘金旭; 张沙沙; 石洁; 赵钟钟; 张朝

    2009-01-01

    [目的] 研究Snapin蛋白与心肌L型钙离子通道Cav1.3的相互作用.[方法]利用免疫共沉淀实验确定Snapin蛋白与Cav1.3在体外表达系统和心房组织存在相互作用.[结果]在过表达Snapin和Cav1.3的HEK293细胞及内源性表达二者的心房肌组织中,Snapin蛋白与Cav1.3钙离子通道存在相互作用.

  9. Phosphorylation of Cav1.2 on S1928 uncouples the L-type Ca2+ channel from the β2 adrenergic receptor.

    Science.gov (United States)

    Patriarchi, Tommaso; Qian, Hai; Di Biase, Valentina; Malik, Zulfiquar A; Chowdhury, Dhrubajyoti; Price, Jennifer L; Hammes, Erik A; Buonarati, Olivia R; Westenbroek, Ruth E; Catterall, William A; Hofmann, Franz; Xiang, Yang K; Murphy, Geoffrey G; Chen, Chao-Ye; Navedo, Manuel F; Hell, Johannes W

    2016-06-15

    Agonist-triggered downregulation of β-adrenergic receptors (ARs) constitutes vital negative feedback to prevent cellular overexcitation. Here, we report a novel downregulation of β2AR signaling highly specific for Cav1.2. We find that β2-AR binding to Cav1.2 residues 1923-1942 is required for β-adrenergic regulation of Cav1.2. Despite the prominence of PKA-mediated phosphorylation of Cav1.2 S1928 within the newly identified β2AR binding site, its physiological function has so far escaped identification. We show that phosphorylation of S1928 displaces the β2AR from Cav1.2 upon β-adrenergic stimulation rendering Cav1.2 refractory for several minutes from further β-adrenergic stimulation. This effect is lost in S1928A knock-in mice. Although AMPARs are clustered at postsynaptic sites like Cav1.2, β2AR association with and regulation of AMPARs do not show such dissociation. Accordingly, displacement of the β2AR from Cav1.2 is a uniquely specific desensitization mechanism of Cav1.2 regulation by highly localized β2AR/cAMP/PKA/S1928 signaling. The physiological implications of this mechanism are underscored by our finding that LTP induced by prolonged theta tetanus (PTT-LTP) depends on Cav1.2 and its regulation by channel-associated β2AR.

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

    Science.gov (United States)

    Zhang, Tan; Taylor, Jackson; Jiang, Yang; Pereyra, Andrea S; Messi, Maria Laura; Wang, Zhong-Min; Hereñú, Claudia; Delbono, Osvaldo

    2015-08-15

    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.

  11. A novel CaV2.2 channel inhibition by piracetam in peripheral and central neurons.

    Science.gov (United States)

    Bravo-Martínez, Jorge; Arenas, Isabel; Vivas, Oscar; Rebolledo-Antúnez, Santiago; Vázquez-García, Mario; Larrazolo, Arturo; García, David E

    2012-10-01

    No mechanistic actions for piracetam have been documented to support its nootropic effects. Voltage-gated calcium channels have been proposed as a promising pharmacological target of nootropic drugs. In this study, we investigated the effect of piracetam on Ca(V)2.2 channels in peripheral neurons, using patch-clamp recordings from cultured superior cervical ganglion neurons. In addition, we tested if Ca(V)2.2 channel inhibition could be related with the effects of piracetam on central neurons. We found that piracetam inhibited native Ca(V)2.2 channels in superior cervical ganglion neurons in a dose-dependent manner, with an IC(50) of 3.4 μmol/L and a Hill coefficient of 1.1. GDPβS dialysis did not prevent piracetam-induced inhibition of Ca(V)2.2 channels and G-protein-coupled receptor activation by noradrenaline did not occlude the piracetam effect. Piracetam altered the biophysical characteristics of Ca(V)2.2 channel such as facilitation ratio. In hippocampal slices, piracetam and ω-conotoxin GVIA diminished the frequency of excitatory postsynaptic potentials and action potentials. Our results provide evidence of piracetam's actions on Ca(V)2.2 channels in peripheral neurons, which might explain some of its nootropic effects in central neurons.

  12. 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.

  13. Deltamethrin Inhibits the Human T-type Voltage-Sensitive Calcium Channel (Cav3.2

    Directory of Open Access Journals (Sweden)

    Steven B. Symington

    2009-01-01

    Full Text Available The goal of this study was to determine the effect of deltamethrin, a pyrethroid insecticide, on CaV3.2, a human T-type voltage-sensitive calcium channel expressed in Xenopus laevis (X.laevis oocytes. Cav3.2 cDNA was transcribed into cRNA; the cRNA was then injected into X.laevis oocytes and electrophysiologically characterized using the two-electrode voltage clamp technique with Ba2+ as a charge carrier. Deltamethrin (10-7 M reduced peak current in a nonreversible manner compared to the untreated control, but had no effect on the voltagedependent activation and inactivation kinetics. These findings confirm that human CaV3.2 is a target for deltamethrin and quite possibly other pyrethroid insecticides. These studies provide insight into the molecular mechanisms of the effect that pyrethroids have on voltage-sensitive calcium channels in general. This information will allow a more complete understanding of the molecular and cellular nature of pyrethroid-induced toxicity and expand our knowledge of the structure-activity relationships of pyrethroids with regard to their action on voltage-sensitive calcium channels.

  14. 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...

  15. Voltage-dependent regulation of CaV2.2 channels by Gq-coupled receptor is facilitated by membrane-localized β subunit.

    Science.gov (United States)

    Keum, Dongil; Baek, Christina; Kim, Dong-Il; Kweon, Hae-Jin; Suh, Byung-Chang

    2014-10-01

    G protein-coupled receptors (GPCRs) signal through molecular messengers, such as Gβγ, Ca(2+), and phosphatidylinositol 4,5-bisphosphate (PIP2), to modulate N-type voltage-gated Ca(2+) (CaV2.2) channels, playing a crucial role in regulating synaptic transmission. However, the cellular pathways through which GqPCRs inhibit CaV2.2 channel current are not completely understood. Here, we report that the location of CaV β subunits is key to determining the voltage dependence of CaV2.2 channel modulation by GqPCRs. Application of the muscarinic agonist oxotremorine-M to tsA-201 cells expressing M1 receptors, together with CaV N-type α1B, α2δ1, and membrane-localized β2a subunits, shifted the current-voltage relationship for CaV2.2 activation 5 mV to the right and slowed current activation. Muscarinic suppression of CaV2.2 activity was relieved by strong depolarizing prepulses. Moreover, when the C terminus of β-adrenergic receptor kinase (which binds Gβγ) was coexpressed with N-type channels, inhibition of CaV2.2 current after M1 receptor activation was markedly reduced and delayed, whereas the delay between PIP2 hydrolysis and inhibition of CaV2.2 current was decreased. When the Gβγ-insensitive CaV2.2 α1C-1B chimera was expressed, voltage-dependent inhibition of calcium current was virtually abolished, suggesting that M1 receptors act through Gβγ to inhibit CaV2.2 channels bearing membrane-localized CaV β2a subunits. Expression of cytosolic β subunits such as β2b and β3, as well as the palmitoylation-negative mutant β2a(C3,4S), reduced the voltage dependence of M1 muscarinic inhibition of CaV2.2 channels, whereas it increased inhibition mediated by PIP2 depletion. Together, our results indicate that, with membrane-localized CaV β subunits, CaV2.2 channels are subject to Gβγ-mediated voltage-dependent inhibition, whereas cytosol-localized β subunits confer more effective PIP2-mediated voltage-independent regulation. Thus, the voltage dependence of

  16. A Frame-Shift Mutation in CAV1 Is Associated with a Severe Neonatal Progeroid and Lipodystrophy Syndrome.

    Science.gov (United States)

    Schrauwen, Isabelle; Szelinger, Szabolcs; Siniard, Ashley L; Kurdoglu, Ahmet; Corneveaux, Jason J; Malenica, Ivana; Richholt, Ryan; Van Camp, Guy; De Both, Matt; Swaminathan, Shanker; Turk, Mari; Ramsey, Keri; Craig, David W; Narayanan, Vinodh; Huentelman, Matthew J

    2015-01-01

    A 3-year-old female patient presenting with an unknown syndrome of a neonatal progeroid appearance, lipodystrophy, pulmonary hypertension, cutis marmorata, feeding disorder and failure to thrive was investigated by whole-genome sequencing. This revealed a de novo, heterozygous, frame-shift mutation in the Caveolin1 gene (CAV1) (p.Phe160X). Mutations in CAV1, encoding the main component of the caveolae in plasma membranes, cause Berardinelli-Seip congenital lipodystrophy type 3 (BSCL). Although BSCL is recessive, heterozygous carriers either show a reduced phenotype of partial lipodystrophy, pulmonary hypertension, or no phenotype. To investigate the pathogenic mechanisms underlying this syndrome in more depth, we performed next generation RNA sequencing of peripheral blood, which showed several dysregulated pathways in the patient that might be related to the phenotypic progeroid features (apoptosis, DNA repair/replication, mitochondrial). Secondly, we found a significant down-regulation of known Cav1 interaction partners, verifying the dysfunction of CAV1. Other known progeroid genes and lipodystrophy genes were also dysregulated. Next, western blotting of lysates of cultured fibroblasts showed that the patient shows a significantly decreased expression of wild-type CAV1 protein, demonstrating a loss-of-function mutation, though her phenotype is more severe that other heterozygotes with similar mutations. This phenotypic variety could be explained by differences in genetic background. Indications for this are supported by additional rare variants we found in AGPAT2 and LPIN1 lipodystrophy genes. CAV1, AGPAT2 and LPIN1 all play an important role in triacylglycerol (TAG) biosynthesis in adipose tissue, and the defective function in different parts of this pathway, though not all to the same extend, could contribute to a more severe lipoatrophic phenotype in this patient. In conclusion, we report, for the first time, an association of CAV1 dysfunction with a syndrome

  17. 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.

  18. 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.

  19. CaV 3.1 and CaV 3.3 account for T-type Ca2+ current in GH3 cells

    Directory of Open Access Journals (Sweden)

    M.A. Mudado

    2004-06-01

    Full Text Available T-type Ca2+ channels are important for cell signaling by a variety of cells. We report here the electrophysiological and molecular characteristics of the whole-cell Ca2+ current in GH3 clonal pituitary cells. The current inactivation at 0 mV was described by a single exponential function with a time constant of 18.32 ± 1.87 ms (N = 16. The I-V relationship measured with Ca2+ as a charge carrier was shifted to the left when we applied a conditioning pre-pulse of up to -120 mV, indicating that a low voltage-activated current may be present in GH3 cells. Transient currents were first activated at -50 mV and peaked around -20 mV. The half-maximal voltage activation and the slope factors for the two conditions are -35.02 ± 2.4 and 6.7 ± 0.3 mV (pre-pulse of -120 mV, N = 15, and -27.0 ± 0.97 and 7.5 ± 0.7 mV (pre-pulse of -40 mV, N = 9. The 8-mV shift in the activation mid-point was statistically significant (P < 0.05. The tail currents decayed bi-exponentially suggesting two different T-type Ca2+ channel populations. RT-PCR revealed the presence of a1G (CaV3.1 and a1I (CaV3.3 T-type Ca2+ channel mRNA transcripts.

  20. Salvileucalin B, a novel diterpenoid with an unprecedented rearranged neoclerodane skeleton from Salvia leucantha Cav.

    Science.gov (United States)

    Aoyagi, Yutaka; Yamazaki, Akira; Nakatsugawa, Chihiro; Fukaya, Haruhiko; Takeya, Koichi; Kawauchi, Susumu; Izumi, Hiroshi

    2008-10-16

    Salvileucalin B (2), having an unprecedented rearranged neoclerodane skeleton, was isolated from the aerial parts of Salvia leucantha Cav. (Labiatae) along with salvileucalin A (1). The absolute structures were elucidated by spectroscopic analysis, X-ray crystallographic analysis, and vibrational circular dichroism. Compound 2 represents a novel neoclerodane, characterized by a tricyclo[3.2.1.0 (2,7)]octane substructure incorporating the exocyclic C-20 methylene of 1. This molecule exerted cytotoxic activity against A549 and HT-29 cells with IC50 values of 5.23 and 1.88 microg/mL, respectively.

  1. Distal C terminus of CaV1.2 channels plays a crucial role in the neural differentiation of dental pulp stem cells.

    Directory of Open Access Journals (Sweden)

    Jianping Ge

    Full Text Available L-type voltage-dependent CaV1.2 channels play an important role in the maintenance of intracellular calcium homeostasis, and influence multiple cellular processes. C-terminal cleavage of CaV1.2 channels was reported in several types of excitable cells, but its expression and possible roles in non-excitable cells is still not clear. The aim of this study was to determine whether distal C-terminal fragment of CaV1.2 channels is present in rat dental pulp stem cells and its possible role in the neural differentiation of rat dental pulp stem cells. We generated stable CaV1.2 knockdown cells via short hairpin RNA (shRNA. Rat dental pulp stem cells with deleted distal C-terminal of CaV1.2 channels lost the potential of differentiation to neural cells. Re-expression of distal C-terminal of CaV1.2 rescued the effect of knocking down the endogenous CaV1.2 on the neural differentiation of rat dental pulp stem cells, indicating that the distal C-terminal of CaV1.2 is required for neural differentiation of rat dental pulp stem cells. These results provide new insights into the role of voltage-gated Ca(2+ channels in stem cells during differentiation.

  2. Critical role of CAV1/caveolin-1 in cell stress responses in human breast cancer cells via modulation of lysosomal function and autophagy.

    Science.gov (United States)

    Shi, Yin; Tan, Shi-Hao; Ng, Shukie; Zhou, Jing; Yang, Na-Di; Koo, Gi-Bang; McMahon, Kerrie-Ann; Parton, Robert G; Hill, Michelle M; Del Pozo, Miguel A; Kim, You-Sun; Shen, Han-Ming

    2015-01-01

    CAV1 (caveolin 1, caveolae protein, 22kDa) is well known as a principal scaffolding protein of caveolae, a specialized plasma membrane structure. Relatively, the caveolae-independent function of CAV1 is less studied. Autophagy is a process known to involve various membrane structures, including autophagosomes, lysosomes, and autolysosomes for degradation of intracellular proteins and organelles. Currently, the function of CAV1 in autophagy remains largely elusive. In this study, we demonstrate for the first time that CAV1 deficiency promotes both basal and inducible autophagy. Interestingly, the promoting effect was found mainly in the late stage of autophagy via enhancing lysosomal function and autophagosome-lysosome fusion. Notably, the regulatory function of CAV1 in lysosome and autophagy was found to be caveolae-independent, and acts through lipid rafts. Furthermore, the elevated autophagy level induced by CAV1 deficiency serves as a cell survival mechanism under starvation. Importantly, downregulation of CAV1 and enhanced autophagy level were observed in human breast cancer cells and tissues. Taken together, our data reveal a novel function of CAV1 and lipid rafts in breast cancer development via modulation of lysosomal function and autophagy.

  3. Apamin Boosting of Synaptic Potentials in CaV2.3 R-Type Ca2+ Channel Null Mice.

    Science.gov (United States)

    Wang, Kang; Kelley, Melissa H; Wu, Wendy W; Adelman, John P; Maylie, James

    2015-01-01

    SK2- and KV4.2-containing K+ channels modulate evoked synaptic potentials in CA1 pyramidal neurons. Each is coupled to a distinct Ca2+ source that provides Ca2+-dependent feedback regulation to limit AMPA receptor (AMPAR)- and NMDA receptor (NMDAR)-mediated postsynaptic depolarization. SK2-containing channels are activated by Ca2+ entry through NMDARs, whereas KV4.2-containing channel availability is increased by Ca2+ entry through SNX-482 (SNX) sensitive CaV2.3 R-type Ca2+ channels. Recent studies have challenged the functional coupling between NMDARs and SK2-containing channels, suggesting that synaptic SK2-containing channels are instead activated by Ca2+ entry through R-type Ca2+ channels. Furthermore, SNX has been implicated to have off target affects, which would challenge the proposed coupling between R-type Ca2+ channels and KV4.2-containing K+ channels. To reconcile these conflicting results, we evaluated the effect of SK channel blocker apamin and R-type Ca2+ channel blocker SNX on evoked excitatory postsynaptic potentials (EPSPs) in CA1 pyramidal neurons from CaV2.3 null mice. The results show that in the absence of CaV2.3 channels, apamin application still boosted EPSPs. The boosting effect of CaV2.3 channel blockers on EPSPs observed in neurons from wild type mice was not observed in neurons from CaV2.3 null mice. These data are consistent with a model in which SK2-containing channels are functionally coupled to NMDARs and KV4.2-containing channels to CaV2.3 channels to provide negative feedback regulation of EPSPs in the spines of CA1 pyramidal neurons.

  4. Altered thalamocortical rhythmicity and connectivity in mice lacking CaV3.1 T-type Ca2+ channels in unconsciousness.

    Science.gov (United States)

    Choi, Soonwook; Yu, Eunah; Lee, Seongwon; Llinás, Rodolfo R

    2015-06-23

    In unconscious status (e.g., deep sleep and anesthetic unconsciousness) where cognitive functions are not generated there is still a significant level of brain activity present. Indeed, the electrophysiology of the unconscious brain is characterized by well-defined thalamocortical rhythmicity. Here we address the ionic basis for such thalamocortical rhythms during unconsciousness. In particular, we address the role of CaV3.1 T-type Ca(2+) channels, which are richly expressed in thalamic neurons. Toward this aim, we examined the electrophysiological and behavioral phenotypes of mice lacking CaV3.1 channels (CaV3.1 knockout) during unconsciousness induced by ketamine or ethanol administration. Our findings indicate that CaV3.1 KO mice displayed attenuated low-frequency oscillations in thalamocortical loops, especially in the 1- to 4-Hz delta band, compared with control mice (CaV3.1 WT). Intriguingly, we also found that CaV3.1 KO mice exhibited augmented high-frequency oscillations during unconsciousness. In a behavioral measure of unconsciousness dynamics, CaV3.1 KO mice took longer to fall into the unconscious state than controls. In addition, such unconscious events had a shorter duration than those of control mice. The thalamocortical interaction level between mediodorsal thalamus and frontal cortex in CaV3.1 KO mice was significantly lower, especially for delta band oscillations, compared with that of CaV3.1 WT mice, during unconsciousness. These results suggest that the CaV3.1 channel is required for the generation of a given set of thalamocortical rhythms during unconsciousness. Further, that thalamocortical resonant neuronal activity supported by this channel is important for the control of vigilance states.

  5. The nociception role of Cav3.2 calcium channel%Cav3.2通道在伤害性感受中的作用

    Institute of Scientific and Technical Information of China (English)

    苏昊; 徐世元

    2015-01-01

    Background Some of the earliest detailed descriptions of biophysical properties of low voltage-activated or transient (T) type Calcium channels were done using in vitro preparation of primary sensory or dorsal root ganglion (DRG) neurons that are known for their functional role in processing pain signals.However, in spite of these early discoveries, T-type calcium channels were not implicated in sensory transmission in general and pain processing (nociception) in particular until recently.Objective To summarize the important role of peripheral T-type Calcium channels in boosting nociceptive transmission.Content T-type calcium channels in peripheral sensory neurons play important role under physiological conditions (e.g.acute nociceptive pain) and to pain processing under pathological conditions.Trend peripheral T-type Calcium channels in nociception and the value of these channels as cellular targets for potential drug developments.%背景 疼痛是危害人类健康,影响人类生存质量的重要因素,但目前可用于临床治疗的手段尚不能满足患者的无痛要求,且存在不同程度的副作用,其主要原因之一是目前对疼痛的产生机制尚未完全明了.最近的研究表明,T型钙通道Cav3.2亚型在疼痛信号转导中起重要作用. 目的 综述T型钙通道Cav3.2亚型在痛觉传导中的作用. 内容 阐述位于背根神经节(dorsal root ganglion,DRG)的T型钙通道参与传递生理性疼痛(疼痛感知)和病理性疼痛(神经病理性疼痛)伤害性刺激信号. 趋向 T型钙离子通道可调控疼痛伤害性刺激信号转导,并可作为可能的疼痛治疗靶点.

  6. Whole exome sequencing identifies de novo heterozygous CAV1 mutations associated with a novel neonatal onset lipodystrophy syndrome.

    Science.gov (United States)

    Garg, Abhimanyu; Kircher, Martin; Del Campo, Miguel; Amato, R Stephen; Agarwal, Anil K

    2015-08-01

    Despite remarkable progress in identifying causal genes for many types of genetic lipodystrophies in the last decade, the molecular basis of many extremely rare lipodystrophy patients with distinctive phenotypes remains unclear. We conducted whole exome sequencing of the parents and probands from six pedigrees with neonatal onset of generalized loss of subcutaneous fat with additional distinctive phenotypic features and report de novo heterozygous null mutations, c.424C>T (p.Q142*) and c.479_480delTT (p.F160*), in CAV1 in a 7-year-old male and a 3-year-old female of European origin, respectively. Both the patients had generalized fat loss, thin mottled skin and progeroid features at birth. The male patient had cataracts requiring extraction at age 30 months and the female patient had pulmonary arterial hypertension. Dermal fibroblasts of the female patient revealed negligible CAV1 immunofluorescence staining compared to control but there were no differences in the number and morphology of caveolae upon electron microscopy examination. Based upon the similarities in the clinical features of these two patients, previous reports of CAV1 mutations in patients with lipodystrophies and pulmonary hypertension, and similar features seen in CAV1 null mice, we conclude that these variants are the most likely cause of one subtype of neonatal onset generalized lipodystrophy syndrome.

  7. Role of protein phosphatases in the run down of guinea pig cardiac Cav1.2 Ca2+ channels.

    Science.gov (United States)

    Yu, Lifeng; Xu, Jianjun; Minobe, Etsuko; Kameyama, Asako; Yang, Lei; Feng, Rui; Hao, Liying; Kameyama, Masaki

    2016-05-15

    This study aimed to investigate protein phosphatases involved in the run down of Cav1.2 Ca(2+) channels. Single ventricular myocytes obtained from adult guinea pig hearts were used to record Ca(2+) channel currents with the patch-clamp technique. Calmodulin (CaM) and ATP were used to restore channel activity in inside-out patches. Inhibitors of protein phosphatases were applied to investigate the role of phosphatases. The specific protein phosphatase type 1 (PP1) inhibitor (PP1 inhibitor-2) and protein phosphatase type 2A (PP2A) inhibitor (fostriecin) abolished the slow run down of Cav1.2 Ca(2+) channels, which was evident as the time-dependent attenuation of the reversing effect of CaM/ATP on the run down. However, protein phosphatase type 2B (PP2B, calcineurin) inhibitor cyclosporine A together with cyclophilin A had no effect on the channel run down. Furthermore, PP1 inhibitor-2 mainly prolonged the open time constants of the channel, specifically, the slow open time. Fostriecin primarily shortened the slow close time constants. Our data suggest that PP1 and PP2A were involved in the slow phase of Cav1.2 Ca(2+) channel run down. In addition, they exerted different effects on the open-close kinetics of the channel. All above support the view that PP1 and PP2A may dephosphorylate distinct phosphorylation sites on the Cav1.2 Ca(2+) channel.

  8. A Frame-Shift Mutation in CAV1 Is Associated with a Severe Neonatal Progeroid and Lipodystrophy Syndrome.

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    Isabelle Schrauwen

    Full Text Available A 3-year-old female patient presenting with an unknown syndrome of a neonatal progeroid appearance, lipodystrophy, pulmonary hypertension, cutis marmorata, feeding disorder and failure to thrive was investigated by whole-genome sequencing. This revealed a de novo, heterozygous, frame-shift mutation in the Caveolin1 gene (CAV1 (p.Phe160X. Mutations in CAV1, encoding the main component of the caveolae in plasma membranes, cause Berardinelli-Seip congenital lipodystrophy type 3 (BSCL. Although BSCL is recessive, heterozygous carriers either show a reduced phenotype of partial lipodystrophy, pulmonary hypertension, or no phenotype. To investigate the pathogenic mechanisms underlying this syndrome in more depth, we performed next generation RNA sequencing of peripheral blood, which showed several dysregulated pathways in the patient that might be related to the phenotypic progeroid features (apoptosis, DNA repair/replication, mitochondrial. Secondly, we found a significant down-regulation of known Cav1 interaction partners, verifying the dysfunction of CAV1. Other known progeroid genes and lipodystrophy genes were also dysregulated. Next, western blotting of lysates of cultured fibroblasts showed that the patient shows a significantly decreased expression of wild-type CAV1 protein, demonstrating a loss-of-function mutation, though her phenotype is more severe that other heterozygotes with similar mutations. This phenotypic variety could be explained by differences in genetic background. Indications for this are supported by additional rare variants we found in AGPAT2 and LPIN1 lipodystrophy genes. CAV1, AGPAT2 and LPIN1 all play an important role in triacylglycerol (TAG biosynthesis in adipose tissue, and the defective function in different parts of this pathway, though not all to the same extend, could contribute to a more severe lipoatrophic phenotype in this patient. In conclusion, we report, for the first time, an association of CAV1 dysfunction

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

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    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.

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

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    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.

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

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    Armando A. Durant

    2013-01-01

    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.

  12. Redetermination of metarossite, CaV5+2O6·2H2O

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    Anaïs Kobsch

    2016-09-01

    Full Text Available The crystal structure of metarossite, ideally CaV2O6·2H2O [chemical name: calcium divanadium(V hexaoxide dihydrate], was first determined using precession photographs, with fixed isotropic displacement parameters and without locating the positions of the H atoms, leading to a reliability factor R = 0.11 [Kelsey & Barnes (1960. Can. Mineral. 6, 448–466]. This communication reports a structure redetermination of this mineral on the basis of single-crystal X-ray diffraction data of a natural sample from the Blue Cap mine, San Juan County, Utah, USA (R1 = 0.036. Our study not only confirms the structural topology reported in the previous study, but also makes possible the refinement of all non-H atoms with anisotropic displacement parameters and all H atoms located. The metarossite structure is characterized by chains of edge-sharing [CaO8] polyhedra parallel to [100] that are themselves connected by chains of alternating [VO5] trigonal bipyramids parallel to [010]. The two H2O molecules are bonded to Ca. Analysis of the displacement parameters show that the [VO5] chains librate around [010]. In addition, we measured the Raman spectrum of metarossite and compared it with IR and Raman data previously reported. Moreover, heating of metarossite led to a loss of water, which results in a transformation to the brannerite-type structure, CaV2O6, implying a possible dehydration pathway for the compounds M2+V2O6·xH2O, with M = Cu, Cd, Mg or Mn, and x = 2 or 4.

  13. Endogenous and exogenous hydrogen sulfide facilitates T-type calcium channel currents in Cav3.2-expressing HEK293 cells.

    Science.gov (United States)

    Sekiguchi, Fumiko; Miyamoto, Yosuke; Kanaoka, Daiki; Ide, Hiroki; Yoshida, Shigeru; Ohkubo, Tsuyako; Kawabata, Atsufumi

    2014-02-28

    Hydrogen sulfide (H2S), a gasotransmitter, is formed from l-cysteine by multiple enzymes including cystathionine-γ-lyase (CSE). We have shown that an H2S donor, NaHS, causes hyperalgesia in rodents, an effect inhibited by knockdown of Cav3.2 T-type Ca(2+) channels (T-channels), and that NaHS facilitates T-channel-dependent currents (T-currents) in NG108-15 cells that naturally express Cav3.2. In the present study, we asked if endogenous and exogenous H2S participates in regulation of the channel functions in Cav3.2-transfected HEK293 (Cav3.2-HEK293) cells. dl-Propargylglycine (PPG), a CSE inhibitor, significantly decreased T-currents in Cav3.2-HEK293 cells, but not in NG108-15 cells. NaHS at 1.5mM did not affect T-currents in Cav3.2-HEK293 cells, but enhanced T-currents in NG108-15 cells. In the presence of PPG, NaHS at 1.5mM, but not 0.1-0.3mM, increased T-currents in Cav3.2-HEK293 cells. Similarly, Na2S, another H2S donor, at 0.1-0.3mM significantly increased T-currents in the presence, but not absence, of PPG in Cav3.2-HEK293 cells. Expression of CSE was detected at protein and mRNA levels in HEK293 cells. Intraplantar administration of Na2S, like NaHS, caused mechanical hyperalgesia, an effect blocked by NNC 55-0396, a T-channel inhibitor. The in vivo potency of Na2S was higher than NaHS. These results suggest that the function of Cav3.2 T-channels is tonically enhanced by endogenous H2S synthesized by CSE in Cav3.2-HEK293 cells, and that exogenous H2S is capable of enhancing Cav3.2 function when endogenous H2S production by CSE is inhibited. In addition, Na2S is considered a more potent H2S donor than NaHS in vitro as well as in vivo.

  14. CACNA1B (Cav2.2 Overexpression and Its Association with Clinicopathologic Characteristics and Unfavorable Prognosis in Non-Small Cell Lung Cancer

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    Xiaoyu Zhou

    2017-01-01

    Full Text Available CACNA1B (Cav2.2 encodes an N-type voltage-gated calcium channel (VGCC ubiquitously expressed in brain and peripheral nervous system that is important for regulating neuropathic pain. Because intracellular calcium concentration is a key player in cell proliferation and apoptosis, VGCCs are implicated in tumorigenesis. Recent studies have identified CACNA1B (Cav2.2 being overexpressed in prostate and breast cancer tissues when compared to adjacent normal tissues; however, its role in non-small cell lung cancer (NSCLC has not been investigated. In this study, we determined the mRNA and protein expression of CACNA1B (Cav2.2 in NSCLC tumorous and adjacent nontumorous tissues by quantitative reverse transcription PCR (qRT-PCR and tissue microarray immunohistochemistry analysis (TMA-IHC, respectively. CACNA1B (Cav2.2 protein expressions in tumorous tissues were correlated with NSCLC patients’ clinical characteristics and overall survival. CACNA1B (Cav2.2 mRNA and protein expression levels were higher in NSCLC tumorous tissues than in nontumorous tissues. High CACNA1B (Cav2.2 protein expression was associated with higher TNM stages, and CACNA1B (Cav2.2 protein expression is an independent prognostic marker in NSCLC. Based on our results, we conclude that CACNA1B (Cav2.2 plays a role in NSCLC development and progression. Elucidating the underlying mechanism may help design novel treatment by specifically targeting the calcium regulation pathway for NSCLC, a devastating disease with increasing incidence and mortality in China.

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

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    Giuseppe eGangarossa

    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.

  16. 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.

  17. RGS12 interacts with the SNARE-binding region of the Cav2.2 calcium channel.

    Science.gov (United States)

    Richman, Ryan W; Strock, Jesse; Hains, Melinda D; Cabanilla, Nory Jun; Lau, King-Kei; Siderovski, David P; Diversé-Pierluissi, María

    2005-01-14

    Activation of GABAB receptors in chick dorsal root ganglion (DRG) neurons inhibits the Cav2.2 calcium channel in both a voltage-dependent and voltage-independent manner. The voltage-independent inhibition requires activation of a tyrosine kinase that phosphorylates the alpha1 subunit of the channel and thereby recruits RGS12, a member of the "regulator of G protein signaling" (RGS) proteins. Here we report that RGS12 binds to the SNARE-binding or "synprint" region (amino acids 726-985) in loop II-III of the calcium channel alpha1 subunit. A recombinant protein encompassing the N-terminal PTB domain of RGS12 binds to the synprint region in protein overlay and surface plasmon resonance binding assays; this interaction is dependent on tyrosine phosphorylation and yet is within a sequence that differs from the canonical NPXY motif targeted by other PTB domains. In electrophysiological experiments, microinjection of DRG neurons with synprint-derived peptides containing the tyrosine residue Tyr-804 altered the rate of desensitization of neurotransmitter-mediated inhibition of the Cav2.2 calcium channel, whereas peptides centered about a second tyrosine residue, Tyr-815, were without effect. RGS12 from a DRG neuron lysate was precipitated using synprint peptides containing phosphorylated Tyr-804. The high degree of conservation of Tyr-804 in the SNARE-binding region of Cav2.1 and Cav2.2 calcium channels suggests that this region, in addition to the binding of SNARE proteins, is also important for determining the time course of the modulation of calcium current via tyrosine phosphorylation.

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

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    Christian Schön

    Full Text Available 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.

  19. CAV1基因沉默增强MCF-7多细胞球对多柔比星的敏感性%CAV1 gene silencing enhances adriamycin sensitivity of breast carcinoma MCF-7 multicellular spheroids

    Institute of Scientific and Technical Information of China (English)

    顾栋桦; 平金良; 董吉顺; 朱荣; 陈琦

    2011-01-01

    背景与目的:肿瘤细胞形成多细胞球后,对许多化疗药物的耐受性明显增强,这种现象被称为“多细胞耐药”.其发生机制可能与细胞黏附引起的信号通路活性的改变有关.细胞质膜微囊蛋白-1 (caveolin-1,CAV1)是一个细胞膜蛋白,在细胞膜信号通路的调节中发挥重要作用.本研究旨在探讨CAV1基因沉默是否能增强MCF-7多细胞球对多柔比星的敏感性,并初步探讨其机制.方法:用liquid overlay技术培养获得MCF-7多细胞球,用脂质体转染法把特异针对CAV1基因的双链小RNA干扰片段导入MCF-7细胞中,采用台盼蓝拒染法检测多柔比星对MCF-7的细胞抑制率,分别用RT-PCR和免疫印迹法检测CAV1 mRNA水平及蛋白水平,用免疫印迹法检测细胞总黏着斑激酶(focal adhesion kinase,FAK)及磷酸化FAK (p-FAK)蛋白水平.结果:与单层细胞相比,多细胞球组的多柔比星细胞抑制率明显降低,FAK相对磷酸化水平增加2.64倍(t=7.32,P<0.01);CAV1基因沉默后,MCF-7多细胞球生长缓慢,球体较松散,与未转染组相比,干扰组多细胞球多柔比星(1 μ mol/L)的细胞抑制率增加了1.07倍(t=5.77,P<0.01),FAK相对磷酸化水平下降68.7%(t=6.57,P<0.01).结论:多细胞球的形成可以增强MCF-7细胞对多柔比星的耐受性;CAV1基因沉默能逆转MCF-7多细胞球对多柔比星的耐药现象,FAK相对磷酸化水平的下调可能是其重要机制.%Background and purpose: Tumor spheroids exhibit decreased sensitivity to anticancer drugs compared with monolayer cells, which is known as multicellular resistance (MCR). Its mechanism underlying the phenomenon is unknown, but the signal transduction modified by cell-cell adhesion may be important in this procedure. Caveolin-l(CAVl) is a membrane protein which can regulate cell signal transduction. This study aimed to investigate the role oiCAVl gene expression on drug resistance of multicellular spheroids (MCS) of breast

  20. Five different profiles of dihydropyridines in blocking T-type Ca(2+) channel subtypes (Ca(v)3.1 (alpha(1G)), Ca(v)3.2 (alpha(1H)), and Ca(v)3.3 (alpha(1I))) expressed in Xenopus oocytes.

    Science.gov (United States)

    Furukawa, Taiji; Nukada, Toshihide; Namiki, Yoshiko; Miyashita, Yoriko; Hatsuno, Kento; Ueno, Yasunari; Yamakawa, Takeshi; Isshiki, Takaaki

    2009-06-24

    1,4-dihydropyridine (DHP) Ca(2+) antagonists have recently been shown to block T-type Ca(2+) channels, which may render favorable actions on cardiovascular systems. However, this evaluation remains to be done systematically for each T-type Ca(2+) channel subtype except for the Ca(v)3.1 (alpha(1G)) subtype. To address this issue at the molecular level, blocking effects of 14 kinds of DHPs (amlodipine, aranidipine, azelnidipine, barnidipine, benidipine, cilnidipine, efonidipine, felodipine, manidipine, nicardipine, nifedipine, nilvadipine, nimodipine, nitrendipine), which are clinically used for treatments of hypertension, on 3 subtypes of T-type Ca(2+) channels [Ca(v)3.2 (alpha(1H)), Ca(v)3.3 (alpha(1I)), and Ca(v)3.1 (alpha(1G))] were investigated in the Xenopus oocyte expression system using the two-microelectrode voltage-clamp technique. These 3 kinds (alpha(1H), alpha(1I) and alpha(1G)) of T-type channels were blocked by amlodipine, manidipine and nicardipine. On the other hand, azelnidipine, barnidipine, benidipine and efonidipine significantly blocked alpha(1H) and alpha(1G), but not alpha(1I) channels, while nilvadipine and nimodipine apparently blocked alpha(1H) and alpha(1I), but not alpha(1G) channels. Moreover, aranidipine blocked only alpha(1H) channels. By contrast, cilnidipine, felodipine, nifedipine and nitrendipine had little effects on these subtypes of T-type channels. The result indicates that the blockade of T-type Ca(2+) channels by derivatives of DHP Ca(2+) antagonist was selective for the channel subtype. Therefore, these selectivities of DHPs in blocking T-type Ca(2+) channel subtypes would provide useful pharmacological and clinical information on the mode of action of the drugs including side-effects and adverse effects.

  1. Ser1928 phosphorylation by PKA stimulates the L-type Ca2+ channel CaV1.2 and vasoconstriction during acute hyperglycemia and diabetes

    Science.gov (United States)

    Nystoriak, Matthew A.; Nieves-Cintrón, Madeline; Patriarchi, Tommaso; Buonarati, Olivia R.; Prada, Maria Paz; Morotti, Stefano; Grandi, Eleonora; Fernandes, Julia Dos Santos; Forbush, Katherine; Hofmann, Franz; Sasse, Kent C.; Scott, John D.; Ward, Sean M.; Hell, Johannes W.; Navedo, Manuel F.

    2017-01-01

    Hypercontractility of arterial myocytes and enhanced vascular tone during diabetes are, in part, attributed to the effects of increased glucose (hyperglycemia) on L-type CaV1.2 channels. In murine arterial myocytes, kinase-dependent mechanisms mediate the increase in CaV1.2 activity in response to increased extracellular glucose. We identified a subpopulation of the CaV1.2 channel pore-forming subunit (α1C) within nanometer proximity of protein kinase A (PKA) at the sarcolemma of murine and human arterial myocytes. This arrangement depended upon scaffolding of PKA by an A-kinase anchoring protein 150 (AKAP150) in mice. Glucose-mediated increases in CaV1.2 channel activity were associated with PKA activity, leading to α1C phosphorylation at Ser1928. Compared to arteries from low-fat diet (LFD)–fed mice and nondiabetic patients, arteries from high-fat diet (HFD)–fed mice and from diabetic patients had increased Ser1928 phosphorylation and CaV1.2 activity. Arterial myocytes and arteries from mice lacking AKAP150 or expressing mutant AKAP150 unable to bind PKA did not exhibit increased Ser1928 phosphorylation and CaV1.2 current density in response to increased glucose or to HFD. Consistent with a functional role for Ser1928 phosphorylation, arterial myocytes and arteries from knockin mice expressing a CaV1.2 with Ser1928 mutated to alanine (S1928A) lacked glucose-mediated increases in CaV1.2 activity and vasoconstriction. Furthermore, the HFD-induced increases in CaV1.2 current density and myogenic tone were prevented in S1928A knockin mice. These findings reveal an essential role for α1C phosphorylation at Ser1928 in stimulating CaV1.2 channel activity and vasoconstriction by AKAP-targeted PKA upon exposure to increased glucose and in diabetes. PMID:28119464

  2. Extension of CAVS coarse-grained model to phospholipid membranes: The importance of electrostatics.

    Science.gov (United States)

    Shen, Hujun; Deng, Mingsen; Zhang, Yachao

    2017-05-15

    It is evident from experiment that electrostatic potential (or dipole potential) is positive inside PC or PE lipid bilayers in the absence of ions. MARTINI coarse-grained (CG) model, which has been widely used in simulating physical properties of lipid bilayers, fails to reproduce the positive value for the dipole potential in the membrane interior. Although the total dipole potential can be correctly described by the BMW/MARTINI model, the contribution from the ester dipoles, playing a nontrivial role in the electrostatic potential across lipid membranes, is neglected by this hybrid approach. In the ELBA CG model, the role of the ester dipoles is considered, but it is overweighed because various atomistic models have consistently shown that water is actually the leading contributor of dipole potential. Here, we present a CG approach by combining the BMW-like water model (namely CAVS model) with the ELBA-like lipid model proposed in this work. Our CG model was designed not only to correctly reproduce the positive values for the dipole potential inside PC and PE lipid bilayers but also to properly balance the individual contributions from the ester dipoles and water, surmounting the limitations of current CG models in the calculations of dipole potential. © 2017 Wiley Periodicals, Inc.

  3. Ciclo de vida de Pellaea ternifolia (Cav. Link subsp. ternifolia (Pteridaceae-Polypodiidae

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    Jorge Huerta-Zavala

    2015-08-01

    Full Text Available Se describe el ciclo de vida dePellaea ternifolia(Cav. Link subsp.ternifolia. Los especímenes fueron recolectados en un bosque dePinusperturbado en San Miguel de los Alcanfores, municipio de Tlaxco, Tlaxcala, México. Las esporas fueron sembradas en vasos de unicel de 262 ml con cerámica (barro molido, piedra de río, tela de mosquitero y tres soportes de cultivo (tierra, maquique y musgo, todo previamente esterilizado y cerrado con plástico y alambre de cobre. El ciclo fue isospórico con desarrollo del protalo tipoAdiantum. En la fase cordada se apreciaron gametófitos con anteridios y arquegonio en una proporción cercana al 70% y el otro 30% fueron gametófitos apogámicos. Los esporófitos jóvenes se obtuvieron a partir de los 63 días.

  4. Chemical Studies of Yellow Tamarillo (Solanum betaceum Cav. Fruit Flavor by Using a Molecular Sensory Approach

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    Juliana María García

    2016-12-01

    Full Text Available The odor-active volatile compounds of yellow tamarillo fruit (S. betaceum Cav. were identified and quantified by using a sensomics approach, combining a gentle volatile extraction (solvent-assisted flavor evaporation (SAFE, gas chromatography-mass spectrometry (GC-MS, and sensory analyses (gas chromatography-olfactometry (GC-O and aroma extract dilution analysis (AEDA. The medium-term purpose of this work is to evaluate the change of odor-active volatiles during processing. Thus, (Z-3-hexenal, hexanal, and ethyl butanoate were identified as key aroma compounds of yellow tamarillo. The C6-aliphatic compounds, aliphatic esters, and terpenols were characterized as the volatiles responsible for the herbal-green, fruity, and fresh-mint odor notes of this variety, respectively. Additionally, one non-volatile compound contributing to the residual bitter taste of this fruit was isolated by a bioguided (taste sensory analyses fractionation. The freeze-dried fruit was sequentially liquid-liquid partitioned with solvents of different polarity, and then the ethyl acetate fraction was submitted to size exclusion chromatography. Then, its structure was elucidated as rosmarinic acid, by using common spectroscopic methods (mass spectrometry (MS and nuclear magnetic resonance (NMR. The amount of rosmarinic acid was quantified as 46.17 ± 1.20 mg/100 g of dried fruit, by the external standard method. Its bitter taste threshold value was determined by using the 3AFC (alternative forced choice method to be 37.00 ± 1.25 mg/L.

  5. 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.

  6. Cav1.3 channels control D2-autoreceptor responses via NCS-1 in substantia nigra dopamine neurons.

    Science.gov (United States)

    Dragicevic, Elena; Poetschke, Christina; Duda, Johanna; Schlaudraff, Falk; Lammel, Stephan; Schiemann, Julia; Fauler, Michael; Hetzel, Andrea; Watanabe, Masahiko; Lujan, Rafael; Malenka, Robert C; Striessnig, Joerg; Liss, Birgit

    2014-08-01

    Dopamine midbrain neurons within the substantia nigra are particularly prone to degeneration in Parkinson's disease. Their selective loss causes the major motor symptoms of Parkinson's disease, but the causes for the high vulnerability of SN DA neurons, compared to neighbouring, more resistant ventral tegmental area dopamine neurons, are still unclear. Consequently, there is still no cure available for Parkinson's disease. Current therapies compensate the progressive loss of dopamine by administering its precursor l-DOPA and/or dopamine D2-receptor agonists. D2-autoreceptors and Cav1.3-containing L-type Ca(2+) channels both contribute to Parkinson's disease pathology. L-type Ca(2+) channel blockers protect SN DA neurons from degeneration in Parkinson's disease and its mouse models, and they are in clinical trials for neuroprotective Parkinson's disease therapy. However, their physiological functions in SN DA neurons remain unclear. D2-autoreceptors tune firing rates and dopamine release of SN DA neurons in a negative feedback loop through activation of G-protein coupled potassium channels (GIRK2, or KCNJ6). Mature SN DA neurons display prominent, non-desensitizing somatodendritic D2-autoreceptor responses that show pronounced desensitization in PARK-gene Parkinson's disease mouse models. We analysed surviving human SN DA neurons from patients with Parkinson's disease and from controls, and detected elevated messenger RNA levels of D2-autoreceptors and GIRK2 in Parkinson's disease. By electrophysiological analysis of postnatal juvenile and adult mouse SN DA neurons in in vitro brain-slices, we observed that D2-autoreceptor desensitization is reduced with postnatal maturation. Furthermore, a transient high-dopamine state in vivo, caused by one injection of either l-DOPA or cocaine, induced adult-like, non-desensitizing D2-autoreceptor responses, selectively in juvenile SN DA neurons, but not ventral tegmental area dopamine neurons. With pharmacological and genetic

  7. Ca2+-binding protein-1 facilitates and forms a postsynaptic complex with Cav1.2 (L-type) Ca2+ channels.

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    Zhou, Hong; Kim, Seong-Ah; Kirk, Elizabeth A; Tippens, Alyssa L; Sun, Hong; Haeseleer, Françoise; Lee, Amy

    2004-05-12

    Ca2+-binding protein-1 (CaBP1) is a Ca2+-binding protein that is closely related to calmodulin (CaM) and localized in somatodendritic regions of principal neurons throughout the brain, but how CaBP1 participates in postsynaptic Ca2+ signaling is not known. Here, we describe a novel role for CaBP1 in the regulation of Ca2+ influx through Ca(v)1.2 (L-type) Ca2+ channels. CaBP1 interacts directly with the alpha1 subunit of Ca(v)1.2 at sites that also bind CaM. CaBP1 binding to one of these sites, the IQ domain, is Ca2+ dependent and competitive with CaM binding. The physiological significance of this interaction is supported by the association of Ca(v)1.2 and CaBP1 in postsynaptic density fractions purified from rat brain. Moreover, in double-label immunofluorescence experiments, CaBP1 and Ca(v)1.2 colocalize in numerous cell bodies and dendrites of neurons, particularly in pyramidal cells in the CA3 region of the hippocampus and in the dorsal cortex. In electrophysiological recordings of cells transfected with Ca(v)1.2, CaBP1 greatly prolonged Ca2+ currents, prevented Ca2+-dependent inactivation, and caused Ca2+-dependent facilitation of currents evoked by step depolarizations and repetitive stimuli. These effects contrast with those of CaM, which promoted strong Ca2+-dependent inactivation of Ca(v)1.2 with these same voltage protocols. Our findings reveal how Ca2+-binding proteins, such as CaM and CaBP1, differentially adjust Ca2+ influx through Ca(v)1.2 channels, which may specify diverse modes of Ca2+ signaling in neurons.

  8. Coexpression of high-voltage-activated ion channels Kv3.4 and Cav1.2 in pioneer axons during pathfinding in the developing rat forebrain.

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    Huang, Chia-Yi; Chu, Dachen; Hwang, Wei-Chao; Tsaur, Meei-Ling

    2012-11-01

    Precise axon pathfinding is crucial for establishment of the initial neuronal network during development. Pioneer axons navigate without the help of preexisting axons and pave the way for follower axons that project later. Voltage-gated ion channels make up the intrinsic electrical activity of pioneer axons and regulate axon pathfinding. To elucidate which channel molecules are present in pioneer axons, immunohistochemical analysis was performed to examine 14 voltage-gated ion channels (Kv1.1-Kv1.3, Kv3.1-Kv3.4, Kv4.3, Cav1.2, Cav1.3, Cav2.2, Nav1.2, Nav1.6, and Nav1.9) in nine axonal tracts in the developing rat forebrain, including the optic nerve, corpus callosum, corticofugal fibers, thalamocortical axons, lateral olfactory tract, hippocamposeptal projection, anterior commissure, hippocampal commissure, and medial longitudinal fasciculus. We found A-type K⁺ channel Kv3.4 in both pioneer axons and early follower axons and L-type Ca²⁺ channel Cav1.2 in pioneer axons and early and late follower axons. Spatially, Kv3.4 and Cav1.2 were colocalized with markers of pioneer neurons and pioneer axons, such as deleted in colorectal cancer (DCC), in most fiber tracts examined. Temporally, Kv3.4 and Cav1.2 were expressed abundantly in most fiber tracts during axon pathfinding but were downregulated beginning in synaptogenesis. By contrast, delayed rectifier Kv channels (e.g., Kv1.1) and Nav channels (e.g., Nav1.2) were absent from these fiber tracts (except for the corpus callosum) during pathfinding of pioneer axons. These data suggest that Kv3.4 and Cav1.2, two high-voltage-activated ion channels, may act together to control Ca²⁺ -dependent electrical activity of pioneer axons and play important roles during axon pathfinding.

  9. Hydrogen sulfide-induced itch requires activation of Cav3.2 T-type calcium channel in mice

    Science.gov (United States)

    Wang, Xue-Long; Tian, Bin; Huang, Ya; Peng, Xiao-Yan; Chen, Li-Hua; Li, Jun-Cheng; Liu, Tong

    2015-01-01

    The contributions of gasotransmitters to itch sensation are largely unknown. In this study, we aimed to investigate the roles of hydrogen sulfide (H2S), a ubiquitous gasotransmitter, in itch signaling. We found that intradermal injection of H2S donors NaHS or Na2S, but not GYY4137 (a slow-releasing H2S donor), dose-dependently induced scratching behavior in a μ-opioid receptor-dependent and histamine-independent manner in mice. Interestingly, NaHS induced itch via unique mechanisms that involved capsaicin-insensitive A-fibers, but not TRPV1-expressing C-fibers that are traditionally considered for mediating itch, revealed by depletion of TRPV1-expressing C-fibers by systemic resiniferatoxin treatment. Moreover, local application of capsaizapine (TRPV1 blocker) or HC-030031 (TRPA1 blocker) had no effects on NaHS-evoked scratching. Strikingly, pharmacological blockade and silencing of Cav3.2 T-type calcium channel by mibefradil, ascorbic acid, zinc chloride or Cav3.2 siRNA dramatically decreased NaHS-evoked scratching. NaHS induced robust alloknesis (touch-evoked itch), which was inhibited by T-type calcium channels blocker mibefradil. Compound 48/80-induced itch was enhanced by an endogenous precursor of H2S (L-cysteine) but attenuated by inhibitors of H2S-producing enzymes cystathionine γ-lyase and cystathionine β-synthase. These results indicated that H2S, as a novel nonhistaminergic itch mediator, may activates Cav3.2 T-type calcium channel, probably located at A-fibers, to induce scratching and alloknesis in mice. PMID:26602811

  10. 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...

  11. 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 larg...

  12. GDM-Induced Macrosomia Is Reversed by Cav-1 via AMPK-Mediated Fatty Acid Transport and GLUT1-Mediated Glucose Transport in Placenta

    Science.gov (United States)

    Wang, Di; Yang, Ruirui; Sang, Hui; Han, Linlin; Zhu, Yuexia; Lu, Yanyan; Tan, Yeke; Shang, Zhanping

    2017-01-01

    Objective To investigate if the role of Cav-1 in GDM-induced macrosomia is through regulating AMPK signaling pathway in placenta. Methods We used diagnostic criteria of gestational diabetes mellitus (GDM) and macrosomia to separate and compare placental protein and mRNA levels from GDM with macrosomia group (GDMM), GDM with normal birth weight group (GDMN) and normal glucose tolerance (NGT) with normal birth weight group (CON). Western blotting was performed to examine differentially expressed proteins of caveolin-1 (Cav-1) and Adenosine monophosphate-activated protein kinase (AMPK) signaling pathway related proteins, including phosphorylated-AMPKα(Thr172), AMPKα, phosphorylated-Acetyl-CoA carboxylase(Ser79) (p-ACC(Ser79)), ACC and glucose transporter 1 (GLUT1) in placenta between the three groups. The mRNA levels of Cav-1, AMPKα, ACC and GLUT1 in placenta were measured by real time-PCR. Results In the GDMM placenta group, both protein and mRNA levels of Cav-1 were down-regulated, while GLUT1 was up-regulated; the phosphorylation and mRNA levels of ACC and AMPKα were decreased, but total ACC protein levels were increased compared to both the GDMN (pGLUT1 protein levels. Besides, in GDMM group placental mRNA levels, NBW had a positive correlation with GLUT1 (pGLUT1 (pGLUT1. Conclusion GDM-induced macrosomias have more severe inhibition of Cav-1 expression in placenta. Cav-1 is associated with placental glucose and fatty acid transport via the induction of AMPK signaling pathway and the reduction of GLUT1 signaling pathway to reverse GDM-induced macrosomia. PMID:28125642

  13. 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

  14. Ultrastructural evidence for pre- and postsynaptic localization of Cav1.2 L-type Ca2+ channels in the rat hippocampus.

    Science.gov (United States)

    Tippens, Alyssa L; Pare, Jean-Francois; Langwieser, Nicole; Moosmang, Sven; Milner, Teresa A; Smith, Yoland; Lee, Amy

    2008-02-01

    In the hippocampal formation, Ca(v)1.2 (L-type) voltage-gated Ca(2+) channels mediate Ca(2+) signals that can trigger long-term alterations in synaptic efficacy underlying learning and memory. Immunocytochemical studies indicate that Ca(v)1.2 channels are localized mainly in the soma and proximal dendrites of hippocampal pyramidal neurons, but electrophysiological data suggest a broader distribution of these channels. To define the subcellular substrates underlying Ca(v)1.2 Ca(2+) signals, we analyzed the localization of Ca(v)1.2 in the hippocampal formation by using antibodies against the pore-forming alpha(1)-subunit of Ca(v)1.2 (alpha(1)1.2). By light microscopy, alpha(1)1.2-like immunoreactivity (alpha(1)1.2-IR) was detected in pyramidal cell soma and dendritic fields of areas CA1-CA3 and in granule cell soma and fibers in the dentate gyrus. At the electron microscopic level, alpha(1)1.2-IR was localized in dendrites, but also in axons, axon terminals, and glial processes in all hippocampal subfields. Plasmalemmal immunogold particles representing alpha(1)1.2-IR were more significant for small- than large-caliber dendrites and were largely associated with extrasynaptic regions in dendritic spines and axon terminals. These findings provide the first detailed ultrastructural analysis of Ca(v)1.2 localization in the brain and support functionally diverse roles of these channels in the hippocampal formation.

  15. Rem, a member of the RGK GTPases, inhibits recombinant CaV1.2 channels using multiple mechanisms that require distinct conformations of the GTPase.

    Science.gov (United States)

    Yang, Tingting; Xu, Xianghua; Kernan, Timothy; Wu, Vincent; Colecraft, Henry M

    2010-05-15

    Rad/Rem/Gem/Kir (RGK) GTPases potently inhibit Ca(V)1 and Ca(V)2 (Ca(V)1-2) channels, a paradigm of ion channel regulation by monomeric G-proteins with significant physiological ramifications and potential biotechnology applications. The mechanism(s) underlying how RGK proteins inhibit I(Ca) is unknown, and it is unclear how key structural and regulatory properties of these GTPases (such as the role of GTP binding to the nucleotide binding domain (NBD), and the C-terminus which contains a membrane-targeting motif) feature in this effect. Here, we show that Rem inhibits Ca(V)1.2 channels by three independent mechanisms that rely on distinct configurations of the GTPase: (1) a reduction in surface density of channels is accomplished by enhancing dynamin-dependent endocytosis, (2) a diminution of channel open probability (P(o)) that occurs without impacting on voltage sensor movement, and (3) an immobilization of Ca(V) channel voltage sensors. The presence of both the Rem NBD and C-terminus (whether membrane-targeted or not) in one molecule is sufficient to reconstitute all three mechanisms. However, membrane localization of the NBD by a generic membrane-targeting module reconstitutes only the decreased P(o) function (mechanism 2). A point mutation that prevents GTP binding to the NBD selectively eliminates the capacity to immobilize voltage sensors (mechanism 3). The results reveal an uncommon multiplicity in the mechanisms Rem uses to inhibit I(Ca), predict new physiological dimensions of the RGK GTPase-Ca(V) channel crosstalk, and suggest original approaches for developing novel Ca(V) channel blockers.

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

    Science.gov (United States)

    Garau, Gianpiero; Magotti, Paola; Heine, Martin; Korotchenko, Svetlana; Lievens, Patricia Marie-Jeanne; Berezin, Vladimir; Dityatev, Alexander

    2015-12-01

    Our previous studies revealed that L-type voltage-dependent Ca(2+) 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-forming domain of Cav1.2α1 subunit bound to heparin. At 25 °C the binding of the peptide P7 (MGKMHKTCYN) was at ~50 μM, and that of the peptide P8 (GHGRQCQNGTVCKPGWDGPKHG) was at ~21 μM. The Cav1.2α1 first pore forming segment that contained both peptides maintained a high affinity for heparin (~23 μM), integrating their enthalpic and entropic binding contributions. Interaction between heparin and recombinant as well as native full-length neuronal Cav1.2α1 channels was confirmed using the heparin-agarose pull down assay. Whole cell patch clamp recordings in HEK293 cells transfected with neuronal Cav1.2 channels revealed that enzymatic digestion of highly sulfated heparan sulfates with heparinase 1 affects neither voltage-dependence of channel activation nor the level of steady state inactivation, but did speed up channel inactivation. Treatment of hippocampal cultures with heparinase 1 reduced the firing rate and led to appearance of long-lasting bursts in the same manner as treatment with the inhibitor of L-VDCC diltiazem. Thus, heparan sulfate proteoglycans may bind to and regulate L-VDCC inactivation and network activity.

  17. Alternative splicing generates a smaller assortment of CaV2.1 transcripts in cerebellar Purkinje cells than in the cerebellum.

    Science.gov (United States)

    Kanumilli, Srinivasan; Tringham, Elizabeth W; Payne, C Elizabeth; Dupere, Jonathan R B; Venkateswarlu, Kanamarlapudi; Usowicz, Maria M

    2006-01-12

    P/Q-type calcium channels control many calcium-driven functions in the brain. The CACNA1A gene encoding the pore-forming CaV2.1 (alpha1A) subunit of P/Q-type channels undergoes alternative splicing at multiple loci. This results in channel variants with different phenotypes. However, the combinatorial patterns of alternative splice events at two or more loci, and hence the diversity of CaV2.1 transcripts, are incompletely defined for specific brain regions and types of brain neurons. Using RT-PCR and splice variant-specific primers, we have identified multiple CaV2.1 transcript variants defined by different pairs of splice events in the cerebellum of adult rat. We have uncovered new splice variations between exons 28 and 34 (some of which predict a premature stop codon) and a new variation in exon 47 (which predicts a novel extended COOH-terminus). Single cell RT-PCR reveals that each individual cerebellar Purkinje neuron also expresses multiple alternative CaV2.1 transcripts, but the assortment is smaller than in the cerebellum. Two of these variants encode different extended COOH-termini which are not the same as those previously reported in Purkinje cells of the mouse. Our patch-clamp recordings show that calcium channel currents in the soma and dendrites of Purkinje cells are largely inhibited by a concentration of omega-agatoxin IVA selective for P-type over Q-type channels, suggesting that the different transcripts may form phenotypic variants of P-type calcium channels in Purkinje cells. These results expand the known diversity of CaV2.1 transcripts in cerebellar Purkinje cells, and propose the selective expression of distinct assortments of CaV2.1 transcripts in different brain neurons and species.

  18. ATP hydrolysis is critically required for function of CaV1.3 channels in cochlear inner hair cells via fueling Ca2+ clearance.

    Science.gov (United States)

    Weiler, Simon; Krinner, Stefanie; Wong, Aaron B; Moser, Tobias; Pangršič, Tina

    2014-05-14

    Sound encoding is mediated by Ca(2+) influx-evoked release of glutamate at the ribbon synapse of inner hair cells. Here we studied the role of ATP in this process focusing on Ca(2+) current through CaV1.3 channels and Ca(2+) homeostasis in mouse inner hair cells. Patch-clamp recordings and Ca(2+) imaging demonstrate that hydrolyzable ATP is essential to maintain synaptic Ca(2+) influx in inner hair cells via fueling Ca(2+)-ATPases to avoid an increase in cytosolic [Ca(2+)] and subsequent Ca(2+)/calmodulin-dependent inactivation of CaV1.3 channels.

  19. 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

  20. Protein Kinase Cα and P-Type Ca2+ Channel CaV2.1 in Red Blood Cell Calcium Signalling

    Directory of Open Access Journals (Sweden)

    Lisa Wagner-Britz

    2013-06-01

    Full Text Available Background/Aims: Protein kinase Cα (PKCα is activated by an increase in cytosolic Ca2+ in red blood cells (RBCs. Previous work has suggested that PKCα directly stimulates the CaV2.1 channel, whereas other studies revealed that CaV2.1 is insensitive to activation by PKC. The aim of this study was to resolve this discrepancy. Methods: We performed experiments based on a single cell read-out of the intracellular Ca2+ concentration in terms of Fluo-4 fluorescence intensity and phosphatidylserine exposure to the external membrane leaflet. Measurement modalities included flow cytometry and live cell imaging. Results: Treatment of RBCs with phorbol 12-myristate 13-acetate (PMA led to two distinct populations of cells with an increase in intracellular Ca2+: a weak-responding and a strong-responding population. The EC50 of PMA for the number of cells with Ca2+ elevation was 2.7±1.2 µM; for phosphatidylserine exposure to the external membrane surface, it was 2.8±0.5 µM; and for RBC haemolysis, it was 2.9±0.5 µM. Using pharmacological manipulation with the CaV2.1 inhibitor ω-agatoxin TK and the broad protein kinase C inhibitor Gö6983, we are able to show that there are two independent PMA-activated Ca2+ entry processes: the first is independent of CaV2.1 and directly PKCα-activated, while the second is associated with a likely indirect activation of CaV2.1. Further studies using lysophosphatidic acid (LPA as a stimulation agent have provided additional evidence that PKCα and CaV2.1 are not directly interconnected in a signalling chain. Conclusion: Although we provide evidence for a lack of interaction between PKCα and CaV2.1 in RBCs, further studies are required to decipher the signalling relationship between LPA, PKCα and CaV2.1.

  1. 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

    M), integrating their enthalpic and entropic binding contributions. Interaction between heparin and recombinant as well as native full-length neuronal Cav1.2α1 channels was confirmed using the heparin–agarose pull down assay. Whole cell patch clamp recordings in HEK293 cells transfected with neuronal Cav1...... rate and led to appearance of long-lasting bursts in the same manner as treatment with the inhibitor of L-VDCC diltiazem. Thus, heparan sulfate proteoglycans may bind to and regulate L-VDCC inactivation and network activity....

  2. CAV_KO: a Simple 1-D Langrangian Hydrocode for MS EXCEL™ with Automatic Generation of X-T Diagrams

    Science.gov (United States)

    Tsembelis, K.; Ramsden, B.; Proud, W. G.; Borg, J.

    2007-12-01

    Hydrocodes are widely used to predict or simulate highly dynamic and transient events such as blast and impact. Codes such as GRIM, CTH or AUTODYN are well developed and involve complex numerical methods and in many cases require a large computing infrastructure. In this paper we present a simple 1-D Langrangian hydrocode developed at the University of Cambridge, called CAV_KO written in Visual Basic. The motivation being to produce a code which, while being relatively simple, is useful for both experimental planning and teaching. The code has been adapted from the original KO code written in FORTRAN by J. Borg, which, in turn, is based on the algorithm developed by Wilkins [1]. The developed GUI within MS Excel™ and the automatic generation of x-t diagrams allow CAV_KO to be a useful tool for quick calculations of plate impact events and teaching purposes. The VB code is licensed under the GNU General Public License and a MS Excel™ spreadsheet containing the code can be downloaded from www.shockphysics.com together with a copy of the user guide.

  3. Age-dependent impact of CaV3.2 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

    .2-dependent and -independent effects. No changes in mRNA expression of several important K(+) and Ca(2+) channel genes were induced by CaV3.2 knock-out. However, the expression of the other T-type channel isoform (CaV3.1) was reduced at the mRNA and protein level in mature adult compared to young WT arteries......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. CaV3.2 T-type calcium channels...... are expressed in both VSMCs and ECs of small arteries. The T-type channels are important drug targets but due to the lack of specific antagonists our understanding of the role of CaV3.2 channels in vasomotor tone at various ages is scarce. We evaluated the myogenic response, flow-mediated vasodilatation...

  4. Combined anti-ages and antioxidant activities of different solvent extracts of Solanum elaeagnifolium Cav (Solanacea) fruits during ripening and related to their phytochemical compositions

    Science.gov (United States)

    Houda, Mejri; Derbré, Séverine; Jedy, Ahmed; Tlili, Nizar; Legault, Jean; Richomme, Pascal; Limam, Ferid; Saidani-Tounsi, Moufida

    2014-01-01

    Oxidative stress and advanced glycation end products (AGEs) are known as key factors for the development of diabetic complications such as retinopathy, cataract as well as atherosclerosis and neurodegenerative diseases, including Alzheimer’s diseases. In this context, natural products have been previously identified as promising sources for antioxidant and anti-glycation compounds. The current study focuses on the evaluation of antioxidant and glycation inhibitory activities of different solvent extracts of Solanum elaeagnifolium Cav (Solanaceae) fruits at different ripening stages. The results showed that antioxidant and anti-AGEs activities were significantly influenced by solvents polarities and ripening stages of S. elaeagnifolium Cav. With one exception, methanolic extract of overripe S. elaeagnifolium Cav fruit showed important protective effects against cellular oxidative stress. The aqueous extract showed the highest ABTS+ scavenging ability. Principal component analysis showed that total phenolic and flavonoid contents correlated well with observed antioxidants and anti-glycation activities. These results bring attention to the possible use of S. elaeagnifolium Cav as a valuable source of bioactive compounds exhibiting antioxidant effects and potentially alleviating diabetic complications. PMID:26417319

  5. 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

    2016-06-03

    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.

  6. 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.

  7. A CACNA1C variant associated with reduced voltage-dependent inactivation, increased CaV1.2 channel window current, and arrhythmogenesis.

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    Jessica A Hennessey

    Full Text Available Mutations in CACNA1C that increase current through the CaV1.2 L-type Ca2+ channel underlie rare forms of long QT syndrome (LQTS, and Timothy syndrome (TS. We identified a variant in CACNA1C in a male child of Filipino descent with arrhythmias and extracardiac features by candidate gene sequencing and performed functional expression studies to electrophysiologically characterize the effects of the variant on CaV1.2 channels. As a baby, the subject developed seizures and displayed developmental delays at 30 months of age. At age 5 years, he displayed a QTc of 520 ms and experienced recurrent VT. Physical exam at 17 years of age was notable for microcephaly, short stature, lower extremity weakness and atrophy with hyperreflexia, spastic diplegia, multiple dental caries and episodes of rhabdomyolysis. Candidate gene sequencing identified a G>C transversion at position 5731 of CACNA1C (rs374528680 predicting a glycine>arginine substitution at residue 1911 (p.G1911R of CaV1.2. The allele frequency of this variant is 0.01 in Malays, but absent in 984 Caucasian alleles and in the 1000 genomes project. In electrophysiological analyses, the variant decreased voltage-dependent inactivation, thus causing a gain of function of CaV1.2. We also observed a negative shift of V1/2 of activation and positive shift of V1/2 of channel inactivation, resulting in an increase of the window current. Together, these suggest a gain-of-function effect on CaV1.2 and suggest increased susceptibility for arrhythmias in certain clinical settings. The p.G1911R variant was also identified in a case of sudden unexplained infant death (SUID, for which an increasing number of clinical observations have demonstrated can be associated with arrhythmogenic mutations in cardiac ion channels. In summary, the combined effects of the CACNA1C variant to diminish voltage-dependent inactivation of CaV1.2 and increase window current expand our appreciation of mechanisms by which a gain of

  8. Two distinct voltage-sensing domains control voltage sensitivity and kinetics of current activation in CaV1.1 calcium channels.

    Science.gov (United States)

    Tuluc, Petronel; Benedetti, Bruno; Coste de Bagneaux, Pierre; Grabner, Manfred; Flucher, Bernhard E

    2016-06-01

    Alternative splicing of the skeletal muscle CaV1.1 voltage-gated calcium channel gives rise to two channel variants with very different gating properties. The currents of both channels activate slowly; however, insertion of exon 29 in the adult splice variant CaV1.1a causes an ∼30-mV right shift in the voltage dependence of activation. Existing evidence suggests that the S3-S4 linker in repeat IV (containing exon 29) regulates voltage sensitivity in this voltage-sensing domain (VSD) by modulating interactions between the adjacent transmembrane segments IVS3 and IVS4. However, activation kinetics are thought to be determined by corresponding structures in repeat I. Here, we use patch-clamp analysis of dysgenic (CaV1.1 null) myotubes reconstituted with CaV1.1 mutants and chimeras to identify the specific roles of these regions in regulating channel gating properties. Using site-directed mutagenesis, we demonstrate that the structure and/or hydrophobicity of the IVS3-S4 linker is critical for regulating voltage sensitivity in the IV VSD, but by itself cannot modulate voltage sensitivity in the I VSD. Swapping sequence domains between the I and the IV VSDs reveals that IVS4 plus the IVS3-S4 linker is sufficient to confer CaV1.1a-like voltage dependence to the I VSD and that the IS3-S4 linker plus IS4 is sufficient to transfer CaV1.1e-like voltage dependence to the IV VSD. Any mismatch of transmembrane helices S3 and S4 from the I and IV VSDs causes a right shift of voltage sensitivity, indicating that regulation of voltage sensitivity by the IVS3-S4 linker requires specific interaction of IVS4 with its corresponding IVS3 segment. In contrast, slow current kinetics are perturbed by any heterologous sequences inserted into the I VSD and cannot be transferred by moving VSD I sequences to VSD IV. Thus, CaV1.1 calcium channels are organized in a modular manner, and control of voltage sensitivity and activation kinetics is accomplished by specific molecular mechanisms

  9. Upregulation of Voltage-Gated Calcium Channel Cav1.3 in Bovine Somatotropes Treated with Ghrelin

    Directory of Open Access Journals (Sweden)

    V. M. Salinas Zarate

    2013-01-01

    the Cav1.2 and Cav1.3 pore-forming subunits of L-type channels. The treatment with Ghrelin significantly increased the Cav1.3 subunit expression, suggeting that the chronic stimulation of the GHS receptor with Ghrelin or GHRP-6 increases the number of voltage-gated Ca2+ channels at the cell surface of BS.

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

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    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.

  11. 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.

  12. CaV3.2 T-type Ca2+ channels mediate the augmented calcium influx in carotid body glomus cells by chronic intermittent hypoxia.

    Science.gov (United States)

    Makarenko, Vladislav V; Ahmmed, Gias U; Peng, Ying-Jie; Khan, Shakil A; Nanduri, Jayasri; Kumar, Ganesh K; Fox, Aaron P; Prabhakar, Nanduri R

    2016-01-01

    Chronic intermittent hypoxia (CIH) is a hallmark manifestation of sleep apnea. A heightened carotid body activity and the resulting chemosensory reflex mediate increased sympathetic nerve activity by CIH. However, the mechanisms underlying heightened carotid body activity by CIH are not known. An elevation of intracellular calcium ion concentration ([Ca(2+)]i) in glomus cells, the primary oxygen-sensing cells, is an essential step for carotid body activation by hypoxia. In the present study, we examined the effects of CIH on the glomus cell [Ca(2+)]i response to hypoxia and assessed the underlying mechanisms. Glomus cells were harvested from adult rats or wild-type mice treated with 10 days of either room air (control) or CIH (alternating cycles of 15 s of hypoxia and 5 min of room air; 9 episodes/h; 8 h/day). CIH-treated glomus cells exhibited an enhanced [Ca(2+)]i response to hypoxia, and this effect was absent in the presence of 2-(4-cyclopropylphenyl)-N-((1R)-1-[5-[(2,2,2-trifluoroethyl)oxo]-pyridin-2-yl]ethyl)acetamide (TTA-A2), a specific inhibitor of T-type Ca(2+) channels, and in voltage-gated calcium channel, type 3.2 (CaV3.2), null glomus cells. CaV3.2 knockout mice exhibited an absence of CIH-induced hypersensitivity of the carotid body. CIH increased reactive oxygen species (ROS) levels in glomus cells. A ROS scavenger prevented the exaggerated TTA-A2-sensitive [Ca(2+)]i response to hypoxia. CIH had no effect on CaV3.2 mRNA levels. CIH augmented Ca(2+) currents and increased CaV3.2 protein in plasma membrane fractions of human embryonic kidney-293 cells stably expressing CaV3.2, and either a ROS scavenger or brefeldin-A, an inhibitor of protein trafficking, prevented these effects. These findings suggest that CIH leads to an augmented Ca(2+) influx via ROS-dependent facilitation of CaV3.2 protein trafficking to the plasma membrane.

  13. Luminal Glucose Does Not Enhance Active Intestinal Calcium Absorption in mice: Evidence Against a Role for Cav1.3 as a Mediator of Calcium Uptake During Absorption

    Science.gov (United States)

    Reyes-Fernandez, Perla C.; Fleet, James C.

    2015-01-01

    Intestinal Ca absorption occurs through a 1,25 dihydroxyvitamin D3 (1,25(OH)2D3)-regulated transcellular pathway, especially when habitual dietary Ca intake is low. Recently the L-type voltage-gated Ca channel, Cav1.3, was proposed to mediate active, transcellular Ca absorption in response to membrane depolarization caused by elevated luminal glucose levels following a meal. We tested the hypothesis that high luminal glucose could reveal a role for Cav1.3 in active intestinal Ca absorption in mice. Nine week-old male C57BL/6J mice were fed AIN93G diets containing either low (0.125%) or high (1%) Ca for 1 week and Ca absorption was examined by an oral gavage method using a 45Ca-transport buffer containing 25 mmol/L of glucose or fructose. Transient receptor potential vanilloid 6 (TRPV6), Calbindin D9k (CaBPD9k) and Cav1.3 mRNA levels were measured in the duodenum, jejunum and ileum. TRPV6 and CaBPD9k expression were highest in the duodenum, where active, 1,25(OH)2D3-regulated Ca absorption occurs while Cav1.3 mRNA levels were similar across the intestinal segments. As expected, the low Ca diet increased renal cytochrome p450-27B1 (CYP27B1) mRNA (p=0.003), serum 1,25(OH)2D3 (p<0.001) and Ca absorption efficiency by 2-fold with the fructose buffer. However, the glucose buffer used to favor Cav1.3 activation did not increase Ca absorption efficiency (p=0.6) regardless of the dietary Ca intake level. Collectively, our results show that glucose did not enhance Ca absorption and they do not support a critical role for Cav1.3 in either basal or vitamin D-regulated intestinal Ca absorption in vivo. PMID:26403486

  14. Actividad biológica de extractos crudos de Larrea divaricata Cav. y Capparis atamisquea Kuntze sobre Sitophilus oryzae (L. Biological activity of crude extracts of Larrea divaricata Cav. and Capparis atamisquea Kuntze on Sitophilus oryzae (L.

    Directory of Open Access Journals (Sweden)

    A.I. Viglianco

    2006-12-01

    Full Text Available Se evaluaron las propiedades repelentes y antialimentarias de extractos crudos de dos plantas difundidas en la provincia de Córdoba (Argentina, jarilla (Larrea divaricata Cav. y atamisqui (Capparis atamisquea Kuntze., sobre Sitophilus oryzae (L.. Se estudiaron los extractos crudos en etanol, cloroformo y hexano de ambas especies. Para cada extracto se determino el coeficiente de disuasión total. Para las pruebas de repelencia se utilizaron papeles de filtro separados en dos mitades iguales; se determinaron los porcentajes de repelencia de cada extracto. Se observó mayor efecto antialimentario de los extractos de atamisqui con respecto a los de jarilla, y de los de hojas o tallos sobre los de frutos. El extracto de mayor efecto antialimentario fue el etanólico de tallos de atamisqui (clase ++++ seguido por los extractos etanólico y clorofórmico de hojas de esta especie (clase +++. Se observó un moderado efecto de repelencia de los extractos de jarilla y atamisqui sobre S. oryzae, destacándose el de hojas de atamisqui en hexano como el de mayor efecto de repelencia (clase IV. El extracto clorofórmico de hojas de atamisqui, los tres de hoja de jarilla y los clorofórmicos de tallos y frutos de jarilla presentaron efecto de repelencia (clase III.Repellent and antifeedant properties of crude extracts of two plants widely distributed in the province of Córdoba (Argentina, jarilla (Larrea divaricata Cav. and atamisqui (Capparis atamisquea Kuntze on Sitophilus oryzae were studied. Crude extracts in ethanol, chloroform and hexane of both species were evaluated. The Total Deterrence Coefficient for each extract was determined. Repellence tests were conducted using filter paper separated in halves. Repellence percentages were determined for each extract. A greater antifeedant effect was observed in the atamisqui extracts than in the jarilla ones; the effect was also greater in extracts of leaves or twigs than in extracts of fruits. The extracts

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

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    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.

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

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    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.

  17. Transcriptional repression of Caveolin-1 (CAV1) gene expression by GATA-6 in bladder smooth muscle hypertrophy in mice and human beings.

    Science.gov (United States)

    Boopathi, Ettickan; Gomes, Cristiano Mendes; Goldfarb, Robert; John, Mary; Srinivasan, Vittala Gopal; Alanzi, Jaber; Malkowicz, S Bruce; Kathuria, Hasmeena; Zderic, Stephen A; Wein, Alan J; Chacko, Samuel

    2011-05-01

    Hypertrophy occurs in urinary bladder wall smooth muscle (BSM) in men with partial bladder outlet obstruction (PBOO) caused by benign prostatic hyperplasia (BPH) and in animal models of PBOO. Hypertrophied BSM from the rabbit model exhibits down-regulation of caveolin-1, a structural and functional protein of caveolae that function as signaling platforms to mediate interaction between receptor proteins and adaptor and effector molecules to regulate signal generation, amplification, and diversification. Caveolin-1 expression is diminished in PBOO-induced BSM hypertrophy in mice and in men with BPH. The proximal promoter of the human and mouse caveolin-1 (CAV1) gene was characterized, and it was observed that the transcription factor GATA-6 binds this promoter, causing reduced expression of caveolin-1. Furthermore, caveolin-1 expression levels inversely correlate with the abundance of GATA-6 in BSM hypertrophy in mice and human beings. Silencing of GATA6 gene expression up-regulates caveolin-1 expression, whereas overexpression of GATA-6 protein sustains the transcriptional repression of caveolin-1 in bladder smooth muscle cells. Together, these data suggest that GATA-6 acts as a transcriptional repressor of CAV1 gene expression in PBOO-induced BSM hypertrophy in men and mice. GATA-6-induced transcriptional repression represents a new regulatory mechanism of CAV1 gene expression in pathologic BSM, and may serve as a target for new therapy for BPH-induced bladder dysfunction in aging men.

  18. The α2δ-1 subunit remodels CaV1.2 voltage sensors and allows Ca2+ influx at physiological membrane potentials

    Science.gov (United States)

    Pantazis, Antonios; Sigg, Daniel; Weiss, James N.; Neely, Alan

    2016-01-01

    Excitation-evoked calcium influx across cellular membranes is strictly controlled by voltage-gated calcium channels (CaV), which possess four distinct voltage-sensing domains (VSDs) that direct the opening of a central pore. The energetic interactions between the VSDs and the pore are critical for tuning the channel’s voltage dependence. The accessory α2δ-1 subunit is known to facilitate CaV1.2 voltage-dependent activation, but the underlying mechanism is unknown. In this study, using voltage clamp fluorometry, we track the activation of the four individual VSDs in a human L-type CaV1.2 channel consisting of α1C and β3 subunits. We find that, without α2δ-1, the channel complex displays a right-shifted voltage dependence such that currents mainly develop at nonphysiological membrane potentials because of very weak VSD–pore interactions. The presence of α2δ-1 facilitates channel activation by increasing the voltage sensitivity (i.e., the effective charge) of VSDs I–III. Moreover, the α2δ-1 subunit also makes VSDs I–III more efficient at opening the channel by increasing the coupling energy between VSDs II and III and the pore, thus allowing Ca influx within the range of physiological membrane potentials. PMID:27481713

  19. Regulation of neuronal high-voltage activated Ca(V)2 Ca(2+) channels by the small GTPase RhoA.

    Science.gov (United States)

    Rousset, Matthieu; Cens, Thierry; Menard, Claudine; Bowerman, Melissa; Bellis, Michel; Brusés, Juan; Raoul, Cedric; Scamps, Frédérique; Charnet, Pierre

    2015-10-01

    High-Voltage-Activated (HVA) Ca(2+) channels are known regulators of synapse formation and transmission and play fundamental roles in neuronal pathophysiology. Small GTPases of Rho and RGK families, via their action on both cytoskeleton and Ca(2+) channels are key molecules for these processes. While the effects of RGK GTPases on neuronal HVA Ca(2+) channels have been widely studied, the effects of RhoA on the HVA channels remains however elusive. Using heterologous expression in Xenopus laevis oocytes, we show that RhoA activity reduces Ba(2+) currents through CaV2.1, CaV2.2 and CaV2.3 Ca(2+) channels independently of CaVβ subunit. This inhibition occurs independently of RGKs activity and without modification of biophysical properties and global level of expression of the channel subunit. Instead, we observed a marked decrease in the number of active channels at the plasma membrane. Pharmacological and expression studies suggest that channel expression at the plasma membrane is impaired via a ROCK-sensitive pathway. Expression of constitutively active RhoA in primary culture of spinal motoneurons also drastically reduced HVA Ca(2+) current amplitude. Altogether our data revealed that HVA Ca(2+) channels regulation by RhoA might govern synaptic transmission during development and potentially contribute to pathophysiological processes when axon regeneration and growth cone kinetics are impaired.

  20. Enhancing VTA Cav1.3 L-type Ca(2+) channel activity promotes cocaine and mood-related behaviors via overlapping AMPA receptor mechanisms in the nucleus accumbens.

    Science.gov (United States)

    Martínez-Rivera, A; Hao, J; Tropea, T F; Giordano, T P; Kosovsky, M; Rice, R C; Lee, A; Huganir, R L; Striessnig, J; Addy, N A; Han, S; Rajadhyaksha, A M

    2017-02-14

    Genetic factors significantly influence susceptibility for substance abuse and mood disorders. Rodent studies have begun to elucidate a role of Cav1.3 L-type Ca(2+) channels in neuropsychiatric-related behaviors, such as addictive and depressive-like behaviors. Human studies have also linked the CACNA1D gene, which codes for the Cav1.3 protein, with bipolar disorder. However, the neurocircuitry and the molecular mechanisms underlying the role of Cav1.3 in neuropsychiatric phenotypes are not well established. In the present study, we directly manipulated Cav1.3 channels in Cav1.2 dihydropyridine insensitive mutant mice and found that ventral tegmental area (VTA) Cav1.3 channels mediate cocaine-related and depressive-like behavior through a common nucleus accumbens (NAc) shell calcium-permeable α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (CP-AMPAR) mechanism that requires GluA1 phosphorylation at S831. Selective activation of VTA Cav1.3 with (±)-BayK-8644 (BayK) enhanced cocaine conditioned place preference and cocaine psychomotor activity while inducing depressive-like behavior, an effect not observed in S831A phospho-mutant mice. Infusion of the CP-AMPAR-specific blocker Naspm into the NAc shell reversed the cocaine and depressive-like phenotypes. In addition, activation of VTA Cav1.3 channels resulted in social behavioral deficits. In contrast to the cocaine- and depression-related phenotypes, GluA1/A2 AMPARs in the NAc core mediated social deficits, independent of S831-GluA1 phosphorylation. Using a candidate gene analysis approach, we also identified single-nucleotide polymorphisms in the CACNA1D gene associated with cocaine dependence in human subjects. Together, our findings reveal novel, overlapping mechanisms through which VTA Cav1.3 mediates cocaine-related, depressive-like and social phenotypes, suggesting that Cav1.3 may serve as a target for the treatment of neuropsychiatric symptoms.Molecular Psychiatry advance online publication, 14

  1. miR-1和miR-133 a对大鼠肥大心肌细胞L-型钙通道Cavβ2和α1 C亚基的调控作用%Regulation of miR-1 and miR-133 a on L-type calcium channel Cavβ2 and α1C subunits in rat cardiomyocyte hypertrophy

    Institute of Scientific and Technical Information of China (English)

    王玉琴; 耿鹏; 吴扬

    2015-01-01

    Objective To investigate the regulation of miR-1 and miR-133 a on L-type calcium channel β2 subunit ( Cavβ2 ) and α1C subunit during rat cardiomyocyte hypertrophy .Methods Cardiomyocyte hypertrophy was in-duced by isoproterenol (ISO, 10μmol/L).The targets of miR-1 and miR-133a were predicted by online database microCosm and Targetscan , respectively .The 3′untranslated region sequences of Cavβ2 andα1C were respectively cloned into reporter vector and then transiently transfected into HEK 293 cells.The luciferase activities of samples were measured for demonstrating the expression of luciferase reporter vector .The protein expression of Cavβ2 andα1C were evaluated by Western blot .The expression levels of Cavβ2 andα1C were inhibited by RNAi to determine theeffectsofCavβ2andα1Concardiomyocytehypertrophy.Results 1)Cavβ2wasoneofpotentialtargetsof miR-1,α1C was the one of potential targets of miR-133a.2) The luciferase activities of HEK293 cells with the plasmid containing widetype Cavβ2 3′UTR sequence or α1 C significantly decreased ( P <0.05 , P <0.01 ) . 3 ) Upregulation of the miR-1 and miR-133 a by miR-1 mimic and miR-133 a mimic transfection suppressed pro-tein expression of Cavβ2 and α1C, respectively(P<0.01, P<0.05).4)Downregulation of Cavβ2 andα1C by RNAi could markedly inhibit the increase of cell surface area ( P<0.01 ) , mRNA expression of ANP andβ-MHC (P<0.05).Conclusions Cavβ2 is the target gene of miR-1 and α1C is the target gene of miR-133a.miR-1 and miR-133a can negatively regulate the expression of L-type calcium channel Cavβ2 andα1C subunit, inhibi-ting cardiomyocyte hypertrophy.%目的:研究miR-1和miR-133a在大鼠心肌细胞肥大中对L-型钙通道β2亚基( Cavβ2)和α1C亚基的调控作用。方法异丙肾上腺素( ISO )诱导大鼠心肌细胞肥大;在线数据库microCosm 和Targetscan 预测miR-1和miR-133a的靶基因;分别构建含有Cavβ23′UTR或α1C 3′UTR的重组质粒,与miR-1

  2. Developmental pathway from leaves to galls induced by a sap-feeding insect on Schinus polygamus (Cav. Cabrera (Anacardiaceae

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    GRACIELA G. DIAS

    2013-03-01

    Full Text Available Galling sap-feeding insects are presumed to cause only minor changes in host plant tissues, because they usually do not require development of nutritive tissues for their own use. This premise was examined through comparison of the histometry, cytometry and anatomical development of non-galled leaves and galls of Calophya duvauae (Scott (Hemiptera: Calophyidae on Schinus polygamus (Cav. Cabrera (Anacardiaceae. Cell fates changed from non-galled leaves to galls during the course of tissue differentiation. C. duvauae caused changes in dermal, ground, and vascular systems of the leaves of S. polygamus. Its feeding activity induced the homogenization of the parenchyma, and the neoformation of vascular bundles and trichomes. The histometric and cytometric data revealed compensatory effects of hyperplasia and cell hypertrophy in the epidermis, with hyperplasia predominating in the adaxial epidermis. There was a balance between these processes in the other tissues. Thus, we found major differences between the developmental pathways of non-galled leaves and galls. These changes were associated with phenotypic alterations related to shelter and appropriate microenvironmental conditions for the gall inducer. The nondifferentiation of a typical nutritive tissue in this case was compared to other non-phylogenetically related arthropod gall systems, and is suggested to result from convergence associated with the piercing feeding apparatus of the corresponding gall-inducer.

  3. A Cytotoxic and Anti-inflammatory Campesterol Derivative from Genetically Transformed Hairy Roots of Lopezia racemosa Cav. (Onagraceae

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    Norma Elizabeth Moreno-Anzúrez

    2017-01-01

    Full Text Available The genetically transformed hairy root line LRT 7.31 obtained by infecting leaf explants of Lopezia racemosa Cav with the Agrobacterium rhizogenes strain ATCC15834/pTDT, was evaluated to identify the anti-inflammatory and cytotoxic compounds reported previously for the wild plant. After several subcultures of the LRT 7.31 line, the bio-guided fractionation of the dichloromethane–methanol (1:1 extract obtained from dry biomass afforded a fraction that showed important in vivo anti-inflammatory, and in vitro cytotoxic activities. Chemical separation of the active fraction allowed us to identify the triterpenes ursolic (1 and oleanolic (2 acids, and (23R-2α,3β,23,28-tetrahydroxy-14,15-dehydrocampesterol (3 as the anti-inflammatory principles of the active fraction. A new molecule 3 was characterized by spectroscopic analysis of its tetraacetate derivative 3a. This compound was not described in previous reports of callus cultures, in vitro germinated seedlings and wild plant extracts of whole L. racemosa plants. The anti-inflammatory and cytotoxic activities displayed by the fraction are associated to the presence of compounds 1–3. The present study reports the obtaining of the transformed hairy roots, the bioguided isolation of the new molecule 3, and its structure characterization.

  4. A role for CaV1 and calcineurin signaling in depolarization-induced changes in neuronal DNA methylation

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    Eilis Hannon

    2015-07-01

    Full Text Available Direct manipulations of neuronal activity have been shown to induce changes in DNA methylation (DNAm, although little is known about the cellular signaling pathways involved. Using reduced representation bisulfite sequencing, we identify DNAm changes associated with moderate chronic depolarization in dissociated rat hippocampal cultures. Consistent with previous findings, these changes occurred primarily in the vicinity of loci implicated in neuronal function, being enriched in intergenic regions and underrepresented in CpG-rich promoter regulatory regions. We subsequently used 2 pharmacological interventions (nifedipine and FK-506 to test whether the identified changes depended on 2 interrelated signaling pathways known to mediate multiple forms of neuronal plasticity. Both pharmacological manipulations had notable effects on the extent and magnitude of depolarization-induced DNAm changes indicating that a high proportion of activity-induced changes are likely to be mediated by calcium entry through L-type CaV1 channels and/or downstream signaling via the calcium-dependent phosphatase calcineurin.

  5. Hepatoprotective Activity of Aqueous Acetone Extract from Cienfuegosia digitata Cav. (Malvaceae Against Alcohol Hepatotoxicity in Albinos Wistar Rats

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    K. Konaté

    2012-06-01

    Full Text Available The present study was conducted to evaluate the hepatoprotective effects of aqueous acetone extract of Cienfuegosia digitata Cav. (Malvaceae an herbal plant used in Burkina Faso to treat traditionally liver disease in albinos Wistar rats. Animals received by gavage 25, 50 and 100 mg/kg body weight of extract daily for a period of 28 days. Control groups received alcohol 35% and water. In vivo administration of 35% ethanol for a period of 28 days in rats showed an activity of liver marker enzymes (AST, ALT, ALP and GGT, triglycerides, total cholesterol, total bilirubin and direct bilirubin in serum compared with rats which received water (control water. However, administration of 35% ethanol along with aqueous acetone extract decreased the activities of liver markers enzyme in serum comparatively to the control water group (p0.05 and p<0.05. This study revealed that Cienfuegosia digitata presents a hepatoprotective potential and this plant could be traditionally exploited in the treatment of liver disease particularly hepatitis B in Burkina Faso.

  6. First-principles investigation of exchange interactions in quasi-one-dimensional antiferromagnet CaV2O4

    Science.gov (United States)

    Pchelkina, Z. V.; Solovyev, I. V.

    2015-01-01

    The effects of orbital degrees of freedom on the exchange interactions in a quasi-one-dimensional spin-1 antiferromagnet CaV2O4 are systematically studied. For this purpose a realistic low-energy electron model with the parameters derived from the first-principles calculations is constructed in the Wannier basis for the t2g bands. The exchange interactions are calculated using both the theory of infinitesimal spin rotations near the mean-field ground state and the superexchange model, which provide a consistent description. The obtained behaviour of exchange interactions differs substantially from the previously proposed phenomenological picture based on magnetic measurements and structural considerations, namely: (i) despite the quasi-one-dimensional character of the crystal structure, consisting of the zigzag chains of the edge-sharing VO6 octahedra, the electronic structure is essentially three-dimensional, that leads to finite interactions between the chains; (ii) the exchange interactions along the legs of the chains appear to dominate; and (iii) there is a substantial difference in exchange interactions in two crystallographically inequivalent chains. The combination of these three factors successfully reproduces the behaviour of experimental magnetic susceptibility.

  7. ÁCIDO GIBERÉLICO EN LA GERMINACIÓN DE SEMILLAS DE Jaltomata procumbens (Cav. J. L. GENTRY

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    Pedro Sald\\u00EDvar-Iglesias

    2010-01-01

    Full Text Available El objetivo del presente trabajo fue evaluar la aplicación de ácido giberélico (AG 3 en la germinación de semillas provenie ntes de frutos silvestres de Jaltomata procumbens (Cav. J. L. Ge ntry. El ensayo se realizó durante los meses de julio-agosto de 2006. Los tratamie ntos utilizados fueron 0; 50; 100; 150; 200 y 250 mg/l de AG 3 con 12 y 24 h de remojo, mismos que fueron analizados bajo un diseño completamente aleatorio en arreglo bifactorial (tiempo de remojo y concentración de ácido giberélico. Las variables fueron: porcentaje, velocidad y periodo de germinación. Se gún los resultados, el tratamie nto 250 mg/l presentó un 87,0% de germinación con 1,7 plantas/ germinadas/día en un periodo de 25,5 días; superior a los valores presentados por los demás tratamie ntos. Se observó que el tiempo de remojo influye menos que la aplicación de ácido giberélico.

  8. Upregulation of the CaV 1.1-ryanodine receptor complex in a rat model of critical illness myopathy.

    Science.gov (United States)

    Kraner, Susan D; Wang, Qingbo; Novak, Kevin R; Cheng, Dongmei; Cool, David R; Peng, Junmin; Rich, Mark M

    2011-06-01

    The processes that trigger severe muscle atrophy and loss of myosin in critical illness myopathy (CIM) are poorly understood. It has been reported that muscle disuse alters Ca(2+) handling by the sarcoplasmic reticulum. Since inactivity is an important contributor to CIM, this finding raises the possibility that elevated levels of the proteins involved in Ca(2+) handling might contribute to development of CIM. CIM was induced in 3- to 5-mo-old rats by sciatic nerve lesion and infusion of dexamethasone for 1 wk. Western blot analysis revealed increased levels of ryanodine receptor (RYR) isoforms-1 and -2 as well as the dihydropyridine receptor/voltage-gated calcium channel type 1.1 (DHPR/Ca(V) 1.1). Immunostaining revealed a subset of fibers with elevation of RYR1 and Ca(V) 1.1 that had severe atrophy and disorganization of sarcomeres. These findings suggest increased Ca(2+) release from the sarcoplasmic reticulum may be an important contributor to development of CIM. To assess the endogenous functional effects of increased intracellular Ca(2+) in CIM, proteolysis of α-fodrin, a well-known target substrate of Ca(2+)-activated proteases, was measured and found to be 50% greater in CIM. There was also selective degradation of myosin heavy chain relative to actin in CIM muscle. Taken together, our findings suggest that increased Ca(2+) release from the sarcoplasmic reticulum may contribute to pathology in CIM.

  9. 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.

  10. KIF13B establishes a CAV1-enriched microdomain at the ciliary transition zone to promote Sonic hedgehog signalling

    Science.gov (United States)

    Schou, Kenneth B.; Mogensen, Johanne B.; Morthorst, Stine K.; Nielsen, Brian S.; Aleliunaite, Aiste; Serra-Marques, Andrea; Fürstenberg, Nicoline; Saunier, Sophie; Bizet, Albane A.; Veland, Iben R.; Akhmanova, Anna; Christensen, Søren T.; Pedersen, Lotte B.

    2017-01-01

    Ciliary membrane composition is controlled by transition zone (TZ) proteins such as RPGRIP1, RPGRIPL and NPHP4, which are vital for balanced coordination of diverse signalling systems like the Sonic hedgehog (Shh) pathway. Activation of this pathway involves Shh-induced ciliary accumulation of Smoothened (SMO), which is disrupted by disease-causing mutations in TZ components. Here we identify kinesin-3 motor protein KIF13B as a novel member of the RPGRIP1N-C2 domain-containing protein family and show that KIF13B regulates TZ membrane composition and ciliary SMO accumulation. KIF13B is upregulated during ciliogenesis and is recruited to the ciliary base by NPHP4, which binds to two distinct sites in the KIF13B tail region, including an RPGRIP1N-C2 domain. KIF13B and NPHP4 are both essential for establishment of a CAV1 membrane microdomain at the TZ, which in turn is required for Shh-induced ciliary SMO accumulation. Thus KIF13B is a novel regulator of ciliary TZ configuration, membrane composition and Shh signalling. PMID:28134340

  11. Molecular Regions Determining The Voltage-dependence of Inactivation for T-type Calcium Channel Cav3.1%调控T型钙通道Cav3.1电压依赖性失活的分子结构域

    Institute of Scientific and Technical Information of China (English)

    贺秉军; 胡芬; 商学良; 韩丽鑫; 吴广彦; 李俊英; 孙金生

    2014-01-01

    T型钙通道(Cav3)广泛分布于各类细胞,其显著的电生理学特点是低电位激活和快速的电压依赖性失活.失活在通道的生理功能调节中起十分重要的作用,但具体参与通道失活的分子基础目前并不完全清楚.为明确Cav3.1通道中调控电压依赖性失活的结构域,用Cav1.2通道(无电压依赖性失活)结构域Ⅰ和Ⅱ中的S1~S4、S5~S6区及Ⅰ和Ⅱ间的联系区替换Cav3.1中的相应区域,构建嵌合通道,并在卵母细胞中表达,用电压钳技术分析通道的电生理学特性.结果表明,替换Ⅰ中的S1~S4或S5~S6区可使Cav3.1的失活特性显著改变,但这种改变主要是由激活-失活偶联所致.Ⅱ的替换使通道的失活曲线参数发生显著改变,表明结构域Ⅱ,包括S1~S4和S5~S6均参与Cav3.1失活过程的调控.Ⅰ、Ⅱ间的联系区及Ⅰ中的S5~S6主要调控Cav3.1的失活速率,Ⅰ和Ⅱ中的S1~S4对通道失活速率无影响.综上所述,结构域Ⅱ是调控Cav3.1电压依赖性失活的关键因素,结构域Ⅰ不参与该通道失活过程的调控.Ⅰ、Ⅱ间的联系区及Ⅰ中的S5~S6主要调控Cav3.1通道的失活速率,Ⅰ、Ⅱ中的S1~S4对通道失活速率无影响.

  12. 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

  13. Composting Phragmites australis Cav. plant material and compost effects on soil and tomato (Lycopersicon esculentum Mill.) growth.

    Science.gov (United States)

    Toumpeli, Anna; Pavlatou-Ve, Athina K; Kostopoulou, Sofia K; Mamolos, Andreas P; Siomos, Anastasios S; Kalburtji, Kiriaki L

    2013-10-15

    Composting organic residues is a friendly to the environment alternative to producing fertilizer. This research was carried out to study the process of composting Phragmites australis Cav. plant material alone or with animal manure on a pilot-scale, to evaluate firstly the quality of the composts produced and secondly, using a pot experiment, the effects of their application on soil physicochemical characteristics and tomato plants development. For the compost production a randomized complete block design was used with five treatments (five compost types) and four replications. For the pot experiment, a completely randomized design was used with 17 treatments (plain soil, soil with synthetic fertilizer and the application of five compost types, at three rates each) and five replications. Compost N increased with composting time, while C/N ratio decreased significantly and by the end it ranged from 43.3 for CM to 22.6 for CY. Compost pH became almost neutral, ranging from 6.73 for CY to 7.21 for CM3Y3AM4 by the end. Compost combinations CY7AM3 and CM7AM3 had a more positive influence on the soil physicochemical characteristics than the others. Soil N, P, Ca and Mg concentrations and the reduction of clay dispersion were the highest when CM7AM3 compost was added. The macro-aggregate stability was the highest for CY7AM3, which also sustained plant growth. The latter compost combination improved most of the soil physicochemical characteristics and plant growth especially, when the application rate was 4% (w/w), which equals to 156 Mg ha(-1).

  14. CaV3.2 T-type Ca²⁺ channels in H₂S-mediated hypoxic response of the carotid body.

    Science.gov (United States)

    Makarenko, Vladislav V; Peng, Ying-Jie; Yuan, Guoxiang; Fox, Aaron P; Kumar, Ganesh K; Nanduri, Jayasri; Prabhakar, Nanduri R

    2015-01-15

    Arterial blood O2 levels are detected by specialized sensory organs called carotid bodies. Voltage-gated Ca(2+) channels (VGCCs) are important for carotid body O2 sensing. Given that T-type VGCCs contribute to nociceptive sensation, we hypothesized that they participate in carotid body O2 sensing. The rat carotid body expresses high levels of mRNA encoding the α1H-subunit, and α1H protein is localized to glomus cells, the primary O2-sensing cells in the chemoreceptor tissue, suggesting that CaV3.2 is the major T-type VGCC isoform expressed in the carotid body. Mibefradil and TTA-A2, selective blockers of the T-type VGCC, markedly attenuated elevation of hypoxia-evoked intracellular Ca(2+) concentration, secretion of catecholamines from glomus cells, and sensory excitation of the rat carotid body. Similar results were obtained in the carotid body and glomus cells from CaV3.2 knockout (Cacna1h(-/-)) mice. Since cystathionine-γ-lyase (CSE)-derived H2S is a critical mediator of the carotid body response to hypoxia, the role of T-type VGCCs in H2S-mediated O2 sensing was examined. Like hypoxia, NaHS, a H2S donor, increased intracellular Ca(2+) concentration and augmented carotid body sensory nerve activity in wild-type mice, and these effects were markedly attenuated in Cacna1h(-/-) mice. In wild-type mice, TTA-A2 markedly attenuated glomus cell and carotid body sensory nerve responses to hypoxia, and these effects were absent in CSE knockout mice. These results demonstrate that CaV3.2 T-type VGCCs contribute to the H2S-mediated carotid body response to hypoxia.

  15. CaV3.2 T-type Ca2+ channels in H2S-mediated hypoxic response of the carotid body

    Science.gov (United States)

    Makarenko, Vladislav V.; Peng, Ying-Jie; Yuan, Guoxiang; Fox, Aaron P.; Kumar, Ganesh K.; Nanduri, Jayasri

    2014-01-01

    Arterial blood O2 levels are detected by specialized sensory organs called carotid bodies. Voltage-gated Ca2+ channels (VGCCs) are important for carotid body O2 sensing. Given that T-type VGCCs contribute to nociceptive sensation, we hypothesized that they participate in carotid body O2 sensing. The rat carotid body expresses high levels of mRNA encoding the α1H-subunit, and α1H protein is localized to glomus cells, the primary O2-sensing cells in the chemoreceptor tissue, suggesting that CaV3.2 is the major T-type VGCC isoform expressed in the carotid body. Mibefradil and TTA-A2, selective blockers of the T-type VGCC, markedly attenuated elevation of hypoxia-evoked intracellular Ca2+ concentration, secretion of catecholamines from glomus cells, and sensory excitation of the rat carotid body. Similar results were obtained in the carotid body and glomus cells from CaV3.2 knockout (Cacna1h−/−) mice. Since cystathionine-γ-lyase (CSE)-derived H2S is a critical mediator of the carotid body response to hypoxia, the role of T-type VGCCs in H2S-mediated O2 sensing was examined. Like hypoxia, NaHS, a H2S donor, increased intracellular Ca2+ concentration and augmented carotid body sensory nerve activity in wild-type mice, and these effects were markedly attenuated in Cacna1h−/− mice. In wild-type mice, TTA-A2 markedly attenuated glomus cell and carotid body sensory nerve responses to hypoxia, and these effects were absent in CSE knockout mice. These results demonstrate that CaV3.2 T-type VGCCs contribute to the H2S-mediated carotid body response to hypoxia. PMID:25377087

  16. Sites of proteolytic processing and noncovalent association of the distal C-terminal domain of CaV1.1 channels in skeletal muscle.

    Science.gov (United States)

    Hulme, Joanne T; Konoki, Keiichi; Lin, Teddy W-C; Gritsenko, Marina A; Camp, David G; Bigelow, Diana J; Catterall, William A

    2005-04-05

    In skeletal muscle cells, voltage-dependent potentiation of Ca2+ channel activity requires phosphorylation by cAMP-dependent protein kinase (PKA) anchored via an A-kinase anchoring protein (AKAP15), and the most rapid sites of phosphorylation are located in the C-terminal domain. Surprisingly, the site of interaction of the complex of PKA and AKAP15 with the alpha1-subunit of Ca(V)1.1 channels lies in the distal C terminus, which is cleaved from the remainder of the channel by in vivo proteolytic processing. Here we report that the distal C terminus is noncovalently associated with the remainder of the channel via an interaction with a site in the proximal C-terminal domain when expressed as a separate protein in mammalian nonmuscle cells. Deletion mapping of the C terminus of the alpha1-subunit using the yeast two-hybrid assay revealed that a distal C-terminal peptide containing amino acids 1802-1841 specifically interacts with a region in the proximal C terminus containing amino acid residues 1556-1612. Analysis of the purified alpha1-subunit of Ca(V)1.1 channels from skeletal muscle by saturation sequencing of the intracellular peptides by tandem mass spectrometry identified the site of proteolytic processing as alanine 1664. Our results support the conclusion that a noncovalently associated complex of the alpha1-subunit truncated at A1664 with the proteolytically cleaved distal C-terminal domain, AKAP15, and PKA is the primary physiological form of Ca(V)1.1 channels in skeletal muscle cells.

  17. Cav2-type calcium channels encoded by cac regulate AP-independent neurotransmitter release at cholinergic synapses in adult Drosophila brain.

    Science.gov (United States)

    Gu, Huaiyu; Jiang, Shaojuan Amy; Campusano, Jorge M; Iniguez, Jorge; Su, Hailing; Hoang, Andy An; Lavian, Monica; Sun, Xicui; O'Dowd, Diane K

    2009-01-01

    Voltage-gated calcium channels containing alpha1 subunits encoded by Ca(v)2 family genes are critical in regulating release of neurotransmitter at chemical synapses. In Drosophila, cac is the only Ca(v)2-type gene. Cacophony (CAC) channels are localized in motor neuron terminals where they have been shown to mediate evoked, but not AP-independent, release of glutamate at the larval neuromuscular junction (NMJ). Cultured embryonic neurons also express CAC channels, but there is no information about the properties of CAC-mediated currents in adult brain nor how these channels regulate transmission in central neural circuits where fast excitatory synaptic transmission is predominantly cholinergic. Here we report that wild-type neurons cultured from late stage pupal brains and antennal lobe projection neurons (PNs) examined in adult brains, express calcium currents with two components: a slow-inactivating current sensitive to the spider toxin Plectreurys toxin II (PLTXII) and a fast-inactivating PLTXII-resistant component. CAC channels are the major contributors to the slow-inactivating PLTXII-sensitive current based on selective reduction of this component in hypomorphic cac mutants (NT27 and TS3). Another characteristic of cac mutant neurons both in culture and in whole brain recordings is a reduced cholinergic miniature excitatory postsynaptic current frequency that is mimicked in wild-type neurons by acute application of PLTXII. These data demonstrate that cac encoded Ca(v)2-type calcium channels regulate action potential (AP)-independent release of neurotransmitter at excitatory cholinergic synapses in the adult brain, a function not predicted from studies at the larval NMJ.

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

    OpenAIRE

    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...

  19. N-type calcium current, Cav2.2, is enhanced in small-diameter sensory neurons isolated from Nf1+/- mice.

    Science.gov (United States)

    Duan, J-H; Hodgdon, K E; Hingtgen, C M; Nicol, G D

    2014-06-13

    Major aspects of neuronal function are regulated by Ca(2+) including neurotransmitter release, excitability, developmental plasticity, and gene expression. We reported previously that sensory neurons isolated from a mouse model with a heterozygous mutation of the Nf1 gene (Nf1+/-) exhibited both greater excitability and evoked release of neuropeptides compared to wildtype mice. Furthermore, augmented voltage-dependent sodium currents but not potassium currents contribute to the enhanced excitability. To determine the mechanisms giving rise to the enhanced release of substance P and calcitonin gene-related peptide in the Nf1+/- sensory neurons, the potential differences in the total voltage-dependent calcium current (ICa) as well as the contributions of individual Ca(2+) channel subtypes were assessed. Whole-cell patch-clamp recordings from small-diameter capsaicin-sensitive sensory neurons demonstrated that the average peak ICa densities were not different between the two genotypes. However, by using selective blockers of channel subtypes, the current density of N-type (Cav2.2) ICa was significantly larger in Nf1+/- neurons compared to wildtype neurons. In contrast, there were no significant differences in L-, P/Q- and R-type currents between the two genotypes. Quantitative real-time polymerase chain reaction measurements made from the isolated but intact dorsal root ganglia indicated that N-type (Cav2.2) and P/Q-type (Cav2.1) Ca(2+) channels exhibited the highest mRNA expression levels although there were no significant differences in the levels of mRNA expression between the genotypes. These results suggest that the augmented N-type (Cav2.2) ICa observed in the Nf1+/- sensory neurons does not result from genomic differences but may reflect post-translational or some other non-genomic modifications. Thus, our results demonstrate that sensory neurons from Nf1+/- mice, exhibit increased N-type ICa and likely account for the increased release of substance P and

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

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    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.

  1. 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

  2. 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

  3. Chemical Constituents of Salvia leucantha CAV.%墨西哥鼠尾草的化学成分研究

    Institute of Scientific and Technical Information of China (English)

    蒋永俊; 朱雅竹; 李洪梅; 李蓉涛

    2015-01-01

    利用硅胶、羟丙基葡聚糖凝胶(Sephadex LH-20)、MCI、中压 ODS 柱色谱及高效液相色谱(HPLC)等分离手段及波谱学方法,从墨西哥鼠尾草(Salvia leucantha CAV.)地上部分中共分离并鉴定了15个化合物,分别为二萜类:salviandulin E (1),dugesin A (2),salvileucantholide (3),isopuberulin (4),dugesin B (5),4-epi-abietinol (6);三萜类:3β-acetoxy-11-ursen-13α,30-olide (7),3β-19α-dihydroxy-30-norurs-12-ene (8),3β-hydroxyolean-12-en-28-al (9),熊果醛(3β-hydroxyurs-12-en-28-al,10);黄酮类:柚皮素(naringenin,11)和橙皮素(hespertin,12)及其他类成分:p-hydroxyphenethyl trans-ferulate (13),vomifoliol (14),pubinernoid A (15).其中2,5和6~15为首次从该植物中分离得到的化合物.%By the column chromatographies of silica gel,Sephadex LH -20,MCI,ODS and HPLC,and spec-troscopic evidences,15 compounds are isolated and identified from the aerial part of Salvia leucantha CAV.,in-cluding 6 diterpenoids,named salviandulin E (1),dugesin A (2),salvileucantholide (3),isopuberulin (4), dugesin B (5),and 4-epi-abietinol (6),4 triterpenoids,named 3β-acetoxy-11-ursen-13α,30-olide (7),3β-19α-dihydroxy-30-norurs-12-ene (8 ), 3β-hydroxyolean-12-en-28-al (9 ), and 3β-hydroxyurs-12-en-28-al (10),2 flavonoids,named naringenin (11)and hesperetin (12),and 3 other constituents,named p-hydroxy-phenethyl trans-ferulate (13),vomifoliol (14),and pubinernoid A (15).Among them,2,5 and 6 15 are i-solated from the title plant for the first time.

  4. 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.

  5. 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.

  6. Cav-1 mRNA、S100A4 mRNA 和 CD31 的表达与前列腺癌转移及生存率关系的研究%Correlation of Cav-1 mRNA, S100A4 mRNA and CD31 expression with prostate cancer tumor metastasis and patient survival rate

    Institute of Scientific and Technical Information of China (English)

    庄黛娜; 闫海强; 杨最素; 丁国芳

    2012-01-01

    Objective To explore the expression of Cav-1 mRNA,S100A4 mRNA and CD31 in prostate cancer (PCa) and their correlation with tumor metastasis and patient survival rate. Methods PCa specimens (n =42) and adjacent tissue specimens ( n =12 ) from radical prostatectomy were obtained from January 2004 to May 2006.The mean age of patient was 71.6 ± 7.6 years ( range 58 - 86 years).According to Gleason scores,prostatectomy specimens were stratified into≤6 (n =17),7 (n =12) and ≥8( n =13 ) groups.Patients were classified as clinical stage T1 ( n =16),stage T2 ( n =9 ),stage T3 ( n =11 )and stage T4 (n =6).Patients were divided into PCa with bone metastasis (n =8 ) and PCa without bone metastasis ( n =34).Preoperative PSA levels of the patients were stratified into three groups: < 4 ( n =4)μg/L,4-10 (n=10)μg/Land >10 μg/L (n=28).12 adjacent tissues 1 -2 cm away from tumor or another lobe of prostate were microscopically verified without cancer cells and were tested for comparison.The expression of Cav-1 mRNA and S100A4 mRNA were detected by Situ hybridization in 42 PCa specimens and 12 adjacent tissues and using CD31 for marking vascular endothelial cells,the tumor microvascular density (MVD) was counted.The correlation of Cav-1 mRNA,S100A4 mRNA and CD31 expression was analyzed in combination.with clinical and pathological fcatures including Gleason score,TNM staging,PSA values and bone metastasis. Results The positive expression rate of Cav-1 mRNA in PCa was 35.7% ( 15/42),while it was 0% (0/12) in controls,P <0.05.The positive expression rate of S100A4 mRNA in PCa was 47.6% (20/42),while it was 8.3% (1/12) in controls,P <0.05.The positive expression rate of Cav-1 mRNA in PCa was positively correlated with Gleason score,TNM stage and bone metastasis.The positive expression rate of S100A4 mRNA in PCa was positively correlated with TNM stage and bone metastasis.The average MVD in patients of negative expression of Cav-1 mRNA was (62.8 ± 10

  7. Initial impacts and field validation of host range for Boreioglycaspis melaleucae Moore (Hemiptera: Psyllidae),a biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) Blake (Myrtales: Myrtaceae: Leptosp

    Science.gov (United States)

    Invasion of south Florida wetlands by the Australian paperbark tree (“melaleuca”), Melaleuca quinquenervia (Cav.) S.T. Blake (melaleuca) has caused adverse economic and environmental impacts. The tree’s biological attributes along with favorable ambient biophysical conditions combine to complicate ...

  8. 正交法在全空气定风量系统中的应用%Application of Orthogonal Method in CAV System

    Institute of Scientific and Technical Information of China (English)

    张航; 邵宗义; 陈红兵; 段良飞

    2015-01-01

    为了研究影响全空气定风量系统的因素,需要进行大量的模拟试验,但这会花费大量的时间和经历。为此,可以通过设计合理的试验,并建立合理的模型,进行模拟计算,从而能得出较为准确的结果。采用正交实验法,减少了模拟次数,得出影响全空气定风量系统的主要因素和次要因素;运用DeST软件进行能耗模拟,根据不同地域、不同建筑功能及不同围护结构的合理搭配,得出一个最适合采用全空气定风量系统的建筑。在今后研究中,还可以根据不同的影响因素,通过模拟计算,得出不同空调系统的能耗,从而选出最合适的系统。%We need to do extensive simulations in order to study the influencing factor of CAV system, but it will cost lots of time and experience. Therefore, we can design the reasonable test and establish properly model to get the accurate results. Using the orthogonal experimental method can reduce simulation times and find the main factors and secondary factors which affect CAV system; using DeST software measuring the energy consumption according to different regions, different functions and different building envelope, we can get the most suitable building for CAV system. In the future research, according to different influencing factor, we can simulate and calculate the test to analyze energy consumption and select the most suitable system.

  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).

    Science.gov (United States)

    Konaté, K; Souza, A; Coulibaly, A Y; Meda, N T R; Kiendrebeogo, M; Lamien-Meda, A; Millogo-Rasolodimby, J; Lamidi, M; Nacoulma, O G

    2010-11-15

    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.

  10. 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.

  11. (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.

  12. Expression of Cav-1, AQP1 and AQP5 in lung of acute pancreatitis-associated lung injury rats and the therapeutic role of Qingyitang%急性胰腺炎肺损伤大鼠肺组织中Cav-1与水通道蛋白1、5的表达及清胰汤的治疗作用

    Institute of Scientific and Technical Information of China (English)

    王钢; 陈海龙; 任凤; 李洁; 李雅琼

    2010-01-01

    Objective To investigate the expression and location of caveolin-1 (Cav-1), aquaporin 1 ( AQP1 ) and AQP5 in the lung of acute pancreatitis-associated lung injury rats and to determine the role of these molecules in the pathologic progress and the potential therapeutic mechanisms of Qingyitang. Methods Forty Wistar rats were randomly divided into sham operation (SHAM) group, acute lung injury (ALI)group, dexamethasone (DEX) group and Qingyitang (QYT) group. ALI was induced by reverse injection of deoxycholate into biliopancreatic duct of rats. Blood and lung tissues were collected after 24 h. Serum amylase, lung wet/dry (W/D) ratio and pathological section were detected to evaluate the degree of lung injury. reverse transcription-polymerase chain reaction was taken to detect the mRNA levels of Cav-1, AQP1 and AQP5. Lipid rafts were prepared for detection of the distribution and expression level of Cav-1, AQP1 and AQP5 proteins by Western blot. Results The concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously increased in ALI rats. Cav-1, AQP1 and AQP5 were present in the lung while the mRNA level decreased in ALI rats. Cav-1 appeared mainly in lipid rafts and less in non-lipid rafts. AQP1 was localized to lipid rafts while AQP5 to non-lipid rafts. The localization of these three molecules in the lung of ALI rats did not change compared with SHAM rats while their protein levels decreased. Compared with ALI rats, the concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously decreased in DEX and QYT rats. The mRNA and the protein expression of Cay-1, AQP1 and AQP5 increased in various degrees by DEX or QYT treatment. Conclusion Cav-1 and AQP1 are enriched in lipid rafts while AQP5 in non-lipid rafts. The down-regulated expression of these three molecules may play important role in acute pancreatitis-associated lung injury. DEX and QYT may relieve lung injury effectively by up

  13. Etnobotánica, micrografía analítica de hojas y tallos y fitoquimica de Cuphea aequipetala Cav. (Lythraceae): una contribución a la Farmacopea Herbolaria de los Estados Unidos Mexicanos (FHEUM)

    OpenAIRE

    Silvia AGUILAR-RODRÍGUEZ; Nayeli L. ECHEVESTE-RAMÍREZ; Ma. Edith LÓPEZ-VILLAFRANCO; Aguilar-Contreras, Abigail; Vega-Ávila, Elisa; Reyes-Chilpa, Ricardo

    2012-01-01

    Cuphea aquipetala Cav. (Hierba del cancer) is used in Mexican traditional medicine. In the present work, its most frequent medicinal applications were determined through interviews, revision of herbarium specimens, and bibliography. The quoted applications were: to treat pounds and wounds. For these purposes, the drug (leaf and stem) is prepared and administered as infusion, or as poultice. In order to provide data for inclusion of this species in the Herbal Pharmacopoeia of the United Mexica...

  14. COOH-terminal association of human smooth muscle calcium channel Ca(v)1.2b with Src kinase protein binding domains: effect of nitrotyrosylation.

    Science.gov (United States)

    Kang, Minho; Ross, Gracious R; Akbarali, Hamid I

    2007-12-01

    The carboxyl terminus of the calcium channel plays an important role in the regulation of calcium entry, signal transduction, and gene expression. Potential protein-protein interaction sites within the COOH terminus of the L-type calcium channel include those for the SH3 and SH2 binding domains of c-Src kinase that regulates calcium currents in smooth muscle. In this study, we examined the binding sites involved in Src kinase-mediated phosphorylation of the human voltage-gated calcium channel (Ca(v)) 1.2b (hCav1.2b) and the effect of nitrotyrosylation. Cotransfection of human embryonic kidney (HEK)-293 cells with hCa(v)1.2b and c-Src resulted in tyrosine phosphorylation of the calcium channel, which was prevented by nitration of tyrosine residues by peroxynitrite. Whole cell calcium currents were reduced by 58 + 5% by the Src kinase inhibitor PP2 and 64 + 6% by peroxynitrite. Nitrotyrosylation prevented Src-mediated regulation of the currents. Glutathione S-transferase fusion protein of the distal COOH terminus of hCa(v)1.2b (1809-2138) bound to SH2 domain of Src following tyrosine phosphorylation, while binding to SH3 required the presence of the proline-rich motif. Site-directed mutation of Y(2134) prevented SH2 binding and resulted in reduced phosphorylation of hCa(v)1.2b. Within the distal COOH terminus, single, double, or triple mutations of Y(1837), Y(1861), and Y(2134) were constructed and expressed in HEK-293 cells. The inhibitory effects of PP2 and peroxynitrite on calcium currents were significantly reduced in the double mutant Y(1837-2134F). These data demonstrate that the COOH terminus of hCa(v)1.2b contains sites for the SH2 and SH3 binding of Src kinase. Nitrotyrosylation of these sites prevents Src kinase regulation and may be importantly involved in calcium influx regulation during inflammation.

  15. Enhanced excitatory transmission at cortical synapses as the basis for facilitated spreading depression in Ca(v)2.1 knockin migraine mice.

    Science.gov (United States)

    Tottene, Angelita; Conti, Rossella; Fabbro, Alessandra; Vecchia, Dania; Shapovalova, Maryna; Santello, Mirko; van den Maagdenberg, Arn M J M; Ferrari, Michel D; Pietrobon, Daniela

    2009-03-12

    Migraine is a common disabling brain disorder. A subtype of migraine with aura (familial hemiplegic migraine type 1: FHM1) is caused by mutations in Ca(V)2.1 (P/Q-type) Ca(2+) channels. Knockin mice carrying a FHM1 mutation show increased neuronal P/Q-type current and facilitation of induction and propagation of cortical spreading depression (CSD), the phenomenon that underlies migraine aura and may activate migraine headache mechanisms. We studied cortical neurotransmission in neuronal microcultures and brain slices of FHM1 mice. We show gain of function of excitatory neurotransmission due to increased action-potential-evoked Ca(2+) influx and increased probability of glutamate release at pyramidal cell synapses but unaltered inhibitory neurotransmission at fast-spiking interneuron synapses. Using an in vitro model of CSD, we show a causative link between enhanced glutamate release and CSD facilitation. The synapse-specific effect of FHM1 mutations points to disruption of excitation-inhibition balance and neuronal hyperactivity as the basis for episodic vulnerability to CSD ignition in migraine.

  16. Production of the Anti-Inflammatory Compound 6-O-Palmitoyl-3-O-β-D-glucopyranosylcampesterol by Callus Cultures of Lopezia racemosa Cav. (Onagraceae

    Directory of Open Access Journals (Sweden)

    Roberta Salinas

    2014-06-01

    Full Text Available Lopezia racemosa Cav. is a plant used in Mexican traditional medicine to heal inflammatory diseases. From this plant we isolated the novel compound 6-O-palmitoyl- 3-O-β-D-glucopyranosylcampesterol (1 and 6-O-palmitoyl-3-O-β-D-glucopyranosyl-β-sitosterol (2, previously reported to have cytotoxic activity on several cancer cell lines. We evaluated the anti-inflammatory activity of 1 in vivo by mouse ear edema induced with 12-O-tetradecanoylphorbol-13-acetate (TPA and 57.14% inhibition was observed. The aim of our study was to obtain callus cultures derived from this plant species with the ability to produce the compounds of interest. Callus cultures were initiated on MS basal medium amended with variable amounts of naphthaleneacetic acid (NAA, or 2,4-dichlorophenoxyacetic acid (2,4-D, combined or not with 6-benzylaminopurine (BAP. Ten treatments with these growth regulators were carried out, using in vitro germinated seedlings as source of three different explants: hypocotyl, stem node, and leaf. Highest yield of 1 was observed on callus derived from leaf explants growing in medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Selected callus lines produced less 1 than wild plants but the in vitro cultured seedlings showed higher production. So we conclude that it could be attractive to further investigate their metabolic potential.

  17. 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.

  18. 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.

  19. Timing of harvest of Phragmites australis (CAV.) Trin. ex Steudel affects subsequent canopy structure and nutritive value of roughage in subtropical highland.

    Science.gov (United States)

    Tanaka, Takashi S T; Irbis, Chagan; Kumagai, Hajime; Inamura, Tatsuya

    2016-01-15

    In recent decades, constructed wetlands dominated by common reeds [Phragmites australis (CAV.) Trin. ex Steudel] have been utilized for treating nitrogen-rich wastewaters. Although plant harvest is a vegetation management in constructed wetlands for the purpose of improving nutrient removal, harvested biomass has become a problem in many places. The reed has attracted increasing interest for its potential as high-quality roughage for ruminants. Therefore, it is crucial to understand the effect of reed harvest timing on subsequent regrowth, reconstruction of canopy structure, and nutritive value of regrown biomass for roughage when defining an appropriate vegetation management in constructed wetlands. The shoots of common reeds were harvested in January (winter), March (spring), and May (early summer) in a free-water surface constructed wetland in southwest China. Harvesting in winter enhanced the shoot regrowth and concentrations of total digestible nutrients (TDN), probably due to vigorous translocations of nonstructural carbohydrates from rhizomes. Harvesting in spring and early summer decreased aboveground biomass, nitrogen (N) standing stock, and concentrations of TDN. From fifty to 110 days after harvest, the TDN had sharply declined to values similar to non-harvested stands. Thus, to obtain high-quality roughage, it is recommended that regrown shoots be harvested again within a year in the early growing stage after the first harvest in winter.

  20. Calcium absorption by Cav1.3 induces terminal web myosin II phosphorylation and apical GLUT2 insertion in rat intestine.

    Science.gov (United States)

    Mace, Oliver J; Morgan, Emma L; Affleck, Julie A; Lister, Norma; Kellett, George L

    2007-04-15

    Glucose absorption in rat jejunum involves Ca(2+)- and PKC betaII-dependent insertion of GLUT2 into the apical membrane. Ca(2+)-induced rearrangement of the enterocyte cytoskeleton is thought to enhance paracellular flow. We have therefore investigated the relationships between myosin II regulatory light chain phosphorylation (RLC(20)), absorption of glucose, water and calcium, and mannitol clearance. ML-7, an inhibitor of myosin light chain kinase, diminished the phloretin-sensitive apical GLUT2 but not the phloretin-insensitive SGLT1 component of glucose absorption in rat jejunum perfused with 75 mM glucose. Western blotting and immunocytochemistry revealed marked decreases in RLC(20) phosphorylation in the terminal web and in the levels of apical GLUT2 and PKC betaII, but not SGLT1. Perfusion with phloridzin or 75 mM mannitol, removal of luminal Ca(2+), or inhibition of unidirectional (45)Ca(2+) absorption by nifedipine exerted similar effects. ML-7 had no effect on the absorption of 10 mM Ca(2+), nor clearance of [(14)C]-mannitol, which was less than 0.7% of the rate of glucose absorption. Water absorption did not correlate with (45)Ca(2+) absorption or mannitol clearance. We conclude that the Ca(2+) necessary for contraction of myosin II in the terminal web enters via an L-type channel, most likely Ca(v)1.3, and is dependent on SGLT1. Moreover, terminal web RLC(20) phosphorylation is necessary for apical GLUT2 insertion. The data confirm that glucose absorption by paracellular flow is negligible, and show further that paracellular flow makes no more than a minimal contribution to jejunal Ca(2+) absorption at luminal concentrations prevailing after a meal.

  1. 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.

  2. Efectividad biológica delazoxystrobin para el control de pyricularia oryzae cav. y cercospora oryzae miyake. en arroz de temporal en Veracruz, México

    Directory of Open Access Journals (Sweden)

    Enrique Becerra

    2001-01-01

    Full Text Available El estado de Veracruz, México, siembra anualmente 22000 ha de arroz de temporal con un rendimiento medio de 3,5 t/ha, debido principalmente a problemas de sequía lo cual favorece la presencia de hongos como Pyricularia oryzae Cav. (quema del arroz y Cercospora oryzae Miyake. (Mancha angosta. Con el objeto de conocer el comportamiento de nuevos fungicidas se evaluó la eficacia del Azoxystrobin en el control de estos hongos. El experimento se estableció en el municipio de Tres Valles, Veracruz, durante el temporal de 1999, con semilla de Milagro Filipino Depurado. El diseño estadístico utilizado fue bloques al azar con cuatro repeticiones. Los tratamientos evaluados fueron: Azoxystrobin a 0,2, 0,4 y 0,6 l/ha vs Tecto60 a 0,5 kg/ha y un testigo sin aplicación. Estos fueron aplicados al presentarse los primeros síntomas de las enfermedades. Se evaluó la incidencia, número de lesiones en 20 plantas, índice de intensidad, el rendimiento de grano y fitotoxicidad. Se encontró que el Azoxystrobin obtuvo mejores resultados que el Tecto 60 y éste a su vez que el testigo no tratado. El mejor control para P. oryzae y C. oryzae y el mayor rendimiento de grano (4432 kg/ha se logró cuando se aplicó Azoxystrobin en dosis de 0,6 l/ha, aunque con 0,2 y 0,4 l/ha de este fungicida tuvo un buen control de estas enfermedades. Ninguno de los fungicidas causó toxicidad al arroz

  3. Correlaciones genotípicas, fenotípicas y ambientales, y análisis de sendero en tomate de árbol (Cyphomandra betacea Cav. Sendt.

    Directory of Open Access Journals (Sweden)

    Liz Katherine Lagos Santander

    2013-07-01

    Full Text Available En el estudio se evaluaron diferentes componentes de producción y se estimaron las correlaciones fenotípicas, genéticas y ambientales, considerando nueve variables relacionadas con el tamaño y la calidad del fruto de tomate de árbol (Cyphomandra betacea Cav. Sendt.; igualmente se establecieron los efectos directos e indirectos de las variables componentes de calidad sobre el peso del fruto. Para el efecto se utilizaron los datos existentes de 81 híbridos (72 híbridos interpoblacionales y nueve testigos dispuestos en tres repeticiones, en condiciones del departamento de Nariño, Colombia. Los resultados obtenidos indicaron que las correlaciones genotípicas fueron superiores a las fenotípicas y ambientales. El peso de fruto (PF presentó las mayores correlaciones genéticas con el peso de semilla por fruto (rG = 0.97 y con el peso de pulpa más semilla (rG = 0.92. El análisis de sendero con base en correlaciones genotípicas mostró que el peso de pulpa más semilla fue la variable que tuvo el mayor efecto directo sobre el PF (1.353. Esto demuestra que una selección por peso de fruto da como resultado un aumento en peso de pulpa más semilla. Teniendo en cuenta las correlaciones fenotípicas, este análisis permitió establecer que los efectos directos de peso de semilla de fruto (PSF y peso de pulpa más semilla (P + S (1.166 y 0.743, respectivamente son los que más contribuyen al PF.

  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

    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 Ca2+-activated KCa2/SK outward current, which eventually reduces the repolarizing force and thereby prolong action potentials duration and 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. PMID:27014060

  5. Identificação dos inibidores de acetilcolinesterase em Tetrapterys mucronata Cav. (Malpighiaceae) e comparação quali e quantitativa dos derivados triptamícos presentes na espécie em estudo e Ayahuasca

    OpenAIRE

    Queiroz, Marcos Marçal Ferreira [UNESP

    2013-01-01

    The present work aims at seeking compounds that inhibit in vitro the enzyme acetylcholinesterase (AChE), and might be useful for the treatment of chronic neuro-degenerative diseases such as Alzheimer's. In this context the species Tetrapterys mucronata Cav. (Malpighiaceae) was selected from NuBBE’s bench of extract to be the focus of this work, due to the positive result for the in vitro inhibition of AChE conducted prior to this study. In the literature this species is also reported as a spe...

  6. 鸡贫血病病毒VP2基因在大肠杆菌中的表达及其特性%Expression of Chicken Anaemia Virus(CAV) VP2 Gene in E. coli and It's Biological Activities

    Institute of Scientific and Technical Information of China (English)

    刘岳龙; 秦爱建; 金文杰; 叶建强; 吉荣; 崔治中; 段玉友

    2003-01-01

    将鸡贫血病病毒(chicken anaemia virus,CAV)VP2基因克隆入表达性载体pGEX-5X-3,在大肠杆菌以谷胱甘肽转移酶(GST)融合蛋白的形式获得了表达.以此表达产物免疫小鼠,制备抗CAV VP2的多克隆抗体.通过对CAV感染的MSB1细胞作间接免疫荧光试验(IFA)检测,结果为阳性,这表明表达产物保留了CAV相关的抗原特性.本研究为进一步探索CAV VP2的生物学特性奠定了基础.

  7. 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.

  8. Familial hemiplegic migraine type 1 mutations W1684R and V1696I alter G protein-mediated regulation of Ca(V)2.1 voltage-gated calcium channels.

    Science.gov (United States)

    Garza-López, Edgar; Sandoval, Alejandro; González-Ramírez, Ricardo; Gandini, María A; Van den Maagdenberg, Arn; De Waard, Michel; Felix, Ricardo

    2012-08-01

    Familial hemiplegic migraine type 1 (FHM-1) is a monogenic form of migraine with aura that is characterized by recurrent attacks of a typical migraine headache with transient hemiparesis during the aura phase. In a subset of patients, additional symptoms such as epilepsy and cerebellar ataxia are part of the clinical phenotype. FHM-1 is caused by missense mutations in the CACNA1A gene that encodes the pore-forming subunit of Ca(V)2.1 voltage-gated Ca(2+) channels. Although the functional effects of an increasing number of FHM-1 mutations have been characterized, knowledge on the influence of most of these mutations on G protein regulation of channel function is lacking. Here, we explored the effects of G protein-dependent modulation on mutations W1684R and V1696I which cause FHM-1 with and without cerebellar ataxia, respectively. Both mutations were introduced into the human Ca(V)2.1α(1) subunit and their functional consequences investigated after heterologous expression in human embryonic kidney 293 (HEK-293) cells using patch-clamp recordings. When co-expressed along with the human μ-opioid receptor, application of the agonist [d-Ala2, N-MePhe4, Gly-ol]-enkephalin (DAMGO) inhibited currents through both wild-type (WT) and mutant Ca(V)2.1 channels, which is consistent with the known modulation of these channels by G protein-coupled receptors. Prepulse facilitation, which is a way to characterize the relief of direct voltage-dependent G protein regulation, was reduced by both FHM-1 mutations. Moreover, the kinetic analysis of the onset and decay of facilitation showed that the W1684R and V1696I mutations affect the apparent dissociation and reassociation rates of the Gβγ dimer from the channel complex, suggesting that the G protein-Ca(2+) channel affinity may be altered by the mutations. These biophysical studies may shed new light on the pathophysiology underlying FHM-1.

  9. Epidemic investigation of the co-infection of MDV, REV, CAV and ARV from commercial broilers flocks%商品肉鸡群中MDV、REV、CAV和ARV多重感染的检测

    Institute of Scientific and Technical Information of China (English)

    颜赟; 刁有祥; 孙杰; 刘霞; 吴焕荣

    2012-01-01

    从山东省东营、日照、潍坊、聊城等地区自然发病和临床健康AA商品肉鸡群中分别采集脏器样品,用特异性核酸探针对样品进行马立克氏病病毒(Marek’s disease virus,MDV)、网状内皮组织增生症病毒(Reticuloendotheliosis virus,REV)、鸡传染性贫血病病毒(Chicken anemia virus,CAV)和禽呼肠孤病毒(Avian reoviruses,ARV)检测。结果显示,自然发病AA商品肉鸡群中MDV、REV、CAV和ARV的检出率均较高,分别为69.30%、57.46%、63.60%和67.11%;临床健康AA商品肉鸡群中MDV、REV、CAV的检出率分别为36.96%、43.48%和30.42%,且自然发病和健康鸡群中均存在不同病毒组合的多重感染,感染率分别为85.96%和43.46%。用x2检验进行分析发现,自然发病商品肉鸡群与临床健康商品肉鸡群中MDV、CAV、MDV+REV、REV+CAV的检出率和未检出的比例差异极显著(P〈0.01);REV、MDV+CAV检出率差异显著(P〈0.05)。对自然发病商品肉鸡的肝脏、脾脏、法氏囊中4种病毒检出率进行X2检验分析发现,MDV在脾脏中检出率显著高于肝脏和法氏囊;REV在法氏囊中检出率显著高于肝脏和脾脏,而CAV和ARV分别在脾脏和肝脏中检出率较高。结果表明,多种免疫抑制性病毒的共感染已普遍存在,是目前AA商品肉鸡易发病且生长缓慢的重要流行病学因素之一。%Detection of Mareks disease virus (MDV), reticuloendotheliosis virus(REV), chicken anemia virus (CAV) and avian reoviruses(ARV) was done with a specific nucleic acid probes on the samples which were collected from natural incidence and clinical health AA commercial broilers flocks at Dongying, Rizhao, Weifang, Liaocheng and other areas of Shandong province. The detected ratios of MDV, REV, CAV, and ARV were all higher in natural incidence AA commercial broilers and they were 69.30%, 57.46 % , 63

  10. 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

  11. Base to Tip and Long-Distance Transport of Sodium in the Root of Common Reed [Phragmites australis (Cav.) Trin. ex Steud.] at Steady State Under Constant High-Salt Conditions.

    Science.gov (United States)

    Fujimaki, Shu; Maruyama, Teppei; Suzui, Nobuo; Kawachi, Naoki; Miwa, Eitaro; Higuchi, Kyoko

    2015-05-01

    We analyzed the directions and rates of translocation of sodium ions (Na(+)) within tissues of a salt-tolerant plant, common reed [Phragmites australis (Cav.) Trin. ex Steud.], and a salt-sensitive plant, rice (Oryza sativa L.), under constant high-salt conditions using radioactive (22)Na tracer and a positron-emitting tracer imaging system (PETIS). First, the test plants were incubated in a nutrient solution containing 50 mM NaCl and a trace level of (22)Na for 24 h (feeding step). Then the original solution was replaced with a fresh solution containing 50 mM NaCl but no (22)Na, in which the test plants remained for >48 h (chase step). Non-invasive dynamic visualization of (22)Na distribution in the test plants was conducted during feeding and chase steps with PETIS. Our results revealed that (22)Na was absorbed in the roots of common reed, but not transported to the upper shoot beyond the shoot base. During the chase step, a basal to distal movement of (22)Na was detected within the root tissue over >5 cm with a velocity of approximately 0.5 cm h(-1). On the other hand, (22)Na that was absorbed in the roots of rice was continuously translocated to and accumulated in the whole shoot. We concluded that the basal roots and the shoot base of common reed have constitutive functions of Na(+) exclusion only in the direction of root tips, even under constant high-salt conditions. This function apparently may contribute to the low Na(+) concentration in the upper shoot and high salt tolerance of common reed.

  12. Blocking the L-type Ca2+ channel (Cav 1.2) is the key mechanism for the vascular relaxing effect of Pterodon spp. and its isolated diterpene methyl-6α-acetoxy-7β-hydroxyvouacapan-17β-oate.

    Science.gov (United States)

    de Fátima Reis, Carolina; de Andrade, Daniela Medeiros Lobo; Junior Neves, Bruno; de Almeida Ribeiro Oliveira, Leandra; Pinho, José Felippe; da Silva, Leidiane Pinha; Dos Santos Cruz, Jader; Bara, Maria Teresa Freitas; Andrade, Carolina Horta; Rocha, Matheus Lavorenti

    2015-10-01

    Pterodon spp. Vogel (Fabaceae), popularly known as "sucupira", has ethnopharmacological application which is described as having antispasmodic and relaxant effects. Hence, it was hypothesized that sucupira oil-resin (SOR) could induce smooth muscle relaxation. So, this study investigated the mechanisms involved in the vasorelaxant effect of SOR and its isolated diterpene (methyl-6α-acetoxy-7β-hydroxyvouacapan-17β-oate). Vascular reactivity experiments were performed using rat aortic rings (n=5-8) with (E+) or without endothelium (E-) in an isolated bath organ. The SOR (0-56 μg/mL) relaxed phenylephrine (E+: 86.7±7.1%; E-: 92.3±4.7%) and KCl contracted rings (E-: 97.1±2.8%). In the same way, diterpene (0-48 μg/mL) also relaxed phenylephrine (E+: 94.5±3.6%; E-: 92.2±3.4%) and KCl contracted rings (E-: 99.7±0.2%). The pre-incubation of arterial rings with cyclopiazonic acid (reticular Ca2+-ATPase inhibitor), tetraethylammonium (K+ channels blocker) or MDL-12,330A (adenylyl cyclesinhibitor) did not modify either SOR- or diterpeneinduced vasorelaxation. However, ODQ (guanylyl cyclase inhibitor) impaired only diterpene-induced vasorelaxation. SOR and diterpene significantly reduced CaCl2-induced contraction stimulated by Bay K8644 (1 μM), phenylephrine (0.1 μM) or KCl solution (40 mM). Computational molecular docking studies demonstrated that the vasodilator effect of diterpene relies on blocking the Cav 1.2 channel, and patch clamp results showed that diterpene substantially decreased the ionic current through Cav 1.2 in freshly dissociated vascular smooth muscle cells. These findings suggest that SOR and its isolated diterpene induce endothelium-independent vascular relaxation by blocking the L-type Ca2+ channel (Cav 1.2).

  13. Analysis of Invasive Potential of Cenchrus calyculatus Cav.in Sandy Habitats%基于外来入侵植物光梗蒺藜草的发生分布特点的潜在入侵生境分析

    Institute of Scientific and Technical Information of China (English)

    贺俊英; 王志新; 哈斯巴根

    2014-01-01

    The invasive potential of Cenchrus calyculatus Cav.in the fixed sand, mobile sand, semi -fixed sand and semi-shifting sand was studied.C.calyculatus was the dominant species in the semi -fixed sand, with a relative coverage and importance value higher than the obtained in the other three types .Based on Shannon diversity indices C. calyculatus led to a reduced species diversity in semi -fixed sand .Further research should pay more attention to the inva-siveness of C.calyculatus in the semi-fixed sand.%通过对光梗蒺藜草在固定沙地、流动沙地、半固定沙地、半流动沙地4种沙地类型中作用的分析,表明光梗蒺藜草为半固定沙地上的优势种,其相对盖度和重要值均高于其他3种样地类型,占主导地位;通过Shannon多样性指数分析表明光梗蒺藜草有导致半固定沙地物种多样性减弱的趋势。进一步的研究应更注重光梗蒺藜草在半固定沙地的入侵趋势。

  14. 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.

  15. VibroCav: Hydrodynamic Vibration and Cavitation Technology

    NARCIS (Netherlands)

    Bakker, T.W.

    2012-01-01

    Vibration and cavitation can be generated in many ways and serve many useful purposes. This study describes physical aspects of useful vibration and cavitation for a broad spectrum of applications at atmospheric or elevated pressures. After a review of available devices, hydrodynamic vibrating-body

  16. Определение флавоноидов и гидроксикоричных кислот в траве Tagetes erecta L. , Tagetes patula L. и таgetes tenuifolia Cav. Методом ВЭЖХ

    OpenAIRE

    МАРЧИШИН СВЕТЛАНА МИХАЙЛОВНА; БЕРДЕЙ ТАТЬЯНА СТЕПАНОВНА; КОЗАЧОК СОЛОМИЯ СТЕПАНОВНА; ДЕМЫДЯК ОЛЬГА ЛЮТОСЛАВОВНА

    2013-01-01

    Методом ВЭЖХ с УФ-детектором определены фенольные соединения в траве трех видов бархатцев (Tagetes erecta L., Tagetes patula L., Tаgetes tenuifolia Cav.). Установлено, что исследованные объекты в наибольшем количестве содержат гликозиды кверцетина и хлорогеновую кислоту.

  17. Comparison of arbutin contents from Bergenia purpurascens in Yunnan%滇产岩白菜中熊果苷含量的比较

    Institute of Scientific and Technical Information of China (English)

    姜洪君; 郭凤根; 张丽梅; 陈严平; 李孙文; 杨芳

    2010-01-01

    目的:分析云南境内不同岩白菜居群的熊果苷含量,筛选出优良种源和最佳利用部位.方法:应用HPLC,色谱柱SB-C18,流动相甲醇-水(15∶85),流速1 mL·min-1,检测波长282 nm,柱温30 ℃.结果:同一居群的不同部位间熊果苷含量差异悬殊,叶片>叶柄>根>根状茎;不同居群的叶片中的熊果苷含量存在极显著差异,介于6.36%~1.51%;不同居群的根状茎中的熊果苷含量也存在极显著的差异,在1.72%~0.40%.结论:叶片是提取熊果苷的最佳部位;兰坪老君山居群是开发利用岩白菜中熊果苷资源的最优种源.

  18. 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.

  19. Postharvest behavior of tamarillo (solanum betaceum cav.) treated with cacl2 under different storage temperatures

    OpenAIRE

    2014-01-01

    The tamarillo, or tree tomato, produced in Colombia, has great potential for commercialization in the global market for tropical exotic fruits, but suffers quality losses during the postharvest phase due to the use of inappropriate technologies. In order to extend the postharvest life of these fruits, the effect of calcium chloride (CaCl2) and different storage temperatures was evaluated. A completely randomized design was used, where the block criterion was the temperature with three treatme...

  20. Antibacterial and Anti-Inflammatory activities of Bunchosia armeniaca (Cav. DC. (Malpighiaceae

    Directory of Open Access Journals (Sweden)

    Gustavo S. Queiroz

    2015-04-01

    Full Text Available Bunchosia armeniaca (B. armeniaca is a native plant to America, and popularly called “cafezinho”, “ciruela” or “falso-guaraná”. In traditional medicine is used to treat different pathologies including infectious and inflammatory diseases. From ethanolic extract of B. armeniaca leaves (Malpighiaceae afforded a flavonoid mixture consisting of rutin 1 (83.5%, isoquercitrin 2 (5.6% and afzelin 5 (10.9%, which were identified and individually quantified as helpful for capillary electrophoresis and 1H and 13C NMR spectroscopy. The ethanolic extract showed an excellent antibacterial activity against Staphylococcus aureus (S.aureus and moderate activity against Escherichia coli (E. coli and Pseudomonas aeruginosa (P. aeruginosa. The flavonoid mixture showed antibacterial activity, mainly against the gram negative bacteria. Moreover, this plant demonstrated significant anti-inflammatory action, inhibiting the leukocyte influx and exudate formation in pleural cavity caused by carrageenan. The inflammation mediators involved in this model study, myeloperoxidase, nitric oxide and tumor necrosis factor alpha were significantly inhibited by ethanolic extract and flavonoid mixture of B. armeniaca. The results show that B. armeniaca has a significant antibacterial and anti-inflammatory effects and that these effects is due, at least in part, to the presence of rutin, isoquercetrin and afzelin in large amounts. Hence, these compounds have potential as novel lead compounds for the future development of therapeutic interventions for the treatment of patients with infectious and inflammatory disorders.

  1. Antibacterial and Anti-Inflammatory activities of Bunchosia armeniaca (Cav.) DC. (Malpighiaceae)

    OpenAIRE

    2015-01-01

    Bunchosia armeniaca (B. armeniaca) is a native plant to America, and popularly called “cafezinho”, “ciruela” or “falso-guaraná”. In traditional medicine is used to treat different pathologies including infectious and inflammatory diseases. From ethanolic extract of B. armeniaca leaves (Malpighiaceae) afforded a flavonoid mixture consisting of rutin 1 (83.5%), isoquercitrin 2 (5.6%) and afzelin 5 (10.9%), which were identified and individually quantified as helpful for capillary electrophoresi...

  2. Complete Genome Sequence of KPC-Producing Klebsiella pneumoniae Strain CAV1193.

    Science.gov (United States)

    Sheppard, Anna E; Stoesser, Nicole; Sebra, Robert; Kasarskis, Andrew; Deikus, Gintaras; Anson, Luke; Walker, A Sarah; Peto, Tim E; Crook, Derrick W; Mathers, Amy J

    2016-01-28

    Carbapenem resistance in Klebsiella pneumoniae, frequently conferred by the blaKPC gene, is a major public health threat. We sequenced a blaKPC-containing strain of K. pneumoniae belonging to the emergent lineage ST941, in order to better understand the evolution of blaKPC within this species.

  3. Roles of CRAC and Cav-like channels in T cells: more than one gatekeeper?

    Science.gov (United States)

    Kotturi, Maya F; Hunt, Simon V; Jefferies, Wilfred A

    2006-07-01

    Ca2+ channels in the plasma membrane of T cells vitally influence Ca2+-dependent signals that lead ultimately to cytokine secretion, cellular proliferation and apoptosis. Conventional models depict the Ca2+ inrush across the T-cell membrane following T-cell receptor engagement as being due to Ca2+-release-activated Ca2+ (CRAC) channels. A poorly understood mechanism detects the lowered Ca2+ concentrations within intracellular stores that open CRAC channels. Mammalian homologs of the Drosophila transient receptor potential Ca2+ channels possibly help to gate the store-operated, Ca2+-borne CRAC current. In this article, we review evidence of a supplementary involvement of other Ca2+ channels, the opening of which does not necessarily reflect intracellular Ca2+-store depletion. We highlight a role for variants of L-type voltage-dependent Ca2+ channels in increasing intracellular Ca2+ concentrations during activation. For more-accurate modeling of lymphocyte activation and possible pharmacological interventions, future research should aim to identify physiologically relevant situations in which such channels help to shape the Ca2+ signal.

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

    Science.gov (United States)

    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 between 0.16 and 0.31 mg/mL while those of Gram-negative bacteria ranged between 0.31 and 0.63 mg/mL. The Gram-positive bacteria were more susceptible to the essential oil than the Gram-negative bacteria. Most of the major components of this oil in other plants have been reported for antimicrobial activities. The antibacterial activity can be attributed to effects of the combination of several components of the oil. The results indicate that the C. bipinnatus might be exploited as natural antibacterial agent and have application in the treatment of several infectious diseases caused by these bacteria. Since this species is endemic to the eastern Free State, the plant could be collected during its bloom and used efficiently in the management of bacterial infections in South Africa.

  5. 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.

  6. LANDSCAPE-SCALE MONITORING OF AN OPPORTUNIST: PHRAGMITES AUSTRALIS (CAV) STEUDEL IN GREAT LAKES COASTAL WETLANDS

    Science.gov (United States)

    Coastal wetlands of the Laurentian Great Lakes (LGL) are among the most fragmented ecosystems in the world, with a long history of human-induced disturbances, primarily as a result of agricultural conversions and hydrologic changes. A substantial number of remnant LGL coastal wet...

  7. 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.

  8. 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

  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

    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.

  11. KIF13B establishes a CAV1-enriched microdomain at the ciliary transition zone to promote Sonic hedgehog signalling

    DEFF Research Database (Denmark)

    Schou, Kenneth Bødtker; Mogensen, Johanne Bay; Morthorst, Stine Kjær

    2017-01-01

    Ciliary membrane composition is controlled by transition zone (TZ) proteins such as RPGRIP1, RPGRIPL and NPHP4, which are vital for balanced coordination of diverse signalling systems like the Sonic hedgehog (Shh) pathway. Activation of this pathway involves Shh-induced ciliary accumulation...

  12. 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.

  13. 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.

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

    OpenAIRE

    Estela Sandoval; Robert A. Bye; 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. Transcriptional and electrophysiological consequences of KChIP2-mediated regulation of CaV1.2

    DEFF Research Database (Denmark)

    Thomsen, Morten B; Foster, Erika; Nguyen, Katherine H;

    2009-01-01

    Potassium channel interacting proteins (KChIP) are Ca(2+)-binding proteins that originally were identified as auxiliary subunits for K(V)4 channels. K(V)4 channels encode the voltage gated A-current (I(A)) in neuronal tissue and the fast, transient outward current (I(to,f)) in cardiac tissue...

  16. Phenolic composition and antioxidant capacity of yellow and purple-red Ecuadorian cultivars of tree tomato (Solanum betaceum Cav.).

    Science.gov (United States)

    Espin, Susana; Gonzalez-Manzano, Susana; Taco, Verónica; Poveda, Cristina; Ayuda-Durán, Begoña; Gonzalez-Paramas, Ana M; Santos-Buelga, Celestino

    2016-03-01

    Tree tomato fruits from the yellow giant, giant purple and New Zealand purple cultivars, cultivated in Ecuador were analysed for their phenolic composition and antioxidant capacity. Twelve hydroxycinnamoyl derivatives and four anthocyanins (in the purple cultivars) were detected and identified. The hydroxycinnamoyl derivatives mostly derived from caffeic acid, being 3-O-caffeoylquinic acid and rosmarinic acid the majority compounds. Furthermore, various rosmarinic acid glucosides, caffeoyl glucoside, feruloyl glucoside and two ferulic acid dehydrodimers were tentatively identified. The presence of rosmarinic acid is particularly relevant as it constituted a majority phenolic compound in the four studied tree tomato cultivars and it had not been reported previously in this fruit. In the purple cultivars main anthocyanins were pelargonidin 3-O-rutinoside and delphinidin 3-O-rutinoside. The New Zealand purple cultivar was by far the richest sample in both hydroxycinnamates (421.6mg/100g dry pulp) and anthocyanins (168.9mg/100g dry pulp). Antioxidant capacity, as determined by FRAP, ABTS and ORAC assays, followed the same pattern as phenolic contents, with the New Zealand purple cultivar being the one with the highest and the yellow giant cultivar with the lowest values.

  17. Use of membrane technology and resins for the separation and purification of polyphenols purple tree tomato (Solanum betaceum Cav

    Directory of Open Access Journals (Sweden)

    Elena Beltrán

    2016-12-01

    Full Text Available The polyphenols fractionation of mesocarp juices, placenta and purple tree tomato peel using the pore size of the membrane as a means of selection was performed using micro and ultrafiltration tangential; tests made with HPLC showed that fractionation of phenolic compounds was not performed. The processes of concentration of the clarified juices of placenta and tree tomato peel by reverse osmosis obtained a volumetric concentration factor of 2 and 2.2 respectively. The polyphenol concentration increased by 1.5 times for placenta and 2.4 times for peel and antioxidant capacity increased by 2.4 times for both juices. The anthocyanins of placenta increased by 2.6 times. The use of the resin XA 5071 FG concentrated phenolic compounds with an increase of antioxidant capacity in a range between 4 and 5.5, and anthocyanins concentration was increased 1.8 times in the placenta juice. In conclusion the process of polyphenols concentration of tree tomato was more efficient using the resin XA 5071 FG than the reverse osmosis applied in this research.

  18. Luz industrial e imagen tecnificada: de Moholy Nagy al C.A.V.S. (Center for Advanced Visual Studies)

    OpenAIRE

    Quiroga Fernández, Sofía

    2016-01-01

    El desarrollo de la tecnología de la luz implicará la transformación de la vida social, cultural y económica. Tanto las consideraciones espaciales del Movimiento Moderno, como los efectos producidos por la segunda Guerra Mundial, tendrán efectos visibles en las nuevas configuraciones espaciales y en la relación simbiótica y recíproca que se dará entre ideología y tecnología. La transformación en la comprensión de la articulación espacial, asociada al desarrollo tecnológico, afectará al modo ...

  19. 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

  20. 岩白菜和厚叶岩白菜不同部位有效成分的比较研究%Comparative studies on content of arbutin,bergenin and catechin in different part of Bergenia purpurascens and B.crassifolia

    Institute of Scientific and Technical Information of China (English)

    孙欣光; 黄文华; 马淼; 郭宝林; 王桂影

    2010-01-01

    目的:建立岩白菜药材中有效成分熊果苷、岩白菜素、儿茶素HPLC含量测定方法,为其药材质量评价和资源合理利用提供科学依据.方法:采用反相C18色谱柱,流动相为甲醇-水,流速为1.0 mL·min-1,检测波长为275 nm,对岩白菜属植物岩白菜和厚叶岩白菜的主根、须根和叶片等不同部位中的熊果苷、岩白菜素JL茶素含量进行HPLC分析比较.结果:岩白菜和厚叶岩白菜中的熊果苷、岩白菜素、儿茶素含量存在一定差异;同植物不同部位中的熊果苷、岩白菜素、儿茶素含量存在明显差异.主根和须根中岩白菜素的含量较高,而叶中熊果苷的含量较高.结论:本研究方法可以同时测定熊果苷、岩白菜索和儿茶素的含量,为岩白菜属植物的科学研究及药材质量评价奠定基础.

  1. Diversity for chemical composition in a collection of different varietal types of tree tomato (Solanum betaceum Cav.), an Andean exotic fruit.

    Science.gov (United States)

    Acosta-Quezada, Pablo G; Raigón, María D; Riofrío-Cuenca, Tania; García-Martínez, María D; Plazas, Mariola; Burneo, Juan I; Figueroa, Jorge G; Vilanova, Santiago; Prohens, Jaime

    2015-02-15

    We evaluated 23 tree tomato (Solanum betaceum) accessions from five cultivar groups and one wild relative (Solanum cajanumense) for 26 composition traits. For all traits we found highly significant differences (Pcomposition traits was matched by a high diversity within each of the cultivar groups. We found that sucrose and citric acid were the most important soluble sugar and organic acid, respectively, in tree tomato. Fruit in the anthocyanin pigmented (purple) group had a carotenoid content similar to that in the yellow-orange cultivar groups. Total phenolic content was significantly correlated (r=0.8607) with antioxidant activity. Analyses of mineral content showed that tree tomato is a good source of K, Mg, and Cu. Multivariate principal components analysis (PCA) confirmed that an important diversity exists within each cultivar group. The results we have obtained indicate that the high diversity found within the tree tomato could be exploited for selection and breeding for developing the tree tomato as a commercial crop.

  2. Aberrant Splicing Promotes Proteasomal Degradation of L-type Ca v 1.2 Calcium Channels by Competitive Binding for CaV β Subunits in Cardiac Hypertrophy

    NARCIS (Netherlands)

    Hu, Zhenyu; Wang, Jiong Wei; Yu, Dejie; Soon, Jia Lin; De Kleijn, Dominique P V; Foo, Roger; Liao, Ping; Colecraft, Henry M.; Soong, Tuck Wah

    2016-01-01

    Decreased expression and activity of Ca V1.2 calcium channels has been reported in pressure overload-induced cardiac hypertrophy and heart failure. However, the underlying mechanisms remain unknown. Here we identified in rodents a splice variant of Ca V1.2 channel, named Ca V1.2 e21+22, that contain

  3. Association of whirlin with Cav1.3 (alpha1D) channels in photoreceptors, defining a novel member of the usher protein network.

    NARCIS (Netherlands)

    Kersten, F.F.J.; Wijk, E. van; Reeuwijk, J. van; Zwaag, B. van der; Marker, T.; Peters, T.A.; Katsanis, N.; Wolfrum, U.; Keunen, J.E.E.; Roepman, R.; Kremer, J.M.J.

    2010-01-01

    PURPOSE: Usher syndrome is the most common form of hereditary deaf-blindness. It is both clinically and genetically heterogeneous. The USH2D protein whirlin interacts via its PDZ domains with other Usher-associated proteins containing a C-terminal type I PDZ-binding motif. These proteins co-localize

  4. STABILITY OF ANTHOCYANINS FROM Rubus glaucus Benth AND Solanum betaceum Cav.dark-red strain AS AFFECTED BY TEMPERATURE, STORAGE TIME AND WATER ACTIVITY

    Directory of Open Access Journals (Sweden)

    C.M. OLAYA

    2009-01-01

    Full Text Available 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 disminuyó, mientras que el contenido fenólico aumentó con el tiempo. La actividad antioxidante de las muestras de mora de Castilla presentó una alta correlación con los contenidos 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 microencapsuladas como colorantes alimenticios.

  5. 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

  6. Characterization of the microbial community in the rhizosphere of Phragmites australis (cav.) trin ex. steudel growing in the Sun Island Wetland.

    Science.gov (United States)

    Ma, Fang; Wu, Jieting; Wang, Li; Yang, Jixian; Li, Shiyang; Li, Zhe; Zhag, Xue

    2014-03-01

    Rhizospheric microorganisms are important for environmental conservancy. The constancy and variability of the microorganisms in the rhizosphere of Phragmites australis in relation to the spatiotemporal variations in wetland ecosystems were studied. During the peak and trough of the vegetative period of the Phragmites australis growing across the hydrologic gradients of the Sun Island Wetland, Biolog and denaturing gradient gel electrophoresis (DGGE) were used to investigate the rhizospheric microbial characteristics. Both methods demonstrated that the microbial activity, richness, and diversity decreased from summer to autumn. However, these properties did not show significant correlation with hydrologic gradient, except that the genetic richness and diversity of the fungi decreased with it. Cluster analysis also demonstrated that the rhizospheric microbial community seemed to be largely affected by a vegetative period. In addition, this research was extended to a broader range of determining the universal microorganisms, which showed notable adaptability.

  7. Response of Arbuscular Mycorrhizal Fungi to Hydrologic Gradients in the Rhizosphere of Phragmites australis (Cav.) Trin ex. Steudel Growing in the Sun Island Wetland.

    Science.gov (United States)

    Wang, Li; Wu, Jieting; Ma, Fang; Yang, Jixian; Li, Shiyang; Li, Zhe; Zhang, Xue

    2015-01-01

    Within the rhizosphere, AM fungi are a sensitive variable to changes of botanic and environmental conditions, and they may interact with the biomass of plant and other microbes. During the vegetative period of the Phragmites australis growing in the Sun Island Wetland (SIW), the variations of AM fungi colonization were studied. Root samples of three hydrologic gradients generally showed AM fungi colonization, suggesting that AM fungi have the ability for adaptation to flooded habitats. There were direct and indirect hydrological related effects with respect to AM fungi biomass, which interacted simultaneously in the rhizosphere. Though water content in soil and reed growth parameters were both positively associated with AM fungi colonization, only the positive correlations between reed biomass parameters and the colonization could be expected, or both the host plant biomass and the AM fungi could be beneficial. The variations in response of host plant to the edaphic and hydrologic conditions may influence the effectiveness of the plant-mycorrhizal association. This study included a hydrologic component to better assess the role and distribution of AM fungi in wetland ecosystems. And because of that, the range of AM fungi was extended, since they actually showed a notable adaptability to hydrologic gradients.

  8. 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.

  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. 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.

  11. SELECCIÓN Y REGENERACIÓN in vitro DE SOMACLONES DE TOMATE DE ÁRBOL (Solanum betacea cav. Sendt UTILIZANDO FILTRADOS DE CULTIVO DE Colletotrichum acutatum CON ACTIVIDAD PECTINASA SELECTION AND in vitro REGENERATION OF SOMACLONES OF TREE TOMATO (Solanum betacea cav. Sendt USING CULTURE FILTRATES OF Colletotrichum acutatum WITH PECTINASA ACTIVITY

    Directory of Open Access Journals (Sweden)

    Carlos Patiño Torres

    2007-12-01

    Full Text Available Se utilizaron filtrados de cultivo de Colletotrichum acutatum, agente causal de la antracnosis del tomate de árbol, para la selección in vitro de variantes somaclonales de ésta planta con resistencia potencial a la enfermedad. El filtrado, utilizado a varias concentraciones, demostró ser un agente de selección efectivo cuando se integró al medio de cultivo, pues causó niveles elevados de mortalidad de los explantes comparado con los tratamientos sin filtrado. Igualmente, se estudió el efecto de la benziladenina (BA ó BAP sobre la respuesta organogénica de explantes de tejido foliar de tomate de árbol.Culture filtrates of Colletotrichum acutatum, causal agent of the anthracnose disease in tree tomato, were used for the in vitro selection of somaclonal variants with potential resistance to this disease. Culture filtrates used at different concentrations, showed to be an effective agent of selection when they were mixed with the growth media. The findings indicated a high level of mortality for the explants growing on the ammended media as compared with these growing on non ammended one. The effect of the benziladenina (BA ó BAP on the organogenic response of leaf explants of tomato tree was also studied.

  12. 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.

  13. 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.

  14. Effects of chicken anemia virus and infectious bursal disease virus in commercial chickens.

    Science.gov (United States)

    Toro, H; van Santen, V L; Hoerr, F J; Breedlove, C

    2009-03-01

    The effects of chicken anemia virus (CAV) and infectious bursal disease virus (IBDV) coinfection in commercial layer-type and meat-type (broiler) chickens with specific maternal immunity were evaluated. In addition, the broiler progeny used had been vaccinated in ovo against IBDV. Layer chickens were inoculated intramuscularly on day 3 of age with CAV and orally on day 7 of age with an IBDV standard strain (APHIS). Broiler chickens were exposed to CAV and/or an IBDV variant strain (AL2) via the drinking water on days 3 and 14 of age. Following CAV and IBDV inoculation neither mortality nor overt clinical disease was observed in any layer or broiler group. In spite of maternal immunity against both IBDV and CAV, mean hematocrits of all layer groups inoculated with CAV (CAV, CAV + APHIS) were lower than uninfected chickens. IBDV APHIS alone or in combination with CAV did not affect the layer weight gain. However, on day 30 of age and concomitantly with maternal antibody decay, bursa lymphocyte depletion became evident in CAV + APHIS-infected layer chickens. These birds (CAV + APHIS) also seroconverted to IBDV on day 35 of age. CAV persisted at low levels in the layer chickens throughout the experimental period in CAV- and CAV+APHIS-infected chickens. Similarly, infected broiler chickens did not show changes in weight gain. Compared to CAV-infected or uninfected controls, CAV+AL2- and AL2-infected broiler chickens showed significant lymphocyte depletion in the bursa as assessed both by bursal indices and histomorphometry. Broilers also seroconverted to IBDV after day 30 of age confirming that bursal lymphocyte depletion was due to IBDV resuming replication. Thymus histomorphometry revealed significant lymphocyte depletion in all infected broiler groups at 30 days of age, but only in CAV+AL2-infected broiler chickens at 41 days of age, suggesting that IBDV infection delayed repopulation of the thymus.

  15. Impact of the loss of caveolin-1 on lung mass and cholesterol metabolism in mice with and without the lysosomal cholesterol transporter, Niemann-Pick type C1.

    Science.gov (United States)

    Mundy, Dorothy I; Lopez, Adam M; Posey, Kenneth S; Chuang, Jen-Chieh; Ramirez, Charina M; Scherer, Philipp E; Turley, Stephen D

    2014-07-01

    Caveolin-1 (Cav-1) is a major structural protein in caveolae in the plasma membranes of many cell types, particularly endothelial cells and adipocytes. Loss of Cav-1 function has been implicated in multiple diseases affecting the cardiopulmonary and central nervous systems, as well as in specific aspects of sterol and lipid metabolism in the liver and intestine. Lungs contain an exceptionally high level of Cav-1. Parameters of cholesterol metabolism in the lung were measured, initially in Cav-1-deficient mice (Cav-1(-/-)), and subsequently in Cav-1(-/-) mice that also lacked the lysosomal cholesterol transporter Niemann-Pick C1 (Npc1) (Cav-1(-/-):Npc1(-/-)). In 50-day-old Cav-1(-/-) mice fed a low- or high-cholesterol chow diet, the total cholesterol concentration (mg/g) in the lungs was marginally lower than in the Cav-1(+/+) controls, but due to an expansion in their lung mass exceeding 30%, whole-lung cholesterol content (mg/organ) was moderately elevated. Lung mass (g) in the Cav-1(-/-):Npc1(-/-) mice (0.356±0.022) markedly exceeded that in their Cav-1(+/+):Npc1(+/+) controls (0.137±0.009), as well as in their Cav-1(-/-):Npc1(+/+) (0.191±0.013) and Cav-1(+/+):Npc1(-/-) (0.213±0.022) littermates. The corresponding lung total cholesterol contents (mg/organ) in mice of these genotypes were 6.74±0.17, 0.71±0.05, 0.96±0.05 and 3.12±0.43, respectively, with the extra cholesterol in the Cav-1(-/-):Npc1(-/-) and Cav-1(+/+):Npc1(-/-) mice being nearly all unesterified (UC). The exacerbation of the Npc1 lung phenotype and increase in the UC level in the Cav-1(-/-):Npc1(-/-) mice imply a regulatory role of Cav-1 in pulmonary cholesterol metabolism when lysosomal sterol transport is disrupted.

  16. Pathological and immunohistochemical studies of subclinical infection of chicken anemia virus in 4-week-old chickens.

    Science.gov (United States)

    Haridy, Mohie; Sasaki, Jun; Ikezawa, Mitsutaka; Okada, Kosuke; Goryo, Masanobu

    2012-06-01

    Subclinical infection of chicken anemia virus (CAV) at 4 to 6 weeks of age, after maternal antibodies have waned, is implicated in several field problems in broiler flocks. In order to understand the pathogenesis of subclinical infection with CAV, an immunopathological study of CAV-inoculated 4-week-old SPF chickens was performed. Sixty 4-week-old SPF chickens were equally divided into CAV and control groups. The CAV group was inoculated intramuscularly with the MSB1-TK5803 strain of CAV. Neither mortality nor anemia was detected in the CAV and control groups. In the CAV group, no signs were observed, except that some chickens were grossly smaller compared with the control group. Sporadic thymus lobes appeared to be reddening and atrophied. Within the first two weeks p.i. of CAV, there was a mild to moderate depletion of lymphocytes in the thymus cortex and spleen in some chickens. Moreover, lymphoid depletion of the bursa of Fabricius, proventriculus and cecal tonsils was observed. Hyperplastic lymphoid foci were observed in the liver, lungs, kidneys and heart at the 4th week p.i. of CAV. Immunohistochemically, a moderate lymphoid depletion of CD4(+)and CD8(+) T cells in the thymus cortex and spleen was observed in some chickens within two weeks p.i. of CAV. CAV inclusions and antigens were detected infrequently in the thymus cortex and spleen. It could be concluded that the immunosuppression in subclinical infection with CAV occurs as a result of reduction of cellular immunity.

  17. Chemotaxonomic analysis of the aroma compounds in essential oils of two different Ocimum basilicum L. varieties from Iran.

    Science.gov (United States)

    Pirmoradi, Mohammad Reza; Moghaddam, Mohammad; Farhadi, Nasrin

    2013-07-01

    Hydrodistilled essential oils of 21 accessions of Ocimum basilicum L. belonging to two different varieties (var. purpurascens and var. dianatnejadii) from Iran were characterized by GC-FID and GC/MS analyses. The oil yield was found to be between 0.6 and 1.1% (v/w). In total, 49 compounds, accounting for 96.6-99.7% of the oil compositions, were identified. Aromatic compounds, represented mainly by methyl chavicol (33.6-49.1%), and oxygenated monoterpenes, represented by linalool (14.4-39.3%), were the main components in all essential oils. Monoterpene hydrocarbons were present in the essential oils of all accessions of the purpurascens variety, whereas they were completely absent in those of the dianatnejadii variety, indicating that monoterpene hydrocarbons might be considered as marker constituents of the purpurascens variety. The chemotaxonomic value of the essential-oil compositions was discussed according to the results of the cluster analysis (CA). The CA showed a clear separation of the O. basilicum var. purpurascens accessions and the O. basilicum var. dianatnejadii accessions, although the data showed no major chemotype variation between the studied varieties. Indeed, the CA revealed only one principal chemotype (methyl chavicol/linalool) for both varieties. In conclusion, GC/MS analyses in combination with CA showed to be a flexible and reliable method for the characterization of the chemical profiles of different varieties of Ocimum basilicum L.

  18. Gclust Server: 27249 [Gclust Server

    Lifescience Database Archive (English)

    Full Text Available 27249 DME_CG6219_28571862 Cluster Sequences - 343 cav: caravaggio CG6219-PE, isofor...cluster sequences Cluster Sequences Link to related sequences - Sequence length 343 Representative annotation cav: caravaggio

  19. 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.

  20. Syntaxin-3 Binds and Regulates Both R- and L-Type Calcium Channels in Insulin-Secreting INS-1 832/13 Cells.

    Directory of Open Access Journals (Sweden)

    Li Xie

    Full Text Available Syntaxin (Syn-1A mediates exocytosis of predocked insulin-containing secretory granules (SGs during first-phase glucose-stimulated insulin secretion (GSIS in part via its interaction with plasma membrane (PM-bound L-type voltage-gated calcium channels (Cav. In contrast, Syn-3 mediates exocytosis of newcomer SGs that accounts for second-phase GSIS. We now hypothesize that the newcomer SG Syn-3 preferentially binds and modulates R-type Cav opening, which was postulated to mediate second-phase GSIS. Indeed, glucose-stimulation of pancreatic islet β-cell line INS-1 induced a predominant increase in interaction between Syn-3 and Cavα1 pore-forming subunits of R-type Cav2.3 and to lesser extent L-type Cavs, while confirming the preferential interactions between Syn-1A with L-type (Cav1.2, Cav1.3 Cavs. Consistently, direct binding studies employing heterologous HEK cells confirmed that Syn-3 preferentially binds Cav2.3, whereas Syn-1A prefers L-type Cavs. We then used siRNA knockdown (KD of Syn-3 in INS-1 to study the endogenous modulatory actions of Syn-3 on Cav channels. Syn-3 KD enhanced Ca2+ currents by 46% attributed mostly to R- and L-type Cavs. Interestingly, while the transmembrane domain of Syn-1A is the putative functional domain modulating Cav activity, it is the cytoplasmic domain of Syn-3 that appears to modulate Cav activity. We conclude that Syn-3 may mimic Syn-1A in the ability to bind and modulate Cavs, but preferring Cav2.3 to perhaps participate in triggering fusion of newcomer insulin SGs during second-phase GSIS.

  1. Functional importance of T-type voltage-gated calcium channels in the cardiovascular and renal system: news from the world of knockout mice.

    Science.gov (United States)

    Hansen, Pernille B L

    2015-02-15

    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 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 Cav3.2, are expressed in blood vessels, the kidney, and the heart. Studies with gene-deficient mice have provided a way to investigate the Cav3.1 and Cav3.2 channels and their role in the cardiovascular system. This review discusses the results from these knockout mice. Evaluation of the literature leads to the conclusion that Cav3.1 and Cav3.2 channels have important, but different, functions in mice. T-type Cav3.1 channels affect heart rate, whereas Cav3.2 channels are involved in cardiac hypertrophy. In the vascular system, Cav3.2 activation leads to dilation of blood vessels, whereas Cav3.1 channels are mainly suggested to affect constriction. The Cav3.1 channel is also involved in neointima formation following vascular damage. In the kidney, Cav3.1 regulates plasma flow and Cav3.2 plays a role setting glomerular filtration rate. In conclusion, Cav3.1 and Cav3.2 are new therapeutic targets in several cardiovascular pathologies, but the use of T-type blockers should be specifically directed to the disease and to the channel subtype.

  2. 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 us...... 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....

  3. Oligomerization of Clostridium perfringens epsilon toxin is dependent upon caveolins 1 and 2.

    Directory of Open Access Journals (Sweden)

    Christine M Fennessey

    Full Text Available Evidence from multiple studies suggests that Clostridium perfringens ε-toxin is a pore-forming toxin, assembling into oligomeric complexes in the plasma membrane of sensitive cells. In a previous study, we used gene-trap mutagenesis to identify mammalian factors contributing to toxin activity, including caveolin-2 (CAV2. In this study, we demonstrate the importance of caveolin-2 and its interaction partner, caveolin-1 (CAV1, in ε-toxin-induced cytotoxicity. Using CAV2-specific shRNA in a toxin-sensitive human kidney cell line, ACHN, we confirmed that cells deficient in CAV2 exhibit increased resistance to ε-toxin. Similarly, using CAV1-specific shRNA, we demonstrate that cells deficient in CAV1 also exhibit increased resistance to the toxin. Immunoprecipitation of CAV1 and CAV2 from ε-toxin-treated ACHN cells demonstrated interaction of both CAV1 and -2 with the toxin. Furthermore, blue-native PAGE indicated that the toxin and caveolins were components of a 670 kDa protein complex. Although ε-toxin binding was only slightly perturbed in caveolin-deficient cells, oligomerization of the toxin was dramatically reduced in both CAV1- and CAV2-deficient cells. These results indicate that CAV1 and -2 potentiate ε-toxin induced cytotoxicity by promoting toxin oligomerization - an event which is requisite for pore formation and, by extension, cell death.

  4. 78 FR 75574 - Agency Information Collection Activities: Submission for OMB Review; Comment Request...

    Science.gov (United States)

    2013-12-12

    ... Program CAC and CAV Reports AGENCY: Federal Emergency Management Agency, DHS. ACTION: Notice. SUMMARY: The... Implementation of the NFIP Community Assistance Program CAC and CAV Reports. OMB Number: 1660-0023. Type of...: Through the use of a Community Assistance Contact (CAC) or Community Assistance Visit (CAV), FEMA can...

  5. 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.

  6. 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.

  7. Investigation of Genotypic Difference of Nitrate Contents in Brassicaceae Eruca vesicaria (L.) Cav. var. sativa%芝麻菜硝酸盐含量基因型差异研究

    Institute of Scientific and Technical Information of China (English)

    雷娟利; 寿伟松; 董文其; 徐志豪

    2007-01-01

    对从国内外引进的170个芝麻菜品种,采用水杨酸-浓硫酸方法进行了硝酸盐含量的测定,结果表明芝麻菜基因型间硝酸盐含量存在显著差异,可划分为硝酸盐含量高和硝酸盐含量低的品种类群.

  8. 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.

  9. 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.

  10. The Actuality and Trend in the Study of Phragmites australis(Cav.)Trin. exSteud. in China%中国芦苇研究现状与趋势

    Institute of Scientific and Technical Information of China (English)

    王振庆; 王丽娜; 吴大千; 张治国

    2006-01-01

    芦苇Phragmites australis为多年生高大草本植物,是世界广布的重要湿地物种,具有广泛的适应性及形态变异,并有极高的生态学和社会经济学价值.本文通过对近10年来中国各地对于芦苇研究的成果文献进行统计分析,揭示出我国芦苇研究的现状与趋势.

  11. "Balsa" Ochroma pyramidale (Cav. ex Lam.) Urb. (Bombacaceae) : Etnobotánica, anatomía, ensayos fitoquímicos y actividades biológicas

    OpenAIRE

    Ramos Corrales, Pablo Cesar

    2016-01-01

    O. pyramidale (Bombacaceae) conocida como "balsa", es una especie maderable que crece silvestre en la selva amazónica y constituye uno de los principales recursos económicos de Ecuador, donde se la cultiva a gran escala. La madera (leño) se utiliza para la construcción de viviendas y embarcaciones, fabricación de numerosos utensilios de uso domestico y artesanías, entre otros. En algunas regiones de Latinoamérica se le asignan también usos medicinales, especialmente a la c...

  12. Cultivation experiment on compound substrate composed in Comos bipinnatus Cav. and Tagetes patula%波丝菊、万寿菊的稻壳复合基质栽培试验

    Institute of Scientific and Technical Information of China (English)

    谢嘉霖; 徐秋华

    2010-01-01

    将稻壳、锯屑、泥炭、蛭石和珍珠岩按不同比例混合制成栽培基质,用以波丝菊、万寿菊为主要组成部分的野花组合做试材进行无土栽培试验.结果表明:波丝菊、万寿菊在处理1(泥炭 :蛭石 :珍珠岩=2 :1 :1)、处理5(稻壳 :泥炭=3 :2)和处理8(稻壳 :锯屑 :珍珠岩=2 :1 :1)中生长和开花较好.其中处理5和处理8以稻壳为栽培基质的主要原料,原料来源丰富,生产成本低,适宜在南方推广.

  13. Un rara avis de la alfareria Prehistórica en Asturias: las sugerencias inducidas del Vaso Neolítico (IV milenio a. de C. TL de la Hoz de los Cavíos (Piloña.

    Directory of Open Access Journals (Sweden)

    De Blas Cortina, Miguel Ángel

    2011-01-01

    Full Text Available La cerámica prehistórica anterior al Edad del Hierro es apenas conocida en Asturias, tal vez por su propia escasez, probablemente sustituida por recipientes corruptibles. El hallazgo solitario de un vaso de gran calidad técnica y cuidada decoración, depositado en una pequeña cueva situada en el fondo umbrío e insalubre de un desfiladero da pie a distintas consideraciones sobre su posible significado. Fechada la vasija por el método TL en el IV milenio a. de C., su ubicación en un claro paraje de conexión y ruptura entre ambientes naturales muy distintos, abona la hipótesis de la anotación simbólica de los lugares de paso y de articulación de universos económicamente complementarios propiciando, además, la revisión de la naturaleza y distribución las huellas de la vida neolítica en un territorio crucial en sector centro-este asturiano.

  14. US Army Order of Battle 1919-1941. Volume 2. The Arms: Calvary, Field Artillery, and Coast Artillery, 1919-41

    Science.gov (United States)

    2010-01-01

    101st Cav. Regt. Brooklyn, NY 102d Cav. Regt. Newark, NJ 121st Cav. Regt. Rochester, NY 51st MG Sq. New York City, NY 52d Cav. Brig. Wayne ...District of Columbia HQ—Hagerstown, MD, 1921–24; Towson , MD, 1924–40; Baltimore, MD, 1940–41 The 62d Cav. Div. was constituted on 15 October 1921, allotted...authorized units had been activated. The division HQ was relocated to York and Allegheny Avenues in Towson , MD, on 30 June 1924. It was moved one final

  15. Immunocytochemical localization of caveolin-3 in the synoviocytes of the rat temporomandibular joint during development.

    Science.gov (United States)

    Niwano, Masahiro; Nozawa-Inoue, Kayoko; Suzuki, Akiko; Ikeda, Nobuyuki; Takagi, Ritsuo; Maeda, Takeyasu

    2008-03-01

    Caveolins -- caveolin-1, -2, -3 (Cav1, 2, 3) -- are major components of caveolae, which have diverse functions. Our recent study on the temporomandibular joint (TMJ) revealed expressions of Cav1 and muscle-specific Cav3 in some synovial fibroblast-like type B cells with well-developed caveolae. However, the involvement of Cav3 expression in the differentiation and maturation of type B cells remains unclear. The present study therefore examined the chronological alterations in the localization of Cav3 in the synovial lining cells of the rat TMJ during postnatal development by immunocytochemical techniques. Observations showed immature type B cells possessed a few caveolae with Cav1 but lacked Cav3 protein at postnatal day 5 (P5). At P14, Cav3-immunopositive type B cells first appeared in the synovial lining layer. They increased in number and immunointensity from P14 to P21 as occlusion became active. In immunoelectron microscopy and double immunolabeling with heat shock protein 25 (Hsp25) and Cav3, coexpressed type B cells developed rough endoplasmic reticulum and numerous caveolae, while the Cav3-immunonegative type B cell with Hsp25 immunoreaction possessed few of these. Results suggest that Cav3 expression, which is closely related to added functional stimuli, reflects the differentiation of the type B synoviocytes.

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

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    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.

  17. Sequence and phylogenetic analysis of chicken anaemia virus obtained from backyard and commercial chickens in Nigeria.

    Science.gov (United States)

    Oluwayelu, D O; Todd, D; Olaleye, O D

    2008-12-01

    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/CI-8 and NGR/CI-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.

  18. Caveolin-1 is necessary for hepatic oxidative lipid metabolism: evidence for crosstalk between caveolin-1 and bile acid signaling.

    Science.gov (United States)

    Fernández-Rojo, Manuel A; Gongora, Milena; Fitzsimmons, Rebecca L; Martel, Nick; Martin, Sheree D; Nixon, Susan J; Brooks, Andrew J; Ikonomopoulou, Maria P; Martin, Sally; Lo, Harriet P; Myers, Stephen A; Restall, Christina; Ferguson, Charles; Pilch, Paul F; McGee, Sean L; Anderson, Robin L; Waters, Michael J; Hancock, John F; Grimmond, Sean M; Muscat, George E O; Parton, Robert G

    2013-07-25

    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.

  19. 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.

  20. 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.

  1. 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.

  2. The different functions and clinical significances of caveolin-1 in human adenocarcinoma and squamous cell carcinoma

    Science.gov (United States)

    Fu, Pin; Chen, Fuchun; Pan, Qi; Zhao, Xianda; Zhao, Chen; Cho, William Chi-Shing; Chen, Honglei

    2017-01-01

    Caveolin-1 (Cav-1), a major structural protein of caveolae, is an integral membrane protein which plays an important role in the progression of carcinoma. However, whether Cav-1 acts as a tumor promoter or a tumor suppressor still remains controversial. For example, the tumor-promoting function of Cav-1 has been found in renal cancer, prostate cancer, tongue squamous cell carcinoma (SCC), lung SCC and bladder SCC. In contrast, Cav-1 also plays an inhibitory role in esophagus adenocarcinoma, lung adenocarcinoma and cutaneous SCC. The role of Cav-1 is still controversial in thyroid cancer, hepatocellular carcinoma, gastric adenocarcinoma, colon adenocarcinoma, breast cancer, pancreas cancer, oral SCC, laryngeal SCC, head and neck SCC, esophageal SCC and cervical SCC. Besides, it has been reported that the loss of stromal Cav-1 might predict poor prognosis in breast cancer, gastric cancer, pancreas cancer, prostate cancer, oral SCC and esophageal SCC. However, the accumulation of stromal Cav-1 has been found to be promoted by the progression of tongue SCC. Taken together, Cav-1 seems playing a different role in different cancer subtypes even of the same organ, as well as acting differently in the same cancer subtype of different organs. Thus, we hereby explore the functions of Cav-1 in human adenocarcinoma and SCC from the perspective of clinical significances and pathogenesis. We envision that novel targets may come with the further investigation of Cav-1 in carcinogenesis. PMID:28243118

  3. Sirtuin1 protects endothelial Caveolin-1 expression and preserves endothelial function via suppressing miR-204 and endoplasmic reticulum stress

    Science.gov (United States)

    Kassan, M.; Vikram, A.; Kim, Y. R.; Li, Q.; Kassan, A.; Patel, H. H.; Kumar, S.; Gabani, M.; Liu, J.; Jacobs, J. S.; Irani, K.

    2017-01-01

    Sirtuin1 (Sirt1) is a class III histone deacetylase that regulates a variety of physiological processes, including endothelial function. Caveolin1 (Cav1) is also an important determinant of endothelial function. We asked if Sirt1 governs endothelial Cav1 and endothelial function by regulating miR-204 expression and endoplasmic reticulum (ER) stress. Knockdown of Sirt1 in endothelial cells, and in vivo deletion of endothelial Sirt1, induced endothelial ER stress and miR-204 expression, reduced Cav1, and impaired endothelium-dependent vasorelaxation. All of these effects were reversed by a miR-204 inhibitor (miR-204 I) or with overexpression of Cav1. A miR-204 mimic (miR-204 M) decreased Cav1 in endothelial cells. In addition, high-fat diet (HFD) feeding induced vascular miR-204 and reduced endothelial Cav1. MiR-204-I protected against HFD-induced downregulation of endothelial Cav1. Moreover, pharmacologic induction of ER stress with tunicamycin downregulated endothelial Cav1 and impaired endothelium-dependent vasorelaxation that was rescued by overexpressing Cav1. In conclusion, Sirt1 preserves Cav1-dependent endothelial function by mitigating miR-204-mediated vascular ER stress. PMID:28181559

  4. Caveolin-1 expression level in cancer associated fibroblasts predicts outcome in gastric cancer.

    Directory of Open Access Journals (Sweden)

    Xianda Zhao

    Full Text Available 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.

  5. Molecular Characterization of Chicken Anemia Virus Circulating in Chicken Flocks in Egypt

    Directory of Open Access Journals (Sweden)

    Mohammed AboElkhair

    2014-01-01

    Full Text Available Introduction. Although many previous studies reported detection of chicken anemia virus (CAV in Egypt since 1990, genomic characterization of this circulating CAV has not been published. In the present study, four nucleotide sequences of detected CAV were genetically characterized. Methods. These nucleotide sequences were obtained from commercial chicken flocks in two different locations of Egypt during 2010. The target region for sequencing was 675 bp nucleotide of partial coding region of VP1 protein. The nucleotide and deduced amino acid sequences of the detected CAV were aligned and compared to worldwide CAV isolates including commonly used vaccine strains. Phylogenetic analysis of these sequences was also carried out. Results. Our results showed that all the Egyptian CAV sequences were grouped in one group with viruses from diverse geographic regions. This group is characterized by amino acids profile 75I, 97L, 139Q, and 144Q in VP1. The phylogenetic and amino acid analyses of deduced amino acid indicated that the detected CAV sequences differ from CAV vaccine strains. Conclusion. This is the first report that describes molecular characterization of circulating CAV in Egypt. The study showed that the detected CAV, in Egypt are field viruses and unrelated to vaccine strains.

  6. Identification, expression pattern, cellular location and potential role of the caveolin-1 gene from Artemia sinica.

    Science.gov (United States)

    Li, Xuejie; Yao, Feng; Zhang, Wei; Cheng, Cheng; Chu, Bing; Liu, Yan; Mei, Yanli; Wu, Yang; Zou, Xiangyang; Hou, Lin

    2014-05-01

    Caveolins are integral membrane proteins that serve as scaffolds to recruit numerous signaling molecules. Caveolins play an important role in membrane trafficking, signal transduction, substrate transport and endocytosis in differentiated cells. In this study, a caveolin-1 gene from Artemia sinica (As-cav-1) was successfully cloned for the first time. The full-length cDNA of As-cav-1 comprises 974 bp, with a 675 bp open reading frame (ORF) that encodes a polypeptide of 224 amino acids with a caveolin scaffolding domain (CSD) and two transmembrane domains. Multiple sequence alignment revealed that the putative As-CAV-1 protein sequence was relatively conserved across species, especially in the CSD domain. Real-time PCR revealed high levels of the As-cav-1 transcript at 0h of embryo development. Furthermore, As-cav-1 transcripts were highly upregulated under high salinity (200‰) and low temperature stresses (15°C). To further characterize As-cav-1, recombinant pET30a-cav-1 protein was expressed using a prokaryotic expression system. The recombinant protein comprised 290 amino acids with a theoretical molecular weight of 32kDa, and a predicted isoelectric point of 5.6. Western blotting of the expression levels of As-CAV-1 during different embryo development stages revealed that As-CAV-1 levels decreased gradually during development stages from 0 h to 40 h, and increased at 3d. Furthermore, western blotting showed that As-CAV-1 was upregulated to its highest expression level by low temperature stress (15°C) and high salinity. Confocal laser microscopy analysis, using antibodies generated against the recombinant As-CAV-1 protein, showed that As-CAV-1 was mostly located in the cell membrane. Our results suggested that As-cav-1 plays a vital role in protecting embryos from high salt damage and low temperature stress, especially during post-diapause embryonic development.

  7. Membrane-localized β-subunits alter the PIP2 regulation of high-voltage activated Ca2+ channels.

    Science.gov (United States)

    Suh, Byung-Chang; Kim, Dong-Il; Falkenburger, Björn H; Hille, Bertil

    2012-02-21

    The β-subunits of voltage-gated Ca(2+) (Ca(V)) channels regulate the functional expression and several biophysical properties of high-voltage-activated Ca(V) channels. We find that Ca(V) β-subunits also determine channel regulation by the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP(2)). When Ca(V)1.3, -2.1, or -2.2 channels are cotransfected with the β3-subunit, a cytosolic protein, they can be inhibited by activating a voltage-sensitive lipid phosphatase to deplete PIP(2). When these channels are coexpressed with a β2a-subunit, a palmitoylated peripheral membrane protein, the inhibition is much smaller. PIP(2) sensitivity could be increased by disabling the two palmitoylation sites in the β2a-subunit. To further test effects of membrane targeting of Ca(V) β-subunits on PIP(2) regulation, the N terminus of Lyn was ligated onto the cytosolic β3-subunit to confer lipidation. This chimera, like the Ca(V) β2a-subunit, displayed plasma membrane localization, slowed the inactivation of Ca(V)2.2 channels, and increased the current density. In addition, the Lyn-β3 subunit significantly decreased Ca(V) channel inhibition by PIP(2) depletion. Evidently lipidation and membrane anchoring of Ca(V) β-subunits compete with the PIP(2) regulation of high-voltage-activated Ca(V) channels. Compared with expression with Ca(V) β3-subunits alone, inhibition of Ca(V)2.2 channels by PIP(2) depletion could be significantly attenuated when β2a was coexpressed with β3. Our data suggest that the Ca(V) currents in neurons would be regulated by membrane PIP(2) to a degree that depends on their endogenous β-subunit combinations.

  8. Caveolin-1 plays a key role in the oleanolic acid-induced apoptosis of HL-60 cells.

    Science.gov (United States)

    Ma, Wei; Wang, Di-Di; Li, Li; Feng, Yu-Kuan; Gu, Hong-Mei; Zhu, Gui-Ming; Piao, Jin-Hua; Yang, Yu; Gao, Xu; Zhang, Peng-Xia

    2014-07-01

    Our previous study found that caveolin-1 (CAV-1) protein expression is upregulated during oleanolic acid (OA)-induced inhibition of proliferation and promotion of apoptosis in HL-60 cells. CAV-1 is the main structural protein component of caveolae, playing important roles in tumorigenesis and tumor development. It has been shown that cav-1 expression is lower in leukemia cancer cell lines SUP-B15, HL-60, THP-1 and K562 and in chronic lymphocytic leukemia primary (CLP) cells when compared with normal white blood cells, with the lowest cav-1 expression level found in HL-60 cells. To study the effects of cav-1 in HL-60 cells and the effects of cav-1 overexpression on OA drug efficacy, cav-1 was overexpressed in HL-60 cells using lentiviral-mediated transfection combined with OA treatment. The results showed that cav-1 overexpression inhibited HL-60 cell proliferation, promoted apoptosis, arrested the cell cycle in the G1 phase and inhibited activation of the PI3K/AKT/mTOR signaling pathway. Overexpression of CAV-1 also increased HL-60 cell sensitivity to OA. To further verify whether OA affects HL-60 cells via the activation of downstream signaling pathways by CAV-1, cav-1 gene expression was silenced using RNAi, and the cells were treated with OA to examine its efficacy. The results showed that after cav-1 silencing, OA had little effect on cell activity, apoptosis, the cell cycle and phosphorylation of HL-60 cells. This study is the first to show that CAV-1 plays a crucial role in the effects of OA on HL-60 cells.

  9. 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...

  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. Fungi causing dying out of heather seedlings

    OpenAIRE

    Maria Kowalik; Agnieszka Wandzel

    2012-01-01

    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 leveille...

  12. Environmental Impact Statement for BOMARC Missile Site, McGuire AFB, New Jersey. Volume 3. Methodology Development. Appendices 3-1 through 3-8

    Science.gov (United States)

    1992-05-01

    Aclepias syriaca Common Milkweed ftis Msvahis Summer Grape Fesuca rubra Red Fescue I Cirsium vulgare Common Thistle Cassiafascicu/ata Partridge Pea...purpurascens Purple Milkweed iburnum recognitum Smooth Arrowroot I Lespedexa cuneaa Sericea Rhexia virginica Meadow Beauty Chrysopsisfawaa Sickle-leaved...incidence of resulting health effects is very small. There have been no direct measurements of increased cancer incidence rates for I low-level radiation

  13. Transcriptional upregulation of α2δ-1 elevates arterial smooth muscle cell voltage-dependent Ca2+ channel surface expression and cerebrovascular constriction in genetic hypertension.

    Science.gov (United States)

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

    2012-10-01

    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 composed of 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 associated with hypertension. Age-dependent development of hypertension in spontaneously hypertensive rats was associated with an unequal elevation in α2δ-1 and CaV1.2α1 mRNA and protein in cerebral artery myocytes, with α2δ-1 increasing more than CaV1.2α1. Other α2δ isoforms did not emerge in hypertension. Myocytes and arteries of hypertensive spontaneously hypertensive rats displayed higher surface-localized α2δ-1 and CaV1.2α1 proteins, surface α2δ-1:CaV1.2α1 ratio, CaV1.2 current density and noninactivating current, and pressure- and depolarization-induced vasoconstriction than those of Wistar-Kyoto controls. Pregabalin, an α2δ-1 ligand, did not alter α2δ-1 or CaV1.2α1 total protein but normalized α2δ-1 and CaV1.2α1 surface expression, surface α2δ-1:CaV1.2α1, CaV1.2 current density and inactivation, and vasoconstriction in myocytes and arteries of hypertensive rats to control levels. Genetic hypertension is associated with an elevation in α2δ-1 expression that promotes surface trafficking of CaV1.2 channels in cerebral artery myocytes. This leads to an increase in CaV1.2 current-density and a reduction in current inactivation that induces vasoconstriction. Data also suggest that α2δ-1 targeting is a novel strategy that may be used to reverse pathological CaV1.2 channel trafficking to induce cerebrovascular dilation in hypertension.

  14. 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.

  15. In MMTV-Her-2/neu transgenic mammary tumors the absence of caveolin-1-/- alters PTEN and NHERF1 but not β-catenin expression.

    Science.gov (United States)

    Cuello-Carrión, F Darío; Cayado-Gutiérrez, Niubys; Natoli, Anthony L; Restall, Christina; Anderson, Robin L; Nadin, Silvina; Alvarez-Olmedo, Daiana; Castro, Gisela N; Gago, Francisco E; Fanelli, Mariel A; Ciocca, Daniel R

    2013-09-01

    In a recent study, we have shown that in mammary tumors from mice lacking the Cav-1 gene, there are alterations in specific heat shock proteins as well as in tumor development. With this in mind, we have now investigated other proteins in the same mammary mouse tumor model (Her-2/neu expressing mammary tumors from Cav-1 wild type and Cav-1 null mice), to further comprehend the complex tumor-stroma mechanisms involved in regulating stress responses during tumor development. In this tumor model the cancer cells always lacked of Cav-1, so the KO influenced the Cav-1 in the stroma. By immunohistochemistry, we have found a striking co-expression of β-catenin and Her-2/neu in the tumor cells. The absence of Cav-1 in the tumor stroma had no effect on expression or localization of β-catenin and Her-2/neu. Both proteins appeared co-localized at the cell surface during tumor development and progression. Since Her-2/neu activation induces MTA1, we next evaluated MTA1 in the mouse tumors. Although this protein was found in numerous nuclei, the absence of Cav-1 did not alter its expression level. In contrast, significantly more PTEN protein was noted in the tumors lacking Cav-1 in the stroma, with the protein localized mainly in the nuclei. P-Akt levels were relatively low in tumors from both Cav-1 WT and Cav-1 KO mice. There was also an increase in nuclear NHERF1 expression levels in the tumors arising from Cav-1 KO mice. The data obtained in the MMTV-neu model are consistent with a role for Cav-1 in adjacent breast cancer stromal cells in modulating the expression and localization of important proteins implicated in tumor cell behavior.

  16. 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.

  17. Loss of caveolin-1 accelerates neurodegeneration and aging.

    Directory of Open Access Journals (Sweden)

    Brian P Head

    Full Text Available BACKGROUND: The aged brain exhibits a loss in gray matter and a decrease in spines and synaptic densities that may represent a sequela for neurodegenerative diseases such as Alzheimer's. Membrane/lipid rafts (MLR, discrete regions of the plasmalemma enriched in cholesterol, glycosphingolipids, and sphingomyelin, are essential for the development and stabilization of synapses. Caveolin-1 (Cav-1, a cholesterol binding protein organizes synaptic signaling components within MLR. It is unknown whether loss of synapses is dependent on an age-related loss of Cav-1 expression and whether this has implications for neurodegenerative diseases such as Alzheimer's disease. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed brains from young (Yg, 3-6 months, middle age (Md, 12 months, aged (Ag, >18 months, and young Cav-1 KO mice and show that localization of PSD-95, NR2A, NR2B, TrkBR, AMPAR, and Cav-1 to MLR is decreased in aged hippocampi. Young Cav-1 KO mice showed signs of premature neuronal aging and degeneration. Hippocampi synaptosomes from Cav-1 KO mice showed reduced PSD-95, NR2A, NR2B, and Cav-1, an inability to be protected against cerebral ischemia-reperfusion injury compared to young WT mice, increased Aβ, P-Tau, and astrogliosis, decreased cerebrovascular volume compared to young WT mice. As with aged hippocampi, Cav-1 KO brains showed significantly reduced synapses. Neuron-targeted re-expression of Cav-1 in Cav-1 KO neurons in vitro decreased Aβ expression. CONCLUSIONS: Therefore, Cav-1 represents a novel control point for healthy neuronal aging and loss of Cav-1 represents a non-mutational model for Alzheimer's disease.

  18. Regulation of presynaptic strength by controlling Ca2+ channel mobility: effects of cholesterol depletion on release at the cone ribbon synapse.

    Science.gov (United States)

    Mercer, Aaron J; Szalewski, Robert J; Jackman, Skyler L; Van Hook, Matthew J; Thoreson, Wallace B

    2012-06-01

    Synaptic communication requires proper coupling between voltage-gated Ca(2+) (Ca(V)) channels and synaptic vesicles. In photoreceptors, L-type Ca(V) channels are clustered close to synaptic ribbon release sites. Although clustered, Ca(V) channels move continuously within a confined domain slightly larger than the base of the ribbon. We hypothesized that expanding Ca(V) channel confinement domains should increase the number of channel openings needed to trigger vesicle release. Using single-particle tracking techniques, we measured the expansion of Ca(V) channel confinement domains caused by depletion of membrane cholesterol with cholesterol oxidase or methyl-β-cyclodextrin. With paired whole cell recordings from cones and horizontal cells, we then determined the number of Ca(V) channel openings contributing to cone Ca(V) currents (I(Ca)) and the number of vesicle fusion events contributing to horizontal cell excitatory postsynaptic currents (EPSCs) following cholesterol depletion. Expansion of Ca(V) channel confinement domains reduced the peak efficiency of release, decreasing the number of vesicle fusion events accompanying opening of each Ca(V) channel. Cholesterol depletion also inhibited exocytotic capacitance increases evoked by brief depolarizing steps. Changes in efficiency were not due to changes in I(Ca) amplitude or glutamate receptor properties. Replenishing cholesterol restored Ca(V) channel domain size and release efficiency to control levels. These results indicate that cholesterol is important for organizing the cone active zone. Furthermore, the finding that cholesterol depletion impairs coupling between channel opening and vesicle release by allowing Ca(V) channels to move further from release sites shows that changes in presynaptic Ca(V) channel mobility can be a mechanism for adjusting synaptic strength.

  19. Matrix remodeling stimulates stromal autophagy, “fueling” cancer cell mitochondrial metabolism and metastasis

    OpenAIRE

    Castello-Cros, Remedios; Bonuccelli, Gloria; Molchansky, Alex; Capozza, Franco; Witkiewicz, Agnieszka K.; Birbe, Ruth C; Howell, Anthony; Pestell, Richard G.; Whitaker-Menezes, Diana; Sotgia, Federica; Lisanti, Michael P

    2011-01-01

    We have previously demonstrated that loss of stromal caveolin-1 (Cav-1) in cancer-associated fibroblasts is a strong and independent predictor of poor clinical outcome in human breast cancer patients. However, the signaling mechanism(s) by which Cav-1 downregulation leads to this tumor-promoting microenvironment are not well understood. To address this issue, we performed an unbiased comparative proteomic analysis of wild-type (WT) and Cav-1-/- null mammary stromal fibroblasts (MSFs). Our res...

  20. Restoration of caveolin-1 expression suppresses growth, membrane-type-4 metalloproteinase expression and metastasis-associated activities in colon cancer cells.

    Science.gov (United States)

    Nimri, Lili; Barak, Hossei; Graeve, Lutz; Schwartz, Betty

    2013-11-01

    Caveolin-1 (cav-1) and flotillin-1 are two major structural proteins associated with lipid rafts in mammalian cells. The membrane-type matrix metalloproteinases (MT-MMPs) are expressed at the cell surface, hydrolyze extracellular matrix, and play an important role in cancer cell migration and metastasis. Expression of cav-1, flotillin-1, and MT4-MMP in lysates and lipid rafts of LS174T and HM-7 colon cancer cells was determined. The impact of restoration of cav-1 expression on proliferation, adhesion, motility in vitro, and growth of implanted tumors in vivo was characterized. Cav-1 is not expressed in lipid rafts of the highly metastatic colon cancer cell line (HM-7), but expressed in cytosolic fractions of the parental lower metastatic cell line (LS174T). In contrast, MT4-MMP was expressed in lipid rafts of HM-7 cells but not in LS174T cells. Overexpression of cav-1 in HM-7 cells down-regulate proliferation, viability, wound closure, adhesion to laminin, invasion, and development of filopodial and lamellipodial structures in a dose-dependent manner. Cav-1 positive HM-7 clones ceased to express MT4-MMP in their lipid rafts. Comparative proteomic analyses of lipid rafts from cav-1 positive and cav-1 negative cells demonstrated de novo expression of flotillin-1 only on the cells expressing cav-1. Xenografting control cells devoid of cav-1 in nude mice induced development of bigger tumors expressing higher levels of proliferating cell nuclear antigen as compared to mice injected with cells expressing the highest cav-1 levels. We conclude that cav-1 orchestrates and reorganize several proteins in lipid rafts, activities directly associated with reduced tumorigenic and metastatic ability of colon cancer cells.

  1. 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

    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...... to the conclusion that Cav3.1 and Cav3.2 channels have important, but different, functions in mice. T-type Cav3.1 channels affect heart rate, whereas Cav3.2 channels are involved in cardiac hypertrophy. In the vascular system, Cav3.2 activation leads to dilation of blood vessels, whereas Cav3.1 channels are mainly.......2, are expressed in blood vessels, the kidney, and the heart. Studies with gene-deficient mice have provided a way to investigate the Cav3.1 and Cav3.2 channels and their role in the cardiovascular system. This review discusses the results from these knockout mice. Evaluation of the literature leads...

  2. 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

  3. Biomechanical remodeling of the microenvironment by stromal Caveolin-1 favors tumor invasion and metastasis

    Science.gov (United States)

    Goetz, Jacky G.; Minguet, Susana; Navarro-Lérida, Inmaculada; Lazcano, Juan José; Samaniego, Rafael; Calvo, Enrique; Tello, Marta; Osteso-Ibáñez, Teresa; Pellinen, Teijo; Echarri, Asier; Cerezo, Ana; Klein-Szanto, Andres J.P.; Garcia, Ricardo; Keely, Patricia J.; Sánchez-Mateos, Paloma; Cukierman, Edna; Del Pozo, Miguel A.

    2011-01-01

    Summary Mechanotransduction, a key determinant of tissue homeostasis and tumor progression, is driven by intercellular adhesions, cell contractility and forces generated with the microenvironment, dependent on extracellular matrix composition, organization and compliance. Caveolin-1 (Cav1) favors cell elongation in 3D cultures and promotes Rho-and force-dependent contraction, matrix alignment and microenvironment stiffening through regulation of p190RhoGAP. In turn, microenvironment remodeling by Cav1-fibroblasts forces cell elongation. Cav1-deficient mice have disorganized stromal tissue architecture. Stroma associated with human carcinomas and melanoma metastases is enriched in Cav1-expressing carcinoma-associated fibroblasts (CAFs). Cav1 expression in breast CAFs correlates with low survival, and Cav1 depletion in CAFs decreases CAF contractility. Consistently, fibroblast expression of Cav1, through p190RhoGAP regulation, favors directional migration and invasiveness of carcinoma cells in vitro. In vivo, stromal Cav1 remodels peri- and intratumoral microenvironments to facilitate tumor invasion, correlating with increased metastatic potency. Thus, Cav1 modulates tissue responses through force-dependent architectural regulation of the microenvironment. PMID:21729786

  4. Time-dependent regulation of muscle caveolin activation and insulin signalling in response to high-fat diet.

    Science.gov (United States)

    Gómez-Ruiz, Ana; de Miguel, Carlos; Campión, Javier; Martínez, J Alfredo; Milagro, Fermín I

    2009-10-06

    We studied the effect of high-fat diet on the expression and activation of the three caveolins in rat skeletal muscle and their association with the insulin signalling cascade. Initial response was characterized by increased signalling through Cav-1 and Cav-3 phosphorylation, suggesting that both participate in an initial acute response to the calorie surplus. Afterwards, Cav-1 signalling was slightly reduced, whereas Cav-3 remained active. Late chronic phase signalling through both proteins was impaired inducing a prediabetic state. Summarizing, caveolins seem to mediate a time-dependent regulation of insulin cascade in response to high-fat diet in muscle.

  5. BARP suppresses voltage-gated calcium channel activity and Ca2+-evoked exocytosis.

    Science.gov (United States)

    Béguin, Pascal; Nagashima, Kazuaki; Mahalakshmi, Ramasubbu N; Vigot, Réjan; Matsunaga, Atsuko; Miki, Takafumi; Ng, Mei Yong; Ng, Yu Jin Alvin; Lim, Chiaw Hwee; Tay, Hock Soon; Hwang, Le-Ann; Firsov, Dmitri; Tang, Bor Luen; Inagaki, Nobuya; Mori, Yasuo; Seino, Susumu; Launey, Thomas; Hunziker, Walter

    2014-04-28

    Voltage-gated calcium channels (VGCCs) are key regulators of cell signaling and Ca(2+)-dependent release of neurotransmitters and hormones. Understanding the mechanisms that inactivate VGCCs to prevent intracellular Ca(2+) overload and govern their specific subcellular localization is of critical importance. We report the identification and functional characterization of VGCC β-anchoring and -regulatory protein (BARP), a previously uncharacterized integral membrane glycoprotein expressed in neuroendocrine cells and neurons. BARP interacts via two cytosolic domains (I and II) with all Cavβ subunit isoforms, affecting their subcellular localization and suppressing VGCC activity. Domain I interacts at the α1 interaction domain-binding pocket in Cavβ and interferes with the association between Cavβ and Cavα1. In the absence of domain I binding, BARP can form a ternary complex with Cavα1 and Cavβ via domain II. BARP does not affect cell surface expression of Cavα1 but inhibits Ca(2+) channel activity at the plasma membrane, resulting in the inhibition of Ca(2+)-evoked exocytosis. Thus, BARP can modulate the localization of Cavβ and its association with the Cavα1 subunit to negatively regulate VGCC activity.

  6. Caveolin-1 protects B6129 mice against Helicobacter pylori gastritis.

    Directory of Open Access Journals (Sweden)

    Ivana Hitkova

    Full Text Available 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.

  7. Integrin alphaVbeta6 is a high-affinity receptor for coxsackievirus A9.

    Science.gov (United States)

    Heikkilä, Outi; Susi, Petri; Stanway, Glyn; Hyypiä, Timo

    2009-01-01

    Coxsackievirus A9 (CAV9), a member of the genus Enterovirus in the family Picornaviridae, possesses an integrin-binding arginine-glycine-aspartic acid (RGD) motif in the C terminus of VP1 capsid protein. CAV9 has been shown to utilize integrins alphaVbeta3 and alphaVbeta6 as primary receptors for cell attachment. While CAV9 RGD-mutants (RGE and RGDdel) are capable of infecting rhabdomyosarcoma (RD) cell line, they grow very poorly in an epithelial lung carcinoma cell line (A549). In this study, the relationships between CAV9 infectivity in A549 and RD cells, receptor expression and integrin binding were analysed. A549 cells were shown to express both integrins alphaVbeta3 and alphaVbeta6, whereas alphaVbeta6 expression was not detected on the RD cells. Native CAV9 but not RGE and RGDdel mutants bound efficiently to immobilized alphaVbeta3 and alphaVbeta6. Adhesion of CAV9 but not RGE/RGDdel to A549 cells was also significantly higher than to RD cells. In contrast, no affinity or adhesion of bacterially produced VP1 proteins to the integrins or to the cells was detected. Function-blocking antibodies against alphaV-integrins blocked CAV9 but not CAV9-RGDdel infectivity, indicating that the viruses use different internalization routes; this may explain the differential infection kinetics of CAV9 and RGDdel. In an affinity assay, soluble alphaVbeta6, but not alphaVbeta3, bound to immobilized CAV9. Similarly, only soluble alphaVbeta6 blocked virus infectivity. These data suggest that CAV9 binding to alphaVbeta6 is a high-affinity interaction, which may indicate its importance in clinical infections; this remains to be determined.

  8. Rab5 is required in metastatic cancer cells for Caveolin-1-enhanced Rac1 activation, migration and invasion.

    Science.gov (United States)

    Díaz, Jorge; Mendoza, Pablo; Ortiz, Rina; Díaz, Natalia; Leyton, Lisette; Stupack, Dwayne; Quest, Andrew F G; Torres, Vicente A

    2014-06-01

    Rab5 is a small GTPase that regulates early endosome trafficking and other cellular processes, including cell adhesion and migration. Specifically, Rab5 promotes Rac1 activation and cancer cell migration, but little is known about the upstream regulators of Rab5. We have previously shown that the scaffolding protein Caveolin-1 (CAV1) promotes Rac1 activation and migration of cancer cells. Here, we hypothesized that CAV1 stimulates Rab5 activation, leading to increased Rac1 activity and cell migration. Expression of CAV1 in B16-F10 mouse melanoma and HT-29(US) human colon adenocarcinoma cells increased the GTP loading of Rab5, whereas shRNA-mediated targeting of endogenous CAV1 in MDA-MB-231 breast cancer cells decreased Rab5-GTP levels. Accordingly, shRNA-mediated downregulation of Rab5 decreased CAV1-mediated Rac1 activation, cell migration and invasion in B16-F10 and HT-29(US) cells. Expression of CAV1 was accompanied by increased recruitment of Tiam1, a Rac1 guanine nucleotide exchange factor (GEF), to Rab5-positive early endosomes. Using the inhibitor NSC23766, Tiam1 was shown to be required for Rac1 activation and cell migration induced by CAV1 and Rab5. Mechanistically, we provide evidence implicating p85α (also known as PIK3R1), a Rab5 GTPase-activating protein (GAP), in CAV1-dependent effects, by showing that CAV1 recruits p85α, precluding p85α-mediated Rab5 inactivation and increasing cell migration. In summary, these studies identify a novel CAV1-Rab5-Rac1 signaling axis, whereby CAV1 prevents Rab5 inactivation, leading to increased Rac1 activity and enhanced tumor cell migration and invasion.

  9. 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.

  10. Effects of eugenol on T-type Ca2+ channel isoforms.

    Science.gov (United States)

    Seo, Haengsoo; Li, Hai Ying; Perez-Reyes, Edward; Lee, Jung-Ha

    2013-11-01

    Eugenol has been used as an analgesic in dentistry. Previous studies have demonstrated that voltage-gated Na(+) channels and high-voltage-activated Ca(2+) channels expressed in trigeminal ganglion (TG) neurons sensing dental pain are molecular targets of eugenol for its analgesic effects. However, it has not been investigated whether eugenol can affect T-type Ca(2+) channels, which are known to be detected in the afferent neurons. In this report, we investigate how eugenol can influence cloned T-type channel isoforms expressed in HEK293 cells, using whole-cell patch clamp. Application of eugenol inhibited Cav3.1, Cav3.2, and Cav3.3 currents in a concentration-dependent manner with IC50 values of 463, 486, and 708 μM, respectively. Eugenol was found to negatively shift the steady-state inactivation curves of the T-type channel isoforms, but it did not shift their activation curves. In addition, eugenol had little effect on the current kinetics of Cav3.1 and Cav3.2, but it accelerated the inactivation kinetics of Cav3.3 currents. Reduction of channel availability enhanced eugenol inhibition sensitivity for Cav3.1 and Cav3.2, but not for Cav3.3. Moreover, eugenol inhibition of T-type channel isoforms was found to be use dependent. Finally, we show that the T-type currents recorded from rat TG neurons were inhibited by eugenol with a similar potency to Cav3.1 and Cav3.2 isoforms. Taken together, our findings suggest that T-type Ca(2+) channels are additional molecular targets for the pain-relieving effects of eugenol.

  11. 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.

  12. Caveolin-1 Promotes the Imbalance of Th17/Treg in Patients with Chronic Obstructive Pulmonary Disease.

    Science.gov (United States)

    Sun, Nina; Wei, Xiaofang; Wang, Jingluan; Cheng, Zhaozhong; Sun, Weihong

    2016-12-01

    The imbalance of Th17/Treg cells plays an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Caveolin-1 (Cav-1) has been regarded as a potential critical regulatory protein in pathological mechanisms of chronic inflammatory respiratory diseases. Therefore, we investigated whether the loss of Cav-1 is involved in the homeostasis of Th17/Treg cells in COPD. We examined the expressions of plasma Cav-1 and circulating Th17, Treg cells, and the related cytokines in patients with COPD. Enzyme-linked immunosorbent assay (ELISA) analyses showed a significant reduction of plasma Cav-1 levels in patients with stable COPD (SCOPD) and acutely exacerbated COPD (AECOPD) compared to smokers without COPD. This loss was associated with an increase in frequency of Treg and decreased in frequency of Th17 cells. To further identify the role of Cav-1, we studied the effects of Cav-1 overexpression or downregulation on frequencies of Treg and Th17 cells in peripheral blood mononuclear cells (PBMCs) from subjects. Interestingly, small interfering RNA (siRNA) downregulation of Cav-1 was accompanied by an augmentation of Treg and reduction of Th17 expression. Together, our study demonstrated that the loss of Cav-1 contributed to the imbalance of Th17/Treg cells in patients with COPD.

  13. Relationship of PON1 192 and 55 gene polymorphisms to calcific valvular aortic stenosis

    DEFF Research Database (Denmark)

    Moura, Luis M; Faria, Susana; Brito, Miguel

    2012-01-01

    Paraoxonases may exert anti-atherogenic action by reducing lipid peroxidation. Previous studies examined associations between polymorphisms in the paraoxonase 1 (PON1) gene and development of coronary artery disease (CAD), with inconsistent results. Given the similarities in clinical and pathophy...... and pathophysiological risk factors of CAD and calcific aortic valve stenosis (CAVS), we postulated a link between PON1 alleles and CAVS progression....

  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. 75 FR 42765 - Agency Information Collection Activities: Proposed Collection; Comment Request, 1660-0023...

    Science.gov (United States)

    2010-07-22

    ... Program CAC and CAV Reports AGENCY: Federal Emergency Management Agency, DHS. ACTION: Notice; 60-day... to meet these objectives are through the Community Assistance Contact (CAC) and Community Assistance... Assistance Program CAC and CAV Reports. Type of Information Collection: Revision of a currently...

  16. 78 FR 51198 - Agency Information Collection Activities: Proposed Collection; Comment Request; Effectiveness of...

    Science.gov (United States)

    2013-08-20

    ... CAC and CAV Reports AGENCY: Federal Emergency Management Agency, DHS. ACTION: Notice. SUMMARY: The... NFIP Community Assistance Program Community Assistance Contact (CAC) and Community Assistance Visit... Program CAC and CAV Reports. OMB Number: 1660-0023. Type of Information Collection: Revision of...

  17. 75 FR 68613 - Agency Information Collection Activities: Submission for OMB Review; Comment Request, OMB No...

    Science.gov (United States)

    2010-11-08

    ... Community Assistance Program CAC and CAV Reports AGENCY: Federal Emergency Management Agency, DHS. ACTION... Assistance Program CAC and CAV Reports. Type of information collection: Revision of a currently approved... Report. Abstract: Through the use of a Community Assistance Contact (CAC) or Community Assistance...

  18. Voltage-Gated R-Type Calcium Channel Inhibition via Human μ-, δ-, and κ-opioid Receptors Is Voltage-Independently Mediated by Gβγ Protein Subunits.

    Science.gov (United States)

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

    2016-01-01

    Elucidating the mechanisms that modulate calcium channels via opioid receptor activation is fundamental to our understanding of both pain perception and how opioids modulate pain. Neuronal voltage-gated N-type calcium channels (Cav2.2) are inhibited by activation of G protein-coupled opioid receptors (ORs). However, inhibition of R-type (Cav2.3) channels by μ- or κ-ORs is poorly defined and has not been reported for δ-ORs. To investigate such interactions, we coexpressed human μ-, δ-, or κ-ORs with human Cav2.3 or Cav2.2 in human embryonic kidney 293 cells and measured depolarization-activated Ba(2+) currents (IBa). Selective agonists of μ-, δ-, and κ-ORs inhibited IBa through Cav2.3 channels by 35%. Cav2.2 channels were inhibited to a similar extent by κ-ORs, but more potently (60%) via μ- and δ-ORs. Antagonists of δ- and κ-ORs potentiated IBa amplitude mediated by Cav2.3 and Cav2.2 channels. Consistent with G protein βγ (Gβγ) interaction, modulation of Cav2.2 was primarily voltage-dependent and transiently relieved by depolarizing prepulses. In contrast, Cav2.3 modulation was voltage-independent and unaffected by depolarizing prepulses. However, Cav2.3 inhibition was sensitive to pertussis toxin and to intracellular application of guanosine 5'-[β-thio]diphosphate trilithium salt and guanosine 5'-[γ-thio]triphosphate tetralithium salt. Coexpression of Gβγ-specific scavengers-namely, the carboxyl terminus of the G protein-coupled receptor kinase 2 or membrane-targeted myristoylated-phosducin-attenuated or abolished Cav2.3 modulation. Our study reveals the diversity of OR-mediated signaling at Cav2 channels and identifies neuronal Cav2.3 channels as potential targets for opioid analgesics. Their novel modulation is dependent on pre-existing OR activity and mediated by membrane-delimited Gβγ subunits in a voltage-independent manner.

  19. Energy Impact of Different Penetrations of Connected and Automated Vehicles: A Preliminary Assessment

    Energy Technology Data Exchange (ETDEWEB)

    Rios-Torres, Jackeline [ORNL; Malikopoulos, Andreas [ORNL

    2016-01-01

    Previous research reported in the literature has shown the benefits of traffic coordination to alleviate congestion, and reduce fuel consumption and emissions. However, there are still many remaining challenges that need to be addressed before a massive deployment of fully automated vehicles. This paper aims to investigate the energy impacts of different penetration rates of connected and automated vehicles (CAVs) and their interaction with human-driven vehicles. We develop a simulation framework for mixed traffic (CAVs interacting with human-driven vehicles) in merging roadways and analyze the energy impact of different penetration rates of CAVs on the energy consumption. The Gipps car following model is used along with heuristic controls to represent the driver decisions in a merging roadways traffic scenario. Using different penetration rates of CAVs, the simulation results indicated that for low penetration rates, the fuel consumption benefits are significant but the total travel time increases. The benefits in travel time are noticeable for higher penetration rates of CAVs.

  20. Current Status of Cardiac Allograft Vasculopathy Diagnosis%移植心脏冠状动脉血管病变的诊断现状

    Institute of Scientific and Technical Information of China (English)

    韩春勇

    2012-01-01

    Cardiac allograft vasculopathy ( CAV) is a complicated coronary artery disease that occurs after heart transplantation. CAV primarily contributes to the patients' high postoperative morbidites and mortality. Due to the denervation of cardiac allografts, CAV rarely presents with an ischemic syndrome. Therefore it is important to screen and diagnose CAV early. Coronary angiography remains the principal screening tool for CAV in most centers, even though its sensitivity is not high enough to detect CAV. Intravascular ultrasound is considered to be the most reliable tool for diagnosing CAV as it can directly determine the intimal thickness and the vessel wall morphology. New approaches to diagnosis include dobutamine stress echocardiography, coronary computed tomography, and cardiac MRI. These diagnostic tools require more testing in relation to CAV.%移植心脏血管病变(CAV)是一种相当复杂的心脏移植后冠状血管病变,是在心脏移植术后患者发病或死亡的首要原因,CAV患者通常无症状,因此CAV的早期筛查诊断是非常重要.大部分心脏中心每年常规行冠状动脉造影筛查CAV,而冠状动脉造影敏感性欠佳.血管内超声被认为是最敏感的早期筛查诊断CAV的方法,因为它可以直接准确测定内膜增厚程度、管壁形态.多巴酚丁胺负荷超声心动图、冠状动脉CT和心脏磁共振成像是较新的CAV诊断方法,但仍需要进一步研究.

  1. Deletion of the Mouse Homolog of CACNA1C Disrupts Discrete Forms of Hippocampal-Dependent Memory and Neurogenesis within the Dentate Gyrus

    Science.gov (United States)

    Bell, Ryan Z.; Fisher, Grace L.

    2016-01-01

    Abstract L-type voltage-gated calcium channels (LVGCCs) have been implicated in various forms of learning, memory, and synaptic plasticity. Within the hippocampus, the LVGCC subtype, CaV1.2 is prominently expressed throughout the dentate gyrus. Despite the apparent high levels of CaV1.2 expression in the dentate gyrus, the role of CaV1.2 in hippocampal- and dentate gyrus-associated forms of learning remain unknown. To address this question, we examined alternate forms of hippocampal-dependent associative and spatial memory in mice lacking the mouse ortholog of CACNA1C (Cacna1c), which encodes CaV1.2, with dentate gyrus function implicated in difficult forms of each task. We found that while the deletion of CaV1.2 did not impair the acquisition of fear of a conditioned context, mice lacking CaV1.2 exhibited deficits in the ability to discriminate between two contexts, one in which the mice were conditioned and one in which they were not. Similarly, CaV1.2 knock-out mice exhibited normal acquisition and recall of the location of the hidden platform in a standard Morris water maze, but were unable to form a memory of the platform location when the task was made more difficult by restricting the number of available spatial cues. Within the dentate gyrus, pan-neuronal deletion of CaV1.2 resulted in decreased cell proliferation and the numbers of doublecortin-positive adult-born neurons, implicating CaV1.2 in adult neurogenesis. These results suggest that CaV1.2 is important for dentate gyrus-associated tasks and may mediate these forms of learning via a role in adult neurogenesis and cell proliferation within the dentate gyrus. PMID:27957527

  2. The Ankrd13 Family of Ubiquitin-interacting Motif-bearing Proteins Regulates Valosin-containing Protein/p97 Protein-mediated Lysosomal Trafficking of Caveolin 1.

    Science.gov (United States)

    Burana, Daocharad; Yoshihara, Hidehito; Tanno, Hidetaka; Yamamoto, Akitsugu; Saeki, Yasushi; Tanaka, Keiji; Komada, Masayuki

    2016-03-18

    Caveolin 1 (Cav-1) is an oligomeric protein that forms flask-shaped, lipid-rich pits, termed caveolae, on the plasma membrane. Cav-1 is targeted for lysosomal degradation in ubiquitination- and valosin-containing protein (VCP)-dependent manners. VCP, an ATPase associated with diverse cellular activities that remodels or segregates ubiquitinated protein complexes, has been proposed to disassemble Cav-1 oligomers on the endosomal membrane, facilitating the trafficking of Cav-1 to the lysosome. Genetic mutations in VCP compromise the lysosomal trafficking of Cav-1, leading to a disease called inclusion body myopathy with Paget disease of bone and/or frontotemporal dementia (IBMPFD). Here we identified the Ankrd13 family of ubiquitin-interacting motif (UIM)-containing proteins as novel VCP-interacting molecules on the endosome. Ankrd13 proteins formed a ternary complex with VCP and Cav-1 and exhibited high binding affinity for ubiquitinated Cav-1 oligomers in an UIM-dependent manner. Mass spectrometric analyses revealed that Cav-1 undergoes Lys-63-linked polyubiquitination, which serves as a lysosomal trafficking signal, and that the UIMs of Ankrd13 proteins bind preferentially to this ubiquitin chain type. The overexpression of Ankrd13 caused enlarged hollow late endosomes, which was reminiscent of the phenotype of the VCP mutations in IBMPFD. Overexpression of Ankrd13 proteins also stabilized ubiquitinated Cav-1 oligomers on the limiting membrane of enlarged endosomes. The interaction with Ankrd13 was abrogated in IMBPFD-associated VCP mutants. Collectively, our results suggest that Ankrd13 proteins cooperate with VCP to regulate the lysosomal trafficking of ubiquitinated Cav-1.

  3. Loss of caveolin-1 in prostate cancer stroma correlates with reduced relapse-free survival and is functionally relevant to tumour progression.

    Science.gov (United States)

    Ayala, Gustavo; Morello, Matteo; Frolov, Anna; You, Sungyong; Li, Rile; Rosati, Fabiana; Bartolucci, Gianluca; Danza, Giovanna; Adam, Rosalyn M; Thompson, Timothy C; Lisanti, Michael P; Freeman, Michael R; Di Vizio, Dolores

    2013-09-01

    Levels of caveolin-1 (Cav-1) in tumour epithelial cells increase during prostate cancer progression. Conversely, Cav-1 expression in the stroma can decline in advanced and metastatic prostate cancer. In a large cohort of 724 prostate cancers, we observed significantly decreased levels of stromal Cav-1 in concordance with increased Gleason score (p = 0.012). Importantly, reduced expression of Cav-1 in the stroma correlated with reduced relapse-free survival (p = 0.009), suggesting a role for stromal Cav-1 in inhibiting advanced disease. Silencing of Cav-1 by shRNA in WPMY-1 prostate fibroblasts resulted in up-regulation of Akt phosphorylation, and significantly altered expression of genes involved in angiogenesis, invasion, and metastasis, including a > 2.5-fold increase in TGF-β1 and γ-synuclein (SNCG) gene expression. Moreover, silencing of Cav-1 induced migration of prostate cancer cells when stromal cells were used as attractants. Pharmacological inhibition of Akt caused down-regulation of TGF-β1 and SNCG, suggesting that loss of Cav-1 in the stroma can influence Akt-mediated signalling in the tumour microenvironment. Cav-1-depleted stromal cells exhibited increased levels of intracellular cholesterol, a precursor for androgen biosynthesis, steroidogenic enzymes, and testosterone. These findings suggest that loss of Cav-1 in the tumour microenvironment contributes to the metastatic behaviour of tumour cells by a mechanism that involves up-regulation of TGF-β1 and SNCG through Akt activation. They also suggest that intracrine production of androgens, a process relevant to castration resistance, may occur in the stroma.

  4. Molecular Basis of Regulating High Voltage-Activated Calcium Channels by S-Nitrosylation.

    Science.gov (United States)

    Zhou, Meng-Hua; Bavencoffe, Alexis; Pan, Hui-Lin

    2015-12-18

    Nitric oxide (NO) is involved in a variety of physiological processes, such as vasoregulation and neurotransmission, and has a complex role in the regulation of pain transduction and synaptic transmission. We have shown previously that NO inhibits high voltage-activated Ca(2+) channels in primary sensory neurons and excitatory synaptic transmission in the spinal dorsal horn. However, the molecular mechanism involved in this inhibitory action remains unclear. In this study, we investigated the role of S-nitrosylation in the NO regulation of high voltage-activated Ca(2+) channels. The NO donor S-nitroso-N-acetyl-DL-penicillamine (SNAP) rapidly reduced N-type currents when Cav2.2 was coexpressed with the Cavβ1 or Cavβ3 subunits in HEK293 cells. In contrast, SNAP only slightly inhibited P/Q-type and L-type currents reconstituted with various Cavβ subunits. SNAP caused a depolarizing shift in voltage-dependent N-type channel activation, but it had no effect on Cav2.2 protein levels on the membrane surface. The inhibitory effect of SNAP on N-type currents was blocked by the sulfhydryl-specific modifying reagent methanethiosulfonate ethylammonium. Furthermore, the consensus motifs of S-nitrosylation were much more abundant in Cav2.2 than in Cav1.2 and Cav2.1. Site-directed mutagenesis studies showed that Cys-805, Cys-930, and Cys-1045 in the II-III intracellular loop, Cys-1835 and Cys-2145 in the C terminus of Cav2.2, and Cys-346 in the Cavβ3 subunit were nitrosylation sites mediating NO sensitivity of N-type channels. Our findings demonstrate that the consensus motifs of S-nitrosylation in cytoplasmically accessible sites are critically involved in post-translational regulation of N-type Ca(2+) channels by NO. S-Nitrosylation mediates the feedback regulation of N-type channels by NO.

  5. Multiple nickel-sensitive targets elicit cardiac arrhythmia in isolated mouse hearts after pituitary adenylate cyclase-activating polypeptide-mediated chronotropy.

    Science.gov (United States)

    Tevoufouet, Etienne E; Nembo, Erastus N; Distler, Fabian; Neumaier, Felix; Hescheler, Jürgen; Nguemo, Filomain; Schneider, Toni

    2017-03-01

    The pituitary adenylate cyclase-activating polypeptide (PACAP)-27 modulates various biological processes, from the cellular level to function specification. However, the cardiac actions of this neuropeptide are still under intense studies. Using control (+|+) and mice lacking (-|-) either R-type (Cav2.3) or T-type (Cav3.2) Ca(2+) channels, we investigated the effects of PACAP-27 on cardiac activity of spontaneously beating isolated perfused hearts. Superfusion of PACAP-27 (20nM) caused a significant increase of baseline heart frequency in Cav2.3(+|+) (156.9±10.8 to 239.4±23.4 bpm; p<0.01) and Cav2.3(-|-) (190.3±26.4 to 270.5±25.8 bpm; p<0.05) hearts. For Cav3.2, the heart rate was significantly increased in Cav3.2(-|-) (133.1±8.5 bpm to 204.6±27.9 bpm; p<0.05) compared to Cav3.2(+|+) hearts (185.7±11.2 bpm to 209.3±22.7 bpm). While the P wave duration and QTc interval were significantly increased in Cav2.3(+|+) and Cav2.3(-|-) hearts following PACAP-27 superfusion, there was no effect in Cav3.2(+|+) and Cav3.2(-|-) hearts. The positive chronotropic effects observed in the four study groups, as well as the effect on P wave duration and QTc interval were abolished in the presence of Ni(2+) (50μM) and PACAP-27 (20nM) in hearts from Cav2.3(+|+) and Cav2.3(-|-) mice. In addition to suppressing PACAP's response, Ni(2+) also induced conduction disturbances in investigated hearts. In conclusion, the most Ni(2+)-sensitive Ca(2+) channels (R- and T-type) may modulate the PACAP signaling cascade during cardiac excitation in isolated mouse hearts, albeit to a lesser extent than other Ni(2+)-sensitive targets.

  6. 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.

  7. 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...

  8. 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.

  9. Purification and characterization of alpha-L-arabinofuranosidase from Arthrobacter sp. MTCC 5214 in solid-state fermentation

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.T.H.; Mukherjee, B.; Satwekar, A.; Bhosle, N.B.

    acetobutylicum [6] and Bacillus pumilus PS213 [16], which were 92, 94 and 60 kDa, respectively. High molecular mass a-L-AFases with multiple subunits isolated from Streptomyces purpurascens [32], Butyrivibrio fibrisolvens GS113 [33] and Thermotoga maritime [39... was not observed with pNPAF tested up to 6.8 mmol l C01 concentration. The efficiency of a-L-AFase (V max /K m ) from Arthro- bacter sp. was less than that shown by the a-L-AFase produced by Bacillus stearothermophilus[9] andB. pumilus PS213[16].However...

  10. Screening of medicinal plants from Suriname for 5-HT(1A) ligands: Bioactive isoquinoline alkaloids from the fruit of Annona muricata.

    Science.gov (United States)

    Hasrat, J A; Pieters, L; De Backer, J P; Vauquelin, G; Vlietinck, A J

    1997-06-01

    Plants from Suriname (South-America) and several Annona species, including A. muricata, A. ckerimolia, A. montana and A. glabra were screened for 5-HT(1A) receptor binding activity by ligand-binding-studies (LBS). Crude extracts of all Annona species and from Hibiscus bifurcatus, Irlbarchia purpurascens and Scoparia dulcis showed high activity. The isoquinoline alkaloids asimilobine (1), nornuciferine (2), and annonaine (3) were isolated as the active principles from the fruit of Annona muricata. These results may partially explain the use of Hibiscus bifurcatus and Annona muricata in traditional medicine in Suriname.

  11. Nuevos registros para la fauna de Saturniidae (Lepidoptera en Argentina

    Directory of Open Access Journals (Sweden)

    Adriana I. ZAPATA

    2012-01-01

    Full Text Available Se brindan nuevas citas de 17 especies de Saturniidae para la Argentina. Estos registros corresponden a las subfamilias: Arsenurinae [Arsenura xanthopus (Walker y Titaea tamerlan (Maassen]; Ceratocampinae: [Almeidella approximans (Schaus, Cicia nettia (Schaus, Citheronia aroa Schaus, Citheronia maureillei Wolfe & Herbin, Citioica anthonilis (Herrich- Schäffer, Eacles ducalis (Walker, Othorene cadmus (Herrich-Schäffer, Othorene purpurascens (Schaus y Ptiloscola photophila (Rothschild] y Hemileucinae [Cerodirphia brunnea (Draudt, Dirphia curitiba Draudt, Hirpida nigrolinea (Druce, Hylesia ebalus (Cramer, Leucanella gibbosa (Conte y Molippa strigosa (Maassen & Weyding].

  12. 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.

  13. Variation of Ursolic Acid Content in Eight Ocimum Species from Northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Selene M. Morais

    2008-10-01

    Full Text Available Ursolic acid is a very important compound due to its biological potential as an anti-inflammatory, trypanocidal, antirheumatic, antiviral, antioxidant and antitumoral agent. This study presents the HPLC analysis of ursolic acid (UA content in eight different Ocimum species: O. americanum L., O. basilicum L, O. basilicum var purpurascens Benth, O. basilicum var. minimum L, O. gratissimum L, O. micranthum Willd, O. selloi Benth. and O. tenuiflorum L. grown in Northeastern Brazil. In these Ocimum species, UA was detected in different yields, with O. tenuiflorum showing the highest content (2.02%. This yield is very significant when compared with other sources of UA.

  14. 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.

  15. Loss of caveolin-1 causes blood-retinal barrier breakdown, venous enlargement, and mural cell alteration.

    Science.gov (United States)

    Gu, Xiaowu; Fliesler, Steven J; Zhao, You-Yang; Stallcup, William B; Cohen, Alex W; Elliott, Michael H

    2014-02-01

    Blood-retinal barrier (BRB) breakdown and related vascular changes are implicated in several ocular diseases. The molecules and mechanisms regulating BRB integrity and pathophysiology are not fully elucidated. Caveolin-1 (Cav-1) ablation results in loss of caveolae and microvascular pathologies, but the role of Cav-1 in the retina is largely unknown. We examined BRB integrity and vasculature in Cav-1 knockout mice and found a significant increase in BRB permeability, compared with wild-type controls, with branch veins being frequent sites of breakdown. Vascular hyperpermeability occurred without apparent alteration in junctional proteins. Such hyperpermeability was not rescued by inhibiting eNOS activity. Veins of Cav-1 knockout retinas exhibited additional pathological features, including i) eNOS-independent enlargement, ii) altered expression of mural cell markers (eg, down-regulation of NG2 and up-regulation of αSMA), and iii) dramatic alterations in mural cell phenotype near the optic nerve head. We observed a significant NO-dependent increase in retinal artery diameter in Cav-1 knockout mice, suggesting that Cav-1 plays a role in autoregulation of resistance vessels in the retina. These findings implicate Cav-1 in maintaining BRB integrity in retinal vasculature and suggest a previously undefined role in the retinal venous system and associated mural cells. Our results are relevant to clinically significant retinal disorders with vascular pathologies, including diabetic retinopathy, uveoretinitis, and primary open-angle glaucoma.

  16. 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.

  17. Evaluation of helper-dependent canine adenovirus vectors in a 3D human CNS model.

    Science.gov (United States)

    Simão, D; Pinto, C; Fernandes, P; Peddie, C J; Piersanti, S; Collinson, L M; Salinas, S; Saggio, I; Schiavo, G; Kremer, E J; Brito, C; Alves, P M

    2016-01-01

    Gene therapy is a promising approach with enormous potential for treatment of neurodegenerative disorders. Viral vectors derived from canine adenovirus type 2 (CAV-2) present attractive features for gene delivery strategies in the human brain, by preferentially transducing neurons, are capable of efficient axonal transport to afferent brain structures, have a 30-kb cloning capacity and have low innate and induced immunogenicity in preclinical tests. For clinical translation, in-depth preclinical evaluation of efficacy and safety in a human setting is primordial. Stem cell-derived human neural cells have a great potential as complementary tools by bridging the gap between animal models, which often diverge considerably from human phenotype, and clinical trials. Herein, we explore helper-dependent CAV-2 (hd-CAV-2) efficacy and safety for gene delivery in a human stem cell-derived 3D neural in vitro model. Assessment of hd-CAV-2 vector efficacy was performed at different multiplicities of infection, by evaluating transgene expression and impact on cell viability, ultrastructural cellular organization and neuronal gene expression. Under optimized conditions, hd-CAV-2 transduction led to stable long-term transgene expression with minimal toxicity. hd-CAV-2 preferentially transduced neurons, whereas human adenovirus type 5 (HAdV5) showed increased tropism toward glial cells. This work demonstrates, in a physiologically relevant 3D model, that hd-CAV-2 vectors are efficient tools for gene delivery to human neurons, with stable long-term transgene expression and minimal cytotoxicity.

  18. Involvement of caveolin-1 in low shear stress-induced breast cancer cell motility and adhesion: Roles of FAK/Src and ROCK/p-MLC pathways.

    Science.gov (United States)

    Xiong, Niya; Li, Shun; Tang, Kai; Bai, Hongxia; Peng, Yueting; Yang, Hong; Wu, Chunhui; Liu, Yiyao

    2017-01-01

    Tumor cells translocating to distant sites are subjected to hemodynamic shear forces during their passage in the blood vessels. Low shear stress (LSS) plays a critical role in the regulation of various aspects of tumor cells functions, including motility and adhesion. Beyond its structural role, caveolin-1 (Cav-1), the important component of caveolae, represents a modulator of several cancer-associated functions as tumor progression and metastasis. However, the role of Cav-1 in regulating tumor cells response to shear stress remains poorly explored. Here, we characterized the role of LSS and Cav-1 in mediating cell motility and adhesion on human breast carcinoma MDA-MB-231 cells. We first showed that LSS exposure promoted cell polarity and focal adhesion (FA) dynamics, thus indicating elevated cell migration. Silencing of Cav-1 leaded to a significantly lower formation of stress fibers. However, LSS exposure was able to rescue it via the alteration of actin-associated proteins expression, including ROCK, p-MLC, cofilin and filamin A. Time-lapse migration assay indicated that Cav-1 expression fostered MDA-MB-231 cells motility and LSS triggered cells to rapidly generate new lamellipodia. Furthermore, Cav-1 and LSS significantly influenced cell adhesion. Taken together, our findings provide insights into mechanisms underlying LSS triggered events mediated by downstream Cav-1, including FAK/Src and ROCK/p-MLC pathways, involved in the reorganization of the cytoskeleton, cell motility, FA dynamics and breast cancer cell adhesion.

  19. Honeybee locomotion is impaired by Am-CaV3 low voltage-activated Ca(2+) channel antagonist.

    Science.gov (United States)

    Rousset, M; Collet, C; Cens, T; Bastin, F; Raymond, V; Massou, I; Menard, C; Thibaud, J-B; Charreton, M; Vignes, M; Chahine, M; Sandoz, J C; Charnet, P

    2017-02-01

    Voltage-gated Ca(2+) channels are key transducers of cellular excitability and participate in several crucial physiological responses. In vertebrates, 10 Ca(2+) channel genes, grouped in 3 families (CaV1, CaV2 and CaV3), have been described and characterized. Insects possess only one member of each family. These genes have been isolated in a limited number of species and very few have been characterized although, in addition to their crucial role, they may represent a collateral target for neurotoxic insecticides. We have isolated the 3 genes coding for the 3 Ca(2+) channels expressed in Apis mellifera. This work provides the first detailed characterization of the honeybee T-type CaV3 Ca(2+) channel and demonstrates the low toxicity of inhibiting this channel. Comparing Ca(2+) currents recorded in bee neurons and myocytes with Ca(2+) currents recorded in Xenopus oocytes expressing the honeybee CaV3 gene suggests native expression in bee muscle cells only. High-voltage activated Ca(2+) channels could be recorded in the somata of different cultured bee neurons. These functional data were confirmed by in situ hybridization, immunolocalization and in vivo analysis of the effects of a CaV3 inhibitor. The biophysical and pharmacological characterization and the tissue distribution of CaV3 suggest a role in honeybee muscle function.

  20. Expression of Caveolin 1 is enhanced by DNA demethylation during adipocyte differentiation. status of insulin signaling.

    Science.gov (United States)

    Palacios-Ortega, Sara; Varela-Guruceaga, Maider; Milagro, Fermín Ignacio; Martínez, José Alfredo; de Miguel, Carlos

    2014-01-01

    Caveolin 1 (Cav-1) is an essential constituent of adipocyte caveolae which binds the beta subunit of the insulin receptor (IR) and is implicated in the regulation of insulin signaling. We have found that, during adipocyte differentiation of 3T3-L1 cells the promoter, exon 1 and first intron of the Cav-1 gene undergo a demethylation process that is accompanied by a strong induction of Cav-1 expression, indicating that epigenetic mechanisms must have a pivotal role in this differentiation process. Furthermore, IR, PKB-Akt and Glut-4 expression are also increased during the differentiation process suggesting a coordinated regulation with Cav-1. Activation of Cav-1 protein by phosphorylation arises during the differentiation process, yet in fully mature adipocytes insulin is no longer able to significantly increase Cav-1 phosphorylation. However, these long-term differentiated cells are still able to respond adequately to insulin, increasing IR and PKB-Akt phosphorylation and glucose uptake. The activation of Cav-1 during the adipocyte differentiation process could facilitate the maintenance of insulin sensitivity by these fully mature adipocytes isolated from additional external stimuli. However, under the influence of physiological conditions associated to obesity, such as chronic inflammation and hypoxia, insulin sensitivity would finally be compromised.

  1. Caveolin-1 interacts with the Gag precursor of murine leukaemia virus and modulates virus production

    Directory of Open Access Journals (Sweden)

    Koester Mario

    2006-09-01

    Full Text Available Abstract Background Retroviral Gag determines virus assembly at the plasma membrane and the formation of virus-like particles in intracellular multivesicular bodies. Thereby, retroviruses exploit by interaction with cellular partners the cellular machineries for vesicular transport in various ways. Results The retroviral Gag precursor protein drives assembly of murine leukaemia viruses (MLV at the plasma membrane (PM and the formation of virus like particles in multivesicular bodies (MVBs. In our study we show that caveolin-1 (Cav-1, a multifunctional membrane-associated protein, co-localizes with Gag in a punctate pattern at the PM of infected NIH 3T3 cells. We provide evidence that Cav-1 interacts with the matrix protein (MA of the Gag precursor. This interaction is mediated by a Cav-1 binding domain (CBD within the N-terminus of MA. Interestingly, the CBD motif identified within MA is highly conserved among most other γ-retroviruses. Furthermore, Cav-1 is incorporated into MLV released from NIH 3T3 cells. Overexpression of a GFP fusion protein containing the putative CBD of the retroviral MA resulted in a considerable decrease in production of infectious retrovirus. Moreover, expression of a dominant-negative Cav-1 mutant affected retroviral titres significantly. Conclusion This study demonstrates that Cav-1 interacts with MLV Gag, co-localizes with Gag at the PM and affects the production of infectious virus. The results strongly suggest a role for Cav-1 in the process of virus assembly.

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

    Science.gov (United States)

    Ibanes, Sandy; Kremer, Eric J

    2013-01-01

    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.

  3. 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.

  4. The key molecular events during Macrobrachium rosenbergii nodavirus (MrNV) infection and replication in Sf9 insect cells.

    Science.gov (United States)

    Somrit, Monsicha; Watthammawut, Atthaboon; Chotwiwatthanakun, Charoonroj; Weerachatyanukul, Wattana

    2016-09-02

    In this study we demonstrated that Macrobrachium rosenbergii nodavirus (MrNV) was able to internalize and replicate in Sf9 insect cells, with levels of infection altered by substances affecting the caveolin-(CAV) mediated endocytosis pathway. The use of Sf9 cells for efficient MrNV replication and propagation was demonstrated by confocal microscopy and PCR amplification, through which early viral binding and internalization were initially detectable at 30min post-infection; whereas at 72h, the distinguishable sign of late-MrNV infection was observable as the gradual accumulation of a cytopathic effect (CPE) in the cells, ultimately resulting in cellular disruption. Moreover, during the early period of infection, the MrNV signals were highly co-localized with CAV1 signals of the CAV-mediated endocytosis pathway. The use of genistein as an inhibitor of the CAV-mediated endocytosis pathway significantly reduced MrNV and CAV1 co-localization, and also reduced the levels of MrNV infection in Sf9 cells as shown by PCR and ELISA. Moreover, the addition of the pathway agonist okadaic acid not only recovered but also augmented both the levels of MrNV co-localization with CAV1 and of Sf9 infection in the presence of genistein inhibition; therefore demonstrating that MrNV infection in Sf9 cells was associated with the CAV-mediated endocytosis pathway machinery.

  5. 阿片物质预处理效应依赖于细胞质膜微囊蛋白-3的表达

    Institute of Scientific and Technical Information of China (English)

    Yasuo M. Tsutsumi; Yoshitaka Kawaraguchi; Ingrid R. Niesman; Hemal H. Patel; David M. Roth; 安博

    2012-01-01

    我们验证了细胞质膜微囊蛋白-3(caveolin-3,Cav-3)是体内阿片物质预处理效应的必要因素的假说.Cav-3过度表达组、Cav-3基因敲除组和对照组的小鼠均在经受心肌缺血/再灌注(ischemia/reperfusion,I/R)时,给予选择性δ-受体激动剂SNC-121 (SNC)或非选择性阿片受体拮抗剂纳洛酮.对照组小鼠因SNC免受I/R损伤;Cav-3基因敲除组中SNC未产生保护作用;而与被纳洛酮拮抗的对照组相比,Cav-3过度表达小鼠表现出对I/R的天然保护作用.我们的研究结果表明阿片物质预处理依赖于Cav-3的表达,并且在Cav-3过度表达的小鼠中这种内源性保护作用有阿片依赖性.

  6. Honeybee locomotion is impaired by Am-CaV3 low voltage-activated Ca2+ channel antagonist

    Science.gov (United States)

    Rousset, M.; Collet, C.; Cens, T.; Bastin, F.; Raymond, V.; Massou, I.; Menard, C.; Thibaud, J.-B.; Charreton, M.; Vignes, M.; Chahine, M.; Sandoz, J. C.; Charnet, P.

    2017-01-01

    Voltage‐gated Ca2+ channels are key transducers of cellular excitability and participate in several crucial physiological responses. In vertebrates, 10 Ca2+ channel genes, grouped in 3 families (CaV1, CaV2 and CaV3), have been described and characterized. Insects possess only one member of each family. These genes have been isolated in a limited number of species and very few have been characterized although, in addition to their crucial role, they may represent a collateral target for neurotoxic insecticides. We have isolated the 3 genes coding for the 3 Ca2+ channels expressed in Apis mellifera. This work provides the first detailed characterization of the honeybee T-type CaV3 Ca2+ channel and demonstrates the low toxicity of inhibiting this channel. Comparing Ca2+ currents recorded in bee neurons and myocytes with Ca2+ currents recorded in Xenopus oocytes expressing the honeybee CaV3 gene suggests native expression in bee muscle cells only. High‐voltage activated Ca2+ channels could be recorded in the somata of different cultured bee neurons. These functional data were confirmed by in situ hybridization, immunolocalization and in vivo analysis of the effects of a CaV3 inhibitor. The biophysical and pharmacological characterization and the tissue distribution of CaV3 suggest a role in honeybee muscle function. PMID:28145504

  7. TIR/BB-loop mimetic AS-1 attenuates cardiac ischemia/reperfusion injury via a caveolae and caveolin-3-dependent mechanism

    Science.gov (United States)

    Hu, Yuanping; Zhang, Meiling; Shen, Xin; Dai, Guoliang; Ren, Danyang; Que, Linli; Ha, Tuanzhu; Li, Chuanfu; Xu, Yong; Ju, Wenzheng; Li, Yuehua

    2017-01-01

    AS-1, the TIR/BB loop mimetic, plays a protective role in cardiac ischemia/reperfusion (I/R) but the molecular mechanism remains unclear. The muscle specific caveolin3 (Cav-3) and the caveolae have been found to be critical for cardioprotection. This study aimed to evaluate our hypothesis that caveolae and Cav-3 are essential for AS-1-induced cardioprotection against myocardial I/R injury. To address these issues, we analyzed the involvement of Cav-3 in AS-1 mediated cardioprotection both in vivo and in vitro. We demonstrate that AS-1 administration significantly decreased infarct size, improved cardiac function after myocardial I/R and modulated membrane caveolae and Cav-3 expression in the myocardium. For in vitro studies, AS-1 treatment prevented Cav-3 re-distribution induced by H/R injury. In contrast, disruption of caveolae by MCD treatment or Cav-3 knockdown abolished the protection against H/R-induced myocytes injury by AS-1. Our findings reveal that AS-1 attenuates myocardial I/R injury through caveolae and Cav-3 dependent mechanism. PMID:28291255

  8. Niflumic acid blocks native and recombinant T-type channels.

    Science.gov (United States)

    Balderas, Enrique; Ateaga-Tlecuitl, Rogelio; Rivera, Manuel; Gomora, Juan C; Darszon, Alberto

    2012-06-01

    Voltage-dependent calcium channels are widely distributed in animal cells, including spermatozoa. Calcium is fundamental in many sperm functions such as: motility, capacitation, and the acrosome reaction (AR), all essential for fertilization. Pharmacological evidence has suggested T-type calcium channels participate in the AR. Niflumic acid (NA), a non-steroidal anti-inflammatory drug commonly used as chloride channel blocker, blocks T-currents in mouse spermatogenic cells and Cl(-) channels in testicular sperm. Here we examine the mechanism of NA blockade and explore if it can be used to separate the contribution of different Ca(V)3 members previously detected in these cells. Electrophysiological patch-clamp recordings were performed in isolated mouse spermatogenic cells and in HEK cells heterologously expressing Ca(V)3 channels. NA blocks mouse spermatogenic cell T-type currents with an IC(50) of 73.5 µM, without major voltage-dependent effects. The NA blockade is more potent in the open and in the inactivated state than in the closed state of the T-type channels. Interestingly, we found that heterologously expressed Ca(V)3.1 and Ca(V)3.3 channels were more sensitive to NA than Ca(V)3.2 channels, and this drug substantially slowed the recovery from inactivation of the three isoforms. Molecular docking modeling of drug-channel binding predicts that NA binds preferentially to the extracellular face of Ca(V)3.1 channels. The biophysical characteristics of mouse spermatogenic cell T-type currents more closely resemble those from heterologously expressed Ca(V)3.1 channels, including their sensitivity to NA. As Ca(V)3.1 null mice maintain their spermatogenic cell T-currents, it is likely that a novel Ca(V)3.2 isoform is responsible for them.

  9. Internalization of coxsackievirus A9 is mediated by {beta}2-microglobulin, dynamin, and Arf6 but not by caveolin-1 or clathrin.

    Science.gov (United States)

    Heikkilä, Outi; Susi, Petri; Tevaluoto, Tuire; Härmä, Heidi; Marjomäki, Varpu; Hyypiä, Timo; Kiljunen, Saija

    2010-04-01

    Coxsackievirus A9 (CAV9) is a member of the human enterovirus B species within the Enterovirus genus of the family Picornaviridae. It has been shown to utilize alphaV integrins, particularly alphaVbeta6, as its receptors. The endocytic pathway by which CAV9 enters human cells after the initial attachment to the cell surface has so far been unknown. Here, we present a systematic study concerning the internalization mechanism of CAV9 to A549 human lung carcinoma cells. The small interfering RNA (siRNA) silencing of integrin beta6 subunit inhibited virus proliferation, confirming that alphaVbeta6 mediates the CAV9 infection. However, siRNAs against integrin-linked signaling molecules, such as Src, Fyn, RhoA, phosphatidylinositol 3-kinase, and Akt1, did not reduce CAV9 proliferation, suggesting that the internalization of the virus does not involve integrin-linked signaling events. CAV9 endocytosis was independent of clathrin or caveolin-1 but was restrained by dynasore, an inhibitor of dynamin. The RNA interference silencing of beta2-microglobulin efficiently inhibited virus infection and caused CAV9 to accumulate on the cell surface. Furthermore, CAV9 infection was found to depend on Arf6 as both silencing of this molecule by siRNA and the expression of a dominant negative construct resulted in decreased virus infection. In conclusion, the internalization of CAV9 to A549 cells follows an endocytic pathway that is dependent on integrin alphaVbeta6, beta2-microglobulin, dynamin, and Arf6 but independent of clathrin and caveolin-1.

  10. 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

  11. Chromatography purification of canine adenoviral vectors.

    Science.gov (United States)

    Segura, María Mercedes; Puig, Meritxell; Monfar, Mercè; Chillón, Miguel

    2012-06-01

    Canine adenovirus vectors (CAV2) are currently being evaluated for gene therapy, oncolytic virotherapy, and as vectors for recombinant vaccines. Despite the need for increasing volumes of purified CAV2 preparations for preclinical and clinical testing, their purification still relies on the use of conventional, scale-limited CsCl ultracentrifugation techniques. A complete downstream processing strategy for CAV2 vectors based on membrane filtration and chromatography is reported here. Microfiltration and ultra/diafiltration are selected for clarification and concentration of crude viral stocks containing both intracellular and extracellular CAV2 particles. A DNase digestion step is introduced between ultrafiltration and diafiltration operations. At these early stages, concentration of vector stocks with good recovery of viral particles (above 80%) and removal of a substantial amount of protein and nucleic acid contaminants is achieved. The ability of various chromatography techniques to isolate CAV2 particles was evaluated. Hydrophobic interaction chromatography using a Fractogel propyl tentacle resin was selected as a first chromatography step, because it allows removal of the bulk of contaminating proteins with high CAV2 yields (88%). An anion-exchange chromatography step using monolithic supports is further introduced to remove the remaining contaminants with good recovery of CAV2 particles (58-69%). The main CAV2 viral structural components are visualized in purified preparations by electrophoresis analyses. Purified vector stocks contained intact icosahedral viral particles, low contamination with empty viral capsids (10%), and an acceptable total-to-infectious particle ratio (below 30). The downstream processing strategy that was developed allows preparation of large volumes of high-quality CAV2 stocks.

  12. 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.

  13. Scleroderma-like properties of skin from caveolin-1-deficient mice: implications for new treatment strategies in patients with fibrosis and systemic sclerosis.

    Science.gov (United States)

    Castello-Cros, Remedios; Whitaker-Menezes, Diana; Molchansky, Alex; Purkins, George; Soslowsky, Louis J; Beason, David P; Sotgia, Federica; Iozzo, Renato V; Lisanti, Michael P

    2011-07-01

    Caveolin-1 (Cav-1), the principal structural component of caveolae, participates in the pathogenesis of several fibrotic diseases, including systemic sclerosis (SSc). Interestingly, affected skin and lung samples from patients with SSc show reduced levels of Cav-1, as compared to normal skin. In addition, restoration of Cav-1 function in skin fibroblasts from SSc patients reversed their pro-fibrotic phenotype. Here, we further investigated whether Cav-1 mice are a useful pre-clinical model for studying the pathogenesis of SSc. For this purpose, we performed quantitative transmission electron microscopy, as well as biochemical and immuno-histochemical analysis, of the skin from Cav-1 (-/-) null mice. Using these complementary approaches, we now show that skin from Cav-1 null mice exhibits many of the same characteristics as SSc skin from patients, including a decrease in collagen fiber diameter, increased tensile strength, and stiffness, as well as mononuclear cell infiltration. Furthermore, an increase in autophagy/mitophagy was observed in the stromal cells of the dermis from Cav-1 (-/-) mice. These findings suggest that changes in cellular energy metabolism (e.g., a shift towards aerobic glycolysis) in these stromal cells may be a survival mechanism in this "hostile" or pro-inflammatory microenvironment. Taken together, our results demonstrate that Cav-1 (-/-) null mice are a valuable new pre-clinical model for studying scleroderma. Most importantly, our results suggest that inhibition of autophagy and/or aerobic glycolysis may represent a new promising therapeutic strategy for halting fibrosis in SSc patients. Finally, Cav-1 (-/-) null mice are also a pre-clinical model for a "lethal" tumor micro-environment, possibly explaining the link between fibrosis, tumor progression, and cancer metastasis.

  14. Hyperglycemia and downregulation of caveolin-1 enhance neuregulin-induced demyelination.

    Science.gov (United States)

    Yu, Cuijuan; Rouen, Shefali; Dobrowsky, Rick T

    2008-06-01

    Neuregulins (NRGs) are growth factors which bind to Erb receptor tyrosine kinases that localize to Schwann cells (SCs). Although NRGs can promote cell survival, mitogenesis, and myelination in undifferentiated SCs, they also induce demyelination of myelinated co-cultures of SCs and dorsal root ganglion (DRG) neurons. We have shown previously that Erb B2 activity increased in premyelinating SCs in response to hyperglycemia, and that this correlated with the downregulation of the protein caveolin-1 (Cav-1). As myelinated SCs undergo substantial degeneration in diabetic neuropathy, we used myelinated SC/DRG neuron co-cultures to determine if hyperglycemia and changes in Cav-1 expression could enhance NRG-induced demyelination. In basal glucose, NRG1 caused a 2.4-fold increase in the number of damaged myelin segments. This damage reached 3.8-fold under hyperglycemic conditions, and was also associated with a robust decrease in the expression of Cav-1 and compact myelin proteins. The loss of Cav-1 and compact myelin proteins following hyperglycemia and NRG treatment was not due to neuronal loss, since the axons remained intact and there was no loss of PGP 9.5, an axonal marker protein. To examine if changes in Cav-1 were sufficient to alter the extent of NRG-induced demyelination, SC/DRG neurons co-cultures were infected with antisense or dominant-negative Cav-1(P132L) adenoviruses. Either antisense-mediated downregulation or mis-localization of endogenous Cav-1 by Cav-1(P132L) resulted in a 1.5- to 2.4-fold increase in NRG-induced degeneration compared to that present in control cultures. These data support that hyperglycemia and changes in Cav-1 are sufficient to sensitize myelinated SC/DRG co-cultures to NRG-induced demyelination.

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

    Directory of Open Access Journals (Sweden)

    Jordan D.T. Engbers

    2013-11-01

    Full Text Available 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 IPSPs. 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 effects on

  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

    2013-11-27

    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 (I T) and hyperpolarization-activated cation current (I H) are activated during trains of inhibitory postsynaptic potentials. These currents have distinct, and yet synergistic, roles in the subthreshold domain with I T generating a rebound burst and I H 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 I H to increase the efficacy of I T 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. H2S and Pain: A Novel Aspect for Processing of Somatic, Visceral and Neuropathic Pain Signals.

    Science.gov (United States)

    Terada, Yuka; Kawabata, Atsufumi

    2015-01-01

    Hydrogen sulfide (H2S) formed by multiple enzymes including cystathionine-γ-lyase (CSE) targets Cav3.2 T-type Ca2+ channels (T-channels) and transient receptor potential ankyrin-1 (TRPA1). Intraplantar and intracolonic administration of H2S donors promotes somatic and visceral pain, respectively, via activation of Cav3.2 and TRPA1 in rats and/or mice. Injection of H2S donors into the plantar tissues, pancreatic duct, colonic lumen, or bladder causes T-channel-dependent excitation of nociceptors, determined as phosphorylation of ERK or expression of Fos in the spinal dorsal horn. Electrophysiological studies demonstrate that exogenous and/or endogenous H2S facilitates membrane currents through T-channels in NG108-15 cells and isolated mouse dorsal root ganglion (DRG) neurons that abundantly express Cav3.2 and also in Cav3.2-transfected HEK293 cells. In mice with cerulein-induced pancreatitis and cyclophosphamide-induced cystitis, visceral pain and/or referred hyperalgesia are inhibited by CSE inhibitors and by pharmacological blockade or genetic silencing of Cav3.2, and CSE protein is upregulated in the pancreas and bladder. In rats with neuropathy induced by L5 spinal nerve cutting or by repeated administration of paclitaxel, an anticancer drug, the neuropathic hyperalgesia is reversed by inhibitors of CSE or T-channels and by silencing of Cav3.2. Upregulation of Cav3.2 protein in DRG is detectable in the former, but not in the latter, neuropathic pain models. Thus, H2S appears to function as a nociceptive messenger by facilitating functions of Cav3.2 and TRPA1, and the enhanced function of the CSE/H2S/Cav3.2 pathway is considered to be involved in the pancreatitis- and cystitis-related pain and in neuropathic pain.

  18. PIP2 in pancreatic β-cells regulates voltage-gated calcium channels by a voltage-independent pathway.

    Science.gov (United States)

    de la Cruz, Lizbeth; Puente, Erika I; Reyes-Vaca, Arturo; Arenas, Isabel; Garduño, Julieta; Bravo-Martínez, Jorge; Garcia, David E

    2016-10-01

    Phosphatidylinositol-4,5-bisphosphate (PIP2) is a membrane phosphoinositide that regulates the activity of many ion channels. Influx of calcium primarily through voltage-gated calcium (CaV) channels promotes insulin secretion in pancreatic β-cells. However, whether CaV channels are regulated by PIP2, as is the case for some non-insulin-secreting cells, is unknown. The purpose of this study was to investigate whether CaV channels are regulated by PIP2 depletion in pancreatic β-cells through activation of a muscarinic pathway induced by oxotremorine methiodide (Oxo-M). CaV channel currents were recorded by the patch-clamp technique. The CaV current amplitude was reduced by activation of the muscarinic receptor 1 (M1R) in the absence of kinetic changes. The Oxo-M-induced inhibition exhibited the hallmarks of voltage-independent regulation and did not involve PKC activation. A small fraction of the Oxo-M-induced CaV inhibition was diminished by a high concentration of Ca(2+) chelator, whereas ≥50% of this inhibition was prevented by diC8-PIP2 dialysis. Localization of PIP2 in the plasma membrane was examined by transfecting INS-1 cells with PH-PLCδ1, which revealed a close temporal association between PIP2 hydrolysis and CaV channel inhibition. Furthermore, the depletion of PIP2 by a voltage-sensitive phosphatase reduced CaV currents in a way similar to that observed following M1R activation. These results indicate that activation of the M1R pathway inhibits the CaV channel via PIP2 depletion by a Ca(2+)-dependent mechanism in pancreatic β- and INS-1 cells and thereby support the hypothesis that membrane phospholipids regulate ion channel activity by interacting with ion channels.

  19. 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.

  20. BIN1 localizes the L-type calcium channel to cardiac T-tubules.

    Directory of Open Access Journals (Sweden)

    Ting-Ting Hong

    2010-02-01

    Full Text Available The BAR domain protein superfamily is involved in membrane invagination and endocytosis, but its role in organizing membrane proteins has not been explored. In particular, the membrane scaffolding protein BIN1 functions to initiate T-tubule genesis in skeletal muscle cells. Constitutive knockdown of BIN1 in mice is perinatal lethal, which is associated with an induced dilated hypertrophic cardiomyopathy. However, the functional role of BIN1 in cardiomyocytes is not known. An important function of cardiac T-tubules is to allow L-type calcium channels (Cav1.2 to be in close proximity to sarcoplasmic reticulum-based ryanodine receptors to initiate the intracellular calcium transient. Efficient excitation-contraction (EC coupling and normal cardiac contractility depend upon Cav1.2 localization to T-tubules. We hypothesized that BIN1 not only exists at cardiac T-tubules, but it also localizes Cav1.2 to these membrane structures. We report that BIN1 localizes to cardiac T-tubules and clusters there with Cav1.2. Studies involve freshly acquired human and mouse adult cardiomyocytes using complementary immunocytochemistry, electron microscopy with dual immunogold labeling, and co-immunoprecipitation. Furthermore, we use surface biotinylation and live cell confocal and total internal fluorescence microscopy imaging in cardiomyocytes and cell lines to explore delivery of Cav1.2 to BIN1 structures. We find visually and quantitatively that dynamic microtubules are tethered to membrane scaffolded by BIN1, allowing targeted delivery of Cav1.2 from the microtubules to the associated membrane. Since Cav1.2 delivery to BIN1 occurs in reductionist non-myocyte cell lines, we find that other myocyte-specific structures are not essential and there is an intrinsic relationship between microtubule-based Cav1.2 delivery and its BIN1 scaffold. In differentiated mouse cardiomyocytes, knockdown of BIN1 reduces surface Cav1.2 and delays development of the calcium transient

  1. Localized Calcineurin Confers Ca2+-Dependent Inactivation Upon Neuronal L-Type Ca2+ Channels

    OpenAIRE

    2012-01-01

    Excitation-driven entry of Ca2+ through L-type voltage-gated Ca2+ channels controls gene expression in neurons and a variety of fundamental activities in other kinds of excitable cells. The probability of opening of CaV1.2 L-type channels is subject to pronounced enhancement by cAMP-dependent protein kinase (PKA), which is scaffolded to CaV1.2 channels by A-kinase anchoring proteins (AKAPs). CaV1.2 channels also undergo negative autoregulation via Ca2+-dependent inactivation (CDI), which stro...

  2. 肺心病患者脑循环动力学的研究%Study of cerebrovascular activity in patients with pulmonary heart disease

    Institute of Scientific and Technical Information of China (English)

    李金双; 左剑; 荆春明; 韩云辉

    2002-01-01

    Objective To investigate cerebrovascular activity (CVA) change of pulmonary heart disease patients.Methods To measure CAV of acute stage and compare with CAV of remission stage and control group with CAV tester.Results Average cerebral blood flow velocity,maximal blood velocity,minimal blood velocity and minimal blood flow of pulmonary heart disease patients decrease apparently (P0.05).Conclusion CVA of pulmonary heart disease patients is disorder in different degree,and this disorder aggravate as aggravating of state of disease.

  3. Nuevos registros para la fauna de Saturniidae (Lepidoptera en Argentina New records for the fauna of Saturniidae (Lepidoptera from Argentina

    Directory of Open Access Journals (Sweden)

    Adriana I. Zapata

    2012-12-01

    Full Text Available Se brindan nuevas citas de 17 especies de Saturniidae para la Argentina. Estos registros corresponden a las subfamilias: Arsenurinae [Arsenura xanthopus (Walker y Titaea tamerlan (Maassen]; Ceratocampinae: [Almeidella approximans (Schaus, Cicia nettia (Schaus, Citheronia aroa Schaus, Citheronia maureillei Wolfe & Herbin, Citioica anthonilis (Herrich-Schäffer, Eacles ducalis (Walker, Othorene cadmus (Herrich-Schäffer, Othorene purpurascens (Schaus y Ptiloscola photophila (Rothschild] y Hemileucinae [Cerodirphia brunnea (Draudt, Dirphia curitiba Draudt, Hirpida nigrolinea (Druce, Hylesia ebalus (Cramer, Leucanella gibbosa (Conte y Molippa strigosa (Maassen & Weyding].New records are provided for 17 species of Saturniidae from Argentina. The new records correpond to the following subfamilies: Arsenurinae [Arsenura xanthopus (Walker and Titaea tamerlan (Maassen]; Ceratocampinae [Almeidella approximans (Schaus, Cicia nettia (Schaus, Citheronia aroa Schaus, Citheronia maureillei Wolfe & Herbin, Citioica anthonilis (Herrich-Schäffer, Eacles ducalis (Walker, Othorene cadmus (Herrich-Schäffer, Othorene purpurascens (Schaus and Ptiloscola photophila (Rothschild]; and Hemileucinae [Cerodirphia brunnea (Draudt, Dirphia curitiba Draudt, Hirpida nigrolinea (Druce, Hylesia ebalus (Cramer, Leucanella gibbosa (Conte and Molippa strigosa (Maassen & Weyding].

  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. Effects of litter on substrate conditions and growth of emergent macrophytes

    NARCIS (Netherlands)

    Van der Putten, W.H.; Peters, B.A.M.; Van den Berg, R.S.

    1997-01-01

    Three successive emergent macrophytes (Typha latifolia L., Phragmites australis (Cav.) Steudel and Glyceria maxima (Hartman) Holmbly) were each grown in substrates collected from three different zones of shoreline vegetation development (non- vegetated sediment, the interface between T. latifolia an

  6. Lectotypification of three Iberian endemic species belonging to monotypic genera described by Cosson

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    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.

  7. 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.

  8. 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

  9. N-type calcium channel blockers: novel therapeutics for the treatment of pain.

    Science.gov (United States)

    Schroeder, C I; Doering, C J; Zamponi, G W; Lewis, R J

    2006-09-01

    Highly selective Ca(v)2.2 voltage-gated calcium channel (VGCC) inhibitors have emerged as a new class of therapeutics for the treatment of chronic and neuropathic pain. Cone snail venoms provided the first drug in class with FDA approval granted in 2005 to Prialt (omega-conotoxin MVIIA, Elan) for the treatment of neuropathic pain. Since this pioneering work, major efforts underway to develop alternative small molecule inhibitors of Ca(v)2.2 calcium channel have met with varied success. This review focuses on the properties of the Ca(v)2.2 calcium channel in different pain states, the action of omega-conotoxins GVIA, MVIIA and CVID, describing their structure-activity relationships and potential as leads for the design of improved Ca(v)2.2 calcium channel therapeutics, and finally the development of small molecules for the treatment of chronic pain.

  10. 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

    .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......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...

  11. 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.

  12. Fetal calcium regulates branching morphogenesis in the developing human and mouse lung: involvement of voltage-gated calcium channels.

    Science.gov (United States)

    Brennan, Sarah C; Finney, Brenda A; Lazarou, Maria; Rosser, Anne E; Scherf, Caroline; Adriaensen, Dirk; Kemp, Paul J; Riccardi, Daniela

    2013-01-01

    Airway branching morphogenesis in utero is essential for optimal postnatal lung function. In the fetus, branching morphogenesis occurs during the pseudoglandular stage (weeks 9-17 of human gestation, embryonic days (E)11.5-16.5 in mouse) in a hypercalcaemic environment (~1.7 in the fetus vs. ~1.1-1.3 mM for an adult). Previously we have shown that fetal hypercalcemia exerts an inhibitory brake on branching morphogenesis via the calcium-sensing receptor. In addition, earlier studies have shown that nifedipine, a selective blocker of L-type voltage-gated Ca(2+) channels (VGCC), inhibits fetal lung growth, suggesting a role for VGCC in lung development. The aim of this work was to investigate the expression of VGCC in the pseudoglandular human and mouse lung, and their role in branching morphogenesis. Expression of L-type (CaV1.2 and CaV1.3), P/Q type (CaV2.1), N-type (CaV2.2), R-type (CaV2.3), and T-type (CaV3.2 and CaV3.3) VGCC was investigated in paraffin sections from week 9 human fetal lungs and E12.5 mouse embryos. Here we show, for the first time, that Cav1.2 and Cav1.3 are expressed in both the smooth muscle and epithelium of the developing human and mouse lung. Additionally, Cav2.3 was expressed in the lung epithelium of both species. Incubating E12.5 mouse lung rudiments in the presence of nifedipine doubled the amount of branching, an effect which was partly mimicked by the Cav2.3 inhibitor, SNX-482. Direct measurements of changes in epithelial cell membrane potential, using the voltage-sensitive fluorescent dye DiSBAC2(3), demonstrated that cyclic depolarisations occur within the developing epithelium and coincide with rhythmic occlusions of the lumen, driven by the naturally occurring airway peristalsis. We conclude that VGCC are expressed and functional in the fetal human and mouse lung, where they play a role in branching morphogenesis. Furthermore, rhythmic epithelial depolarisations evoked by airway peristalsis would allow for branching to match

  13. 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

  14. 17β-Estradiol Regulation of the mRNA Expression of T-type Calcium Channel subunits: Role of Estrogen Receptor α and Estrogen Receptor β

    Science.gov (United States)

    Bosch, Martha A.; Hou, Jingwen; Fang, Yuan; Kelly, Martin J.; Rønnekleiv., Oline K.

    2009-01-01

    Low voltage-activated (T-type) calcium channels are responsible for burst firing and transmitter release in neurons and are important for exocytosis and hormone secretion in pituitary cells. T-type channels contain an α1 subunit, of which there are three subtypes, Cav3.1, 3.2 and 3.3, and each subtype has distinct kinetic characteristics. Although 17β-estradiol modulates T-type calcium channel expression and function, little is known about the molecular mechanisms involved. Presently, we used real-time PCR quantification of RNA extracted from hypothalamic nuclei and pituitary in vehicle and E2-treated C57BL/6 mice to elucidate E2-mediated regulation of Cav3.1, 3.2 and 3.3 subunits. The three subunits were expressed in both the hypothalamus and the pituitary. E2 treatment increased the mRNA expression of Cav3.1 and 3.2, but not Cav3.3, in the medial preoptic area and the arcuate nucleus. In the pituitary, Cav3.1 was increased with E2-treatment and Cav3.2 and 3.3 were decreased. In order to examine whether the classical estrogen receptors (ERs) were involved in the regulation, we used ERα- and ERβ-deficient C57BL/6 mice and explored the effects of E2 on T-type channel subtypes. Indeed, we found that the E2-induced increase in Cav3.1 in the hypothalamus was dependent on ERα, whereas the E2 effect on Cav3.2 was dependent on both ERα and ERβ. However, the E2-induced effects in the pituitary were dependent on only the expression of ERα. The robust E2-regulation of the T-type calcium channels could be an important mechanism by which E2 increases the excitability of hypothalamic neurons and modulates pituitary secretion. PMID:19003958

  15. PKC enhances the capacity for secretion by rapidly recruiting covert voltage-gated Ca2+ channels to the membrane.

    Science.gov (United States)

    Groten, Christopher J; Magoski, Neil S

    2015-02-11

    It is unknown whether neurons can dynamically control the capacity for secretion by promptly changing the number of plasma membrane voltage-gated Ca(2+) channels. To address this, we studied peptide release from the bag cell neurons of Aplysia californica, which initiate reproduction by secreting hormone during an afterdischarge. This burst engages protein kinase C (PKC) to trigger the insertion of a covert Ca(2+) channel, Apl Cav2, alongside a basal channel, Apl Cav1. The significance of Apl Cav2 recruitment to secretion remains undetermined; therefore, we used capacitance tracking to assay secretion, along with Ca(2+) imaging and Ca(2+) current measurements, from cultured bag cell neurons under whole-cell voltage-clamp. Activating PKC with the phorbol ester, PMA, enhanced Ca(2+) entry, and potentiated stimulus-evoked secretion. This relied on channel insertion, as it was occluded by preventing Apl Cav2 engagement with prior whole-cell dialysis or the cytoskeletal toxin, latrunculin B. Channel insertion reduced the stimulus duration and/or frequency required to initiate secretion and strengthened excitation-secretion coupling, indicating that Apl Cav2 accesses peptide release more readily than Apl Cav1. The coupling of Apl Cav2 to secretion also changed with behavioral state, as Apl Cav2 failed to evoke secretion in silent neurons from reproductively inactive animals. Finally, PKC also acted secondarily to enhance prolonged exocytosis triggered by mitochondrial Ca(2+) release. Collectively, our results suggest that bag cell neurons dynamically elevate Ca(2+) channel abundance in the membrane to ensure adequate secretion during the afterdischarge.

  16. Hepatic caveolin-1 is enhanced in Cyp27a1/ApoE double knockout mice.

    Science.gov (United States)

    Zurkinden, Line; Mansour, Yosef T; Rohrbach, Beatrice; Vogt, Bruno; Mistry, Hiten D; Escher, Geneviève

    2016-10-01

    Sterol 27-hydroxylase (CYP27A1) is involved in bile acid synthesis and cholesterol homoeostasis. Cyp27a1((-/-))/Apolipoprotein E((-/-)) double knockout mice (DKO) fed a western diet failed to develop atherosclerosis. Caveolin-1 (CAV-1), the main component of caveolae, is associated with lipid homoeostasis and has regulatory roles in vascular diseases. We hypothesized that liver CAV-1 would contribute to the athero-protective mechanism in DKO mice. Cyp27a1((+/+))/ApoE((-/-)) (ApoE KO), Cyp27a1((+/-))/ApoE((-/-)) (het), and DKO mice were fed a western diet for 2 months. Atherosclerotic plaque and CAV-1 protein were quantified in aortas. Hepatic Cav-1 mRNA was assessed using qPCR, CAV-1 protein by immunohistochemistry and western blotting. Total hepatic and plasma cholesterol was measured using chemiluminescence. Cholesterol efflux was performed in RAW264.7 cells, using mice plasma as acceptor. CAV-1 protein expression in aortas was increased in endothelial cells of DKO mice and negatively correlated with plaque surface (P < 0.05). In the liver, both CAV-1 protein and mRNA expression doubled in DKO, compared to ApoE KO and het mice (P < 0.001 for both) and was negatively correlated with total hepatic cholesterol (P < 0.05). Plasma from DKO, ApoE KO and het mice had the same efflux capacity. In the absence of CYP27A1, CAV-1 overexpression might have an additional athero-protective role by partly overcoming the defect in CYP27A1-mediated cholesterol efflux.

  17. Phorbol Ester Modulation of Ca2+ Channels Mediates Nociceptive Transmission in Dorsal Horn Neurones

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    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.

  18. Eps15 Homology Domain-containing Protein 3 Regulates Cardiac T-type Ca2+ Channel Targeting and Function in the Atria*

    Science.gov (United States)

    Curran, Jerry; Musa, Hassan; Kline, Crystal F.; Makara, Michael A.; Little, Sean C.; Higgins, John D.; Hund, Thomas J.; Band, Hamid; Mohler, Peter J.

    2015-01-01

    Proper trafficking of membrane-bound ion channels and transporters is requisite for normal cardiac function. Endosome-based protein trafficking of membrane-bound ion channels and transporters in the heart is poorly understood, particularly in vivo. In fact, for select cardiac cell types such as atrial myocytes, virtually nothing is known regarding endosomal transport. We previously linked the C-terminal Eps15 homology domain-containing protein 3 (EHD3) with endosome-based protein trafficking in ventricular cardiomyocytes. Here we sought to define the roles and membrane protein targets for EHD3 in atria. We identify the voltage-gated T-type Ca2+ channels (CaV3.1, CaV3.2) as substrates for EHD3-dependent trafficking in atria. Mice selectively lacking EHD3 in heart display reduced expression and targeting of both Cav3.1 and CaV3.2 in the atria. Furthermore, functional experiments identify a significant loss of T-type-mediated Ca2+ current in EHD3-deficient atrial myocytes. Moreover, EHD3 associates with both CaV3.1 and CaV3.2 in co-immunoprecipitation experiments. T-type Ca2+ channel function is critical for proper electrical conduction through the atria. Consistent with these roles, EHD3-deficient mice demonstrate heart rate variability, sinus pause, and atrioventricular conduction block. In summary, our findings identify CaV3.1 and CaV3.2 as substrates for EHD3-dependent protein trafficking in heart, provide in vivo data on endosome-based trafficking pathways in atria, and implicate EHD3 as a key player in the regulation of atrial myocyte excitability and cardiac conduction. PMID:25825486

  19. Caveolin-1 enhances resveratrol-mediated cytotoxicity and transport in a hepatocellular carcinoma model

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    Yang Hui-ling

    2009-03-01

    Full Text Available Abstract Background Resveratrol (RES, an estrogen analog, is considered as a potential cancer chemo-preventive agent. However, it remains unclear how RES is transported into cells. In this study, we observed that Caveolin-1(CAV1 expression can increase the cytotoxic and pro-apoptotic activity of RES in a dose- and time-dependent manner both in vitro and in vivo in a Hepatocellular Carcinoma animal model. Methods High performance liquid chromatography (HPLC demonstrated that RES intra-cellular concentration is increased about 2-fold in cells stably expressing CAV1 or CAVM1 (a scaffolding domain (81-101AA-defective CAV1 mutant compared to the untransduced human Hepatoblastoma cell line (HepG2 or after transduction with the green fluorescent protein (GFP control vector. The increased intra-cellular transport of RES was abolished in cells stably expressing CAVM2 (a cholesterol shuttle domain (143-156AA-defective CAV1 mutant or CAVRNAi. In order to further characterize CAV1-dependent RES transport, we synthesized RES-dansyl chloride derivatives as fluorescent probes to visualize the transport process, which demonstrated a distribution consistent with that of CAV1 in HepG2 cells. Results In addition, RES endocytosis was not mediated by estrogen receptor (ER α and β, as suggested by lack of competitive inhibition by estrogen or Tamoxifen. Pathway analysis showed that RES can up-regulate the expression of endogenous CAV1; this activates further the MAPK pathway and caspase-3 expression. Discussion This study provides novel insights about the role played by CAV1 in modulating cellular sensitivity to RES through enhancement of its internalization and trafficking.

  20. Subthreshold membrane potential oscillations in inferior olive neurons are dynamically regulated by P/Q- and T-type calcium channels: a study in mutant mice.

    Science.gov (United States)

    Choi, Soonwook; Yu, Eunah; Kim, Daesoo; Urbano, Francisco J; Makarenko, Vladimir; Shin, Hee-Sup; Llinás, Rodolfo R

    2010-08-15

    The role of P/Q- and T-type calcium channels in the rhythmic oscillatory behaviour of inferior olive (IO) neurons was investigated in mutant mice. Mice lacking either the CaV2.1 gene of the pore-forming alpha1A subunit for P/Q-type calcium channel, or the CaV3.1 gene of the pore-forming alpha1G subunit for T-type calcium channel were used. In vitro intracellular recording from IO neurons reveals that the amplitude and frequency of sinusoidal subthreshold oscillations (SSTOs) were reduced in the CaV2.1-/- mice. In the CaV3.1-/- mice, IO neurons also showed altered patterns of SSTOs and the probability of SSTO generation was significantly lower (15%, 5 of 34 neurons) than that of wild-type (78%, 31 of 40 neurons) or CaV2.1-/- mice (73%, 22 of 30 neurons). In addition, the low-threshold calcium spike and the sustained endogenous oscillation following rebound potentials were absent in IO neurons from CaV3.1-/- mice. Moreover, the phase-reset dynamics of oscillatory properties of single neurons and neuronal clusters in IO were remarkably altered in both CaV2.1-/- and CaV3.1-/- mice. These results suggest that both alpha1A P/Q- and alpha1G T-type calcium channels are required for the dynamic control of neuronal oscillations in the IO. These findings were supported by results from a mathematical IO neuronal model that incorporated T and P/Q channel kinetics.

  1. The evaluation of the caveolin-1 and AT-rich interactive domain 1 alpha expressions in uterine smooth muscle tumors

    OpenAIRE

    2016-01-01

    Objectives: This retrospective study was designed to evaluate the importance of tissue expressions of caveolin-1 (Cav-1) and AT-rich interactive domain 1 alpha (ARID-1A) which are known as signal regulator and tumor suppressor in differential diagnosis of uterine smooth muscle tumors (SMTs). Materials and Methods: Thirty patients recently diagnosed as uterine SMTs at the Tepecik Training and Research Hospital were identified using pathology databases. Immunohistochemical stains for Cav-1 and ...

  2. The evaluation of the caveolin-1 and AT-rich interactive domain 1 alpha expressions in uterine smooth muscle tumors

    Directory of Open Access Journals (Sweden)

    Duygu Ayaz

    2016-01-01

    Full Text Available Objectives: This retrospective study was designed to evaluate the importance of tissue expressions of caveolin-1 (Cav-1 and AT-rich interactive domain 1 alpha (ARID-1A which are known as signal regulator and tumor suppressor in differential diagnosis of uterine smooth muscle tumors (SMTs. Materials and Methods: Thirty patients recently diagnosed as uterine SMTs at the Tepecik Training and Research Hospital were identified using pathology databases. Immunohistochemical stains for Cav-1 and ARID-1A were performed. Results: In this series, there were 10 leiomyosarcomas (LMSs, 10 uterine smooth muscle tumors of uncertain malignant potentials (STUMPs, and 10 leiomyomas (LMs. Cav-1 expression located cytoplasmic or perivascular area. Cytoplasmic Cav-1 expression was determined in 5 LMSs and 2 STUMPs while perivascular Cav-1 expression was determined in 9 LMSs and 2 STUMPs. Statistically, it was determined that if the tumor becomes malignant and more invasive, it gains the perivascular Cav-1 expression (P = 0.029. On the other hand, the mean nuclear staining rate for ARID-1A in LMSs (63 ± 23.4% was higher than both STUMPs (60 ± 18.5% and LMs (34.5 ± 16.5%. Statistically, it was determined that the expression of ARID-1A was significantly downregulated in LMs when compared with STUMPs and LMSs (P = 0.004. Conclusions: Our findings were demonstrated that perivascular Cav-1 expression was seen to be a marker for malignancy of uterine SMTs. Similarly, we found to link of ARID-1A expression and the aggressiveness of SMTs. Therefore, it may be suggested that Cav-1 and ARID-1A may act as predictive biomarkers in uterine SMTs.

  3. H2S does not regulate proliferation via T-type Ca2+ channels.

    Science.gov (United States)

    Elies, Jacobo; Johnson, Emily; Boyle, John P; Scragg, Jason L; Peers, Chris

    2015-06-12

    T-type Ca(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(2+) channels and, in so doing, attenuates proliferation of VSMC. We have also shown that the T-type Ca(2+) channel Cav3.2 is selectively inhibited by hydrogen sulfide (H2S) whilst the other channel isoforms (Cav3.1 and Cav3.3) are unaffected. Here, we explored whether inhibition of Cav3.2 by H2S could account for the anti-proliferative effects of this gasotransmitter. H2S suppressed proliferation in HEK293 cells expressing Cav3.2, as predicted by our previous observations. However, H2S was similarly effective in suppressing proliferation in wild type (non-transfected) HEK293 cells and those expressing the H2S insensitive channel, Cav3.1. Further studies demonstrated that T-type Ca(2+) channels in the smooth muscle cell line A7r5 and in human coronary VSMCs strongly influenced proliferation. In both cell types, H2S caused a concentration-dependent inhibition of proliferation, yet by far the dominant T-type Ca(2+) channel isoform was the H2S-insensitive channel, Cav3.1. Our data indicate that inhibition of T-type Ca(2+) channel-mediated proliferation by H2S is independent of the channels' sensitivity to H2S.

  4. Physiology and evolution of voltage-gated calcium channels in early diverging animal phyla: Cnidaria, Placozoa, Porifera and Ctenophora

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    Adriano Senatore

    2016-11-01

    Full Text Available Voltage-gated calcium (Cav channels serve dual roles in the cell, where they can both depolarize the membrane potential for electrical excitability, and activate transient cytoplasmic Ca2+ signals. In animals, Cav channels play crucial roles including driving muscle contraction (excitation-contraction coupling, gene expression (excitation-transcription coupling, pre-synaptic and neuroendocrine exocytosis (excitation-secretion coupling, regulation of flagellar/ciliary beating, and regulation of cellular excitability, either directly or through modulation of other Ca2+-sensitive ion channels. In recent years, genome sequencing has provided significant insights into the molecular evolution of Cav channels. Furthermore, expanded gene datasets have permitted improved inference of the species phylogeny at the base of Metazoa, providing clearer insights into the evolution of complex animal traits which involve Cav channels, including the nervous system. For the various types of metazoan Cav channels, key properties that determine their cellular contribution include: ion selectivity, pore gating, and, importantly, cytoplasmic protein-protein interactions that direct sub-cellular localization and functional complexing. It is unclear when many of these defining features, many of which are essential for nervous system function, evolved. In this review, we highlight some experimental observations that implicate Cav channels in the physiology and behavior of the most early-diverging animals from the phyla Cnidaria, Placozoa, Porifera and Ctenophora. Given our limited understanding of the molecular biology of Cav channels in these basal animal lineages, we infer insights from better-studied vertebrate and invertebrate animals. We also highlight some apparently conserved cellular functions of Cav channels, which might have emerged very early on during metazoan evolution, or perhaps predated it.

  5. Nerve growth factor blocks the glucose-induced down-regulation of caveolin-1 expression in Schwann cells via p75 neurotrophin receptor signaling.

    Science.gov (United States)

    Tan, Wenbin; Rouen, Shefali; Barkus, Kristin M; Dremina, Yelena S; Hui, Dongwei; Christianson, Julie A; Wright, Douglas E; Yoon, Sung Ok; Dobrowsky, Rick T

    2003-06-20

    Altered neurotrophism in diabetic peripheral neuropathy (DPN) is associated in part with substantial degenerative changes in Schwann cells (SCs) and an increased expression of the p75 neurotrophin receptor (p75NTR). Caveolin-1 (Cav-1) is highly expressed in adult SCs, and changes in its expression can regulate signaling through Erb B2, a co-receptor that mediates the effects of neuregulins in promoting SC growth and differentiation. We examined the hypothesis that hyperglycemia-induced changes in Cav-1 expression and p75NTR signaling may contribute to altered neurotrophism in DPN by modulating SC responses to neuregulins. In an animal model of type 1 diabetes, hyperglycemia induced a progressive decrease of Cav-1 in SCs of sciatic nerve that was reversed by insulin therapy. Treatment of primary neonatal SCs with 20-30 mm d-glucose, but not l-glucose, was sufficient to inhibit transcription from the Cav-1 promoter and decrease Cav-1 mRNA and protein expression. Hyperglycemia prolonged the kinetics of Erb B2 phosphorylation and significantly enhanced the mitogenic response of SCs to neuregulin1-beta1, and this effect was mimicked by the forced down-regulation of Cav-1. Intriguingly, nerve growth factor antagonized the enhanced mitogenic response of SCs to neuregulin1-beta1 and inhibited the glucose-induced down-regulation of Cav-1 transcription, mRNA, and protein expression through p75NTR-dependent activation of JNK. Our data suggest that Cav-1 down-regulation may contribute to altered neurotrophism in DPN by enhancing the response of SCs to neuregulins and that p75NTR-mediated JNK activation may provide a mechanism for the neurotrophic modulation of hyperglycemic stress.

  6. PCB 126 toxicity is modulated by cross-talk between caveolae and Nrf2 signaling.

    Science.gov (United States)

    Petriello, Michael C; Han, Sung Gu; Newsome, Bradley J; Hennig, Bernhard

    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.

  7. Physiology and Evolution of Voltage-Gated Calcium Channels in Early Diverging Animal Phyla: Cnidaria, Placozoa, Porifera and Ctenophora

    Science.gov (United States)

    Senatore, Adriano; Raiss, Hamad; Le, Phuong

    2016-01-01

    Voltage-gated calcium (Cav) channels serve dual roles in the cell, where they can both depolarize the membrane potential for electrical excitability, and activate transient cytoplasmic Ca2+ signals. In animals, Cav channels play crucial roles including driving muscle contraction (excitation-contraction coupling), gene expression (excitation-transcription coupling), pre-synaptic and neuroendocrine exocytosis (excitation-secretion coupling), regulation of flagellar/ciliary beating, and regulation of cellular excitability, either directly or through modulation of other Ca2+-sensitive ion channels. In recent years, genome sequencing has provided significant insights into the molecular evolution of Cav channels. Furthermore, expanded gene datasets have permitted improved inference of the species phylogeny at the base of Metazoa, providing clearer insights into the evolution of complex animal traits which involve Cav channels, including the nervous system. For the various types of metazoan Cav channels, key properties that determine their cellular contribution include: Ion selectivity, pore gating, and, importantly, cytoplasmic protein-protein interactions that direct sub-cellular localization and functional complexing. It is unclear when these defining features, many of which are essential for nervous system function, evolved. In this review, we highlight some experimental observations that implicate Cav channels in the physiology and behavior of the most early-diverging animals from the phyla Cnidaria, Placozoa, Porifera, and Ctenophora. Given our limited understanding of the molecular biology of Cav channels in these basal animal lineages, we infer insights from better-studied vertebrate and invertebrate animals. We also highlight some apparently conserved cellular functions of Cav channels, which might have emerged very early on during metazoan evolution, or perhaps predated it. PMID:27867359

  8. 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

  9. Persistence of chicken anemia virus antigen and inclusions in spontaneous cases of Marek's disease visceral lymphomas in broiler chickens at slaughterhouses.

    Science.gov (United States)

    Ahmed, Mohamed Sabry; Ono, Hiroki; Sasaki, Jun; Ochiai, Kenji; Goryo, Masanobu

    2016-06-01

    The chicken anemia virus (CAV) and Marek's disease virus (MDV) infect chickens worldwide; a single or dual infection by these viruses has a great impact on poultry production. In the present study, we examined the existence of CAV antigen and its inclusions in Marek's disease (MD) lymphomas in chickens in the slaughterhouses of Iwate prefecture, Japan. Forty-nine spleens and 13 livers with different degrees of nodular lesions were histopathologically examined at our laboratory. Grossly, the tested organs showed various sizes and anatomical architectures. Based on the cellular morphology and the infiltrative nature of the neoplastic lymphocytes, MD was confirmed in 76% (37/49) of the spleens and 92% (12/13) of the livers. The lesions of MD, according to the pattern of lymphocytic accumulation in the affected organs, were divided into multifocal, coalesced and diffuse. CAV intranuclear inclusion bodies were detected within the small and the large bizarre lymphocytes of the MD lymphomas in 2 livers and 9 spleens, and the immunostaining test for CAV confirmed the persistence of CAV antigens and inclusions in the neoplastic cells. This study demonstrated the persistence of CAV infection within the neoplastic cells of naturally occurring MD lymphomas in chickens.

  10. Up-regulation of L-type high voltage-gated calcium channel subunits by sustained exposure to 1,4- and 1,5-benzodiazepines in cerebrocortical neurons.

    Science.gov (United States)

    Katsura, Masashi; Shibasaki, Masahiro; Kurokawa, Kazuhiro; Tsujimura, Atsushi; Ohkuma, Seitaro

    2007-12-01

    The aim of this study is to examine how sustained exposure to two 1,4-benzodiazepines (BZDs) with different action period, diazepam and brotizolam, and a 1,5-BZD, clobazam, affects L-type high voltage-gated calcium channel (HVCC) functions and its mechanisms using primary cultures of mouse cerebral cortical neurons. The sustained exposure to these three BZDs increased [(45)Ca2+] influx, which was due to the enhanced [(45)Ca2+] entry through L-type HVCCs but not through of Cav2.1 and Cav2.2. Increase in [(3)H]diltiazem binding after the exposure to these three BZDs was due to the increase in the binding sites of [(3)H]diltiazem. Western blot analysis showed increase of Cav1.2 and Cav1.3 in association with the increased expression of alpha2/delta1 subunit. Similar changes in [(3)H]diltiazem binding and L-type HVCC subunit expression were found in the cerebral cortex from mouse with BZD physical dependence. These results indicate that BZDs examined here have the potential to increase L-type HVCC functions mediated via the enhanced expression of not only Cav1.2 and Cav1.3 but also alpha2/delta1 subunit after their sustained exposure, which may participate in the development of physical dependence by these BZDs.

  11. Crystal structure of dimeric cardiac L-type calcium channel regulatory domains bridged by Ca[superscript 2+]·calmodulins

    Energy Technology Data Exchange (ETDEWEB)

    Fallon, Jennifer L.; Baker, Mariah R.; Xiong, Liangwen; Loy, Ryan E.; Yang, Guojun; Dirksen, Robert T.; Hamilton, Susan L.; Quiocho, Florante A.; (Baylor); (Rochester-Med)

    2009-11-10

    Voltage-dependent calcium channels (Ca(V)) open in response to changes in membrane potential, but their activity is modulated by Ca(2+) binding to calmodulin (CaM). Structural studies of this family of channels have focused on CaM bound to the IQ motif; however, the minimal differences between structures cannot adequately describe CaM's role in the regulation of these channels. We report a unique crystal structure of a 77-residue fragment of the Ca(V)1.2 alpha(1) subunit carboxyl terminus, which includes a tandem of the pre-IQ and IQ domains, in complex with Ca(2+).CaM in 2 distinct binding modes. The structure of the Ca(V)1.2 fragment is an unusual dimer of 2 coiled-coiled pre-IQ regions bridged by 2 Ca(2+).CaMs interacting with the pre-IQ regions and a canonical Ca(V)1-IQ-Ca(2+).CaM complex. Native Ca(V)1.2 channels are shown to be a mixture of monomers/dimers and a point mutation in the pre-IQ region predicted to abolish the coiled-coil structure significantly reduces Ca(2+)-dependent inactivation of heterologously expressed Ca(V)1.2 channels.

  12. Human melanoma cells express FGFR/Src/Rho signaling that entails an adhesion-independent caveolin-1 membrane association.

    Science.gov (United States)

    Fecchi, Katia; Travaglione, Sara; Spadaro, Francesca; Quattrini, Adriano; Parolini, Isabella; Piccaro, Giovanni; Raggi, Carla; Fabbri, Alessia; Felicetti, Federica; Carè, Alessandra; Fiorentini, Carla; Sargiacomo, Massimo

    2012-03-15

    Caveolae have been indicated as a center of cytoskeleton regulation for Src kinase/Rho GTPase signaling. In addition, Src recruitment on intact cortical actin cytoskeleton appears to be required for bFGF/FGFR signal activation. Recently, we established a relationship between caveolin-1 (Cav-1) expression and cell migration in human malignant melanoma, constitutively activated by a bFGF autoregulatory loop. This work intends to investigate whether caveolae's asset, through bFGF/FGFR/c-Src/Rho signaling, could be related to melanoma cell anchorage. Accordingly, we revealed the existence of a FGFR/Src kinase pathway in Cav-1 enriched detergent-resistant membranes (DRMs) of Me665/1 metastatic melanoma cells, as confirmed by FGFR silencing. Moreover, we determined the expression and phosphorylation levels of Cav-1/Src/Erk signal pathway as a function of FGFR activation and cell density. A sucrose density gradient ultracentrifugation was employed to monitor Cav-1 membrane association and buoyancy in Me665/1 cells treated for actin fragmentation or for altered phosphorylation signals. As a result, melanoma cells show remarkable resistance to Cav-1 disassembly, together with persisting cell signal activity, being Src and Cav-1 crucial modulators of Rho GTPases. In conclusion, our study primarily highlights, in a metastatic melanoma cell line expressing caveolin, the circumstances whereby caveola structural and functional endurance enables the FGFR/Src/Rho GTPases pathway to keep on cell progression.

  13. Caveolin-1, caveolin-3 and VEGF expression in the masticatory muscles of mdx mice

    Directory of Open Access Journals (Sweden)

    Alexander Spassov

    2011-07-01

    Full Text Available Duchenne muscular dystrophy (DMD and murine X-linked muscular dystrophy (mdx, its murine model, are characterized by muscle damage and muscle weakness associated with inflammation and new vessel formation. Caveolins, dystrophin-associated proteins, are involved in the pathogenesis of DMD, because increased numbers of caveolae are found in DMD and mdx hindlimb muscles. Caveolae influence angiogenesis due to their content of vascular endothelial growth factor (VEGF receptors. Orofacial muscles in mdx mice undergo muscle necrosis followed by muscle regeneration. To ascertain the role of caveolins and VEGF in the pathogenesis of dystrophic masticatory muscles, we examined the expression of caveolin-1 (cav-1, caveolin-3 (cav-3 and VEGF in control and mdx mice. In mdx masticatory muscles, no changes in transcript and protein levels of VEGF were found, whereas cav-1 and cav-3 expression was increased. Using immunohistochemistry, a strong sarcolemmal staining of caveolin-3 in regenerated muscle fibers was found. Furthermore, immunohistochemistry with the caveolin-1 antibody showed an increase in the amount of blood vessels in areas with regenerating muscle fibers. Dystrophic masticatory muscles showed changes comparable to those of hindlimb muscles in the expression of cav-1 and cav-3. The angiogenesis seems to be unaffected in the jaw muscles of mdx mice. We speculate that the increased caveolin expression could cause extensive and efficient muscle regeneration. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 2, pp. 291–298

  14. Analysis of Morphological Characters and Agronomic Traits of Different Subspecies of Yunjie%芸芥(Eruca Mill.)不同亚种的形态特征及农艺性状分析

    Institute of Scientific and Technical Information of China (English)

    刘雅丽; 孙万仓; 叶剑; 曾军; 张亚宏; 郭秀娟; 杨杰; 魏文惠

    2008-01-01

    在盆栽条件下,研究Eruca vesicaria (L.) Cav. subsps. Vesicaria, Eruca vesicaria (L.) Cav. subsps. sativa和Eruca vesicaria (L.) Cav. subsps. pinnatifida 3个不同芸芥亚种的植物学形态特征和农艺性状.结果表明3个亚种形态特征存在较大差异.Eruca vesicaria (L.) Cav. subsps. vesicaria的叶、薹茎、角果上都密被刺毛,成熟期较一致,且不易落粒,生育期90~100d;而Eruca vesicaria (L.) Cav. subsps. pinatifida薹茎基部密被刺毛而上部较少,种子极小,千粒重0.25g,且易落粒;Eruca vesicaria (L.) Cav. subsps. sativa亚种,角果较长,表面被刺毛或无刺毛,不同材料间生育期差异较大.不同亚种的农艺性状也存在较大差异.

  15. G protein-gated IKACh channels as therapeutic targets for treatment of sick sinus syndrome and heart block.

    Science.gov (United States)

    Mesirca, Pietro; Bidaud, Isabelle; Briec, François; Evain, Stéphane; Torrente, Angelo G; Le Quang, Khai; Leoni, Anne-Laure; Baudot, Matthias; Marger, Laurine; Chung You Chong, Antony; Nargeot, Joël; Striessnig, Joerg; Wickman, Kevin; Charpentier, Flavien; Mangoni, Matteo E

    2016-02-16

    Dysfunction of pacemaker activity in the sinoatrial node (SAN) underlies "sick sinus" syndrome (SSS), a common clinical condition characterized by abnormally low heart rate (bradycardia). If untreated, SSS carries potentially life-threatening symptoms, such as syncope and end-stage organ hypoperfusion. The only currently available therapy for SSS consists of electronic pacemaker implantation. Mice lacking L-type Cav1.3 Ca(2+) channels (Cav1.3(-/-)) recapitulate several symptoms of SSS in humans, including bradycardia and atrioventricular (AV) dysfunction (heart block). Here, we tested whether genetic ablation or pharmacological inhibition of the muscarinic-gated K(+) channel (IKACh) could rescue SSS and heart block in Cav1.3(-/-) mice. We found that genetic inactivation of IKACh abolished SSS symptoms in Cav1.3(-/-) mice without reducing the relative degree of heart rate regulation. Rescuing of SAN and AV dysfunction could be obtained also by pharmacological inhibition of IKACh either in Cav1.3(-/-) mice or following selective inhibition of Cav1.3-mediated L-type Ca(2+) (ICa,L) current in vivo. Ablation of IKACh prevented dysfunction of SAN pacemaker activity by allowing net inward current to flow during the diastolic depolarization phase under cholinergic activation. Our data suggest that patients affected by SSS and heart block may benefit from IKACh suppression achieved by gene therapy or selective pharmacological inhibition.

  16. Caveolin-1-Mediated Expression and Secretion of Kallikrein 6 in Colon Cancer Cells

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    Rebecca S. Henkhaus

    2008-02-01

    Full Text Available Kallikreins are secreted proteases that may play a functional role and/or serve as a serum biomarker for the presence or progression of certain types of cancers. Kallikrein 6 (KLK6 has been shown to be upregulated in several types of cancers, including colon. The aims of this study were to elucidate pathways that influence KLK6 gene expression and KLK6 protein secretion in the HCT116 human colon cancer cells. Our data indicate a central role for caveolin-1 (CAV-1, the main structural protein of caveolae, in both KLK6 gene expression and protein secretion. Sucrose gradient subcellular fractionation reveals that CAV-1 and KLK6 colocalize to lipid raft domains in the plasma membrane of HCT116 cells. Furthermore, we show that CAV-1, although it does not directly interact with the KLK6 molecule, enhances KLK6 secretion from the cells. Deactivation of CAV-1, through SRC-mediated phosphorylation, decreased KLK6 secretion. We also demonstrate that, in colon cancer cells, CAV-1 increased the amount of phosphorylated AKT in cells by inhibiting the activity of the AKT-negative regulators PP1 and PP2A. This study demonstrates that proteins such as CAV-1 and AKT, which are known to be altered in colon cancer, affect KLK6 expression and KLK6 secretion.

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

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    Oya Bulut

    2013-01-01

    Full Text Available 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.

  18. Is two better than one? Limb Activation Treatment combined with Contralesional Arm Vibration to ameliorate signs of left neglect

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    Marco ePitteri

    2013-08-01

    Full Text Available In the present study, we evaluated the effects of the Limb Activation Treatment (LAT alone and in combination with the Contralateral Arm Vibration (CAV on left neglect (LN rehabilitation. We conceived them as techniques that both prompt the activation of the lesioned right hemisphere because of the activation (with the LAT as an active technique and the stimulation (with the CAV as a passive technique of the left hemibody. To test the effect of the simultaneous use of these two techniques (i.e., LAT and CAV on visuo-spatial aspects of LN, we described the case of a LN patient (GR, who showed high intra-individual variability (IIV in performance. Given the high IIV of GR, we used an ABAB repeated-measures design to better define the effectiveness of the combined application of LAT and CAV, as a function of time. The results showed an improvement of GR’s performance on the Bells test following the combined application of LAT and CAV, with respect to the application of LAT alone. We did not find, however, significant effects of treatment on two other LN tests (i.e., Line bisection and Picture scanning. We propose that the combined application of LAT and CAV can be beneficial for some aspects of LN.

  19. 腹泻型肠易激综合征大鼠结肠L-型钙通道α1C及α1D亚基的表达变化%Altered expression of L-type calcium channel α1C and α1D subunits in colon of rats with diarrhea-predominant irritable bowel syndrome

    Institute of Scientific and Technical Information of China (English)

    祝捷; 罗和生; 陈玲; 周婷

    2009-01-01

    Objective To investigate the molecular identities of L-type calcium channel α1C subunit(Cav1.2)and α1D subunit(Cav1.3)responsible for motor dysfunction of diarrhea-predominant imtable bowel syndrome(D-IBS). Methods A total of 30 male Wistar rats were randomly divided into two groups. The traditional method for irritable bowel syndrome in a cold environment and intragagtric administration of Folium Cassiae were combined to develop the D-IBS model. The fecaJ particles of rats and the water content in feces were meagured. Then the expression of Cav1.2 and Cav1.3 mRNA wag detected by reverse transcription polymerasc chain reaction. The expressions of Cav1.2 and Car1.3 were examined by immunohistochemistry in colonic tissues from D-IBS model rats and matched tissues. Results The fecal particles and the water content in feces of D-IBS model rats significantly increaged as compared with the normal rats(6.8±1.4 vs 3.2 ±0.8.P=0.032.80%±4%vs 47%±5%, P=0.018). Cav1.2 and Car1.3 were positively expressed in colon of both model group and control group rats. The immunohistochemical scores of Cav1.2 and Car1.3 expression inereaged in colon of D-IBS model rats as compared with those in normal control rats(3.43 ±0.92 vs 2.82 ±0.60, P=0.034, 4.32 ±0.51 vs 3.75 ±1.05.P=0.039). The immunohistochemical scores of Cav1.3 expression were significantly higher than Cav1.2 in colon of both two group rats(P=0.003, 0.005). Similarly, the expression of Cav1.2 and Cav1.3 mRNA increased in colon of D-IBS model rats as compared with those in normal control rats(1.18 ±0.15 vs 1.06 ±0.12, P=0.023, 1.32 ±0.13 vs 1.23 ±0.13, P=0.033). The expression of Cav1.3 mRNA wag significantly higher than Cav1.2 in colon of both two group rats(P=0.038, 0.012). Conclusions The traditional modeling of irritable bowel syndrome in rats alters the expression of Cav1.2 and Car1.3.It may be directly related to the generation of enhanced colonic contraction in D-IBS. In addition, Cav1.3 may play a more

  20. Концентрация тяжелых металлов в Phragmites australis (Cav. ) Trin. En Steud озера кенон (Забайкальский край)

    OpenAIRE

    Базарова, Б.

    2014-01-01

    Представлены результаты исследования концентрации тяжелых металлов в Phragmites australis озера Кенон, использующегося в качестве водоема охладителя Читинской ТЭЦ-1.

  1. 小窝蛋白-1在膜雌激素受体介导的内皮祖细胞增殖中的作用%Role of caveolin-I on membrane estrogen receptor mediated proliferation of endothelial progenitor cells

    Institute of Scientific and Technical Information of China (English)

    胡飞雪; 王庭槐; 谈智

    2011-01-01

    Objective To investigate the potential role of caveolin-1 ( CAV-1 ) on membrane estrogen receptor (mER) mediated proliferation of endothelial progenitor cells (EPCs).Methods Bone marrow (BM) -derived EPCs were cultured.The proliferation of EPCs induced by estradiol ( E2 ) -BSA in the absence or presence of ICI 182,780 (a pure ER inhibitor),MβCD and CAV-1 siRNA was determined by [3H]-thymidine incorporation.The expression of CAV-1 was detected by Western blot.Results Proliferation of EPC peaked after 10-8 mol/L E2-BSA culture for 24 h (87.5% increase vs.control),and this effect could be inhibited by estrogen receptor blocker ICI 182,780,indicating that mER-initiated membrane signaling pathways was involved in the proliferation effect of estrogen on EPC.Both cholesterol depletion and CAV-1 siRNA significantly attenuated E2-BSA induced [3H ]-thymidine incorporation.Western blot result confirmed that cholesterol depletion or CAV-1 siRNA significantly decreased CAV-1 protein expression ( - 18.6% or -41.2% vs.10-8 mol/L E2-BSA alone).Conclusion Our results suggested that estradiol promoted EPC proliferation through activating CAV-1 pathway.%目的 探讨微囊结构关键蛋白——小窝蛋白-1( caveolin-1,CAV-1)在膜雌激素受体介导的内皮祖细胞(endothelial progenitor cells,EPC)增殖中的作用.方法 培养的EPC分别用不同浓度( 10-9~10-6 mol/L)雌二醇-牛血清白蛋白复合物(E2-BSA)作用24h或10-8 mol/L E2-BSA作用不同时间,或加雌激素受体阻断剂ICI 182,780、环糊精(MβCD)以及CAV-1 siRNA处理,在DMDM培养基中培养的EPC(不加任何试剂)作为对照,使用3H-脱氧胸苷掺入法检测其对EPC增殖的影响.CAV-1 siRNA干扰的效果利用免疫印迹法检测CAV-1蛋白表达来验证.结果 10-8 mol/L E2-BSA作用24h促进EPC增殖作用最大(比对照组高了约87.5%),ICI 182,780可以抑制其增殖作用,表明膜雌激素受体介导的信号通路参与了雌激素对EPC的增殖作用.用环

  2. Co-localization of Caveolin-1 and Extracellular Ca2+-sensing Receptor in Caveolae in Human Umbilical Vein Endothelial Cells%小凹蛋白-1和细胞外钙敏感受体在人脐静脉内皮细胞小凹共定位研究

    Institute of Scientific and Technical Information of China (English)

    龚艳; 胡清华; 钟华; 梁霄; 罗小林; 何芳

    2011-01-01

    前期研究发现小凹蛋白-1(caveolin-1,Cav-1)和细胞外钙敏感受体(extracellular Ca2+-sensing receptor,CaR)在人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)膜上有共定位,且Cav-1下调CaR介导的钙内流.本实验探讨HUVECs中Cav-1与CaR的亚细胞定位,可为进一步机制研究提供理论依据.以3~4代HUVECs随机分为2组:(1)对照组;(2)甲基-β-环糊精(MβCD)处理组,分别采用蔗糖密度梯度离心和Western blot 技术提取含小凹膜碎片和检测Cav-1和CaR蛋白的表达.结果显示,(1) MβCD处理组HUVECs细胞中Cav-1和CaR蛋白的表达与对照组相比无差异(P>0.05).(2)MβCD处理组小凹区Cav-1和CaR蛋白的表达低于对照组(P<0.05).由此可知,Cav-1和CaR蛋白共定位于HUVECs的小凹区,MβCD可抑制两者在小凹的共定位.%Our previous results demonstrated that extracellular Ca2+-sensing receptor (CaR) and caveolin-l(Cav-l) co-localized on the plasma localization of Cav-1 and CaR in HUVECs, which provide a theoretical basis for further functional studies. Methods: The third or fourth passage of HUVECs were randomly divided into two groups:(l) Control groupi (2)methyl-|J-cyelo-dextrin(MpCD) group. We isolated caveolae-enriched membrane fractions from MpCD-treated HUVECs by TritonX-100 sucrose density gradient fractionation and detected the expressions of Cav-1 and CaR protein by Western blot.respectively. Results! (1) The expression of Cav-1 and CaR proteins between control group and MpCD group was no difference (P>0. 05). (2)The expressions of Cav-1 and CaR proteins in caveolae after MpCD treated was significantly reduced(P<0. 05). Conclusion, The present study suggests that CaR and Cav-1 co-localize in caveolae in HUVECS. MpCD reduces the co-localization of Cav-1 and CaR in caveolae.

  3. Upregulation of caveolin-1 and SR-B1 in mice with non-alcoholic fatty liver disease

    Institute of Scientific and Technical Information of China (English)

    Yan Qiu; Shan Liu; Hong-Tan Chen; Chao-Hui Yu; Xiao-Dong Teng; Hong-Tian Yao and Guo-Qiang Xu

    2013-01-01

    BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is one of the most frequent causes of liver diseases, with markedly increased prevalence. However, its mechanisms are not clear. The present study was undertaken to illustrate the role of caveolin-1 (cav1) and the scavenger receptor class B type 1 (SR-B1) in NAFLD. METHODS: Adult male C57BL/6 mice were fed with a normal diet  or  high  fat  and  cholesterol  (HFC)  diet  for  14  weeks.  The mice  were  sacrificed  to  collect  plasma  and  harvest  the  liver; their  plasma  lipid  concentration  was  measured.  Hepatic  cav1 and SR-B1 mRNA and protein expression were determined by real-time  quantitative  polymerase  chain  reaction  (qPCR)  and Western blotting, respectively. In order to study cav1 and SR-B1 distribution and change in hepatocytes, immunohistochemical analysis was performed. RESULTS: HFC diet increased plasma lipids, induced NAFLD and  increased  the  liver/body  weight  ratio.  Compared  to  the control mice (n=6), the mRNA and protein levels of cav1 and SR-B1  in  liver  tissue  of  the  NAFLD  mice  (n=12)  increased significantly (cav1 mRNA: 1.536±0.226 vs 0.980±0.272, P CONCLUSION: NAFLD  is  associated  with  increased  concen-tration  of  plasma  lipids  and  upregulation  of  hepatic  cav1  and SR-B1 gene and protein expressions, which indicate that cav1 and SR-B1 might play crucial roles in the pathogenesis of NAFLD.

  4. Effect of TNF-Alpha on Caveolin-1 Expression and Insulin Signaling During Adipocyte Differentiation and in Mature Adipocytes

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    Sara Palacios-Ortega

    2015-07-01

    Full Text Available Background/Aims: Tumor necrosis factor-α (TNF-α-mediated chronic low-grade inflammation of adipose tissue is associated with obesity and insulin resistance. Caveolin-1 (Cav-1 is the central component of adipocyte caveolae and has an essential role in the regulation of insulin signaling. The effects of TNF-α on Cav-1 expression and insulin signaling during adipocyte differentiation and in mature adipocytes were studied. Methods: 3T3-L1 cells were differentiated (21 days in the presence TNF-α (10 ng/mL and mature adipocytes were also treated with TNF-α for 48 hours. Cav-1 and insulin receptor (IR gene methylation were determined as well as Cav-1, IR, PKB/AKT-2 and Glut-4 expression and activation by real time RT-PCR and western blot. Baseline and insulin-induced glucose uptake was measured by the 2-[C14]-deoxyglucose uptake assay. Results: TNF-α slowed down the differentiation program, hindering the expression of some insulin signaling intermediates without fully eliminating insulin-mediated glucose uptake. In mature adipocytes, TNF-α did not compromise lipid-storage capacity, but downregulated the expression of the insulin signaling intermediates, totally blocking insulin-mediated glucose uptake. Insulin sensitivity correlated with the level of activated phospho-Cav-1 in both situations, strongly suggesting the direct contribution of Cav-1 to the maintenance of this physiological response. Conclusion: Cav-1 activation by phosphorylation seems to be essential for the maintenance of an active and insulin-sensitive glucose uptake.

  5. Evaluation of helper-dependent canine adenovirus vectors in a 3D human CNS model

    Science.gov (United States)

    Simão, Daniel; Pinto, Catarina; Fernandes, Paulo; Peddie, Christopher J.; Piersanti, Stefania; Collinson, Lucy M.; Salinas, Sara; Saggio, Isabella; Schiavo, Giampietro; Kremer, Eric J.; Brito, Catarina; Alves, Paula M.

    2017-01-01

    Gene therapy is a promising approach with enormous potential for treatment of neurodegenerative disorders. Viral vectors derived from canine adenovirus type 2 (CAV-2) present attractive features for gene delivery strategies in the human brain, by preferentially transducing neurons, are capable of efficient axonal transport to afferent brain structures, have a 30-kb cloning capacity and have low innate and induced immunogenicity in pre-clinical tests. For clinical translation, in-depth pre-clinical evaluation of efficacy and safety in a human setting is primordial. Stem cell-derived human neural cells have a great potential as complementary tools by bridging the gap between animal models, which often diverge considerably from human phenotype, and clinical trials. Herein, we explore helper-dependent CAV-2 (hd-CAV-2) efficacy and safety for gene delivery in a human stem cell-derived 3D neural in vitro model. Assessment of hd-CAV-2 vector efficacy was performed at different multiplicities of infection, by evaluating transgene expression and impact on cell viability, ultrastructural cellular organization and neuronal gene expression. Under optimized conditions, hd-CAV-2 transduction led to stable long-term transgene expression with minimal toxicity. hd-CAV-2 preferentially transduced neurons, while human adenovirus type 5 (HAdV5) showed increased tropism towards glial cells. This work demonstrates, in a physiologically relevant 3D model, that hd-CAV-2 vectors are efficient tools for gene delivery to human neurons, with stable long-term transgene expression and minimal cytotoxicity. PMID:26181626

  6. 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.

  7. Caveolin-1 influences human influenza A virus (H1N1 multiplication in cell culture

    Directory of Open Access Journals (Sweden)

    Hemgård Gun-Viol

    2010-05-01

    Full Text Available Abstract Background The threat of recurring influenza pandemics caused by new viral strains and the occurrence of escape mutants necessitate the search for potent therapeutic targets. The dependence of viruses on cellular factors provides a weak-spot in the viral multiplication strategy and a means to interfere with viral multiplication. Results Using a motif-based search strategy for antiviral targets we identified caveolin-1 (Cav-1 as a putative cellular interaction partner of human influenza A viruses, including the pandemic influenza A virus (H1N1 strains of swine origin circulating from spring 2009 on. The influence of Cav-1 on human influenza A/PR/8/34 (H1N1 virus replication was determined in inhibition and competition experiments. RNAi-mediated Cav-1 knock-down as well as transfection of a dominant-negative Cav-1 mutant results in a decrease in virus titre in infected Madin-Darby canine kidney cells (MDCK, a cell line commonly used in basic influenza research as well as in virus vaccine production. To understand the molecular basis of the phenomenon we focussed on the putative caveolin-1 binding domain (CBD located in the lumenal, juxtamembranal portion of the M2 matrix protein which has been identified in the motif-based search. Pull-down assays and co-immunoprecipitation experiments showed that caveolin-1 binds to M2. The data suggest, that Cav-1 modulates influenza virus A replication presumably based on M2/Cav-1 interaction. Conclusion As Cav-1 is involved in the human influenza A virus life cycle, the multifunctional protein and its interaction with M2 protein of human influenza A viruses represent a promising starting point for the search for antiviral agents.

  8. 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.

  9. Electrical coupling between the human serotonin transporter and voltage-gated Ca(2+) channels.

    Science.gov (United States)

    Ruchala, Iwona; Cabra, Vanessa; Solis, Ernesto; Glennon, Richard A; De Felice, Louis J; Eltit, Jose M

    2014-07-01

    Monoamine transporters have been implicated in dopamine or serotonin release in response to abused drugs such as methamphetamine or ecstasy (MDMA). In addition, monoamine transporters show substrate-induced inward currents that may modulate excitability and Ca(2+) mobilization, which could also contribute to neurotransmitter release. How monoamine transporters modulate Ca(2+) permeability is currently unknown. We investigate the functional interaction between the human serotonin transporter (hSERT) and voltage-gated Ca(2+) channels (CaV). We introduce an excitable expression system consisting of cultured muscle cells genetically engineered to express hSERT. Both 5HT and S(+)MDMA depolarize these cells and activate the excitation-contraction (EC)-coupling mechanism. However, hSERT substrates fail to activate EC-coupling in CaV1.1-null muscle cells, thus implicating Ca(2+) channels. CaV1.3 and CaV2.2 channels are natively expressed in neurons. When these channels are co-expressed with hSERT in HEK293T cells, only cells expressing the lower-threshold L-type CaV1.3 channel show Ca(2+) transients evoked by 5HT or S(+)MDMA. In addition, the electrical coupling between hSERT and CaV1.3 takes place at physiological 5HT concentrations. The electrical coupling between monoamine neurotransmitter transporters and Ca(2+) channels such as CaV1.3 is a novel mechanism by which endogenous substrates (neurotransmitters) or exogenous substrates (like ecstasy) could modulate Ca(2+)-driven signals in excitable cells.

  10. Reduced caveolin-1 promotes hyper-inflammation due to abnormal heme oxygenase-1 localizationin LPS challenged macrophages with dysfunctional CFTR

    Science.gov (United States)

    Zhang, Ping-Xia; Murray, Thomas S.; Villella, Valeria Rachela; Ferrari, Eleonora; Esposito, Speranza; D'Souza, Anthony; Raia, Valeria; Maiuri, Luigi; Krause, Diane S.; Egan, Marie E.; Bruscia, Emanuela M.

    2013-01-01

    We have previously reported that TLR4 signaling is increased in lipopolysaccharide (LPS) -stimulated Cystic Fibrosis (CF) macrophages (MΦs), contributing to the robust production of pro-inflammatory cytokines. The heme oxygenase (HO-1)/carbon monoxide (CO) pathway modulates cellular redox status, inflammatory responses, and cell survival. The HO-1 enzyme, together with the scaffold protein caveolin 1 (CAV-1), also acts as a negative regulator of TLR4 signaling in MΦs. Here, we demonstrate that in LPS-challenged CF MΦs, HO-1 does not compartmentalize normally to the cell surface and instead accumulates intracellularly. The abnormal HO-1 localization in CF MΦs in response to LPS is due to decreased CAV-1 expression, which is controlled by the cellular oxidative state, and is required for HO-1 delivery to the cell surface. Overexpression of HO-1 or stimulating the pathway with CO-releasing molecules (CORM2)enhancesCAV-1 expression in CF MΦs, suggesting a positive-feed forward loop between HO-1/CO induction and CAV-1 expression. These manipulations reestablished HO-1 and CAV-1 cell surface localization in CF MΦ's. Consistent with restoration of HO-1/CAV-1 negative regulation of TLR4 signaling, genetic or pharmacological (CORM2)-induced enhancement of this pathway decreased the inflammatory response of CF MΦs and CF mice treated with LPS. In conclusion, our results demonstrate that the counter-regulatory HO-1/CO pathway, which is critical in balancing and limiting the inflammatory response, is defective in CF MΦs through a CAV-1-dependent mechanism, exacerbating the CF MΦ's response to LPS. This pathway could be a potential target for therapeutic intervention for CF lung disease. PMID:23606537

  11. Study on expression and methylation of caveolin 1 gene in non-small cell lung cancers%非小细胞肺癌中窖蛋白1基因的蛋白表达和甲基化及其意义

    Institute of Scientific and Technical Information of China (English)

    张玉霞; 陈洪雷; 叶波; 杨飞; 喻伦银

    2008-01-01

    目的 检测窖蛋白1(Car-1)在非小细胞肺癌(NSCLC)中的蛋白表达以及启动子的甲基化状况,探讨Cav-1基因在NSCLC中的作用及其临床意义.方法 应用免疫组织化学(sP法)和量子点Qd600染色检测123例NSCLC组织、17例良性病变肺组织中Cav一1蛋白表达和亚细胞定位.亚硫酸氢钠处理DNA,甲基化特异性PCR(MSP)检测Cav-1基因启动子区域的甲基化水平.结果 Cav-1蛋白在肺支气管黏膜上皮细胞、肺泡上皮细胞、毛细血管内皮细胞、成纤维细胞、平滑肌细胞的胞质和胞膜高表达.癌旁组织(对照)组和肺癌组中Cav-1蛋白的阳性率分别为17/17、43.1%(53/123),两组间差异有统计学意义(P=0.001);Cav-1蛋白在NSCLC不同的组织学类型(P=0.552)和分化程度(P=0.160)中差异均无统计学意义.Cav-1蛋白阳性率与NSCLC的TNM分期(P=0.001)以及淋巴结转移(P=0.001)均相关.在40例Cav-1蛋白表达为阴性的肺癌组织和12例癌旁肺组织,MSP法均未检测到Cav-1因启动子区域的甲基化.结论 Cav-1蛋白失表达的机制可能与启动子区是否甲基化无关.Cav-1蛋白高表达预示NSCLC恶化进展和高侵袭性.%Objective To study the methylation of CpG islands in the promoter region, expression of caveolin 1(Cav-1)gene and their clinical significance in non. small cell lung cancers(NSCLC). Methods Immunohistochemistry and quanta Qd600 staining were used to detect the expression of Cav-1 in tissues from benign lung lesions(n=17)and NSCLC(n=123). DNA was treated with sodium bisulfite and the Cav-1 promoter region was screened using methylation. specmc polymerase chain reaction for the possible methylation sites. Results Cav. 1 protein was highly expressed in cytoplasm and cell membrane of normal bronchial epithelium, alveolar epithelium, endothelial cells. fibroblasts and smooth muscle cells. The expression rates of Cav-1 protein were 100%(17/17)in the control group and 43. 1%(53/123)in the NSCLC group(P=0. 001

  12. Role of caveolin-1 in down-regulating extracellular Ca2+-sensing receptor-mediated Ca2+ influx in human umbilical vein endothelial cells%小凹蛋白-1下调人脐静脉内皮细胞外钙敏感受体介导的钙内流的作用机制

    Institute of Scientific and Technical Information of China (English)

    钟华; 龚艳; 吴玲玲; 赵慧; 王静; 孙志萍; 邓峰美; 何芳

    2013-01-01

    AIM: To study the role of caveolin-1 (Cav-1) in down-regulating the extracellular Ca2+-sensing receptor ( CaR)-mediated Ca2+ influx in human umbilical vein endothelial cells (HUVECs) and its mechanisms. METHODS : HUVECs were collected and cultured to the second or third passage. Filipin was used to induce acute caveolae disruption. Methyl-p-cyclodextrin (M[$CD) or shRNA targeting Cav-1 combined with CaR agonist spermine and negative al-losteric modulator Calhex 231 was also used in HUVECs. Intracellular concentration of Ca2+ ([ Ca2+ ] i) was measured by Fura-2/AM loading. The protein expression of Cav-1 and CaR was examined by Western blotting. The interaction and co-localization of Cav-1 and CaR were determined by the method of co-immunoprecipitation (Co-IP). Caveolae-enriched membrane (CEM) fractions were isolated and identified by detergent-free (Na2CO3) sucrose density gradient centrifugation. The protein levels of Cav-1, CaR, flotillin-1,β-coat protein (β-COP) , β-actin and transferrin receptor (TfR) were detec- ted by Western blotting. Noncaveolar fraction I (NCF I) and noncaveolar fraction II (NCF II) in the CEM fractions were separated. RESULTS; Using extracellular buffer with Ca2+ , the increase in [Ca2+ ]i induced by spermine in HUVECs was abolished after inhibition of CaR by its negative allosteric modulator calhex231. Conversely, the effect of spermine on the increased [ Ca 2+], in HUVECs was further augmented after acute caveolae disruption by MβCD. No significant difference of the protein levels of CaR and Cav-1 in HUVECs among treating with different concentrations of MβCD was observed. The results of Co-IP showed that the protein levels of CaR and Cav-1 in every group of HUVECs were not significantly different. Compared with control group, the protein expression of CaR and Cav-1 in CEM was decreased in spermine + Ca2+ group, filipin + spermine + Ca2 + group and MfiCD + spermine + Ca2 + group, and that in NCF I was increased. However, the

  13. 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

  14. 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.

  15. 17β-estradiol regulation of T-type calcium channels in gonadotropin-releasing hormone neurons

    Science.gov (United States)

    Zhang, Chunguang; Bosch, Martha A.; Rick, Elizabeth A.; Kelly, Martin J.; Ronnekleiv, Oline K.

    2009-01-01

    T-type calcium channels are responsible for generating low-threshold spikes that facilitate burst firing and neurotransmitter release in neurons. GnRH neurons exhibit burst firing, but the underlying conductances are not known. Previously, we have found that 17β-estradiol (E2) increases T-type channel expression and excitability of hypothalamic arcuate nucleus neurons. Therefore, we used ovariectomized oil- or E2-treated EGFP-GnRH mice to explore the expression and E2-regulation of T-type channels in GnRH neurons. Based on single cell RT-PCR and real-time PCR quantification of the T-type channel α1-subunits, we found that all three subunits were expressed in GnRH neurons with Cav3.3≥Cav3.2>Cav3.1. The mRNA expression of the three subunits was increased with surge-inducing levels of E2 during the morning. During the afternoon, Cav3.3 mRNA expression remained elevated, whereas Cav3.1 and Cav3.2 were decreased. The membrane estrogen receptor agonist STX increased the expression of Cav3.3, but not Cav3.2 in GnRH neurons. Whole-cell patch recordings in GnRH neurons revealed that E2 treatment significantly augmented T-type current density at both time-points, and increased the rebound excitation during the afternoon. Although E2 regulated the mRNA expression of all three subunits in GnRH neurons, the increased expression combined with the slower inactivation kinetics of the T-type current indicates that Cav3.3 may be the most important for bursting activity associated with the GnRH/LH surge. The E2-induced increase in mRNA expression, which depends in part on membrane-initiated signaling, leads to increased channel function and neuronal excitability, and could be a mechanism by which E2 facilitates burst firing and cyclic GnRH neurosecretion. PMID:19710308

  16. 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

  17. Rem uncouples excitation–contraction coupling in adult skeletal muscle fibers

    Science.gov (United States)

    Beqollari, Donald; Romberg, Christin F.; Filipova, Dilyana; Meza, Ulises; Papadopoulos, Symeon

    2015-01-01

    In skeletal muscle, excitation–contraction (EC) coupling requires depolarization-induced conformational rearrangements in L-type Ca2+ channel (CaV1.1) to be communicated to the type 1 ryanodine-sensitive Ca2+ release channel (RYR1) of the sarcoplasmic reticulum (SR) via transient protein–protein interactions. Although the molecular mechanism that underlies conformational coupling between CaV1.1 and RYR1 has been investigated intensely for more than 25 years, the question of whether such signaling occurs via a direct interaction between the principal, voltage-sensing α1S subunit of CaV1.1 and RYR1 or through an intermediary protein persists. A substantial body of evidence supports the idea that the auxiliary β1a subunit of CaV1.1 is a conduit for this intermolecular communication. However, a direct role for β1a has been difficult to test because β1a serves two other functions that are prerequisite for conformational coupling between CaV1.1 and RYR1. Specifically, β1a promotes efficient membrane expression of CaV1.1 and facilitates the tetradic ultrastructural arrangement of CaV1.1 channels within plasma membrane–SR junctions. In this paper, we demonstrate that overexpression of the RGK protein Rem, an established β subunit–interacting protein, in adult mouse flexor digitorum brevis fibers markedly reduces voltage-induced myoplasmic Ca2+ transients without greatly affecting CaV1.1 targeting, intramembrane gating charge movement, or releasable SR Ca2+ store content. In contrast, a β1a-binding–deficient Rem triple mutant (R200A/L227A/H229A) has little effect on myoplasmic Ca2+ release in response to membrane depolarization. Thus, Rem effectively uncouples the voltage sensors of CaV1.1 from RYR1-mediated SR Ca2+ release via its ability to interact with β1a. Our findings reveal Rem-expressing adult muscle as an experimental system that may prove useful in the definition of the precise role of the β1a subunit in skeletal-type EC coupling. PMID:26078055

  18. Early Inflammatory Markers Are Independent Predictors of Cardiac Allograft Vasculopathy in Heart-Transplant Recipients

    Science.gov (United States)

    Labarrere, Carlos A.; Woods, John R.; Hardin, James W.; Jaeger, Beate R.; Zembala, Marian; Deng, Mario C.; Kassab, Ghassan S.

    2014-01-01

    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 improves

  19. Gene splicing of an invertebrate beta subunit (LCavβ in the N-terminal and HOOK domains and its regulation of LCav1 and LCav2 calcium channels.

    Directory of Open Access Journals (Sweden)

    Taylor F Dawson

    Full Text Available The accessory beta subunit (Ca(vβ of calcium channels first appear in the same genome as Ca(v1 L-type calcium channels in single-celled coanoflagellates. The complexity of this relationship expanded in vertebrates to include four different possible Ca(vβ subunits (β1, β2, β3, β4 which associate with four Ca(v1 channel isoforms (Ca(v1.1 to Ca(v1.4 and three Ca(v2 channel isoforms (Ca(v2.1 to Ca(v2.3. Here we assess the fundamentally-shared features of the Ca(vβ subunit in an invertebrate model (pond snail Lymnaea stagnalis that bears only three homologous genes: (LCa(v1, LCa(v2, and LCa(vβ. Invertebrate Ca(vβ subunits (in flatworms, snails, squid and honeybees slow the inactivation kinetics of Ca(v2 channels, and they do so with variable N-termini and lacking the canonical palmitoylation residues of the vertebrate β2a subunit. Alternative splicing of exon 7 of the HOOK domain is a primary determinant of a slow inactivation kinetics imparted by the invertebrate LCa(vβ subunit. LCa(vβ will also slow the inactivation kinetics of LCa(v3 T-type channels, but this is likely not physiologically relevant in vivo. Variable N-termini have little influence on the voltage-dependent inactivation kinetics of differing invertebrate Ca(vβ subunits, but the expression pattern of N-terminal splice isoforms appears to be highly tissue specific. Molluscan LCa(vβ subunits have an N-terminal "A" isoform (coded by exons: 1a and 1b that structurally resembles the muscle specific variant of vertebrate β1a subunit, and has a broad mRNA expression profile in brain, heart, muscle and glands. A more variable "B" N-terminus (exon 2 in the exon position of mammalian β3 and has a more brain-centric mRNA expression pattern. Lastly, we suggest that the facilitation of closed-state inactivation (e.g. observed in Ca(v2.2 and Ca(vβ3 subunit combinations is a specialization in vertebrates, because neither snail subunit (LCa(v2 nor LCa(vβ appears to be compatible

  20. Prevalence and characterization of enterovirus infections among pediatric patients with hand foot mouth disease, herpangina and influenza like illness in Thailand, 2012.

    Science.gov (United States)

    Puenpa, Jiratchaya; Mauleekoonphairoj, John; Linsuwanon, Piyada; Suwannakarn, Kamol; Chieochansin, Thaweesak; Korkong, Sumeth; Theamboonlers, Apiradee; Poovorawan, Yong

    2014-01-01

    Hand, foot, and mouth disease (HFMD) and herpangina are common infectious diseases caused by several genotypes of human enterovirus species A and frequently occurring in young children. This study was aimed at analyzing enteroviruses from patients with these diseases in Thailand in 2012. Detection and genotype determination of enteroviruses were accomplished by reverse transcription-polymerase chain reaction and sequencing of the VP1 region. Enterovirus-positive samples were differentiated into 17 genotypes (coxsackievirus A4 (CAV4), A5, A6, A8, A9, A10, A12, A16, A21, B1, B2, B4, B5, echovirus 7, 16, 25 and Enterovirus 71). The result showed CAV6 (33.5%), followed by CAV16 (9.4%) and EV71 (8.8%) as the most frequent genotypes in HFMD, CAV8 (19.3%) in herpangina and CAV6 (1.5%) in influenza like illness. Enterovirus infections were most prevalent during July with 34.4% in HFMD, 39.8% in herpangina and 1.6% in ILI. The higher enterovirus infection associated with HFMD and herpangina occurred in infants over one year-old. This represents the first report describing the circulation of multiple enteroviruses in Thailand.

  1. 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.

  2. 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.

  3. An optimized polymerase chain reaction assay to identify avian virus vaccine contamination with Chicken anemia virus.

    Science.gov (United States)

    Amer, Haitham M; Elzahed, Hanan M; Elabiare, Elham A; Badawy, Ahmed A; Yousef, Ausama A

    2011-01-01

    The use of embryonating chicken eggs in preparation of avian virus vaccines is the principle cause for contamination with Chicken anemia virus (CAV). Identification of CAV in contaminated vaccines relies on the expensive, tedious, and time-consuming practice of virus isolation in lymphoblastoid cell lines. The experience of the last 2 decades indicates that polymerase chain reaction is extending to replace most of the classic methods for detection of infectious agents. In the present report, a simple, rapid, and accurate polymerase chain reaction method for detection of CAV in poultry vaccines is described. Oligonucleotide primers homologous to highly conserved sequences of the VP1 gene were used to amplify a fragment of 676 bp. The developed assay was specific for detecting CAV from different sources, with no cross reactivity with many avian viruses. No inter- and intra-assay variations were observed. The analytical sensitivity of the test was high enough to detect 5 TCID(50) (50% tissue culture infective dose) of the virus per reaction; however, different factors related to the vaccine matrix showed considerable effects on the detection limit. In conclusion, this method may represent a suitable alternative to virus isolation for identification of CAV contamination of poultry virus vaccines.

  4. Vesicular transport of a ribonucleoprotein to mitochondria

    Directory of Open Access Journals (Sweden)

    Joyita Mukherjee

    2014-10-01

    Full Text Available Intracellular trafficking of viruses and proteins commonly occurs via the early endosome in a process involving Rab5. The RNA Import Complex (RIC-RNA complex is taken up by mammalian cells and targeted to mitochondria. Through RNA interference, it was shown that mito-targeting of the ribonucleoprotein (RNP was dependent on caveolin 1 (Cav1, dynamin 2, Filamin A and NSF. Although a minor fraction of the RNP was transported to endosomes in a Rab5-dependent manner, mito-targeting was independent of Rab5 or other endosomal proteins, suggesting that endosomal uptake and mito-targeting occur independently. Sequential immunoprecipitation of the cytosolic vesicles showed the sorting of the RNP away from Cav1 in a process that was independent of the endosomal effector EEA1 but sensitive to nocodazole. However, the RNP was in two types of vesicle with or without Cav1, with membrane-bound, asymmetrically orientated RIC and entrapped RNA, but no endosomal components, suggesting vesicular sorting rather than escape of free RNP from endosomes. In vitro, RNP was directly transferred from the Type 2 vesicles to mitochondria. Live-cell imaging captured spherical Cav1− RNP vesicles emerging from the fission of large Cav+ particles. Thus, RNP appears to traffic by a different route than the classical Rab5-dependent pathway of viral transport.

  5. The activity of the epithelial sodium channels is regulated by caveolin-1 via a Nedd4-2-dependent mechanism.

    Science.gov (United States)

    Lee, Il-Ha; Campbell, Craig R; Song, Sung-Hee; Day, Margot L; Kumar, Sharad; Cook, David I; Dinudom, Anuwat

    2009-05-08

    It has recently been shown that the epithelial Na(+) channel (ENaC) is compartmentalized in caveolin-rich lipid rafts and that pharmacological depletion of membrane cholesterol, which disrupts lipid raft formation, decreases the activity of ENaC. Here we show, for the first time, that a signature protein of caveolae, caveolin-1 (Cav-1), down-regulates the activity and membrane surface expression of ENaC. Physical interaction between ENaC and Cav-1 was also confirmed in a coimmunoprecipitation assay. We found that the effect of Cav-1 on ENaC requires the activity of Nedd4-2, a ubiquitin protein ligase of the Nedd4 family, which is known to induce ubiquitination and internalization of ENaC. The effect of Cav-1 on ENaC requires the proline-rich motifs at the C termini of the beta- and gamma-subunits of ENaC, the binding motifs that mediate interaction with Nedd4-2. Taken together, our data suggest that Cav-1 inhibits the activity of ENaC by decreasing expression of ENaC at the cell membrane via a mechanism that involves the promotion of Nedd4-2-dependent internalization of the channel.

  6. Triptolide inhibits the migration and invasion of human prostate cancer cells via Caveolin-1/CD147/MMPs pathway.

    Science.gov (United States)

    Yuan, Shiqi; Wang, Liping; Chen, Xixi; Fan, Bo; Yuan, Qingmin; Zhang, Han; Yang, Deyong; Wang, Shujing

    2016-12-01

    Prostate cancer (PCa) is the second most common type of carcinoma and the 5th leading cause of cancer-related death in males. Triptolide, is a main and effective component of Tripterygium wilfordii Hook F, which exerts an broad-spectrum anti-malignant tumor function. However, the effect of triptolide on migration and invasion of human prostate cancer cells is still poorly understood. In this study, we demonstrated that triptolide significantly inhibited the proliferation, migration and invasion of prostate cancer cells in a time- and dose-dependent manner. Caveolin-1 (Cav-1) is regarded as a major structural protein of caveolae and participated in lipid transport, signal transduction and tumor progression. Triptolide treatment inhibited the expression of tumor promoter Cav-1 and reduced CD147 and MMPs activities at both mRNA and protein levels. Meanwhile, triptolide treatment combined with Cav-1 knockdown in PCa cells enhanced the effects of anti-migration and anti-invasion, and those effects were restored following Cav-1-rescued. Together, our research indicates that triptolide represses the migration and invasion through Cav-1/CD147/MMPs pathway in PCa cells, which gives a better understanding of triptolide in clinical aggressive prostate cancer therapy.

  7. Exposure to extremely low-frequency electromagnetic fields inhibits T-type calcium channels via AA/LTE4 signaling pathway.

    Science.gov (United States)

    Cui, Yujie; Liu, Xiaoyu; Yang, Tingting; Mei, Yan-Ai; Hu, Changlong

    2014-01-01

    Extremely low-frequency electromagnetic fields (ELF-EMF) causes various biological effects through altering intracellular calcium homeostasis. The role of high voltage-gated (HVA) calcium channels in ELF-EMF induced effects has been extensively studied. However, the effect of ELF-EMF on low-voltage-gated (LVA) T-type calcium channels has not been reported. In this study, we test the effect of ELF-EMF (50Hz) on human T-type calcium channels transfected in HEK293 cells. Conversely to its stimulant effects on HVA channels, ELF-EMF exposure inhibited all T-type (Cav3.1, Cav3.2 and Cav3.3) channels. Neither the protein expression nor the steady-state activation and inactivation kinetics of Cav3.2 channels were altered by ELF-EMF (50Hz, 0.2mT) exposure. Exposure to ELF-EMF increased both arachidonic acid (AA) and leukotriene E4 (LTE4) levels in HEK293 cells. CAY10502 and bestatin, which block the increase of AA and LTE4 respectively, abrogated the ELF-EMF inhibitory effect on Cav3.2 channels. Exogenous LTE4 mimicked the ELF-EMF inhibition of T-type calcium channels. ELF-EMF (50Hz) inhibits native T-type calcium channels in primary cultured mouse cortical neurons via LTE4. We conclude that 50Hz ELF-EMF inhibits T-type calcium channels through AA/LTE4 signaling pathway.

  8. High Mobility Group Box Protein 1 Boosts Endothelial Albumin Transcytosis through the RAGE/Src/Caveolin-1 Pathway

    Science.gov (United States)

    Shang, Dan; Peng, Tao; Gou, Shanmiao; Li, Yiqing; Wu, Heshui; Wang, Chunyou; Yang, Zhiyong

    2016-01-01

    High-mobility group box protein 1 (HMGB1), an inflammatory mediator, has been reported to destroy cell-cell junctions, resulting in vascular endothelial hyperpermeability. Here, we report that HMGB1 increases the endothelial transcytosis of albumin. In mouse lung vascular endothelial cells (MLVECs), HMGB1 at a concentration of 500 ng/ml or less did not harm cell-cell junctions but rapidly induced endothelial hyperpermeability to 125I-albumin. HMGB1 induced an increase in 125I-albumin and AlexaFluor 488-labeled albumin internalization in endocytosis assays. Depletion of receptor for advanced glycation end products (RAGE), but not TLR2 or TLR4, suppressed HMGB1-induced albumin transcytosis and endocytosis. Genetic and pharmacological destruction of lipid rafts significantly inhibited HMGB1-induced albumin endocytosis and transcytosis. HMGB1 induced the rapid phosphorylation of caveolin (Cav)-1 and Src. Either RAGE gene silencing or soluble RAGE suppressed Cav-1 Tyr14 phosphorylation and Src Tyr418 phosphorylation. The Src inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d] pyrimidine (PP2) blocked HMGB1-induced Cav-1 Tyr14 phosphorylation. PP2 and overexpression of Cav-1 with a T14F mutation significantly inhibited HMGB1-induced transcytosis and albumin endocytosis. Our findings suggest that HMGB1 induces the transcytosis of albumin via RAGE-dependent Src phosphorylation and Cav-1 phosphorylation. These studies revealed a new mechanism of HMGB1-induced endothelial hyperpermeability. PMID:27572515

  9. Caveolin-1 and ATP binding cassette transporter A1 and G1-mediated cholesterol efflux.

    Science.gov (United States)

    Wang, Faqi; Gu, Hong-mei; Zhang, Da-wei

    2014-01-01

    Atherosclerosis is one major cause of cardiovascular diseases, the leading cause of death in industrialized countries. Reverse cholesterol transport (RCT) is thought to be one primary pathway to protect against atherosclerosis. The first and rate-limiting step of RCT is ATP-binding cassette transport A1 (ABCA1) and ABCG1-mediated cholesterol efflux from the cells. Recently, caveolin-1 (CAV1), a scaffolding protein that organizes and concentrates certain caveolin-interacting signaling molecules and receptors within caveolae membranes, has been shown to regulate ABCA1 and ABCG1-mediated cholesterol efflux probably via interacting with them. In the present review, we summarize the current knowledge and views on the regulatory role of CAV1 on the cholesterol homeostasis with emphasis on the association of CAV1 with ABCA1 and ABCG1. We conclude that the dominance of the positive regulation by CAV1 on the ABCA1 and ABCG1-mediated cholesterol efflux is depending on the species, cell types, as well as the levels of CAV1 expression.

  10. Science Signaling Podcast for 24 January 2017: Tissue-specific regulation of L-type calcium channels.

    Science.gov (United States)

    Hell, Johannes W; Navedo, Manuel F; VanHook, Annalisa M

    2017-01-24

    This Podcast features an interview with Johannes Hell and Manuel Navedo, senior authors of two Research Articles that appear in the 24 January 2017 issue of Science Signaling, about tissue-specific regulation of the L-type calcium channel CaV1.2. This channel is present in many tissues, including the heart, vasculature, and brain, and allows calcium to flow into cells when it is activated. Signaling through the β-adrenergic receptor (βAR) stimulates CaV1.2 activity in heart cells and neurons to accelerate heart rate and increase neuronal excitability, respectively. Using mouse models, Qian et al found that βAR-mediated enhancement of CaV1.2 activity in the brain required phosphorylation of Ser(1928), whereas βAR-mediated enhancement of CaV1.2 activity in the heart did not require phosphorylation of this residue. In a related study, Nystoriak et al demonstrated that phosphorylation of Ser(1928) in arterial myocytes was required for vasoconstriction during acute hyperglycemia and in diabetic mice. These findings demonstrate tissue-specific differences in CaV1.2 regulation and suggest that it may be possible to design therapies to target this channel in specific tissues.Listen to Podcast.

  11. Volumetry of Artificial Pulmonary Nodules in Ex Vivo Porcine Lungs: Comparison of Semi-automated Volumetry and Radiologists' Performance

    Energy Technology Data Exchange (ETDEWEB)

    Jeong, Ju Hyeon; Kim, Jin Hwan; Kim, Song Soo [Chungnam National University Hospital, Daejeon (Korea, Republic of); Jeon, Ho Sang [Pusan National University Yangsan Hospital, Yangsan (Korea, Republic of); Lee, Hyun Ju [Seoul National University Hospital, Seoul (Korea, Republic of); Park, Noh Hyuck [Kwandong University College of Medicine, Myungji Hospital, Goyang (Korea, Republic of); Cho, Gyu Seong [KAIST, Daejeon (Korea, Republic of)

    2010-10-15

    With the advent of MSCT, the detection rate of small pulmonary nodules is markedly greater. However, there is no definite diagnostic clue to differentiate between malignant and benign nodules, except for the interval growth in small nodule less than 1 cm in diameter. We evaluated the accuracy of computer aided volumetry (CAV) and compared it with 4 radiologists' measurement. Fifteen artificial nodules that were embedded in the ex vivo porcine lung were scanned by MSCT. The diameters and volumes of nodules were independently measured three times, at 5-day intervals, and by four radiologists as well as by CAV. We evaluated the accuracy of the measurements on the basis of the true diameter and volume of the nodules. Using a paired t-test and a Bland-Altman plot, we evaluated whether there was a statistically significant difference between the radiologists' measurements and the CAV. The accuracy of the manual measurements by radiologists revealed a statistically significant difference from the true diameter and volume of the artificial nodules (p<0.01). Conversely, the accuracy of CAV did not show a statistically significant difference with the true nodule diameter and volume (p>0.01) The results of this study suggest that CAV is an accurate and useful tool to evaluate the volume of pulmonary nodules and can eventually be used to differentiate malignant and benign nodules as well as evaluate the therapeutic response of lung cancer.

  12. Nitroglycerin tolerance in caveolin-1 deficient mice.

    Directory of Open Access Journals (Sweden)

    Mao Mao

    Full Text Available Nitrate tolerance developed after persistent nitroglycerin (GTN exposure limits its clinical utility. Previously, we have shown that the vasodilatory action of GTN is dependent on endothelial nitric oxide synthase (eNOS/NOS3 activity. Caveolin-1 (Cav-1 is known to interact with NOS3 on the cytoplasmic side of cholesterol-enriched plasma membrane microdomains (caveolae and to inhibit NOS3 activity. Loss of Cav-1 expression results in NOS3 hyperactivation and uncoupling, converting NOS3 into a source of superoxide radicals, peroxynitrite, and oxidative stress. Therefore, we hypothesized that nitrate tolerance induced by persistent GTN treatment results from NOS3 dysfunction and vascular toxicity. Exposure to GTN for 48-72 h resulted in nitrosation and depletion (>50% of Cav-1, NOS3 uncoupling as measured by an increase in peroxynitrite production (>100%, and endothelial toxicity in cultured cells. In the Cav-1 deficient mice, NOS3 dysfunction was accompanied by GTN tolerance (>50% dilation inhibition at low GTN concentrations. In conclusion, GTN tolerance results from Cav-1 modification and depletion by GTN that causes persistent NOS3 activation and uncoupling, preventing it from participating in GTN-medicated vasodilation.

  13. Nitroglycerin tolerance in caveolin-1 deficient mice.

    Science.gov (United States)

    Mao, Mao; Varadarajan, Sudhahar; Fukai, Tohru; Bakhshi, Farnaz R; Chernaya, Olga; Dudley, Samuel C; Minshall, Richard D; Bonini, Marcelo G

    2014-01-01

    Nitrate tolerance developed after persistent nitroglycerin (GTN) exposure limits its clinical utility. Previously, we have shown that the vasodilatory action of GTN is dependent on endothelial nitric oxide synthase (eNOS/NOS3) activity. Caveolin-1 (Cav-1) is known to interact with NOS3 on the cytoplasmic side of cholesterol-enriched plasma membrane microdomains (caveolae) and to inhibit NOS3 activity. Loss of Cav-1 expression results in NOS3 hyperactivation and uncoupling, converting NOS3 into a source of superoxide radicals, peroxynitrite, and oxidative stress. Therefore, we hypothesized that nitrate tolerance induced by persistent GTN treatment results from NOS3 dysfunction and vascular toxicity. Exposure to GTN for 48-72 h resulted in nitrosation and depletion (>50%) of Cav-1, NOS3 uncoupling as measured by an increase in peroxynitrite production (>100%), and endothelial toxicity in cultured cells. In the Cav-1 deficient mice, NOS3 dysfunction was accompanied by GTN tolerance (>50% dilation inhibition at low GTN concentrations). In conclusion, GTN tolerance results from Cav-1 modification and depletion by GTN that causes persistent NOS3 activation and uncoupling, preventing it from participating in GTN-medicated vasodilation.

  14. Ca2+ influx through L-type Ca2+ channels and Ca2+-induced Ca2+ release regulate cAMP accumulation and Epac1-dependent ERK 1/2 activation in INS-1 cells.

    Science.gov (United States)

    Pratt, Evan P S; Salyer, Amy E; Guerra, Marcy L; Hockerman, Gregory H

    2016-01-05

    We previously reported that INS-1 cells expressing the intracellular II-III loop of the L-type Ca(2+) channel Cav1.2 (Cav1.2/II-III cells) are deficient in Ca(2+)-induced Ca(2+) release (CICR). Here we show that glucose-stimulated ERK 1/2 phosphorylation (GSEP) is slowed and reduced in Cav1.2/II-III cells compared to INS-1 cells. This parallels a decrease in glucose-stimulated cAMP accumulation (GS-cAMP) in Cav1.2/II-III cells. Influx of Ca(2+) via L-type Ca(2+) channels and CICR play roles in both GSEP and GS-cAMP in INS-1 cells since both are inhibited by nicardipine or ryanodine. Further, the Epac1-selective inhibitor CE3F4 abolishes glucose-stimulated ERK activation in INS-1 cells, as measured using the FRET-based sensor EKAR. The non-selective Epac antagonist ESI-09 but not the Epac2-selective antagonist ESI-05 nor the PKA antagonist Rp-cAMPs inhibits GSEP in both INS-1 and Cav1.2/II-III cells. We conclude that L-type Ca(2+) channel-dependent cAMP accumulation, that's amplified by CICR, activates Epac1 and drives GSEP in INS-1 cells.

  15. Voltage-gated calcium channels and their auxiliary subunits: physiology and pathophysiology and pharmacology.

    Science.gov (United States)

    Dolphin, Annette C

    2016-10-01

    Voltage-gated calcium channels are essential players in many physiological processes in excitable cells. There are three main subdivisions of calcium channel, defined by the pore-forming α1 subunit, the CaV 1, CaV 2 and CaV 3 channels. For all the subtypes of voltage-gated calcium channel, their gating properties are key for the precise control of neurotransmitter release, muscle contraction and cell excitability, among many other processes. For the CaV 1 and CaV 2 channels, their ability to reach their required destinations in the cell membrane, their activation and the fine tuning of their biophysical properties are all dramatically influenced by the auxiliary subunits that associate with them. Furthermore, there are many diseases, both genetic and acquired, involving voltage-gated calcium channels. This review will provide a general introduction and then concentrate particularly on the role of auxiliary α2 δ subunits in both physiological and pathological processes involving calcium channels, and as a therapeutic target.

  16. Transcriptional regulation of voltage-gated Ca(2+) channels.

    Science.gov (United States)

    González-Ramírez, Ricardo; Felix, Ricardo

    2017-03-31

    The transcriptional regulation of voltage-gated Ca(2+) (CaV ) channels is an emerging research area that promises to improve our understanding of how many relevant physiological events are shaped in the central nervous system, the skeletal muscle, and other tissues. Interestingly, a picture of how transcription of CaV channel subunit genes is controlled is evolving with the identification of the promoter regions required for tissue-specific expression, and the identification of transcription factors that control their expression. These promoters share several characteristics that include multiple transcriptional start sites, lack of a TATA box, and the presence of elements conferring tissue-selective expression. Likewise, changes in CaV channel expression occur throughout development, following ischemia, seizures, or chronic drug administration. This review focuses on insights achieved regarding the control of CaV channel gene expression. To further understand the complexities of expression and to increase the possibilities of detecting CaV channel alterations causing human disease, a deeper knowledge on the structure of the 5' upstream regions of the genes encoding these remarkable proteins will be necessary. This article is protected by copyright. All rights reserved.

  17. 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.

  18. Eupenicillium saturniforme, a new species discovered from Northeast China.

    Science.gov (United States)

    Wang, Long; Zhuang, Wen-Ying

    2009-06-01

    A new Eupenicillium species, E. saturniforme was isolated from soil in Jilin Province, northeast China. Morphologically, it resembled E. shearii and E. tropicum, but is distinguished from them by slowly maturing cleistothecia, lenticular ascospores with nearly smooth-walled convex surfaces, strictly velutinous colony texture with abundant conidiogenesis, robust biverticillate penicilli, apically vesiculate metulae and rough-walled stipes and conidia. The partial beta-tubulin gene sequence of the new species (EU644080) showed relationship with Penicillium glabrum in the BLAST search in GenBank. Further analyses of partial calmodulin and ribosomal DNA internally transcribed spacer 1-5.8S-internally transcribed spacer 2 (rDNA ITS1-5.8S-ITS2) sequence data confirmed that E. saturniforme fell in the clade with P. glabrum, P. lividum, P. purpurascens, P. spinulosum and P. thomii of Subgenus Aspergilloides. However, E. saturniforme is a distinctive species lacking close relatives among described species of penicillia.

  19. NUEVAS ESPECIES AMERICANAS DE TURNERA (TURNERACEAE

    Directory of Open Access Journals (Sweden)

    María Mercedes Arbo

    2010-11-01

    Full Text Available El estudio de material de herbario para la revisión del género Turnera, y los viajes de recolección hechos al Brasil en los últimos tres años han permitido el hallazgo de algunas nuevas especies, que se describen aquí: T.aromatica, T.candida, T.cipoensis, T.gardneriana, T.joelii, T.prancei, T.purpurascens, T.rubrobracteata y T.urbanii. Se propone una nueva combinación: T.arenaria (Urban Arbo.

  20. Distribution of drimane sesquiterpenoids and tocopherols in liverworts, ferns and higher plants: Polygonaceae, Canellaceae and Winteraceae species.

    Science.gov (United States)

    Asakawa, Yoshinori; Ludwiczuk, Agnieszka; Harinantenaina, Liva; Toyota, Masao; Nishiki, Mayumi; Bardon, Alicia; Nii, Kaeko

    2012-06-01

    The liverwort, Porella vernicosa complex produces a very hot tasting polygodial, a drimane-type sesquiterpene dialdehyde. The same compound has been isolated from two ferns, Thelypteris hispidula and Blechnum fluviatile, as well as from the higher plants Polygonum hydropiper, P. hydropiper f. purpurascens (Polygonaceae), Cinnamosma, Caspicodendron, Canella and Warburgia species (Canellaceae), and Pseudowintera colorata, Tasmannia lanceolata, Drimys and Zygogynum species (Winteraceae). In addition, the liverworts and higher plants which elaborate polygodial and its related pungent drimane dials contain a small amount of alpha-tocopherol, gamma-tocopherol or delta-tocotrienol. The present paper gives the results of a comparative study on the drimane-type sesquiterpenoids in some liverworts, ferns and higher plants, and the role of tocopherols in these plant groups.

  1. Chemical and genetic relationships among sage ( Salvia officinalis L.) cultivars and Judean sage ( Salvia judaica Boiss.).

    Science.gov (United States)

    Böszörményi, Andrea; Héthelyi, Eva; Farkas, Agnes; Horváth, Györgyi; Papp, Nóra; Lemberkovics, Eva; Szoke, Eva

    2009-06-10

    The essential oil composition and genetic variability of common sage ( Salvia officinalis L.) and its three ornamental cultivars ('Purpurascens', 'Tricolor', and 'Kew Gold') as well as Judean sage ( Salvia judaica Boiss.) were analyzed by GC-FID, GC-MS, and random amplified polymorphic DNA (RAPD). Common sage and its cultivars contained the same volatile compounds; only the ratio of compounds differed. The main compounds were the sesquiterpene alpha-humulene and the monoterpenes beta-pinene, eucalyptol, and camphor. Judean sage contained mainly the sesquiterpenes beta-cubebene and ledol. All of the samples exhibited characteristic RAPD patterns that allowed their identification. Cluster analyses based on oil composition and RAPD markers corresponded very well to each other, suggesting that there is a strong relationship between the chemical profile and the genetic variability.

  2. Estimated Bounds and Important Factors for Fuel Use and Consumer Costs of Connected and Automated Vehicles

    Energy Technology Data Exchange (ETDEWEB)

    Stephens, T. S. [Argonne National Lab. (ANL), Argonne, IL (United States); Gonder, Jeff [National Renewable Energy Lab. (NREL), Golden, CO (United States); Chen, Yuche [National Renewable Energy Lab. (NREL), Golden, CO (United States); Lin, Z. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Liu, C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Gohlke, D. [US Dept. of Energy, Washington, DC (United States)

    2016-11-01

    This report details a study of the potential effects of connected and automated vehicle (CAV) technologies on vehicle miles traveled (VMT), vehicle fuel efficiency, and consumer costs. Related analyses focused on a range of light-duty CAV technologies in conventional powertrain vehicles -- from partial automation to full automation, with and without ridesharing -- compared to today's base-case scenario. Analysis results revealed widely disparate upper- and lower-bound estimates for fuel use and VMT, ranging from a tripling of fuel use to decreasing light-duty fuel use to below 40% of today's level. This wide range reflects uncertainties in the ways that CAV technologies can influence vehicle efficiency and use through changes in vehicle designs, driving habits, and travel behavior. The report further identifies the most significant potential impacting factors, the largest areas of uncertainty, and where further research is particularly needed.

  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...... 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 structural changes...... 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...

  4. Everolimus-Eluting Bioresorbable Vascular Scaffold System in the Treatment of Cardiac Allograft Vasculopathy: the CART (Cardiac Allograft Reparative Therapy) Prospective Multicenter Pilot Study.

    Science.gov (United States)

    Pighi, Michele; Tomai, Fabrizio; Petrolini, Alessandro; de Luca, Leonardo; Tarantini, Giuseppe; Barioli, Alberto; Colombo, Paola; Klugmann, Silvio; Ferlini, Marco; Ormezzano, Maurizio Ferrario; Loi, Bruno; Calabrò, Paolo; Bianchi, Renato Maria; Faggian, Giuseppe; Forni, Alberto; Vassanelli, Corrado; Valgimigli, Marco; Ribichini, Flavio

    2016-02-01

    Cardiac allograft vasculopathy (CAV) is a form of accelerated atherosclerosis, which represents the leading cause of late morbidity and mortality after heart transplantation. The recent bioresorbable vascular scaffold (BVS) technology represents a potential novel therapeutic tool, in the context of CAV, by allowing transient scaffolding and concomitant vessel healing. Eligible subjects will be treated by using the Absorb Everolimus-Eluting BVS (Abbott Vascular, Santa Clara, CA, USA), and evaluated at pre-determined time points, up to 3 years since the index procedure. Both clinical and imaging data will be collected in dedicated case report forms (CRF). All imaging data will be analyzed in an independent core laboratory. The primary aim of the study is to evaluate the angiographic performance at 1 year of second-generation Absorb BVS, in heart transplant recipients affected by CAV.

  5. Antibody titers for canine parvovirus type-2, canine distemper virus, and canine adenovirus type-1 in adult household dogs.

    Science.gov (United States)

    Taguchi, Masayuki; Namikawa, Kazuhiko; Maruo, Takuya; Orito, Kensuke; Lynch, Jonathan; Sahara, Hiroeki

    2011-09-01

    Serum antibody titers for canine parvovirus type-2 (CPV-2), canine distemper virus (CDV) and canine adenovirus type-1 (CAV-1) were investigated in 1031 healthy adult household dogs (2 to 18 years old) given an annual inoculation in the previous 11 to 13 months. The number of dogs retaining significant titers of antibodies against CPV-2, CDV, and CAV-1 were 888 (86%), 744 (72%), and 732 (71%), respectively. There were no differences between males and females in antibody titers against the 3 viruses. Antibody titer for CPV-2 was significantly higher in younger dogs than in older dogs, CDV antibody was significantly higher in older dogs than in younger dogs, and CAV titer was not associated with age.

  6. Structural analysis of coxsackievirus A7 reveals conformational changes associated with uncoating.

    Science.gov (United States)

    Seitsonen, Jani J T; Shakeel, Shabih; Susi, Petri; Pandurangan, Arun P; Sinkovits, Robert S; Hyvönen, Heini; Laurinmäki, Pasi; Ylä-Pelto, Jani; Topf, Maya; Hyypiä, Timo; Butcher, Sarah J

    2012-07-01

    Coxsackievirus A7 (CAV7) is a rarely detected and poorly characterized serotype of the Enterovirus species Human enterovirus A (HEV-A) within the Picornaviridae family. The CAV7-USSR strain has caused polio-like epidemics and was originally thought to represent the fourth poliovirus type, but later evidence linked this strain to the CAV7-Parker prototype. Another isolate, CAV7-275/58, was also serologically similar to Parker but was noninfectious in a mouse model. Sequencing of the genomic region encoding the capsid proteins of the USSR and 275/58 strains and subsequent comparison with the corresponding amino acid sequences of the Parker strain revealed that the Parker and USSR strains are nearly identical, while the 275/58 strain is more distant. Using electron cryomicroscopy and three-dimensional image reconstruction, the structures of the CAV7-USSR virion and empty capsid were resolved to 8.2-Å and 6.1-Å resolutions, respectively. This is one of the first detailed structural analyses of the HEV-A species. Using homology modeling, reconstruction segmentation, and flexible fitting, we constructed a pseudoatomic T = 1 (pseudo T = 3) model incorporating the three major capsid proteins (VP1 to VP3), addressed the conformational changes of the capsid and its constituent viral proteins occurring during RNA release, and mapped the capsid proteins' variable regions to the structure. During uncoating, VP4 and RNA are released analogously to poliovirus 1, the interfaces of VP2 and VP3 are rearranged, and VP1 rotates. Variable regions in the capsid proteins were predicted to map mainly to the surface of VP1 and are thus likely to affect the tropism and pathogenicity of CAV7.

  7. Cd(2+) sensitivity and permeability of a low voltage-activated Ca(2+) channel with CatSper-like selectivity filter.

    Science.gov (United States)

    Garza-López, Edgar; Chávez, Julio César; Santana-Calvo, Carmen; López-González, Ignacio; Nishigaki, Takuya

    2016-07-01

    CatSper is a sperm-specific Ca(2+) channel that plays an essential role in the male fertility. However, its biophysical properties have been poorly characterized mainly due to its deficient heterologous expression. As other voltage-gated Ca(2+) channels (CaVs), CatSper possesses a conserved Ca(2+)-selective filter motif ([T/S]x[D/E]xW) in the pore region. Interestingly, CatSper conserves four aspartic acids (DDDD) as the negatively charged residues in this motif while high voltage-activated CaVs have four glutamic acids (EEEE) and low voltage-activated CaVs possess two glutamic acids and two aspartic acids (EEDD). Previous studies based on site-directed mutagenesis of L- and T-type channels showed that the number of D seems to have a negative correlation with their cadmium (Cd(2+)) sensitivity. These results suggest that CatSper (DDDD) would have low sensitivity to Cd(2+). To explore Cd(2+)-sensitivity and -permeability of CatSper, we performed two types of experiments: 1) Electrophysiological analysis of heterologously expressed human CaV3.1 channel and three pore mutants (DEDD, EDDD and DDDD), 2) Cd(2+) imaging of human spermatozoa with FluoZin-1. Electrophysiological studies showed a significant increase in Cd(2+) and manganese (Mn(2+)) currents through the CaV3.1 mutants as well as a reduction in the inhibitory effect of Cd(2+) on the Ca(2+) current. In fluorescence imaging with human sperm, we observed an increase in Cd(2+) influx potentiated by progesterone, a potent activator of CatSper. These results support our hypothesis, namely that Cd(2+)-sensitivity and -permeability are related to the absolute number of D in the Ca(2+)-selective filter independently to the type of the Cav channels.

  8. The Involvement of Ser1898 of the Human L-Type Calcium Channel in Evoked Secretion

    Directory of Open Access Journals (Sweden)

    Niv Bachnoff

    2011-01-01

    Full Text Available A PKA consensus phosphorylation site S1928 at the α11.2 subunit of the rabbit cardiac L-type channel, CaV1.2, is involved in the regulation of CaV1.2 kinetics and affects catecholamine secretion. This mutation does not alter basal CaV1.2 current properties or regulation of CaV1.2 current by PKA and the beta-adrenergic receptor, but abolishes CaV1.2 phosphorylation by PKA. Here, we test the contribution of the corresponding PKA phosphorylation site of the human α11.2 subunit S1898, to the regulation of catecholamine secretion in bovine chromaffin cells. Chromaffin cells were infected with a Semliki-Forest viral vector containing either the human wt or a mutated S1898A α11.2 subunit. Both subunits harbor a T1036Y mutation conferring nifedipine insensitivity. Secretion evoked by depolarization in the presence of nifedipine was monitored by amperometry. Depolarization-triggered secretion in cells infected with either the wt α11.2 or α11.2/S1898A mutated subunit was elevated to a similar extent by forskolin. Forskolin, known to directly activate adenylyl-cyclase, increased the rate of secretion in a manner that is largely independent of the presence of S1898. Our results are consistent with the involvement of additional PKA regulatory site(s at the C-tail of α11.2, the pore forming subunit of CaV1.2.

  9. PrP genotype frequencies and risk evaluation for scrapie in dairy sheep breeds from southern Italy.

    Science.gov (United States)

    Martemucci, Giovanni; Iamartino, Daniela; Blasi, Michele; D'Alessandro, Angela Gabriella

    2015-12-01

    Concerns regarding scrapie in sheep breeding have increased in the last few decades. The present study was carried out in dairy sheep breeds from southern Italy. In order to find breeding animals resistant to scrapie, the PrP genes of 1,205 animals from entire flocks of dairy native Apulian Leccese and Altamurana breeds, and Sicilian Comisana breed, were analysed for polymorphisms at codons 136, 154, and 171 related to scrapie resistance/susceptibility. The Altamurana breed was considered as two populations (Alt-Cav and Alt-Cra-Zoe), based on presumed cross-breeding. A total of five alleles and ten different genotypes were found. The ARQ allele was predominant for all breeds followed by ARR, the most resistant allele to scrapie, which was highly prevalent in Comisana (50%) and in native Alt-Cav (42.4%). The VRQ allele, associated with the highest susceptibility to scrapie, was detected at not negligeable levels in allocthonous Comisana (3.5%), at a low frequency (0.2%) in native Leccese and Alt-Cra-Zoe, while it was absent in Alt-Cav. The frequencies of PrP genotypes with a very low susceptibility risk to scrapie (R1) was higher in Comisana and Alt-Cav. The most susceptible genotype, ARQ/VRQ, was found only in Comisana. Within the Altamurana breed, there were notable differences between Alt-Cav and Alt-Cra-Zoe sheep. The Alt-Cav was characterised by the absence of VRQ and AHQ alleles and by the higher frequency of the ARR/ARR genotype (18.7%). Breeding programs, mainly in endangered breeds such as Altamurana, should be conducted gradually, combining resistance to scrapie, maintenance of genetic variability, and production.

  10. Canine Recombinant Adenovirus Vector Induces an Immunogenicity-Related Gene Expression Profile in Skin-Migrated CD11b+ -Type DCs

    Science.gov (United States)

    Jouneau, Luc; Bourge, Mickael; Bouet-Cararo, Coraline; Bonneau, Michel; Zientara, Stephan; Klonjkowski, Bernard; Schwartz-Cornil, Isabelle

    2012-01-01

    Gene expression profiling of the blood cell response induced early after vaccination has previously been demonstrated to predict the immunogenicity of vaccines. In this study, we evaluated whether the analysis of the gene expression profile of skin-migrated dendritic cells (DCs) could be informative for the in vitro prediction of immunogenicity of vaccine, using canine adenovirus serotype 2 (CAV2) as vaccine vector. CAV2 has been shown to induce immunity to transgenes in several species including sheep and is an interesting alternative to human adenovirus-based vectors, based on the safety records of the parental strain in dogs and the lack of pre-existing immunity in non-host species. Skin-migrated DCs were collected from pseudo-afferent lymph in sheep. Both the CD11b+ -type and CD103+ -type skin-migrated DCs were transduced by CAV2. An analysis of the global gene response to CAV2 in the two skin DC subsets showed that the gene response in CD11b+ -type DCs was far higher and broader than in the CD103+ -type DCs. A newly released integrative analytic tool from Ingenuity systems revealed that the CAV2-modulated genes in the CD11b+ -type DCs clustered in several activated immunogenicity-related functions, such as immune response, immune cell trafficking and inflammation. Thus gene profiling in skin-migrated DC in vitro indicates that the CD11b+ DC type is more responsive to CAV2 than the CD103+ DC type, and provides valuable information to help in evaluating and possibly improving viral vector vaccine effectiveness. PMID:23300693

  11. Canine recombinant adenovirus vector induces an immunogenicity-related gene expression profile in skin-migrated CD11b⁺ -type DCs.

    Directory of Open Access Journals (Sweden)

    Vanessa Contreras

    Full Text Available Gene expression profiling of the blood cell response induced early after vaccination has previously been demonstrated to predict the immunogenicity of vaccines. In this study, we evaluated whether the analysis of the gene expression profile of skin-migrated dendritic cells (DCs could be informative for the in vitro prediction of immunogenicity of vaccine, using canine adenovirus serotype 2 (CAV2 as vaccine vector. CAV2 has been shown to induce immunity to transgenes in several species including sheep and is an interesting alternative to human adenovirus-based vectors, based on the safety records of the parental strain in dogs and the lack of pre-existing immunity in non-host species. Skin-migrated DCs were collected from pseudo-afferent lymph in sheep. Both the CD11b(+ -type and CD103(+ -type skin-migrated DCs were transduced by CAV2. An analysis of the global gene response to CAV2 in the two skin DC subsets showed that the gene response in CD11b(+ -type DCs was far higher and broader than in the CD103(+ -type DCs. A newly released integrative analytic tool from Ingenuity systems revealed that the CAV2-modulated genes in the CD11b(+ -type DCs clustered in several activated immunogenicity-related functions, such as immune response, immune cell trafficking and inflammation. Thus gene profiling in skin-migrated DC in vitro indicates that the CD11b(+ DC type is more responsive to CAV2 than the CD103(+ DC type, and provides valuable information to help in evaluating and possibly improving viral vector vaccine effectiveness.

  12. Dietary sodium intake regulates angiotensin II type 1, mineralocorticoid receptor, and associated signaling proteins in heart.

    Science.gov (United States)

    Ricchiuti, Vincent; Lapointe, Nathalie; Pojoga, Luminita; Yao, Tham; Tran, Loc; Williams, Gordon H; Adler, Gail K

    2011-10-01

    Liberal or high-sodium (HS) intake, in conjunction with an activated renin-angiotensin-aldosterone system, increases cardiovascular (CV) damage. We tested the hypothesis that sodium intake regulates the type 1 angiotensin II receptor (AT(1)R), mineralocorticoid receptor (MR), and associated signaling pathways in heart tissue from healthy rodents. HS (1.6% Na(+)) and low-sodium (LS; 0.02% Na(+)) rat chow was fed to male healthy Wistar rats (n=7 animals per group). Protein levels were assessed by western blot and immunoprecipitation analysis. Fractionation studies showed that MR, AT(1)R, caveolin-3 (CAV-3), and CAV-1 were located in both cytoplasmic and membrane fractions. In healthy rats, consumption of an LS versus a HS diet led to decreased cardiac levels of AT(1)R and MR. Decreased sodium intake was also associated with decreased cardiac levels of CAV-1 and CAV-3, decreased immunoprecipitation of AT(1)R-CAV-3 and MR-CAV-3 complexes, but increased immunoprecipitation of AT(1)R/MR complexes. Furthermore, decreased sodium intake was associated with decreased cardiac extracellular signal-regulated kinase (ERK), phosphorylated ERK (pERK), and pERK/ERK ratio; increased cardiac striatin; decreased endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (peNOS), but increased peNOS/eNOS ratio; and decreased cardiac plasminogen activator inhibitor-1. Dietary sodium restriction has beneficial effects on the cardiac expression of factors associated with CV injury. These changes may play a role in the cardioprotective effects of dietary sodium restriction.

  13. Regulation of high-voltage-activated Ca(2+) channel function, trafficking, and membrane stability by auxiliary subunits.

    Science.gov (United States)

    Felix, Ricardo; Calderón-Rivera, Aida; Andrade, Arturo

    2013-09-01

    Voltage-gated Ca(2+) (CaV) channels mediate Ca(2+) ions influx into cells in response to depolarization of the plasma membrane. They are responsible for initiation of excitation-contraction and excitation-secretion coupling, and the Ca(2+) that enters cells through this pathway is also important in the regulation of protein phosphorylation, gene transcription, and many other intracellular events. Initial electrophysiological studies divided CaV channels into low-voltage-activated (LVA) and high-voltage-activated (HVA) channels. The HVA CaV channels were further subdivided into L, N, P/Q, and R-types which are oligomeric protein complexes composed of an ion-conducting CaVα1 subunit and auxiliary CaVα2δ, CaVβ, and CaVγ subunits. The functional consequences of the auxiliary subunits include altered functional and pharmacological properties of the channels as well as increased current densities. The latter observation suggests an important role of the auxiliary subunits in membrane trafficking of the CaVα1 subunit. This includes the mechanisms by which CaV channels are targeted to the plasma membrane and to appropriate regions within a given cell. Likewise, the auxiliary subunits seem to participate in the mechanisms that remove CaV channels from the plasma membrane for recycling and/or degradation. Diverse studies have provided important clues to the molecular mechanisms involved in the regulation of CaV channels by the auxiliary subunits, and the roles that these proteins could possibly play in channel targeting and membrane Stabilization.

  14. Experimental Study of Air Vessel Behavior for Energy Storage or System Protection in Water Hammer Events

    Directory of Open Access Journals (Sweden)

    Mohsen Besharat

    2017-01-01

    Full Text Available An experimental assessment of an air pocket (AP, confined in a compressed air vessel (CAV, has been investigated under several different water hammer (WH events to better define the use of protection devices or compressed air energy storage (CAES systems. This research focuses on the size of an AP within an air vessel and tries to describe how it affects important parameters of the system, i.e., the pressure in the pipe, stored pressure, flow velocity, displaced volume of water and water level in the CAV. Results present a specific range of air pockets based on a dimensionless parameter extractable for other real systems.

  15. Distribution of voltage-dependent and intracellular Ca2+ channels in submucosal neurons from rat distal colon.

    Science.gov (United States)

    Rehn, Matthias; Bader, Sandra; Bell, Anna; Diener, Martin

    2013-09-01

    We recently observed a bradykinin-induced increase in the cytosolic Ca2+ concentration in submucosal neurons of rat colon, an increase inhibited by blockers of voltage-dependent Ca2+ (Ca(v)) channels. As the types of Ca(v) channels used by this part of the enteric nervous system are unknown, the expression of various Ca(v) subunits has been investigated in whole-mount submucosal preparations by immunohistochemistry. Submucosal neurons, identified by a neuronal marker (microtubule-associated protein 2), are immunoreactive for Ca(v)1.2, Ca(v)1.3 and Ca(v)2.2, expression being confirmed by reverse transcription plus the polymerase chain reaction. These data agree with previous observations that the inhibition of L- and N-type Ca2+ currents strongly inhibits the response to bradykinin. However, whole-cell patch-clamp experiments have revealed that bradykinin does not enhance Ca2+ inward currents under voltage-clamp conditions. Consequently, bradykinin does not directly interact with Ca(v) channels. Instead, the kinin-induced Ca2+ influx is caused indirectly by the membrane depolarization evoked by this peptide. As intracellular Ca2+ channels on Ca(2+)-storing organelles can also contribute to Ca2+ signaling, their expression has been investigated by imaging experiments and immunohistochemistry. Inositol 1,4,5-trisphosphate (IP3) receptors (IP3R) have been functionally demonstrated in submucosal neurons loaded with the Ca(2+)-sensitive fluorescent dye, fura-2. Histamine, a typical agonist coupled to the phospholipase C pathway, induces an increase in the fura-2 signal ratio, which is suppressed by 2-aminophenylborate, a blocker of IP3 receptors. The expression of IP3R1 has been confirmed by immunohistochemistry. In contrast, ryanodine, tested over a wide concentration range, evokes no increase in the cytosolic Ca2+ concentration nor is there immunohistochemical evidence for the expression of ryanodine receptors in these neurons. Thus, rat submucosal neurons are equipped

  16. A calcium channel mutant mouse model of hypokalemic periodic paralysis

    OpenAIRE

    Wu, Fenfen; Mi, Wentao; Hernández-Ochoa, Erick O.; Burns, Dennis K.; Fu, Yu; Gray, Hillery F; Struyk, Arie F.; Martin F Schneider; Cannon, Stephen C.

    2012-01-01

    Hypokalemic periodic paralysis (HypoPP) is a familial skeletal muscle disorder that presents with recurrent episodes of severe weakness lasting hours to days associated with reduced serum potassium (K+). HypoPP is genetically heterogeneous, with missense mutations of a calcium channel (CaV1.1) or a sodium channel (NaV1.4) accounting for 60% and 20% of cases, respectively. The mechanistic link between CaV1.1 mutations and the ictal loss of muscle excitability during an attack of weakness in Hy...

  17. Calcium channels, neuromuscular synaptic transmission and neurological diseases.

    Science.gov (United States)

    Urbano, Francisco J; Pagani, Mario R; Uchitel, Osvaldo D

    2008-09-15

    Voltage-dependent calcium channels are essential in neuronal signaling and synaptic transmission, and their functional alterations underlie numerous human disorders whether monogenic (e.g., ataxia, migraine, etc.) or autoimmune. We review recent work on Ca(V)2.1 or P/Q channelopathies, mostly using neuromuscular junction preparations, and focus specially on the functional hierarchy among the calcium channels recruited to mediate neurotransmitter release when Ca(V)2.1 channels are mutated or depleted. In either case, synaptic transmission is greatly compromised; evidently, none of the reported functional replacements with other calcium channels compensates fully.

  18. 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.

  19. 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

  20. Caveolin-1 expression in cancer-associated fibroblasts and its correlation with molecular typing of invasive ductal breast carcinoma%Caveolin-1在乳腺浸润性导管癌间质内癌相关成纤维细胞中的表达水平与乳腺癌分子亚型的相关性

    Institute of Scientific and Technical Information of China (English)

    王善伟; 徐侃伦; 赵莉莉; 陈丽荣

    2012-01-01

    Objective To investigate the expression of caveolin-l (Cav-1) in breast cancer-associated fibroblasts(CAFs) and to indicate its correlation with molecular typing and gene copy number of HER-2/neu in invasive ductal breast carcinoma (IDC). Methods One hundred and forty-five cases of breast cancer molecular subtypes were enrolled in the study. En Vision immunohisto-chemical method was used to detect Cav-1 expression of 168 cases with breast cancer in CAFs situation. Fluorescence in situ hybridization ( FISH) was used to detect HER-2 gene amplification status of breast cancer. HER-2 gene amplification in breast cancer and the correlation with stromal expression of Cav-1 was analyzed. Results Cav-1 expression in CAFs in breast cancer with pathological closely related to molecular typing. The positive rates in HER-2 + type and Luminal B type groups were 83. 3% (35/42) ,83. 3% (20/24), significantly higher than that in Luminal A group(58. 1% ) and Basal-like type group(35. 3% ), the differences were statistically significant (P < 0. 05). Cav-1 expression in CAFs was positively correlated with HER-2 protein expression and HER-2 gene status. HER-2 gene status were detected by FISH, stromal Cav-1 positive rate of HER-2 gene amplification group was 83. 3% (55/66), higher than those without amplification group55. 9% (57/102), the differences were statistically significant (P <0.05). Cav-1 expression in CAFs was positively correlated with molecular typing, HER-2 protein expression and HER-2 gene status. Stromal caveolin-1 expression was significantly associated with prognosis (P = 0. 041). Conclusion Cav-1 expression in CAFs is positively correlated with molecular typing, HER-2 protein expression and HER-2 gene status. The over expression of Cav-1 in stromal CAFs of invasive breast cancer predicts good prognostic outcome.%目的 检测Caveolin-1(Gay-1)在乳腺浸润性导管癌间质内癌相关成纤维细胞(CAFs)中的表达,分析Cav-1与乳腺癌分子亚型、HER-2基

  1. Effects of microRNA-1 on negatively regulating L-type calcium channel β2 subunit gene expression during cardiac hypertrophy%MicroRNA-1在心肌肥大中对L-型钙通道β2亚基的负性调控作用

    Institute of Scientific and Technical Information of China (English)

    吴扬; 耿鹏; 王玉琴; 刘艳

    2012-01-01

    Objective: To investigate the negative regulation of microRNA-1 (nriR-1) on L-type calcium channel ft subunit(Cavft) during cardiomyocyte hypertrophy and its mechanism. Methods: Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (HJ2000). The targets of miR-1 were predicted by online database microCoan. The 3' untrans-laJed region sequence of Cavβ2 was cloned into luciferaee reporter vector and then transiently transfected into HEK293 cells. The luciferase activities of samples were measured to verify the expression of luciferase reporter vector. The expression of atrial natriuretic peptide( ANP) , p1-myosin heavy chain(β-MHC), miR-1 and the Cavft mRNA were detected by qRT-PCR. The protein expression of Cavft was detected by Western blot. The level of miR-1 was up-regukted by miR-1 mimic transfection and the expression level of Cavft was down-regulated by RNAi, then effects of which on cardioniyocyte hypertrophy were investigated. Results: ①The expression of miR-1 was significantly reduced in cardiomyocyte hypertrophy. Upregulating the miR-1 level could suppress the increase of cell surface area, the expression of ANP and βMHC mR-NA( P < 0.05). ②Cavβ2 was the one of potential targets of miR-1 by prediction using online database mieroCosm. The luciferase acnvites of HEK293 cells with the plasmid containing miR-1 and widetype Cavft 3' UTR sequence was significantly decreased when compared with that of control group(P<0.01). Up-regulation of the miR-1 level could suppress the protein expression of Cavβ2. ③The expression of Cavβ2 was significantly increased in cardioniyocyte hypertrophy induced by ISO. Downregulation of Cavβ2 by RNAi could markedly inhibit the increase of cell surface area, the expression of ANP and β-MHC mRNA. Condnaon: Cavft is one of potential targets of miR-1 by bioinformatics prediction. The experiment data confirms that Cavβ2 is truly the target of miR-1. MiR-1

  2. Role of caveolin-1 in adult rat CFs proliferation induced by AVP and the intervention of Simvastatin%小窝蛋白-1反义寡核苷酸在AVP诱导大鼠心肌成纤维细胞增殖erk1/2信号转导中的作用及辛伐他汀的干预效应

    Institute of Scientific and Technical Information of China (English)

    何燕萍; 杨军岭; 赵连友; 刘少伟

    2011-01-01

    Objective To study the effect of caveolin-1 antisense oligonucleotides (cav1-AS ODN) on cardiac fibroblasts (CFs) proliferation induced by arginine vasopressin (AVP) and the involvement of simvastatin (Sim) in the modulation of cholesterol. Methods CFs were isolated from adult SD rats and cultured. Phosphorothioate modified oligonucleotides and AVP were added to the CFs. CFs proliferation was assessed by 3H-TdR incorporation. Protein expressions of cav1 ,erk1/2, p21 and cyclin A were evaluated by western blotting. Results The levels of 3H-TdR incorporation and erk1/2 protein expression were (212 ± 6)% and (7.9 ± 0. 3) relative to control,respectively,in cav1-AS ODN + AVP group, being different from those of ( 172 ± 4) % and (5. 7 ± 0. 2) in AVP group ( both P < 0. 0 1 ). Protein expression of p21 was decreased and cyclin A increased in AS ODN ± AVP group. The presence of β-cyclodextrin or progesterone or Sim decreased cav1 protein expression. The levels of cav1 protein expression were (86 ± 3 )%, (91 ± 4 )% and (94 ± 5 )% relative to control, respectively,in the presence of 10,15 or 20 μg/ml cholesterol,being different from that of Sim group (66 ± 4)% relative to control (P <0. 05,P <0. 01 ). Conclusions cav1-AS ODN potentiates the proliferation-inducing effect of AVP on adult rat CFs,and the intervention of cholesterollowing effect of statins may provide new insights into the anti-fibrosis effect of these drugs.%目的 研究小窝蛋白-1反义寡核苷酸(cav1-AS ODN)在血管加压素(AVP)诱导的成年大鼠心肌成纤维细胞(CFs)增殖erk1/2信号转导中的作用及辛伐他汀(Sim)对cav1表达的干预效应.方法 采用脂质体导入法将cav1-AS ODN导入离体培养的成年大鼠CFs,用3H-TdR掺入法定量观察CFs增殖,蛋白免疫印迹法分析cav1-AS ODN对AVP诱导大鼠CFs增殖后磷酸化erk1/2 (p-erk1/2)、p21和细胞周期蛋白A(cyclin A)表达变化及Sim对cav1表达的影响.结果 cav1-AS ODN与10

  3. 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

  4. Chicken anemia virus and avian gyrovirus 2 as contaminants in poultry vaccines.

    Science.gov (United States)

    Varela, Ana Paula Muterle; Dos Santos, Helton Fernandes; Cibulski, Samuel Paulo; Scheffer, Camila Mengue; Schmidt, Candice; Sales Lima, Francisco Esmaile; Silva, Alessandra D'Avila; Esteves, Paulo Augusto; Franco, Ana Cláudia; Roehe, Paulo Michel

    2014-11-01

    This study focuses on the detection of chicken anemia virus (CAV) and avian gyrovirus 2 (AGV2) genomes in commercially available poultry vaccines. A duplex quantitative real-time PCR (dqPCR), capable of identifying genomes of both viruses in a single assay, was employed to determine the viral loads of these agents in commercially available vaccines. Thirty five vaccines from eight manufacturers (32 prepared with live and 3 with inactivated microorganisms) were examined. Genomes of CAV were detected as contaminants in 6/32 live vaccines and in 1/3 inactivated vaccines. The CAV genome loads ranged from 6.4 to 173.4 per 50 ng of vaccine DNA (equivalent to 0.07 to 0.69 genome copies per dose of vaccine). Likewise, AGV2 genomes were detected in 9/32 live vaccines, with viral loads ranging from 93 to 156,187 per 50 ng of vaccine DNA (equivalent to 0.28-9176 genome copies per dose of vaccine). These findings provide evidence for the possibility of contamination of poultry vaccines with CAV and AGV2 and they also emphasize the need of searching for these agents in vaccines in order to ensure the absence of such potential contaminants.

  5. ERRATUM to: Taxonomy, chromosome numbers, clonal diversity and population dynamics of Phragmites australis (vol 64, pg 185, 1999)

    NARCIS (Netherlands)

    Clevering, O.A.; Lissner, J.

    2000-01-01

    Phragmites australis (Cav.) Trin. ex Steud. (common reed) is one of the most widespread plant species in the world. The species has a high phenotypic variation in morphology and life-history traits. This high phenotypic variation can be related to variance in chromosome numbers, clonal diversity, pl

  6. AcEST: BP917517 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 98SV6|Q98SV6_SCOCA Myostatin (Fragment) OS=Scomberomorus cav... 33 5.4 tr|Q4UAA3|Q4UAA3_THEAN Putative uncha...TQQFPNQPASIQGNHHTM 1898 >tr|Q98SV6|Q98SV6_SCOCA Myostatin (Fragment) OS=Scomberom

  7. 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

    -mediated properties such as metabotropic receptor-induced oscillations and plateau potentials failed to be induced in culture. This study shows that changes specifically affecting CaV1.3 channel contribution to intrinsic oscillatory property expression may occur in culture. The results contribute also...... to understanding further the potential for plasticity of mature vertebrate neurons....

  8. 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

  9. The role of melaleuca control in Everglades restoration: accomplishments and future plans

    Science.gov (United States)

    The invasive tree Melaleuca quinquenervia (Cav.) S.T. Blake, melaleuca, was first introduced to Florida in 1886 for ornamental use. It was later planted for bank stabilization and as a forestry crop. Melaleuca now invades a variety of habitats, changing the hydroperiod and fire regime, reducing plan...

  10. 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.

  11. Role of Ryanodine Receptor Subtypes in Initiation and Formation of Calcium Sparks in Arterial Smooth Muscle: Comparison with Striated Muscle

    Directory of Open Access Journals (Sweden)

    Maik Gollasch

    2009-01-01

    Full Text Available Calcium sparks represent local, rapid, and transient calcium release events from a cluster of ryanodine receptors (RyRs in the sarcoplasmic reticulum. In arterial smooth muscle cells (SMCs, calcium sparks activate calcium-dependent potassium channels causing decrease in the global intracellular [Ca2+] and oppose vasoconstriction. This is in contrast to cardiac and skeletal muscle, where spatial and temporal summation of calcium sparks leads to global increases in intracellular [Ca2+] and myocyte contraction. We summarize the present data on local RyR calcium signaling in arterial SMCs in comparison to striated muscle and muscle-specific differences in coupling between L-type calcium channels and RyRs. Accordingly, arterial SMC Cav1.2 L-type channels regulate intracellular calcium stores content, which in turn modulates calcium efflux though RyRs. Downregulation of RyR2 up to a certain degree is compensated by increased SR calcium content to normalize calcium sparks. This indirect coupling between Cav1.2 and RyR in arterial SMCs is opposite to striated muscle, where triggering of calcium sparks is controlled by rapid and direct cross-talk between Cav1.1/Cav1.2 L-type channels and RyRs. We discuss the role of RyR isoforms in initiation and formation of calcium sparks in SMCs and their possible molecular binding partners and regulators, which differ compared to striated muscle.

  12. L-type Ca(2+) channels in mood, cognition and addiction: integrating human and rodent studies with a focus on behavioural endophenotypes.

    Science.gov (United States)

    Kabir, Z D; Lee, A S; Rajadhyaksha, A M

    2016-10-15

    Brain Cav 1.2 and Cav 1.3 L-type Ca(2+) channels play key physiological roles in various neuronal processes that contribute to brain function. Genetic studies have recently identified CACNA1C as a candidate risk gene for bipolar disorder (BD), schizophrenia (SCZ), major depressive disorder (MDD) and autism spectrum disorder (ASD), and CACNA1D for BD and ASD, suggesting a contribution of Cav 1.2 and Cav 1.3 Ca(2+) signalling to the pathophysiology of neuropsychiatric disorders. Once considered sole clinical entities, it is now clear that BD, SCZ, MDD and ASD share common phenotypic features, most likely due to overlapping neurocircuitry and common molecular mechanisms. A major future challenge lies in translating the human genetic findings to pathological mechanisms that are translatable back to the patient. One approach for tackling such a daunting scientific endeavour for complex behaviour-based neuropsychiatric disorders is to examine intermediate biological phenotypes in the context of endophenotypes within distinct behavioural domains. This will better allow us to integrate findings from genes to behaviour across species, and improve the chances of translating preclinical findings to clinical practice.

  13. Negative modulation of the chicken infectious anemia virus promoter by COUP-TF1 and an E box-like element at the transcription start site binding dEF1

    Science.gov (United States)

    Expression of enhanced green fluorescent protein (EGFP) under control of the promoter-enhancer of chicken infectious anemia virus (CAV) is increased in an estrogen receptor-enhanced cell line when treated with estrogen. This promoter-enhancer also binds unidentified proteins that recognize a consens...

  14. Apigenin, a natural flavonoid, inhibits glutamate release in the rat hippocampus.

    Science.gov (United States)

    Chang, Chia Ying; Lin, Tzu Yu; Lu, Cheng Wei; Wang, Chia Chuan; Wang, Ying Chou; Chou, Shang Shing Peter; Wang, Su Jane

    2015-09-05

    The purpose of this study was to examine the effect and mechanism of apigenin, a natural flavonoid, on glutamate release in the rat hippocampus. In rat hippocampal nerve terminals (synaptosomes), apigenin inhibited glutamate release and the elevation of the cytosolic free Ca(2+) concentration evoked by 4-aminopyridine, whereas it had no effect on 4-aminopyridine-mediated depolarization and Na(+) influx. The apigenin-mediated inhibition of evoked glutamate release was prevented by chelating the extracellular Ca(2+) ions and blocking Cav2.2 (N-type) and Cav2.1 (P/Q-type) channel activity. Furthermore, we determined that gamma-aminobutyric acid type A (GABAA) receptors are present in the hippocampal nerve terminals because they are colocalized with the presynaptic marker synaptophysin. However, the effect of apigenin on 4-aminopyridine-evoked glutamate release from synaptosomes was unaffected by the GABAA receptor antagonists SR95531 and bicuculline. Furthermore, in slice preparations, whole-cell patch-clamp experiments showed that apigenin reduced the frequency of spontaneous excitatory postsynaptic currents without affecting their amplitude, suggesting a presynaptic mechanism. On the basis of these results, we suggested that apigenin exerts its presynaptic inhibition probably by reducing Ca(2+) entry mediated by the Cav2.2 (N-type) and Cav2.1 (P/Q-type) channels, thereby inhibiting glutamate release from the rat hippocampal nerve terminals.

  15. Acquired natural enemies of the weed biological control agent Oxyops vitiosa (Coleoptera: Curculionidae)

    Science.gov (United States)

    The Australian curculionid Oxyops vitiosa Pascoe was introduced into Florida during 1997 as a biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) S.T. Blake. Populations of the weevil increased rapidly and became widely distributed throughout much of the invasive tree’s adve...

  16. Research in Biological and Medical Sciences, Including Biochemistry, Communicable Disease and Immunology, Internal Medicine, Nuclear Medicine, Physiology, Psychiatry, Surgery and Veterinary Medicine. Volume 2

    Science.gov (United States)

    1975-07-01

    efficacy of all 3 agents (Group VI vs Group IV; Group IX vs Group VII; Group XII vs Group X). WR 211,537 WR 2975 significant was not demon- this...Rattua rattua Cave aouth, slao far Inside Rattua nallll ap. nov. ( Tabla 3) LlMoton« cliffs, including aouth of cav« Falco aavarua A falcon

  17. Compositional Verification with Abstraction, Learning, and SAT Solving

    Science.gov (United States)

    2015-05-01

    Amla and Kenneth L. McMillan. A Hybrid of Counterexample-Based and Proof-Based Abstraction. In FMCAD, pages 260–274, 2004. 203 [13] Nina Amla and...Verification. In CAV, 2010. [91] Kenneth L. McMillan and Nina Amla . Automatic Abstraction without Coun- terexamples. In TACAS, 2003. [92] Kenneth L. McMillan

  18. Establishment, population increase, spread, and ecological host range of Lophodiplosis trifida (Diptera: Cecidomyiidae), a biological control agent of the invasive tree Melaleuca quinquenervia (Myrtales:Myrtaceae).

    Science.gov (United States)

    The Australian tree Melaleuca quinquenervia (Cav.) Blake is an invasive weed in wetland systems of Florida, USA. A biological control program targeting M. quinquenervia has culminated in the release of the gall forming midge Lophodiplosis trifida Gagné (Cecidomyiidae). Populations of the introduced ...

  19. Physiological host range of a mutualistic pair: Fergusonina turneri and its obligate nematode Fergusobia quinquenerviae, potential biological control agents of Melaleuca quinquenervia

    Science.gov (United States)

    In Australia, galls develop on Melaleuca quinquenervia (Cav.) S.T. Blake (Myrtales: Myrtaceae) as a result of the mutualistic association between the fly Fergusonina turneri Taylor (Diptera: Fergusoninidae) and its obligate nematode Fergusobia quinquenerviae Davies & Giblin-Davis (Tylenchida: Sphaer...

  20. 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...

  1. 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.

  2. The genetic background affects the vascular response in T-type calcium channels 3.2 deficient mice

    DEFF Research Database (Denmark)

    Svenningsen, Per; Hansen, Pernille B L

    2016-01-01

    Voltage-gated calcium channels (Cav ) are important regulators of vascular tone and are attractive targets for pharmacological treatment of hypertension. The clinical used calcium blockers are often not selective for one channel but affect several types of calcium channels (Hansen 2015). L...

  3. Calcium release near l-type calcium channels promotes beat-to-beat variability in ventricular myocytes from the chronic AV block dog

    NARCIS (Netherlands)

    Antoons, G.; Johnson, Daniel M; Dries, Eef; Santiago, Demetrio J; Ozdemir, Semir; Lenaerts, Ilse; Beekman, Jet D M; Houtman, Marien J C; Sipido, Karin R; Vos, Marc A

    2015-01-01

    Beat-to-beat variability of ventricular repolarization (BVR) has been proposed as a strong predictor of Torsades de Pointes (TdP). BVR is also observed at the myocyte level, and a number of studies have shown the importance of calcium handling in influencing this parameter. The chronic AV block (CAV

  4. DJ-1 deficiency impairs glutamate uptake into astrocytes via the regulation of flotillin-1 and caveolin-1 expression

    Science.gov (United States)

    Kim, Jin-Mo; Cha, Seon-Heui; Choi, Yu Ree; Jou, Ilo; Joe, Eun-Hye; Park, Sang Myun

    2016-01-01

    Parkinson’s disease (PD) is a common chronic and progressive neurodegenerative disorder. Although the cause of PD is still poorly understood, mutations in many genes including SNCA, parkin, PINK1, LRRK2, and DJ-1 have been identified in the familial forms of PD. It was recently proposed that alterations in lipid rafts may cause the neurodegeneration shown in PD. Here, we observe that DJ-1 deficiency decreased the expression of flotillin-1 (flot-1) and caveolin-1 (cav-1), the main protein components of lipid rafts, in primary astrocytes and MEF cells. As a mechanism, DJ-1 regulated flot-1 stability by direct interaction, however, decreased cav-1 expression may not be a direct effect of DJ-1, but rather as a result of decreased flot-1 expression. Dysregulation of flot-1 and cav-1 by DJ-1 deficiency caused an alteration in the cellular cholesterol level, membrane fluidity, and alteration in lipid rafts-dependent endocytosis. Moreover, DJ-1 deficiency impaired glutamate uptake into astrocytes, a major function of astrocytes in the maintenance of CNS homeostasis, by altering EAAT2 expression. This study will be helpful to understand the role of DJ-1 in the pathogenesis of PD, and the modulation of lipid rafts through the regulation of flot-1 or cav-1 may be a novel therapeutic target for PD. PMID:27346864

  5. Field Surveys, IOC Valleys. Volume III, Part I. Cultural Resources Survey, Dry Lake Valley, Nevada.

    Science.gov (United States)

    1981-08-01

    Artemisia nova) but also include cliffrose (Cowania mexicana ) and broom snakeweed (Gutierrezia sarothreae) as dominant species. Other species include... CULTURA Ale ~~REOUC SURVEYa AREASczCAvE L CU 11U CUUI 3-2 E-TR-48-III-I 69 was used because it is considered intensive by the Bureau of Land Management and

  6. Mapping of dihydropyridine binding residues in a less sensitive invertebrate L-type calcium channel (LCa v 1).

    Science.gov (United States)

    Senatore, Adriano; Boone, Adrienne; Lam, Stanley; Dawson, Taylor F; Zhorov, Boris; Spafford, J David

    2011-01-01

    Invertebrate L-type calcium channel, LCa(v) 1, isolated from the pond snail Lymnaea stagnalis is nearly indistinguishable from mammalian Ca(v) 1.2 (α1C) calcium channel in biophysical characteristics observed in vitro. These L-type channels are likely constrained within a narrow range of biophysical parameters to perform similar functions in the snail and mammalian cardiovascular systems. What distinguishes snail and mammalian L-type channels is a difference in dihydropyridine sensitivity: 100 nM isradipine exhibits a significant block of mammalian Ca(v) 1.2 currents without effect on snail LCa(v)1 currents. The native snail channel serves as a valuable surrogate for validating key residue differences identified from previous experimental and molecular modeling work. As predicted, three residue changes in LCa(v)1 (N_3o18, F_3i10, and I_4i12) replaced with DHP-sensing residues in respective positions of Ca(v) 1.2, (Q_3o18, Y_3i10, and M_4i12) raises the potency of isradipine block of LCa(v)1 channels to that of mammalian Ca(v) 1.2. Interestingly, the single N_3o18_Q mutation in LCa(v) 1 channels lowers DHP sensitivity even further and the triple mutation bearing enhanced isradipine sensitivity, still retains a reduced potency of agonist, (S)-Bay K8644.

  7. 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.

  8. 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.

  9. Cerebral vascular findings in PAPA syndrome: cerebral arterial vasculopathy or vasculitis and a posterior cerebral artery dissecting aneurysm.

    Science.gov (United States)

    Khatibi, Kasra; Heit, Jeremy J; Telischak, Nicholas A; Elbers, Jorina M; Do, Huy M

    2016-08-01

    A young patient with PAPA (pyogenic arthritis, pyoderma gangrenosum, and acne) syndrome developed an unusual cerebral arterial vasculopathy/vasculitis (CAV) that resulted in subarachnoid hemorrhage from a ruptured dissecting posterior cerebral artery (PCA) aneurysm. This aneurysm was successfully treated by endovascular coil sacrifice of the affected segment of the PCA. The patient made an excellent recovery with no significant residual neurologic deficit.

  10. Wetland Plants of Specialized Habitats in the Arid West

    Science.gov (United States)

    2007-06-01

    FACW FACW NO FACW Lactuca serriola L. 1 FAC FAC FACU FAC- FAC Larrea divaricata auct. non Cav. 1,7 Larrea tridentata (Sessé & Moc. ex DC.) Coville... sativa L. UPL UPL UPL UPL UPL Melilotus alba Medik. Melilotus officinalis (L.) Lam. FACU FACU+ FACU FACU FACU+ Melilotus indica (L.) All. FACU

  11. CAR-associated vesicular transport of an adenovirus in motor neuron axons.

    Science.gov (United States)

    Salinas, Sara; Bilsland, Lynsey G; Henaff, Daniel; Weston, Anne E; Keriel, Anne; Schiavo, Giampietro; Kremer, Eric J

    2009-05-01

    Axonal transport is responsible for the movement of signals and cargo between nerve termini and cell bodies. Pathogens also exploit this pathway to enter and exit the central nervous system. In this study, we characterised the binding, endocytosis and axonal transport of an adenovirus (CAV-2) that preferentially infects neurons. Using biochemical, cell biology, genetic, ultrastructural and live-cell imaging approaches, we show that interaction with the neuronal membrane correlates with coxsackievirus and adenovirus receptor (CAR) surface expression, followed by endocytosis involving clathrin. In axons, long-range CAV-2 motility was bidirectional with a bias for retrograde transport in nonacidic Rab7-positive organelles. Unexpectedly, we found that CAR was associated with CAV-2 vesicles that also transported cargo as functionally distinct as tetanus toxin, neurotrophins, and their receptors. These results suggest that a single axonal transport carrier is capable of transporting functionally distinct cargoes that target different membrane compartments in the soma. We propose that CAV-2 transport is dictated by an innate trafficking of CAR, suggesting an unsuspected function for this adhesion protein during neuronal homeostasis.

  12. TAME: A Specialized Specification and Verification System for Timed Automata

    Science.gov (United States)

    1996-12-01

    Lecture Notes in Computer Science . Springer...Courcoubetis, edi- tor, Computer Aided Veri cation, CAV 󈨡, vol- ume 697 of Lecture Notes in Computer Science , pages 292{304. Springer-Verlag, 1993. [10...automata. In J. C. M. Baeten and J. F. Goote, editors, CONCUR󈨟: 2nd Intern. Conference on Concurrency Theory, volume 527 of Lecture Notes in Computer Science ,

  13. A Faster-than Relation for Asynchronous Processes

    Science.gov (United States)

    2001-01-01

    Lecture Notes in Computer Science , Lisbon, Portugal, June 1998, Springer... Lecture Notes in Computer Science , Warsaw, Poland, July 1997, Springer-Verlag, pp. 166{180. [12] R. Cleaveland and S. Sims, The NCSU Concurrency...Workbench, in Computer Aided Veri cation (CAV 󈨤), R. Alur and T. Henzinger, eds., Vol. 1102 of Lecture Notes in Computer Science , New Brunswick, NJ,

  14. Saturation: An Efficient Iteration Strategy for Symbolic State-space Generation

    Science.gov (United States)

    2001-02-01

    Lecture Notes in Computer Science , Berlin, Germany, March/April 2000, Springer...in 9th International Conference on Computer-Aided Veri cation (CAV 󈨥), O. Grumberg, ed., Vol. 1254 of Lecture Notes in Computer Science , Hai...Conference on Tools and Algorithms for the Construction and Analysis of Systems (TACAS 󈨧), R. Cleaveland, ed., Vol. 1579 of Lecture Notes in Computer Science ,

  15. Lessons Learned Model Checking an Industrial Communications Library

    Science.gov (United States)

    2005-09-01

    Lecture Notes in Computer Science [LNCS], volume 2057). Toronto, Canada, May 19-20, 2001. Berlin...Model Checking,” 128-147. Integrated Formal Methods: The 4th International Conference (IFM 2004) ( Lecture Notes in Computer Science [LNCS], volume...on Computer Aided Verification (CAV 1990). ( Lecture Notes in Computer Science [LNCS], volume 531). New Brunswick, NJ, June 18-21, 1990.

  16. Overview of ComFoRT: A Model Checking Reasoning Framework

    Science.gov (United States)

    2004-04-01

    Lecture Notes in Computer Science [LNCS...Programs Workshop (in Lecture Notes in Computer Science [LNCS], volume 131). Yorktown Heights, NY, May 1981. New York, NY: Springer-Verlag, 1982...Refinement," 154-169. Proceedings of the Computer Aided Verification 12 h International Conference (CAV2000) (in Lecture Notes in Computer Science

  17. COVERT: A Framework for Finding Buffer Overflows in C Programs via Software Verification

    Science.gov (United States)

    2010-08-01

    Lecture Notes in Computer Science . Toronto...Engineering (FASE ’05), volume 3442 of Lecture Notes in Computer Science . Edinburgh, UK: Springer-Verlag, April 2005. [CERT CC 2001] CERT CC. “Code...Conference on Computer Aided Verification (CAV ’05), volume 3576 of Lecture Notes in Computer Science . Edinburgh, Scotland, July 6–10, 2005. New

  18. Nanoscale distribution of ryanodine receptors and caveolin-3 in mouse ventricular myocytes: dilation of t-tubules near junctions.

    Science.gov (United States)

    Wong, Joseph; Baddeley, David; Bushong, Eric A; Yu, Zeyun; Ellisman, Mark H; Hoshijima, Masahiko; Soeller, Christian

    2013-06-04

    We conducted super-resolution light microscopy (LM) imaging of the distribution of ryanodine receptors (RyRs) and caveolin-3 (CAV3) in mouse ventricular myocytes. Quantitative analysis of data at the surface sarcolemma showed that 4.8% of RyR labeling colocalized with CAV3 whereas 3.5% of CAV3 was in areas with RyR labeling. These values increased to 9.2 and 9.0%, respectively, in the interior of myocytes where CAV3 was widely expressed in the t-system but reduced in regions associated with junctional couplings. Electron microscopic (EM) tomography independently showed only few couplings with caveolae and little evidence for caveolar shapes on the t-system. Unexpectedly, both super-resolution LM and three-dimensional EM data (including serial block-face scanning EM) revealed significant increases in local t-system diameters in many regions associated with junctions. We suggest that this regional specialization helps reduce ionic accumulation and depletion in t-system lumen during excitation-contraction coupling to ensure effective local Ca²⁺ release. Our data demonstrate that super-resolution LM and volume EM techniques complementarily enhance information on subcellular structure at the nanoscale.

  19. Caveolin-3 Mutations in Rippling Muscle Disease

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2003-05-01

    Full Text Available Two unrelated patients with novel homozygous missense mutations (L86P and A92T in caveolin-3 gene (CAV3, presenting with a severe form of rippling muscle disease (RMD, are reported from the University of Bonn, and other centers in Germany.

  20. A mutation in CABP2, expressed in cochlear hair cells, causes autosomal-recessive hearing impairment

    NARCIS (Netherlands)

    Schrauwen, I.; Helfmann, S.; Inagaki, A.; Predoehl, F.; Tabatabaiefar, M.A.; Picher, M.M.; Sommen, M.; Seco, C.Z.; Oostrik, J.; Kremer, J.M.J.; Dheedene, A.; Claes, C.; Fransen, E.; Chaleshtori, M.H.; Coucke, P.; Lee, A.; Moser, T.; Camp, G. van

    2012-01-01

    CaBPs are a family of Ca(2+)-binding proteins related to calmodulin and are localized in the brain and sensory organs, including the retina and cochlea. Although their physiological roles are not yet fully elucidated, CaBPs modulate Ca(2+) signaling through effectors such as voltage-gated Ca(v) Ca(2

  1. Simulations of the potential revenue from investment in improved indoor air quality in an office building

    DEFF Research Database (Denmark)

    Wargocki, Pawel; Djukanovic, Rade

    2005-01-01

    of improved worker performance; benefits from reduced health costs and sickness absence were not included. The building was simulated in a cold, a moderate and a hot climate. It was ventilated by a constant air volume (CAV) system with heat recovery and by a variable air volume (VAV) system with an economizer...

  2. Effect of everolimus initiation and early calcineurin inhibitor withdrawal on myocardial FOXP3+ regulatory T cells in heart transplantation

    DEFF Research Database (Denmark)

    Mirza, Kiran; Gustafsson, Finn; Gullestad, Lars

    2016-01-01

    hypothesized treatment with everolimus reduced the risk of CAV by modulating myocardial FoxP3 levels. METHODS: 15 patients from the Schedule trial comparing everolimus, MMF, steroid and early CNI (Everolimus, n=8) withdrawal to conventional CNI based immunosuppression (Controls, n=7) after de novo HTx were...

  3. New Role of P/Q-type Voltage-gated Calcium Channels

    DEFF Research Database (Denmark)

    Hansen, Pernille B L

    2015-01-01

    Voltage-gated calcium channels are important for the depolarization-evoked contraction of vascular smooth muscle cells (SMCs), with L-type channels being the classical channel involved in this mechanism. However, it has been demonstrated that the CaV2.1 subunit, which encodes a neuronal isoform o...

  4. Army Network-Enabled Operations: Expectations, Performance, and Opportunities for Future Improvements

    Science.gov (United States)

    2012-01-01

    the time, Bruce Held, and RAND colleagues xxviii Army Network-Enabled Operations Leland Joe, Louis Moore, Isaac Porche, Daniel Gonzales , and Lionel...November 8, 2006. Lanza, COL Stephen R., MAJ Robert L. Menti, CPT Luis M. Alvarez, and 1LT Michael R. Dalton, “1st Cav Div Arty as a Maneuver BCT

  5. Sodium current and potassium transient outward current genes in brugada syndrome

    DEFF Research Database (Denmark)

    Holst, Anders Gaarsdal; 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....

  6. Chronic cardiac allograft rejection: critical role of ED-A(+) fibronectin and implications for targeted therapy strategies.

    Science.gov (United States)

    Franz, Marcus; Neri, Dario; Berndt, Alexander

    2012-03-01

    Chronic cardiac allograft rejection is characterized by cardiac allograft vasculopathy (CAV) and cardiac interstitial fibrosis (CIF) causing severe long-term complications after heart transplantation and determining allograft function and patients' prognosis. Until now, there have been no sufficient preventive or therapeutic strategies. CAV and CIF are accompanied by changes in the extracellular matrix, including re-expression of the fetal fibronectin splice variant known as ED-A(+) fibronectin. This molecule has been shown to be crucial for the development of myofibroblasts (MyoFbs) as the main cell type in CIF and for the activation of vascular smooth muscle cells (VSMCs) as the main cell type in CAV. Relevant re-expression and protein deposition of ED-A(+) fibronectin has been demonstrated in animal models of chronic rejection, with spatial association to CAV and CIF, and a quantitative correlation to the rejection grade. The paper by Booth et al published in this issue of The Journal of Pathology could prove for the first time the functional importance of ED-A(+) fibronectin for the development of CIF as a main component of chronic cardiac rejection. Thus, promising conclusions for the development of new diagnostic, preventive, and therapeutic strategies for chronic cardiac rejection focusing on ED-A(+) fibronectin can be suggested.

  7. A genetic screen for dihydropyridine (DHP-resistant worms reveals new residues required for DHP-blockage of mammalian calcium channels.

    Directory of Open Access Journals (Sweden)

    Trevor C Y Kwok

    2008-05-01

    Full Text Available Dihydropyridines (DHPs are L-type calcium channel (Ca(v1 blockers prescribed to treat several diseases including hypertension. Ca(v1 channels normally exist in three states: a resting closed state, an open state that is triggered by membrane depolarization, followed by a non-conducting inactivated state that is triggered by the influx of calcium ions, and a rapid change in voltage. DHP binding is thought to alter the conformation of the channel, possibly by engaging a mechanism similar to voltage dependent inactivation, and locking a calcium ion in the pore, thereby blocking channel conductance. As a Ca(v1 channel crystal structure is lacking, the current model of DHP action has largely been achieved by investigating the role of candidate Ca(v1 residues in mediating DHP-sensitivity. To better understand DHP-block and identify additional Ca(v1 residues important for DHP-sensitivity, we screened 440,000 randomly mutated Caenorhabditis elegans genomes for worms resistant to DHP-induced growth defects. We identified 30 missense mutations in the worm Ca(v1 pore-forming (alpha(1 subunit, including eleven in conserved residues known to be necessary for DHP-binding. The remaining polymorphisms are in eight conserved residues not previously associated with DHP-sensitivity. Intriguingly, all of the worm mutants that we analyzed phenotypically exhibited increased channel activity. We also created orthologous mutations in the rat alpha(1C subunit and examined the DHP-block of current through the mutant channels in culture. Six of the seven mutant channels examined either decreased the DHP-sensitivity of the channel and/or exhibited significant residual current at DHP concentrations sufficient to block wild-type channels. Our results further support the idea that DHP-block is intimately associated with voltage dependent inactivation and underscores the utility of C. elegans as a screening tool to identify residues important for DHP interaction with mammalian

  8. 远端型小窝蛋白病患者的临床、病理及基因研究%Clinical, myopathological and genetic research of a patient with distal myopathy caused by caveolin-3 deficiency

    Institute of Scientific and Technical Information of China (English)

    陈涓涓; 曾文双; 韩春锡; 吴军; 童晓欣; 张海鸥

    2015-01-01

    Objective To report the clinical,myopathological and genetic features of a patient with distal myopathy caused by caveolin-3 (CAV3) deficiency.Methods The patient was a 27-year-old female.She had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdle muscles were involved subsequently.Clinical data of this patient were collected.The leg muscle magnetic resonance imaging (MRI) and an open biopsy of left tibialis anterior muscle were performed.In addition to histological,enzyme histochemical staining and ultrastructural examination,immunohistochemical staining with antibody against CAV3 was done.CAV3 gene was analyzed in the patient and her parents.Results Tl-weighted enhanced skeletal muscle MRI of the lower limbs showed the abnormal signal in distal and proximal muscles.Muscle biopsy showed moderate dystrophic changes and immunostaining for CAV3 showed reduced plasmalemma in the muscle fibers.Gene analysis disclosed a heterozygous c.136G > A (p.Ala46Thr)mutation in the CAV3 gene,and the patient's parents did not have this mutation.Conclusions We report a distal myopathy case caused by c.136G > A (p.Ala46Thr) mutation in the CAV3 gene,who had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdal muscles were also involved.This would help clinical doctors to know more about this rare myopathy.%目的 报道1例远端型小窝蛋白病患者,并研究其临床、病理及基因特点.方法 患者为27岁女性,以双下肢非对称起病,逐渐累及近端肢带肌.收集并总结该患者临床资料,行双侧下肢MRI检查.对患者左侧胫前肌行活体组织病理检查.同时对患者及其父母行小窝蛋白-3(caveolin-3,CAV3)基因检测.结果 患者双下肢MRI T1WI加权像增强结果提示远端肌群及近端肌群均存在异常信号影.骨骼肌病理检查发现光镜下呈中度肌营养不良样改变,免疫组织化学染色可见CAV3蛋白表达下降.患者存在CAV3基因c.136G

  9. Systematic Review of topotecan (Hycamtin in relapsed small cell lung cancer

    Directory of Open Access Journals (Sweden)

    Riemsma Rob

    2010-08-01

    Full Text Available Abstract Background To undertake a systematic review of the available data for oral and intravenous topotecan in adults with relapsed small cell lung cancer (SCLC for whom re-treatment with the first line regimen is not considered appropriate. Methods We searched six databases from 1980 up to March 2009 for relevant trials regardless of language or publication status. Relevant studies included any randomised trial of any chemotherapeutic treatment against any comparator in this licensed indication. Where possible we used apposite quantitative methods. Where meta-analysis was considered unsuitable for some or all of the data, we employed a narrative synthesis method. For indirect comparisons we used the method of Bucher et al., where available data allowed it, otherwise we used narrative descriptions. Results Seven unique studies met the inclusion criteria, four of which could be used in our analyses. These included one study comparing oral topotecan plus best supportive care (BSC to BSC alone, one study comparing intravenous topotecan to cyclophosphamide, adriamycin and vincristine (CAV, and two studies comparing oral topotecan with intravenous topotecan. All four studies appear to be well conducted and with low risk of bias. Oral topotecan plus BSC has advantages over BSC alone in terms of survival (hazard ratio = 0.61; 95% CI, 0.43 to 0.87 and quality of life (EQ-5 D difference: 0.15; 95% CI, 0.05 to 0.25. Intravenous topotecan was at least as effective as CAV in the treatment of patients with recurrent small-cell lung cancer and resulted in improved quality-of-life with respect to several symptoms. CAV was associated with significantly less grade 4 thrombocytopenia compared with IV topotecan (risk ratio = 5.83; 95% CI, 2.35 to 14.42. Survival (hazard ratio = 0.98; 95% CI, 0.77 to 1.25 and response (pooled risk ratio = 1.04; 95% CI, 0.58 to 1.85 data were similar for the oral and IV topotecan groups. Symptom control was also very similar

  10. 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

  11. TRIM72 modulates caveolar endocytosis in repair of lung cells.

    Science.gov (United States)

    Nagre, Nagaraja; Wang, Shaohua; Kellett, Thomas; Kanagasabai, Ragu; Deng, Jing; Nishi, Miyuki; Shilo, Konstantin; Oeckler, Richard A; Yalowich, Jack C; Takeshima, Hiroshi; Christman, John; Hubmayr, Rolf D; Zhao, Xiaoli

    2016-03-01

    Alveolar epithelial and endothelial cell injury is a major feature of the acute respiratory distress syndrome, in particular when in conjunction with ventilation therapies. Previously we showed [Kim SC, Kellett T, Wang S, Nishi M, Nagre N, Zhou B, Flodby P, Shilo K, Ghadiali SN, Takeshima H, Hubmayr RD, Zhao X. Am J Physiol Lung Cell Mol Physiol 307: L449-L459, 2014.] that tripartite motif protein 72 (TRIM72) is essential for amending alveolar epithelial cell injury. Here, we posit that TRIM72 improves cellular integrity through its interaction with caveolin 1 (Cav1). Our data show that, in primary type I alveolar epithelial cells, lack of TRIM72 led to significant reduction of Cav1 at the plasma membrane, accompanied by marked attenuation of caveolar endocytosis. Meanwhile, lentivirus-mediated overexpression of TRIM72 selectively increases caveolar endocytosis in rat lung epithelial cells, suggesting a functional association between these two. Further coimmunoprecipitation assays show that deletion of either functional domain of TRIM72, i.e., RING, B-box, coiled-coil, or PRY-SPRY, abolishes the physical interaction between TRIM72 and Cav1, suggesting that all theoretical domains of TRIM72 are required to forge a strong interaction between these two molecules. Moreover, in vivo studies showed that injurious ventilation-induced lung cell death was significantly increased in knockout (KO) TRIM72(KO) and Cav1(KO) lungs compared with wild-type controls and was particularly pronounced in double KO mutants. Apoptosis was accompanied by accentuation of gross lung injury manifestations in the TRIM72(KO) and Cav1(KO) mice. Our data show that TRIM72 directly and indirectly modulates caveolar endocytosis, an essential process involved in repair of lung epithelial cells through removal of plasma membrane wounds. Given TRIM72's role in endomembrane trafficking and cell repair, we consider this molecule an attractive