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Sample records for al gen cftr

  1. Mutations in CFTR gene and clinical correlation in Argentine patients with congenital bilateral absence of the vas deferens Correlación de las características clínicas con mutaciones del gen CFTR en pacientes argentinos con ausencia bilateral congénita de vasos deferentes

    Directory of Open Access Journals (Sweden)

    Estrella M Levy

    2004-06-01

    Full Text Available Congenital bilateral absence of the vas deferens (CBAVD is a form of male infertility in which mutations in the cystic fibrosis transmembrane conductance regulator (CFTR gene have been identified. Here we identify different mutations of CFTR and the poly-T variant of intron 8 (IVS8 in Argentine patients and analyze sweat test values and clinical characteristic related to Cystic Fibrosis (CF. For counseling purposes the two most frequent mutations in Argentine CF population: DF508 and G542X were screened in wives. In all cases, it was possible to reduce the risk of CF/CBAVD descendants in these couples because none of the mutation were found in the 36 samples. Eight patients (23% showed abnormal chloride values (> 60 mmol/l. A second group of 6 patients (18% had borderline values of sweat chloride (40-59 mmol/l. We defined another group with 6 patients (18%, with normal sweat chloride levels (30-39 mmo/l and a fourth group of 14 (41% patients with sweat chloride below 30 mmol/l. DF508, the most frequent CF mutation in the Argentine population, was found on 15 of the 72 chromosomes (21%, R117H mutation was detected on 2 of 62 chromosomes (3%. Only one R347P allele was found on 28 chromosomes analyzed (2%. On a sample of 27 patients, IVS8 analysis showed a frequency of 6/56 chromosomes (11% of 5T allele. Even though these findings present an improvement in the detection of mutations related to clinical correlations in Argentine CBAVD population, the search for other common and uncommon mutations should be continued.La ausencia bilateral congénita de vasos deferentes (CBAVD es una forma de infertilidad masculina en la que se han identificado mutaciones en el gen de la conductancia transmembrana de la fibrosis quística (CFTR. Hemos estudiado en pacientes argentinos diferentes mutaciones en el CFTR y la variante poli T del intron 8 (IVS8 y analizado los valores de test del sudor y las características clínicas relacionadas a la Fibrosis Qu

  2. Factores genéticos de pacientes con ictus asociados al pronóstico

    OpenAIRE

    Giralt Casellas, Dolors

    2016-01-01

    El objetivo de este trabajo es, utilizando una base de datos de pacientes que han padecido un ictus, encontrar tendencias entre pronóstico al alta y medicamentos que toman los pacientes previamente al ictus. Seguidamente, cruzar los factores genéticos asociados anteriormente con bases de datos del genoma humano para ver los genes asociados a las tendencias encontradas y ver la drogabilidad de los genes. L'objectiu d'aquest treball és, utilitzant una base de dades de pacients que han patit ...

  3. Estudio de polimorfismos genéticos y respuesta al tratamiento en pacientes con migraña cronica

    OpenAIRE

    Navarro Ortega, Nicolás

    2015-01-01

    Se han llevado a cabo diversos estudios de asociación genómicos tipo GWAS (Genome Wide Association Studies) utilizando polimorfismos de un solo nucleótido (SNP) con el objeto de identificar regiones genómicas que estuvieran relacionadas tanto con la migraña, como con los tratamientos y los efectos de los mismos, así como el uso de polimorfismos que han sido publicados por su relación con la migraña (Anttila et al., 2010; Chasman et al., 2011; De Vries et al., 2009; Freilinger et al., 2012; Li...

  4. Optimized Gen-II FeCrAl cladding production in large quantity for campaign testing

    Energy Technology Data Exchange (ETDEWEB)

    Yamamoto, Yukinori [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Sun, Zhiqian [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Pint, Bruce A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Terrani, Kurt A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2016-06-03

    There are two major objectives in this report; (1) to optimize microstructure control of ATF FeCrAl alloys during tube drawing processes, and (2) to provide an update on the progress of ATF FeCrAl tube production via commercial manufacturers. Experimental efforts have been made to optimize the process parameters balancing the tube fabricability, especially for tube drawing processes, and microstructure control of the final tube products. Lab-scale sheet materials of Gen II FeCrAl alloys (Mo-containing and Nb-containing FeCrAl alloys) were used in the study, combined with a stepwise warm-rolling process and intermediate annealing, aiming to simulate the tube drawing process in a commercial tube manufacturer. The intermediate annealing at 650ºC for 1h was suggested for the tube-drawing process of Mo-containing FeCrAl alloys because it successfully softened the material by recovering the work hardening introduced through the rolling step, without inducing grain coarsening due to recrystallization. The final tube product is expected to have stabilized deformed microstructure providing the improved tensile properties with sufficient ductility. Optimization efforts on Nb-containing FeCrAl alloys focused on the effect of alloying additions and annealing conditions on the stability of deformed microstructure. Relationships between the second-phase precipitates (Fe2Nb-Laves phase) and microstructure stability are discussed. FeCrAl tube production through commercial tube manufacturers is currently in progress. Three different manufacturers, Century Tubes, Inc. (CTI), Rhenium Alloys, Inc. (RAI), and Superior Tube Company, Inc. (STC), are providing capabilities for cold-drawing, warm-drawing, and HPTR cold-pilgering, respectively. The first two companies are currently working on large quantity tube production (expected 250 ft length) of Gen I model FeCrAl alloy (B136Y3, at CTI) and Gen II (C35M4, at RAI), with the process parameters obtained from the experimental

  5. Mutations in CFTR gene and clinical correlation in Argentine patients with congenital bilateral absence of the vas deferens Correlación de las características clínicas con mutaciones del gen CFTR en pacientes argentinos con ausencia bilateral congénita de vasos deferentes

    OpenAIRE

    Levy, Estrella M; Patricia Granados; Vanesa Rawe; Santiago Brugo Olmedo; María C Luna; Eduardo Cafferata; Omar H Pivetta

    2004-01-01

    Congenital bilateral absence of the vas deferens (CBAVD) is a form of male infertility in which mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been identified. Here we identify different mutations of CFTR and the poly-T variant of intron 8 (IVS8) in Argentine patients and analyze sweat test values and clinical characteristic related to Cystic Fibrosis (CF). For counseling purposes the two most frequent mutations in Argentine CF population: DF508 and G542...

  6. Development of CFTR structure

    Directory of Open Access Journals (Sweden)

    AnnaEPatrick

    2012-09-01

    Full Text Available Cystic fibrosis is a lethal genetic disease caused by lack of functional cystic fibrosis transmembrane conductance regulator (CFTR proteins at the apical surface of secretory epithelia. CFTR is a multidomain protein, containing five domains, and its functional structure is attained in a hierarchical folding process. Most CF-causing mutations in CFTR, including the most common mutation, a deletion of phenylalanine at position 508 (ΔF508, are unable to properly fold into this functional native three dimensional structure. Currently, no high resolution structural information about full length CFTR exists. However, insight has been gained through examining homologous ABC transporter structures, molecular modeling, and high resolution structures of individual, isolated CFTR domains. Taken together, these studies indicate that the prevalent ΔF508 mutation disrupts two essential steps during the development of the native structure: folding of the first nucleotide binding domain (NBD1 and its later association with the fourth intracellular loop (ICL4 in the second transmembrane domain (TMD2. Therapeutics to rescue ΔF508 and other mutants in CFTR can be targeted to correct defects that occur during the complex folding process. This article reviews the structural relationships between CFTR and ABC transporters and current knowledge about how CFTR attains its structure--with a focus on how this process is altered by CF-causing mutations in a manner targetable by therapeutics.

  7. Funciones de los canales iónicos CFTR y ENAC en la fibrosis quística

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    Alejandra G. Palma

    2014-04-01

    Full Text Available La fibrosis quística se debe a la ausencia o defecto del canal transmembrana regulador de la fibrosis quística (CFTR, un canal de cloruro codificado en el gen cftr que juega un papel clave en la homeostasis del agua e iones. El CFTR es activado por el AMPc y se localiza en las membranas apicales y basolaterales de las vías aéreas, intestino y glándulas exocrinas. Una de sus funciones primarias en los pulmones es mantener la capa de líquido superficial a través de su función de canal y regular el canal epitelial de sodio sensible al amiloride (ENaC. Se han identificado más de 1900 mutaciones en el gen cftr. La enfermedad se caracteriza por secreciones viscosas en las glándulas exocrinas y por niveles elevados de cloruro de sodio en el sudor. En la fibrosis quística el CFTR no funciona y el ENaC está desregulado; el resultado es un aumento en la reabsorción de sodio y agua con la formación de un líquido viscoso. En las glándulas sudoríparas tanto el Na+ como el Cl- se retienen en el lumen causando una pérdida de electrolitos durante la sudoración y el NaCl se elimina al sudor. Así, los niveles elevados de NaCl son la base del test del sudor inducido por pilocarpina, un método de diagnóstico para la enfermedad. En esta revisión se discuten los movimientos de Cl- y Na+ en las glándulas sudoríparas y pulmón así como el papel del ENaC en la patogénesis de la enfermedad.

  8. GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update

    OpenAIRE

    Peakall, Rod; Smouse, Peter E.

    2012-01-01

    Summary: GenAlEx: Genetic Analysis in Excel is a cross-platform package for population genetic analyses that runs within Microsoft Excel. GenAlEx offers analysis of diploid codominant, haploid and binary genetic loci and DNA sequences. Both frequency-based (F-statistics, heterozygosity, HWE, population assignment, relatedness) and distance-based (AMOVA, PCoA, Mantel tests, multivariate spatial autocorrelation) analyses are provided. New features include calculation of new estimators of popula...

  9. Cabells blancs, un destí també escrit als gens

    OpenAIRE

    Bueno i Torrens, David, 1965-

    2016-01-01

    Un dels aspectes més evidents del pas del temps és l'emblanquiment dels cabells. Tanmateix, aquest procés únicament es produeix en les persones d'ascendència europea. En les d'origen asiàtic, africà i amerindi, és un fet rar i excepcional. Andrés Ruiz-Linares i els seus col·laboradors, de diverses universitats i centres de recerca europeus, sud-americans i australians, han aïllat el gen que el provoca, i també han identificat disset gens més que contribueixen a definir les característiques ge...

  10. Funciones de los canales iónicos CFTR y ENAC en la fibrosis quística

    OpenAIRE

    Alejandra G. Palma; Basilio A. Kotsias; Gabriela I. Marino

    2014-01-01

    La fibrosis quística se debe a la ausencia o defecto del canal transmembrana regulador de la fibrosis quística (CFTR), un canal de cloruro codificado en el gen cftr que juega un papel clave en la homeostasis del agua e iones. El CFTR es activado por el AMPc y se localiza en las membranas apicales y basolaterales de las vías aéreas, intestino y glándulas exocrinas. Una de sus funciones primarias en los pulmones es mantener la capa de líquido superficial a través de su función de canal y regula...

  11. Mutationen des Androgenrezeptor-Gens als mögliche Ursache der Antiandrogenresistenz beim Prostatakarzinom

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    Röpke A

    2004-01-01

    Full Text Available Der Androgenrezeptor (AR ist ein nukleärer Transkriptionsfaktor, der die intrazelluläre Wirkung der Androgene vermittelt. Das Prostatakarzinom ist in Deutschland das häufigste Krebsleiden beim Mann. Bei fortgeschrittenem Prostatakarzinom ist eine radikale Entfernung der Prostata nicht immer möglich. In diesen Fällen wird auf eine palliative endokrine Behandlung zurückgegriffen. Da primäre Prostatakarzinome in ihrem Wachstum meist androgenabhängig sind, ist die Ansprechrate auf eine Antiandrogentherapie entsprechend hoch und liegt bei ca. 75 %. Mit der Antiandrogentherapie wird versucht, die Konzentration von freizirkulierenden Androgenen herabzusetzen oder die transkriptionelle Eigenschaft des AR zu blockieren. Bei den meisten Patienten kommt es jedoch trotz einer zunächst erfolgreichen Antiandrogentherapie sekundär unter der Therapie zu einer erneuten Proliferation oder Metastasierung des Prostatakarzinoms. Bei ca. 25 % der Patienten wird bereits primär eine Resistenz gegenüber der Antiandrogentherapie beobachtet. Für das Versagen der Antiandrogentherapie können Veränderungen des AR im Tumor verantwortlich sein. Bei Prostatakarzinomen mit sekundärer Antiandrogenresistenz besteht häufig eine Amplifikation des AR-Gens. Dagegen zeigen Prostatakarzinome ohne Antiandrogentherapie sehr selten (1 % eine Amplifikation des AR-Genlokus, jedoch in 11 % eine Polysomie des X-Chromosoms, einschließlich des AR-Gens. Mutationen im AR-Gen werden bei metastasierten oder hormonrefraktären, seltener bei primären Prostatakarzinomen beschrieben. Es konnte in einer Vielzahl von Studien festgestellt werden, daß verschiedene Steroidhormone eine höhere Affinität zu mutierten AR aufweisen, dadurch zu einer höheren transkriptionellen Aktivität führen und so die zelluläre Proliferation stimulieren.

  12. El comentario de textos literarios en Secundaria: del modelo genérico al intertextual

    OpenAIRE

    Caro Valverde, María Teresa; González García, María

    2009-01-01

    El documento presenta un modelo de aprendizaje significativo del comentario de textos, tanto desde el tradicional punto de vista genérico como desde el innovador punto de vista intertextual.Por medio de guías de comentario para ambos casos, seguidas de ejemplos comentados y de una selección de páginas Web escogidas, el lector dispone de una visión estructural, pragmática y heurística sobre este tema didáctico.

  13. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal

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    Jaime Eduardo Muñoz Flórez

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.

  14. Intervista al GEN. B. Mariano MOSSA, Comandante Carabinieri Tutela Patrimonio Culturale

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    Luca Papi

    2014-09-01

    Full Text Available In the framework of the cultural heritage, several records are recognized to our Country. With regard to them, it is worth to mention the establishment of a selected body of investigators Carabinieri Tutela Patrimonio Culturale specifically dedicated to combating crimes against cultural heritage.The Interview to Gen. B. Mariano Mossa is mainly focused to highlight the experience gained over more than 40 years of activity by the mentioned body. In particular, it will be illustrated the features and services which can enjoy a citizen by downloading the mobile app developed by Carabinieri Tutela Patrimonio Culturale.

  15. CFTR activity and mitochondrial function

    OpenAIRE

    Angel Gabriel Valdivieso; Santa-Coloma, Tomás A.

    2013-01-01

    Cystic Fibrosis (CF) is a frequent and lethal autosomal recessive disease, caused by mutations in the gene encoding the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). Before the discovery of the CFTR gene, several hypotheses attempted to explain the etiology of this disease, including the possible role of a chloride channel, diverse alterations in mitochondrial functions, the overexpression of the lysosomal enzyme α-glucosidase and a deficiency in the cytosolic enzyme glucose 6-p...

  16. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal

    Directory of Open Access Journals (Sweden)

    Muñoz Flórez Jaime Eduardo

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.

  17. Análisis comparativo de la aplicación de distintos operadores genéticos al problema de job shop scheduling

    OpenAIRE

    Alfonso, Hugo; Salto, Carolina; Gallard, Raúl Hector

    2000-01-01

    Este trabajo describe un estudio de distintas soluciones al problema del job shop scheduling basado en algoritmos genéticos. La característica fundamental es la utilización de decodificadores incorporada a la representación del cromosuma. El objetivo del trabajo es comparar la performance de los distintos tipos de operadores de crossovers que se pueden usar con esta representación.

  18. CFTR activity and mitochondrial function

    Directory of Open Access Journals (Sweden)

    Angel Gabriel Valdivieso

    2013-01-01

    Full Text Available Cystic Fibrosis (CF is a frequent and lethal autosomal recessive disease, caused by mutations in the gene encoding the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR. Before the discovery of the CFTR gene, several hypotheses attempted to explain the etiology of this disease, including the possible role of a chloride channel, diverse alterations in mitochondrial functions, the overexpression of the lysosomal enzyme α-glucosidase and a deficiency in the cytosolic enzyme glucose 6-phosphate dehydrogenase. Because of the diverse mitochondrial changes found, some authors proposed that the affected gene should codify for a mitochondrial protein. Later, the CFTR cloning and the demonstration of its chloride channel activity turned the mitochondrial, lysosomal and cytosolic hypotheses obsolete. However, in recent years, using new approaches, several investigators reported similar or new alterations of mitochondrial functions in Cystic Fibrosis, thus rediscovering a possible role of mitochondria in this disease. Here, we review these CFTR-driven mitochondrial defects, including differential gene expression, alterations in oxidative phosphorylation, calcium homeostasis, oxidative stress, apoptosis and innate immune response, which might explain some characteristics of the complex CF phenotype and reveals potential new targets for therapy.

  19. La Comunidad Andina frente al reto del acceso a los recursos genéticos y la distribución de beneficios

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    Martha Isabel Gómez Lee

    2012-11-01

    Full Text Available Este artículo estudia las medidas jurídicas y políticas que ha adoptado la Comunidad Andina (CAN sobre el acceso a los recursos genéticos y la distribución de beneficios a la luz de los estudios de equidad en la distribución de beneficios de Bram de Jonge y Niels Louwaar (2009. Según estos autores, el principio de Acceso y Distribución de Beneficios (ADB se justifica por tres asimetrías que están presentes en este asunto. Por un lado, la doble asimetría entre la asignación y la explotación de los recursos y, por otro, la asimetría entre los derechos de propiedad intelectual y los derechos legales de los recursos genéticos y los conocimientos tradicionales. En este contexto, la CAN ratifica los principios de soberanía de los recursos genéticos y de Acceso y Distribución de Beneficios del Convenio sobre Diversidad Biológica. Las principales respuestas que ha dado la CAN al reto del acceso a los recursos genéticos y distribución de beneficios son las Decisiones 391, 486 y 523. En este contexto, el Programa BIOCAN está trabajando en los temas de especial relevancia para la región, dentro de los cuales están los desafíos de asignarle valor a los recursos genéticos y el de la biopiratería.

  20. Plantes, gens, salut i ecologia

    OpenAIRE

    Bueno i Torrens, David, 1965-

    2008-01-01

    Als articles Por als gens i Manipulant els gens vaig parlar de la iniciativa per declarar Catalunya lliure de transgènics i vaig dir que totes les plantes que conreem han estat modificades en el decurs del temps. Els contraris a les plantes transgèniques parlen de possibles efectes negatius sobre la salut i el medi ambient [...].

  1. Microsatélites amplificados al azar (RAM) en estudios de diversidad genética vegetal Random amplified microsatellites (RAM´s) in plant genetic diversity studies

    OpenAIRE

    Jaime Eduardo Muñoz Flórez; Ana Cruz Morillo Coronado; Yacenia Morillo Coronado

    2008-01-01

    Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM) en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se difere...

  2. Base Molecular del asesoramiento genético de fibrosis quística

    OpenAIRE

    Muñoz Domínguez, Carlos

    1997-01-01

    La Fibrosis Quística (FQ) es una enfermedad autosómica recesiva producida por mutaciones en el gen CFTR. Dicho gen produce una proteína (CFTR) implicada en el transporte de iones Cl- a través de la membrana plasmática. La alteración ... La consecuencia de estas secreciones afecta principalmente a pulmones, páncreas exocrino, aparato reproductor masculino y glándulas sudoríparas, provocando un cuadro clínico muy variable de unos pacientes a otros. Tanto la elevada concentración iónica en el su...

  3. Endocytic Trafficking of CFTR in Health and Disease

    OpenAIRE

    Ameen, Nadia; Silvis, Mark; Bradbury, Neil A.

    2006-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl-selective anion channel expressed in epithelial tissues. Mutations in CFTR lead to the debilitating genetic disease cystic fibrosis (CF). Within each epithelial cell, CFTR interacts with a large number of transient macromolecular complexes, many of which are involved in the trafficking and targeting of CFTR. Understanding how these complexes regulate the trafficking and fate of CFTR, provides a singular insight not only in...

  4. A truncated CFTR protein rescues endogenous ΔF508-CFTR and corrects chloride transport in mice

    OpenAIRE

    Cormet-Boyaka, Estelle; Jeong S Hong; Berdiev, Bakhram K.; James A Fortenberry; Rennolds, Jessica; Clancy, J. P.; Benos, Dale J.; Boyaka, Prosper N.; Sorscher, Eric J.

    2009-01-01

    Cystic fibrosis (CF) is most frequently associated with deletion of phenylalanine at position 508 (ΔF508) in the CF transmembrane conductance regulator (CFTR) protein. The ΔF508-CFTR mutant protein exhibits a folding defect that affects its processing and impairs chloride-channel function. This study aimed to determine whether CFTR fragments approximately half the size of wild-type CFTR and complementary to the portion of CFTR bearing the mutation can specifically rescue the processing of end...

  5. Array CGH com a primera opció per al diagnòstic genètic postnatal

    OpenAIRE

    Castells Sarret, Neus

    2015-01-01

    La citogenètica convencional detecta un 3-5% dels pacients amb retard global del desenvolupament / discapacitat intel·lectual (RGD / DI) i / o malformacions congènites (MC). L'amplificació de sondes múltiples dependents de lligació (MLPA) permet incrementar la taxa diagnòstica entre 2,4-5,8%. Actualment els arrays d'hibridació genòmica comparada (aCGH) constitueixen l'eina diagnòstica amb major rendiment en pacients amb RGD / DI, MC i trastorns de l'espectre autista (TEA). L'objectiu del p...

  6. Activation of CFTR-mediated Cl- Transport by Magnolin

    Institute of Scientific and Technical Information of China (English)

    JIN Ling-ling; LIU Xin; SUN Yan; LIN Sen; ZHOU Na; XU Li-na; YU BO; HOU Shu-guang; YANG Hong

    2008-01-01

    Magnolin is a herbal compound from Magnolia biondii Pamp.It possesses numerous biological activities.Cystic fibrosis transmembrane conductance regulator(CFTR)is all epithelial chloride channel that plays a key role in the fluid secretion of various exocrine organs.In the present study,the activation of CFTR-mediated chloride transport by magnolin is indentified and characterized.In CFTR stably trailsfected FRT cells.magnolin increases CFTR Cl- currents in a concentration-dependent manner.The activation of magnolin on CFTR is rapid,reversible,and cAMP-dependent.Magnolin does not elevate cellular cAMP level.indicating that it activates CFTR by direct binding and interaction with CFTR protein.Magnolin selectively activates wildtype CFTR rather than mutant CFTIL Magnolin may present a novel class of therapeutic lead compound for the treatment of diseases associated with reduced CFTR function such as keratoconjunctivitis sicca,idiopathic chronic pancreatiti,and chromc constipation.

  7. DEL GEN EGOISTA AL GEN ALTRUISTA

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    Oscar José Fernández Galíndez

    2010-06-01

    Full Text Available The Darwinian thought derived from the interpretations and / or rereadings done on Charles Darwin's work, they have generated many controversy. Even the same Darwin demonstrated in many occasions her, doubts on her, before raised ideas, of there that I dress the evolutionism as paradigm, it suggests a serious review in the frame of the emergent thought and in the most igalitarian search of a society and less fragmented.

  8. Evaluation of CFTR gene mutations in Adana

    OpenAIRE

    Ozlem Goruroglu Ozturk; Filiz Kibar; Esin Damla Ziyanoglu Karacor; Salih Cetiner; Gulhan Sahin; Akgun Yaman

    2013-01-01

    ABSTRACT Objective: Cystic fibrosis is the most common autosomal recessive inherited disorder seen in the white populations. It develops in result of mutations of cystic fibrosis transmembrane regulator (CFTR) gene. Rate of these mutations vary in different geographical regions. In this study, we aimed to determine the frequency of CFTR gene mutations in Adana. Methods: DNA samples of 63 subjects (21 women, 42 men) who were diagnosed as cystic fibrosis at Balcali Hospital of Cukurova Universi...

  9. Evaluation of CFTR gene mutations in Adana

    OpenAIRE

    Öztürk, Özlem Görüroğlu; Filiz KIBAR; Karaçor, Esin Damla Ziyanoğlu; Çetiner, Salih; Şahin, Gülhan; Yaman, Akgün

    2013-01-01

    ABSTRACT Objective: Cystic fibrosis is the most common autosomal recessive inherited disorder seen in the white populations. It develops in result of mutations of cystic fibrosis transmembrane regulator (CFTR) gene. Rate of these mutations vary in different geographical regions. In this study, we aimed to determine the frequency of CFTR gene mutations in Adana. Methods: DNA samples of 63 subjects (21 women, 42 men) who were diagnosed as cystic fibrosis at Balcalı Hospital of Çukurova Unive...

  10. Detección de Leishmania spp. en base al gen que codifica la proteína HSP20

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    Ana M Montalvo

    2014-10-01

    Full Text Available Objetivos. Explorar una nueva diana para el diagnóstico molecular de Leishmania. Materiales y métodos. Se evaluó la utilidad del gen que codifica la proteína de choque térmico de 20kDa (hsp20 para la detección de Leishmania por medio de la reacción en cadena de la polimerasa (PCR.Se normalizó la PCR y se determinaron los parámetros analíticos, así como la validez y seguridad diagnóstica y la concordancia con la PCR-18S. Se evaluó la PCR-hsp20 con ADN obtenido de un grupo de muestras clínicas de distinta procedencia. Resultados. Los parámetros analíticos resultaron adecuados. La sensibilidad obtenida fue de 86% y la especificidad del 100%, la concordancia con el método de referencia resultó buena (κ = 0,731, lo que apoya su posible uso para el diagnóstico. La posibilidad de identificación posterior de la especie mediante secuenciación del producto amplificado le confiere una ventaja adicional. Conclusiones. Se demuestra la utilidad de este gen como una nueva diana para la detección del género Leishmania. Debido a su potencial, se recomienda mejorar la sensibilidad del procedimiento y realizar su evaluación en diversas regiones endémicas.

  11. Detección de Leishmania spp. en base al gen que codifica la proteína HSP20

    Directory of Open Access Journals (Sweden)

    Ana M. Montalvo

    2014-12-01

    Full Text Available Objetivos. Explorar una nueva diana para el diagnóstico molecular de Leishmania. Materiales y métodos. Se evaluó la utilidad del gen que codifica la proteína de choque térmico de 20kDa (hsp20 para la detección de Leishmania por medio de la reacción en cadena de la polimerasa (PCR.Se normalizó la PCR y se determinaron los parámetros analíticos, así como la validez y seguridad diagnóstica y la concordancia con la PCR-18S. Se evaluó la PCR-hsp20 con ADN obtenido de un grupo de muestras clínicas de distinta procedencia. Resultados. Los parámetros analíticos resultaron adecuados. La sensibilidad obtenida fue de 86% y la especificidad del 100%, la concordancia con el método de referencia resultó buena (κ = 0,731, lo que apoya su posible uso para el diagnóstico. La posibilidad de identificación posterior de la especie mediante secuenciación del producto amplificado le confiere una ventaja adicional. Conclusiones. Se demuestra la utilidad de este gen como una nueva diana para la detección del género Leishmania. Debido a su potencial, se recomienda mejorar la sensibilidad del procedimiento y realizar su evaluación en diversas regiones endémicas.

  12. Parámetros genéticos y fenotípicos para peso al año, circunferencia escrotal y talla en ganado Simmental y Simbrah en México

    OpenAIRE

    José Antonio Torres-Vázquez; Coralia Inés Valentina Manzanilla Pech; Aurelio Borrayo Zepeda; Ángel Ríos-Utrera; Vicente Eliezer Vega-Murillo; Guillermo Martínez-Velázquez; Juan José Baeza Rodríguez; Moisés Montaño-Bermúdez

    2012-01-01

    El objetivo fue estimar los componentes de (co)varianza, heredabilidades y correlaciones genéticas y fenotípicas para peso al año (YW), circunferencia escrotal (SC) y talla (FS) en toros jóvenes Simmental y Simbrah de México. Todas las características se ajustaron a 305 días acorde a los lineamientos descritos por la Federación para el Mejoramiento Genético de los Bovinos de Carne (BIF). El archivo final de datos incluyó 1,949 registros Simmental y 1,259 Simbrah. Para estimar los compones de ...

  13. Modulation of CFTR gating by permeant ions.

    Science.gov (United States)

    Yeh, Han-I; Yeh, Jiunn-Tyng; Hwang, Tzyh-Chang

    2015-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is unique among ion channels in that after its phosphorylation by protein kinase A (PKA), its ATP-dependent gating violates microscopic reversibility caused by the intimate involvement of ATP hydrolysis in controlling channel closure. Recent studies suggest a gating model featuring an energetic coupling between opening and closing of the gate in CFTR's transmembrane domains and association and dissociation of its two nucleotide-binding domains (NBDs). We found that permeant ions such as nitrate can increase the open probability (Po) of wild-type (WT) CFTR by increasing the opening rate and decreasing the closing rate. Nearly identical effects were seen with a construct in which activity does not require phosphorylation of the regulatory domain, indicating that nitrate primarily affects ATP-dependent gating steps rather than PKA-dependent phosphorylation. Surprisingly, the effects of nitrate on CFTR gating are remarkably similar to those of VX-770 (N-(2,4-Di-tert-butyl-5-hydroxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide), a potent CFTR potentiator used in clinics. These include effects on single-channel kinetics of WT CFTR, deceleration of the nonhydrolytic closing rate, and potentiation of the Po of the disease-associated mutant G551D. In addition, both VX-770 and nitrate increased the activity of a CFTR construct lacking NBD2 (ΔNBD2), indicating that these gating effects are independent of NBD dimerization. Nonetheless, whereas VX-770 is equally effective when applied from either side of the membrane, nitrate potentiates gating mainly from the cytoplasmic side, implicating a common mechanism for gating modulation mediated through two separate sites of action. PMID:25512598

  14. Métodos y usos agrícolas de la ingeniería genética aplicada al cultivo de arroz

    Directory of Open Access Journals (Sweden)

    Cristina DiazGranados D.

    2012-08-01

    Full Text Available Methods and agricultural uses of genetic engineering applied to rice crop Resumen: En biotecnología de arroz se han logrado avances en transformación genética, con importantes resultados en el mejoramiento genético de variedades elite de las subespecies japónica e índica. Con el propósito de revisar los métodos y los usos agrícolas de la ingeniería genética aplicada al cultivo del arroz, se usaron varias palabras claves en idioma inglés en algunas de las bases de datos de revistas científicas indexadas, disponibles en el Sistema Nacional de Bibliotecas de la Universidad Nacional de Colombia (SINAB, seleccionando documentos publicados entre 2000 y 2011. La base de esta revisión inicial, se complementó con artículos publicados en fechas anteriores, que se consideraron relevantes, debido a que implicaban cambios metodológicos importantes. Desde que se logró producir la primera planta transgénica de arroz a finales de los 80´s, varios protocolos para la transferencia de genes se han empleado con éxito logrando la modificación genética de más de 60 cultivares de arroz. Para ello se han empleado sistemas de transformación tanto directos como indirectos.  Se han realizado modificaciones de rasgos importantes en el cultivo, tales como la resistencia a factores bióticos (insectos, hongos, bacterias, virus, nematodos, tolerancia a factores abióticos (salinidad, sequía, altas y bajas temperaturas, inmersión, y mejoramiento de características agronómicas (calidad nutricional, rendimiento, uso de nutrientes, tolerancia a herbicidas. Palabras claves: Arroz; cultivos transgénicos; factores bióticos; factores abióticos. Abstract: In rice biotechnology advances have been made in genetic transformation, with significant results in breeding elite varieties of japonica and indica subspecies. In order to review the methods and agricultural uses of genetic engineering applied to rice, calves were used several words in English in

  15. Evaluation of CFTR gene mutations in Adana

    Directory of Open Access Journals (Sweden)

    Ozlem Goruroglu Ozturk

    2013-04-01

    Full Text Available ABSTRACT Objective: Cystic fibrosis is the most common autosomal recessive inherited disorder seen in the white populations. It develops in result of mutations of cystic fibrosis transmembrane regulator (CFTR gene. Rate of these mutations vary in different geographical regions. In this study, we aimed to determine the frequency of CFTR gene mutations in Adana. Methods: DNA samples of 63 subjects (21 women, 42 men who were diagnosed as cystic fibrosis at Balcali Hospital of Cukurova University, were studied for 19 different CFTR mutations by the strip assay method which is based on reverse hybridization. Results: In cystic fibrosis diagnosed patients, 19 mutations were observed of which 9 were homozygous and 10 were heterozygous. ∆F508 frequency was found as 11.9%, and rate of homozygous was found as 66.7%. Mutation frequencies of W1282X and N1303K were found as 2.40% and 4.80% respectively and rate of homozygous mutations were 50% for both. I148T mutation frequency was found as 3.20% and all were heterozygous. For the whole 19 mutations, frequency of mutation in 63 subjects was 22.3%. Conclusion: Detection of CFTR gene mutations by the strip assay method by reverse hybridization is an easy, fast and informative method. However, due to improvability of the common mutations in probable cystic fibrosis patients because of heterogenity in this region, it is still a major problem and does not exclude cystic fibrosis diagnosis. But this problematic issue can be overcome by evaluating the whole exons of CFTR mutations by advanced molecular tecniques. Key words: CFTR, cystic fibrosis, molecular diagnosis, reverse hibridisation [Cukurova Med J 2013; 38(2.000: 202-208

  16. Islet-intrinsic effects of CFTR mutation.

    Science.gov (United States)

    Koivula, Fiona N Manderson; McClenaghan, Neville H; Harper, Alan G S; Kelly, Catriona

    2016-07-01

    Cystic fibrosis-related diabetes (CFRD) is the most significant extra-pulmonary comorbidity in cystic fibrosis (CF) patients, and accelerates lung decline. In addition to the traditional view that CFRD is a consequence of fibrotic destruction of the pancreas as a whole, emerging evidence may implicate a role for cystic fibrosis transmembrane-conductance regulator (CFTR) in the regulation of insulin secretion from the pancreatic islet. Impaired first-phase insulin responses and glucose homeostasis have also been reported in CF patients. CFTR expression in both human and mouse beta cells has been confirmed, and recent studies have shown differences in endocrine pancreatic morphology from birth in CF. Recent experimental evidence suggests that functional CFTR channels are required for insulin exocytosis and the regulation of membrane potential in the pancreatic beta cell, which may account for the impairments in insulin secretion observed in many CF patients. These novel insights suggest that the pathogenesis of CFRD is more complicated than originally thought, with implications for diabetes treatment and screening in the CF population. This review summarises recent emerging evidence in support of a primary role for endocrine pancreatic dysfunction in the development of CFRD. Summary • CF is an autosomal recessive disorder caused by mutations in the CFTR gene • The vast majority of morbidity and mortality in CF results from lung disease. However CFRD is the largest extra-pulmonary co-morbidity and rapidly accelerates lung decline • Recent experimental evidence shows that functional CFTR channels are required for normal patterns of first phase insulin secretion from the pancreatic beta cell • Current clinical recommendations suggest that insulin is more effective than oral glucose-lowering drugs for the treatment of CFRD. However, the emergence of CFTR corrector and potentiator drugs may offer a personalised approach to treating diabetes in the CF population

  17. Manipular els gens

    OpenAIRE

    Bueno i Torrens, David, 1965-

    2008-01-01

    l'article Por als gens vaig parlar de la iniciativa per declarar Catalunya lliure de transgènics, i vaig dir que els motius que esgrimeixen els promotors no tenen base científica. Aquests motius es poden resumir en tres punts: la negativa a manipular organismes vius, la seguretat alimentària i l'afectació ecològica [...].

  18. Microsatélites amplificados al azar (RAM en estudios de diversidad genética vegetal Random amplified microsatellites (RAM´s in plant genetic diversity studies

    Directory of Open Access Journals (Sweden)

    Jaime Eduardo Muñoz Flórez

    2008-12-01

    Full Text Available Se revisó el uso e importancia, ventajas, desventajas y características de la técnica Microsatélites Amplificados al Azar (RAM en uchuva Physalis peruviana, mora Rubus spp, guayaba Psidium guajava y heliconias Heliconia spp. En mora se diferenciaron las especies R. glaucus, R. robustus y R. urticifolius, se detectaron duplicados y se encontró alta variabilidad genética en R. glaucus, la especie más importante. En uchuva se encontró alta diversidad y dos accesiones de fruto rojo que se diferenciaron genéticamente de las amarillas y una región geográfica con alta variabilidad. En guayaba los cebadores fueron altamente polimórficos y se encontró alta variabilidad en el Valle del Cauca. En heliconias y especies relacionadas se diferenciaron las familias del orden Zingiberales, algunos subgéneros y variaciones en la especie. La técnica es de bajo costo, utiliza un cebador, no requiere información previa, es altamente polimórfica y diferencia especies en los taxones evaluados.The use and importance, advantages, disadvantages and features of the Random Amplified Microsatellites RAMs technique, were reviewed in Cape gooseberry Physalis peruviana, blackberry Rubus spp, guava Psidium guajava and heliconias Heliconia spp. In blackberry, we differentiated the species R. glaucus, R. robustus y R. urticifolius, detected duplicated accessions and found high genetic diversity in R. glaucus, the most important specie. In cape gooseberry we found high diversity and two red fruit accessions genetically differentiated from the yellow fruit ones and a geographical region with high variability. In guava, primers were highly polymorphic and found high variability in Valle del Cauca region. In Heliconia and related species we differentiated families belonging to Zingiberal order, between some sub genera and variation among specie. The technique has low cost of implementation, use a single primer, do not require previous information, is highly

  19. GEN 105 Courses/sanptutorial

    OpenAIRE

    ncbdf

    2015-01-01

    GEN 105 Assignment: Reading and Retention GEN 105 Assignment: Elevator Speech GEN 105 CheckPoint: Technological Tools GEN 105 CheckPoint: Distance Learning I GEN 105 CheckPoint: Distance Learning II GEN 105 CheckPoint: Communicating in Forums GEN 105 Week 2 Discussion Questions GEN 105 Week 4 Discussion Questions GEN 105 Week 6 Discussion Questions GEN 105 Week 8 Discussion Questions GEN 105 Assignment: University Library Article Search GEN 105 Chec...

  20. Regulated trafficking of the CFTR chloride channel

    NARCIS (Netherlands)

    Braakman, L.J.; Kleizen, B.; Jonge, H.R. de

    2000-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR), the ABC transporter encoded by the cystic fibrosis gene, is localized in the apical membrane of epithelial cells where it functions as a cyclic AMP-regulated chloride channel and as a regulator of other ion channels and transporters. Wh

  1. Caracterización de la diversidad genética en naranja y comparación del polimorfismo de microsatélites amplificados al azar (RAMs usando electroforesis de poliacrilamida y agarosa

    Directory of Open Access Journals (Sweden)

    Muñoz Flores Jaime Eduardo

    2009-12-01

    Full Text Available Se compararon las eficiencias de tres métodos de electroforesis en agarosa y poliacrilamida, usando la cámara pequeña de DNA Sequencing System y cámara grande OWL Sequi-Gen Sequencing Cell, en la detección del polimorfismo en 21 accesiones de naranja (Citrus sinensis con empleo del cebador CGA. El gel de poliacrilamida dio mejor resolución de los productos amplificados vía PCR producidos por RAMs. Este permitió una mejor detección de bandas de ADN polimórficas, lo que facilitó la identificación de la variabilidad genética. La electroforesis en agarosa puede ser más conveniente en otras aplicaciones, debido al bajo costo y fácil aplicación. El estudio de diversidad genética en naranja usando microsatélites RAMs diferenció 51 accesiones en siete grupos con 0.75 de similaridad y 0.25 de heterocigosidad, lo que revela bajo polimorfismo genético. La técnica RAMs permitió agrupar las accesiones en Comunes o Blancas, Navel y Pigmentadas o Sanguinas.

  2. CFTR and Wnt/beta-catenin signaling in lung development

    Directory of Open Access Journals (Sweden)

    Love Damon

    2008-07-01

    Full Text Available Abstract Background Cystic fibrosis transmembrane conductance regulator (CFTR was shown previously to modify stretch induced differentiation in the lung. The mechanism for CFTR modulation of lung development was examined by in utero gene transfer of either a sense or antisense construct to alter CFTR expression levels. The BAT-gal transgenic reporter mouse line, expressing β-galactosidase under a canonical Wnt/β-catenin-responsive promoter, was used to assess the relative roles of CFTR, Wnt, and parathyroid hormone-related peptide (PTHrP in lung organogenesis. Adenoviruses containing full-length CFTR, a short anti-sense CFTR gene fragment, or a reporter gene as control were used in an intra-amniotic gene therapy procedure to transiently modify CFTR expression in the fetal lung. Results A direct correlation between CFTR expression levels and PTHrP levels was found. An inverse correlation between CFTR and Wnt signaling activities was demonstrated. Conclusion These data are consistent with CFTR participating in the mechanicosensory process essential to regulate Wnt/β-Catenin signaling required for lung organogenesis.

  3. N-Alpha-Acetyltransferases and Regulation of CFTR Expression.

    Science.gov (United States)

    Vetter, Ali J; Karamyshev, Andrey L; Patrick, Anna E; Hudson, Henry; Thomas, Philip J

    2016-01-01

    The majority of cystic fibrosis (CF)-causing mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) lead to the misfolding, mistrafficking, and degradation of the mutant protein. Inhibition of degradation does not effectively increase the amount of trafficking competent CFTR, but typically leads to increased ER retention of misfolded forms. Thus, the initial off pathway steps occur early in the processing of the protein. To identify proteins that interact with these early forms of CFTR, in vitro crosslink experiments identified cotranslational partners of the nascent chain of the severe misfolded mutant, G85E CFTR. The mutant preferentially interacts with a subunit of an N-alpha-acetyltransferase A. Based on recent reports that acetylation of the N-termini of some N-end rule substrates control their ubiquitination and subsequent degradation, a potential role for this modification in regulation of CFTR expression was assessed. Knockdown experiments identified two complexes, which affect G85E CFTR proteins levels, NatA and NatB. Effects of the knockdowns on mRNA levels, translation rates, and degradation rates established that the two complexes regulate G85E CFTR through two separate mechanisms. NatA acts indirectly by regulating transcription levels and NatB acts through a previously identified, but incompletely understood posttranslational mechanism. This regulation did not effect trafficking of G85E CFTR, which remains retained in the ER, nor did it alter the degradation rate of CFTR. A mutation predicted to inhibit N-terminal acetylation of CFTR, Q2P, was without effect, suggesting neither system acts directly on CFTR. These results contradict the prediction that N-terminal acetylation of CFTR determines its fitness as a proteasome substrate, but rather NatB plays a role in the conformational maturation of CFTR in the ER through actions on an unidentified protein. PMID:27182737

  4. GEN 105 Course tutorial/ indigohelp

    OpenAIRE

    SCD

    2015-01-01

    For more classes visit www.indigohelp.com   GEN 105 Assignment: Reading and Retention GEN 105 Assignment: Elevator Speech GEN 105 CheckPoint: Technological Tools GEN 105 CheckPoint: Distance Learning I GEN 105 CheckPoint: Distance Learning II GEN 105 CheckPoint: Communicating in Forums GEN 105 Week 2 Discussion Questions GEN 105 Week 4 Discussion Questions GEN 105 Week 6 Discussion Questions GEN 105 Week 8 Discussion Questions GEN 105 A...

  5. Molecular Motors and Apical CFTR Traffic in Epithelia

    Directory of Open Access Journals (Sweden)

    Dmitri V. Kravtsov

    2013-05-01

    Full Text Available Intracellular protein traffic plays an important role in the regulation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR chloride channels. Microtubule and actin-based motor proteins direct CFTR movement along trafficking pathways. As shown for other regulatory proteins such as adaptors, the involvement of protein motors in CFTR traffic is cell-type specific. Understanding motor specificity provides insight into the biology of the channel and opens opportunity for discovery of organ-specific drug targets for treating CFTR-mediated diseases.

  6. Divergent signaling via SUMO modification: potential for CFTR modulation.

    Science.gov (United States)

    Ahner, Annette; Gong, Xiaoyan; Frizzell, Raymond A

    2016-02-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is generally responsible for the cAMP/PKA regulated anion conductance at the apical membranes of secretory epithelial cells. Mutations in CFTR underlie cystic fibrosis (CF), in which the most common variant, F508del, causes protein misfolding and its proteasome-mediated degradation. A new pathway that contributes to mutant CFTR degradation is mediated by the small heat shock protein, Hsp27, which cooperates with Ubc9, the E2 enzyme for SUMOylation, to selectively conjugate mutant CFTR with SUMO-2/3. This SUMO paralog can form polychains, which are recognized by the ubiquitin E3 enzyme, RNF4, leading to CFTR ubiquitylation and recognition by the proteasome. We found also that F508del CFTR could be modified by SUMO-1, a paralog that does not support SUMO polychain formation. The use of different SUMO paralogs to modify and target a single substrate for divergent purposes is not uncommon. In this short review we discuss the possibility that conjugation with SUMO-1 could protect mutant CFTR from disposal by RNF4 and similar ubiquitin ligases. We hypothesize that such a pathway could contribute to therapeutic efforts to stabilize immature mutant CFTR and thereby enhance the action of therapeutics that correct CFTR trafficking to the apical membranes. PMID:26582473

  7. Problemática legal generada por la falta de regulación respecto al manejo de bases de datos genéticos de uso forense

    OpenAIRE

    Paz Karaman, Paola Andrea

    2012-01-01

    En Colombia, el INMLCF1administra la base de datos genéticos con fines de investigación criminal, en ella se almacenan perfiles de ADN (datos de carácter personal) de víctimas (de desaparición forzada y familiares de los desaparecidos), de personas relacionadas con la autoría de un delito y de vestigios encontrados en el lugar de los hechos. Sin embargo el marco regulatorio actual no delimita los criterios de uso de la base de datos genéticos con fines de investigación criminal de cara a la p...

  8. The ΔF508-CFTR mutation inhibits wild-type CFTR processing and function when co-expressed in human airway epithelia and in mouse nasal mucosa

    Directory of Open Access Journals (Sweden)

    Tucker Torry A

    2012-09-01

    Full Text Available Abstract Background Rescue or correction of CFTR function in native epithelia is the ultimate goal of CF therapeutics development. Wild-type (WT CFTR introduction and replacement is also of particular interest. Such therapies may be complicated by possible CFTR self-assembly into an oligomer or multimer. Results Surprisingly, functional CFTR assays in native airway epithelia showed that the most common CFTR mutant, ΔF508-CFTR (ΔF-CFTR, inhibits WT-CFTR when both forms are co-expressed. To examine more mechanistically, both forms of CFTR were transfected transiently in varying amounts into IB3-1 CF human airway epithelial cells and HEK-293 human embryonic kidney cells null for endogenous CFTR protein expression. Increasing amounts of ΔF-CFTR inhibited WT-CFTR protein processing and function in CF human airway epithelial cells but not in heterologous HEK-293 cells. Stably expressed ΔF-CFTR in clones of the non-CF human airway epithelial cell line, CALU-3, also showed reduction in cAMP-stimulated anion secretion and in WT-CFTR processing. An ultimate test of this dominant negative-like effect of ΔF-CFTR on WT-CFTR was the parallel study of two different CF mouse models: the ΔF-CFTR mouse and the bitransgenic CFTR mouse corrected in the gut but null in the lung and airways. WT/ΔF heterozygotes had an intermediate phenotype with regard to CFTR agonist responses in in vivo nasal potential difference (NPD recordings and in Ussing chamber recordings of short-circuit current (ISC in vitro on primary tracheal epithelial cells isolated from the same mice. In contrast, CFTR bitransgenic +/− heterozygotes had no difference in their responses versus +/+ wild-type mice. Conclusions Taken altogether, these data suggest that ΔF-CFTR and WT-CFTR co-assemble into an oligomeric macromolecular complex in native epithelia and share protein processing machinery and regulation at the level of the endoplasmic reticulum (ER. As a consequence, ΔF-CFTR slows WT-CFTR

  9. Phosphatase inhibitors activate normal and defective CFTR chloride channels.

    OpenAIRE

    Becq, F; Jensen, T J; Chang, X B; Savoia, A.; Rommens, J M; Tsui, L C; Buchwald, M; Riordan, J R; Hanrahan, J W

    1994-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is regulated by phosphorylation and dephosphorylation at multiple sites. Although activation by protein kinases has been studied in some detail, the dephosphorylation step has received little attention. This report examines the mechanisms responsible for the dephosphorylation and spontaneous deactivation ("rundown") of CFTR chloride channels excised from transfected Chinese hamster ovary (CHO) and human airway epi...

  10. Algoritmos genéticos

    OpenAIRE

    Martínez Páez, José Jesús

    2004-01-01

    Esta técnica se basa en el concepto de evolución a través de selección de los mejores individuos, y de los operadores genéticos de selección, reproducción y mutación. Se trata entonces, de definir un espacio de soluciones para el problema que se quiere solucionar, en una cadena de bits. A esto se le conoce como la codificación del cromosoma, donde cada bit, denominado gen  tiene cierto significado especial. Inicialmente el algoritmo genera al azar muchas de estas cadenas o seres, es decir, un...

  11. Marcadores genéticos dopaminérgicos, serotoninérgicos y de la monoamino-oxidasa (MAO) relacionados con la adicción a los opiáceos, a la cocaína y al alcohol

    OpenAIRE

    Mateu Hernández, César Andrés

    2013-01-01

    Introducción: la genética está experimentando un avance espectacular y se postula como una herramienta importante para aportar nuevos descubrimientos en trastornos complejos como son las adicciones. En el presente trabajo se ha realizado un estudio de asociación genética de polimorfismos VNTR, un estudio familiar y un estudio mixto de asociación genético-familiar en una muestra de policonsumidores con adicción al alcohol, la cocaína y los opiáceos. Material y métodos: en una muestra de 302 po...

  12. Identificació de noves dianes terapèutiques i miRNAs implicats en la resistència al metotrexat mitjançant genòmica funcional

    OpenAIRE

    Mencía Trinchant, Núria

    2013-01-01

    Una de les accepcions de la farmacogenòmica es l'estudi dels efectes d'un tractament farmacològic sobre els nivells d'expressió gènica. Aquesta estratègia inclou la identificació de les variacions de gens individuals, l'avaluació de les interaccions entre els seus productes i la caracterització dels fenotips derivats de la resposta al fàrmac El metotrexat (MTX) és un fàrmac inhibidor de l'enzim dihidrofolat reductasa (DHFR) utilitzat en el tractament del càncer. El treball presentat en...

  13. Potentiators exert distinct effects on human, murine, and Xenopus CFTR.

    Science.gov (United States)

    Cui, Guiying; Khazanov, Netaly; Stauffer, Brandon B; Infield, Daniel T; Imhoff, Barry R; Senderowitz, Hanoch; McCarty, Nael A

    2016-08-01

    VX-770 (Ivacaftor) has been approved for clinical usage in cystic fibrosis patients with several CFTR mutations. Yet the binding site(s) on CFTR for this compound and other small molecule potentiators are unknown. We hypothesize that insight into this question could be gained by comparing the effect of potentiators on CFTR channels from different origins, e.g., human, mouse, and Xenopus (frog). In the present study, we combined this comparative molecular pharmacology approach with that of computer-aided drug discovery to identify and characterize new potentiators of CFTR and to explore possible mechanism of action. Our results demonstrate that 1) VX-770, NPPB, GlyH-101, P1, P2, and P3 all exhibited ortholog-specific behavior in that they potentiated hCFTR, mCFTR, and xCFTR with different efficacies; 2) P1, P2, and P3 potentiated hCFTR in excised macropatches in a manner dependent on the degree of PKA-mediated stimulation; 3) P1 and P2 did not have additive effects, suggesting that these compounds might share binding sites. Also 4) using a pharmacophore modeling approach, we identified three new potentiators (IOWH-032, OSSK-2, and OSSK-3) that have structures similar to GlyH-101 and that also exhibit ortholog-specific potentiation of CFTR. These could potentially serve as lead compounds for development of new drugs for the treatment of cystic fibrosis. The ortholog-specific behavior of these compounds suggest that a comparative pharmacology approach, using cross-ortholog chimeras, may be useful for identification of binding sites on human CFTR. PMID:27288484

  14. Insereixen material genètic al nucli cel·lular per a teràpia gènica amb nanodiscs

    OpenAIRE

    Villaverde Corrales, Antonio; Vázquez Gómez, Esther

    2011-01-01

    Investigadors de la UAB han aconseguit encapsular material genètic i alliberar-lo directament dins el nucli de les cèl·lules, per tal de dur a terme teràpia gènica, mitjançant partícules amb forma de disc de la grandària de només un pocs nanòmetres. Els nanodiscs, tal i com els han batejat els investigadors, travessen ràpidament l'interior de la cèl·lula i es concentren en el nucli, de manera que incrementarien l'eficiència del procés de transferència genètica.

  15. Mutationsstudie des Dihydropyrimidin-Dehydrogenase-Gens (DPYD) einer Kontrollpopulation zur Abklärung von DPD-Insuffizienz als genetische Ursache von Fluoropyrimidin-Intoleranz.

    OpenAIRE

    Seck, Katharina

    2006-01-01

    Mithilfe der denaturierenden HPLC wurde eine Mutationsanalyse des gesamten kodierenden Abschnitts des DPYD-Gens durchgeführt. Für jedes Exon und teilweise für einzelne Fragmente eines Exons wurden die optimale DHPLC-Temperatur und der Elutionsgradient anhand von Berechnung sowie experimentell ermittelt, so dass durch das jeweilige Chromatogramm eine definitive Zuordnung zu Wildtyp oder zu spezifischen bekannten Mutanten möglich wurde; stichprobenartige Kontrollen bestätigten in der Sequenzier...

  16. Predominant constitutive CFTR conductance in small airways

    Directory of Open Access Journals (Sweden)

    Lytle Christian

    2005-01-01

    Full Text Available Abstract Background The pathological hallmarks of chronic obstructive pulmonary disease (COPD are inflammation of the small airways (bronchiolitis and destruction of lung parenchyma (emphysema. These forms of disease arise from chronic prolonged infections, which are usually never present in the normal lung. Despite the fact that primary hygiene and defense of the airways presumably requires a well controlled fluid environment on the surface of the bronchiolar airway, very little is known of the fluid and electrolyte transport properties of airways of less than a few mm diameter. Methods We introduce a novel approach to examine some of these properties in a preparation of minimally traumatized porcine bronchioles of about 1 mm diameter by microperfusing the intact bronchiole. Results In bilateral isotonic NaCl Ringer solutions, the spontaneous transepithelial potential (TEP; lumen to bath of the bronchiole was small (mean ± sem: -3 ± 1 mV; n = 25, but when gluconate replaced luminal Cl-, the bionic Cl- diffusion potentials (-58 ± 3 mV; n = 25 were as large as -90 mV. TEP diffusion potentials from 2:1 NaCl dilution showed that epithelial Cl- permeability was at least 5 times greater than Na+ permeability. The anion selectivity sequence was similar to that of CFTR. The bionic TEP became more electronegative with stimulation by luminal forskolin (5 μM+IBMX (100 μM, ATP (100 μM, or adenosine (100 μM, but not by ionomycin. The TEP was partially inhibited by NPPB (100 μM, GlyH-101* (5–50 μM, and CFTRInh-172* (5 μM. RT-PCR gave identifying products for CFTR, α-, β-, and γ-ENaC and NKCC1. Antibodies to CFTR localized specifically to the epithelial cells lining the lumen of the small airways. Conclusion These results indicate that the small airway of the pig is characterized by a constitutively active Cl- conductance that is most likely due to CFTR.

  17. Nasal Potential Difference in Cystic Fibrosis considering Severe CFTR Mutations

    OpenAIRE

    2015-01-01

    The gold standard for diagnosing cystic fibrosis (CF) is a sweat chloride value above 60 mEq/L. However, this historical and important tool has limitations; other techniques should be studied, including the nasal potential difference (NPD) test. CFTR gene sequencing can identify CFTR mutations, but this method is time-consuming and too expensive to be used in all CF centers. The present study compared CF patients with two classes I-III CFTR mutations (10 patients) (G1), CF patients with class...

  18. Identificación genómica de loci y rutas génicas que afectan al crecimiento y deposición grasa en porcino

    OpenAIRE

    Pérez Montarelo, Dafne

    2013-01-01

    El crecimiento y la deposición grasa son algunos de los caracteres económicos más relevantes en la producción porcina, además, el cerdo es utilizado como especie modelo en el estudio de la obesidad y patologías humanas con ella relacionadas, por lo que el estudio de estos caracteres suscita un gran interés. El objetivo general de esta tesis ha sido profundizar en el conocimiento de la base genética de la regulación del crecimiento y la deposición grasa en porcino, identificando genes, rutas g...

  19. GenBank

    Data.gov (United States)

    U.S. Department of Health & Human Services — GenBank is the NIH genetic sequence database, an annotated collection of all publicly available DNA sequences. GenBank is designed to provide and encourage access...

  20. La relación entre los riesgos, la precaución y la responsabilidad en los daños al medio ambiente por la liberación de organismos genéticamente modificados.

    Directory of Open Access Journals (Sweden)

    Sebastián Rebolledo Aguirre

    2012-05-01

    Full Text Available ResumenEl presente trabajo tiene por objeto mostrar la relación teórica y práctica entre la compleja noción de los riesgos; el principio precautorio como norma jurídico-ambiental, y la institución fundamental en todo sistema jurídico de la responsabilidad por daños.de los daños al medio ambiente por la liberación de organismos genéticamente modificados es una utilísima herramienta para determinar el régimen de responsabilidad aplicable, la interpretación de las normas jurídicas involucradas y controlar la discrecionalidad administrativa.Se explica que los riesgos inciertos son el fundamento y la razón que dan operatividad al principio precautorio, y que la funcionalidad de éste en el contexto de los daños al medio ambiente por la liberación de organismos genéticamente modificados es una utilísima herramienta para determinar el régimen de responsabilidad aplicable, la interpretación de las normas jurídicas involucradas y controlar la discrecionalidad administrativa.AbstractThis paper aims to show the theoretical and practical relationship between the complex notion of risk, the precautionary principle as an environmental legal standard, and the legal doctrine of liability for damages.It explains that uncertain risks are the foundation and the reason that makes the precautionary principle operative, and that, in the context of environmental damages caused by the release of GMOs, it is a very useful tool to determine the liability regime and the interpretation of the rules involved, and to control administrative discretion.

  1. Regulatory crosstalk by protein kinases on CFTR trafficking and activity

    Science.gov (United States)

    Farinha, Carlos Miguel; Swiatecka-Urban, Agnieszka; Brautigan, David; Jordan, Peter

    2016-01-01

    Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e. channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease.

  2. Assessing the Disease-Liability of Mutations in CFTR

    OpenAIRE

    Ferec, Claude; Cutting, Garry R

    2012-01-01

    Over 1900 mutations have been reported in the cystic fibrosis transmembrane conductance regulator (CFTR), the gene defective in patients with cystic fibrosis. These mutations have been discovered primarily in individuals who have features consistent with the diagnosis of CF. In some cases, it has been recognized that the mutations are not causative of cystic fibrosis but are responsible for disorders with features similar to CF, and these conditions have been termed CFTR-related disorders or ...

  3. Targeted therapies to improve CFTR function in cystic fibrosis.

    Science.gov (United States)

    Brodlie, Malcolm; Haq, Iram J; Roberts, Katie; Elborn, J Stuart

    2015-01-01

    Cystic fibrosis is the most common genetically determined, life-limiting disorder in populations of European ancestry. The genetic basis of cystic fibrosis is well established to be mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene that codes for an apical membrane chloride channel principally expressed by epithelial cells. Conventional approaches to cystic fibrosis care involve a heavy daily burden of supportive treatments to combat lung infection, help clear airway secretions and maintain nutritional status. In 2012, a new era of precision medicine in cystic fibrosis therapeutics began with the licensing of a small molecule, ivacaftor, which successfully targets the underlying defect and improves CFTR function in a subgroup of patients in a genotype-specific manner. Here, we review the three main targeted approaches that have been adopted to improve CFTR function: potentiators, which recover the function of CFTR at the apical surface of epithelial cells that is disrupted in class III and IV genetic mutations; correctors, which improve intracellular processing of CFTR, increasing surface expression, in class II mutations; and production correctors or read-through agents, which promote transcription of CFTR in class I mutations. The further development of such approaches offers great promise for future therapeutic strategies in cystic fibrosis. PMID:26403534

  4. Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis.

    Science.gov (United States)

    Dekkers, Johanna F; Berkers, Gitte; Kruisselbrink, Evelien; Vonk, Annelotte; de Jonge, Hugo R; Janssens, Hettie M; Bronsveld, Inez; van de Graaf, Eduard A; Nieuwenhuis, Edward E S; Houwen, Roderick H J; Vleggaar, Frank P; Escher, Johanna C; de Rijke, Yolanda B; Majoor, Christof J; Heijerman, Harry G M; de Winter-de Groot, Karin M; Clevers, Hans; van der Ent, Cornelis K; Beekman, Jeffrey M

    2016-06-22

    Identifying subjects with cystic fibrosis (CF) who may benefit from cystic fibrosis transmembrane conductance regulator (CFTR)-modulating drugs is time-consuming, costly, and especially challenging for individuals with rare uncharacterized CFTR mutations. We studied CFTR function and responses to two drugs-the prototypical CFTR potentiator VX-770 (ivacaftor/KALYDECO) and the CFTR corrector VX-809 (lumacaftor)-in organoid cultures derived from the rectal epithelia of subjects with CF, who expressed a broad range of CFTR mutations. We observed that CFTR residual function and responses to drug therapy depended on both the CFTR mutation and the genetic background of the subjects. In vitro drug responses in rectal organoids positively correlated with published outcome data from clinical trials with VX-809 and VX-770, allowing us to predict from preclinical data the potential for CF patients carrying rare CFTR mutations to respond to drug therapy. We demonstrated proof of principle by selecting two subjects expressing an uncharacterized rare CFTR genotype (G1249R/F508del) who showed clinical responses to treatment with ivacaftor and one subject (F508del/R347P) who showed a limited response to drug therapy both in vitro and in vivo. These data suggest that in vitro measurements of CFTR function in patient-derived rectal organoids may be useful for identifying subjects who would benefit from CFTR-correcting treatment, independent of their CFTR mutation. PMID:27334259

  5. Assessment of cystic fibrosis transmembrane conductance regulator (CFTR) activity in CFTR-null mice after bone marrow transplantation

    OpenAIRE

    Bruscia, Emanuela M.; Price, Joanna E.; Cheng, Ee-chun; Weiner, Scott; Caputo, Christina; Ferreira, Elisa C.; Egan, Marie E.; Krause, Diane S.

    2006-01-01

    Several studies have demonstrated that bone marrow (BM)-derived cells give rise to rare epithelial cells in the gastrointestinal (GI) and respiratory tracts after BM transplantation into myeloablated recipients. We investigate whether, after transplantation of cystic fibrosis transmembrane conductance regulator (CFTR)-positive BM-derived cells, BM-derived GI and airway epithelial cells can provide CFTR activity in the GI tract and nasal epithelium of recipient cystic fibrosis mice. CFTR−/− mi...

  6. Physiological Adaptation of the Bacterium Lactococcus lactis in Response to the Production of Human CFTR

    NARCIS (Netherlands)

    Steen, Anton; Wiederhold, Elena; Gandhi, Tejas; Breitling, Rainer; Slotboom, Dirk Jan

    2011-01-01

    Biochemical and biophysical characterization of CFTR (the cystic fibrosis transmembrane conductance regulator) is thwarted by difficulties to obtain sufficient quantities of correctly folded and functional protein. Here we have produced human CFTR in the prokaryotic expression host Lactococcus lacti

  7. Characterizing responses to CFTR-modulating drugs using rectal organoids derived from subjects with cystic fibrosis

    NARCIS (Netherlands)

    Dekkers, Johanna F; Berkers, Gitte; Kruisselbrink, Evelien; Vonk, Annelotte; de Jonge, Hugo R; Janssens, Hettie M; Bronsveld, Inez; van de Graaf, Eduard A; Nieuwenhuis, Edward E S; Houwen, Roderick H J; Vleggaar, Frank P; Escher, Johanna C; de Rijke, Yolanda B; Majoor, Christof J; Heijerman, Harry G M; de Winter-de Groot, Karin M; Clevers, Hans; van der Ent, Cornelis K; Beekman, Jeffrey M

    2016-01-01

    Identifying subjects with cystic fibrosis (CF) who may benefit from cystic fibrosis transmembrane conductance regulator (CFTR)-modulating drugs is time-consuming, costly, and especially challenging for individuals with rare uncharacterized CFTR mutations. We studied CFTR function and responses to tw

  8. 21 CFR 866.5900 - Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation detection system.

    Science.gov (United States)

    2010-04-01

    ... regulator (CFTR) gene mutation detection system. 866.5900 Section 866.5900 Food and Drugs FOOD AND DRUG... DEVICES Immunological Test Systems § 866.5900 Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation detection system. (a) Identification. The CFTR gene mutation detection system is a...

  9. Activation of G551D-CFTR by Bicyclooctane Compounds Is cAMP-dependent and Exhibits Low Sensitivity to Thiazolidinone CFTR Inhibitor CFTRinh-172

    Institute of Scientific and Technical Information of China (English)

    WANG Ying; ZHAO Lu; HE Cheng-yan; XU Li-na; YANG Hong

    2005-01-01

    The G551D-CFTR mutation causing cystic fibrosis (CF) results from a missense mutation at codon 551(G551D) in the gene encoding of the cystic fibrosis transmembrane conductance regulator (CFTR). The G551D mutation in CFTR results in a reduced functional channel but G551D-CFTR is appropriately inserted in the apical membrane. In previous studies we discovered a class of high-affinity bicyclooctane (BCO)G551D-CFTR activators(G551DBCOs) with Kd down to 1μmol/L. In this study, we analyzed the pharmacological activation of G551D-CFTR by the G551DBcos by means of short circuit current analysis and cell-based fluorescence quenching assay. The G551DBCOs-induced G551D-CFTR activation is cAMP-dependent and is less sensitive to thiazolidinone CFTR inhibitor CFTRinh-172. These data suggest that (1) the phosphorylation of G551D-CFTR by protein kinase A is required for the activation by G551DBcos; (2) G551DBCos and CFTRinh-172 may act at the same site on the G551D-CFTR molecule.

  10. Rattlesnake Phospholipase A2 Increases CFTR-Chloride Channel Current and Corrects ∆F508CFTR Dysfunction: Impact in Cystic Fibrosis.

    Science.gov (United States)

    Faure, Grazyna; Bakouh, Naziha; Lourdel, Stéphane; Odolczyk, Norbert; Premchandar, Aiswarya; Servel, Nathalie; Hatton, Aurélie; Ostrowski, Maciej K; Xu, Haijin; Saul, Frederick A; Moquereau, Christelle; Bitam, Sara; Pranke, Iwona; Planelles, Gabrielle; Teulon, Jacques; Herrmann, Harald; Roldan, Ariel; Zielenkiewicz, Piotr; Dadlez, Michal; Lukacs, Gergely L; Sermet-Gaudelus, Isabelle; Ollero, Mario; Corringer, Pierre-Jean; Edelman, Aleksander

    2016-07-17

    Deletion of Phe508 in the nucleotide binding domain (∆F508-NBD1) of the cystic fibrosis transmembrane regulator (CFTR; a cyclic AMP-regulated chloride channel) is the most frequent mutation associated with cystic fibrosis. This mutation affects the maturation and gating of CFTR protein. The search for new high-affinity ligands of CFTR acting as dual modulators (correctors/activators) presents a major challenge in the pharmacology of cystic fibrosis. Snake venoms are a rich source of natural multifunctional proteins, potential binders of ion channels. In this study, we identified the CB subunit of crotoxin from Crotalus durissus terrificus as a new ligand and allosteric modulator of CFTR. We showed that CB interacts with NBD1 of both wild type and ∆F508CFTR and increases their chloride channel currents. The potentiating effect of CB on CFTR activity was demonstrated using electrophysiological techniques in Xenopus laevis oocytes, in CFTR-HeLa cells, and ex vivo in mouse colon tissue. The correcting effect of CB was shown by functional rescue of CFTR activity after 24-h ΔF508CFTR treatments with CB. Moreover, the presence of fully glycosylated CFTR was observed. Molecular docking allowed us to propose a model of the complex involving of the ABCβ and F1-like ATP-binding subdomains of ΔF508-NBD1. Hydrogen-deuterium exchange analysis confirmed stabilization in these regions, also showing allosteric stabilization in two other distal regions. Surface plasmon resonance competition studies showed that CB disrupts the ∆F508CFTR-cytokeratin 8 complex, allowing for the escape of ∆F508CFTR from degradation. Therefore CB, as a dual modulator of ΔF508CFTR, constitutes a template for the development of new anti-CF agents. PMID:27241308

  11. Steviol reduces MDCK Cyst formation and growth by inhibiting CFTR channel activity and promoting proteasome-mediated CFTR degradation.

    Directory of Open Access Journals (Sweden)

    Chaowalit Yuajit

    Full Text Available Cyst enlargement in polycystic kidney disease (PKD involves cAMP-activated proliferation of cyst-lining epithelial cells and transepithelial fluid secretion into the cyst lumen via cystic fibrosis transmembrane conductance regulator (CFTR chloride channel. This study aimed to investigate an inhibitory effect and detailed mechanisms of steviol and its derivatives on cyst growth using a cyst model in Madin-Darby canine kidney (MDCK cells. Among 4 steviol-related compounds tested, steviol was found to be the most potent at inhibiting MDCK cyst growth. Steviol inhibition of cyst growth was dose-dependent; steviol (100 microM reversibly inhibited cyst formation and cyst growth by 72.53.6% and 38.2±8.5%, respectively. Steviol at doses up to 200 microM had no effect on MDCK cell viability, proliferation and apoptosis. However, steviol acutely inhibited forskolin-stimulated apical chloride current in MDCK epithelia, measured with the Ussing chamber technique, in a dose-dependent manner. Prolonged treatment (24 h with steviol (100 microM also strongly inhibited forskolin-stimulated apical chloride current, in part by reducing CFTR protein expression in MDCK cells. Interestingly, proteasome inhibitor, MG-132, abolished the effect of steviol on CFTR protein expression. Immunofluorescence studies demonstrated that prolonged treatment (24 h with steviol (100 microM markedly reduced CFTR expression at the plasma membrane. Taken together, the data suggest that steviol retards MDCK cyst progression in two ways: first by directly inhibiting CFTR chloride channel activity and second by reducing CFTR expression, in part, by promoting proteasomal degradation of CFTR. Steviol and related compounds therefore represent drug candidates for treatment of polycystic kidney disease.

  12. Hanstruepera neustonica gen. nov., sp. nov., a zeaxanthin-producing member of the family Flavobacteriaceae isolated from estuarine water, and emendation of Sediminibacter furfurosus Khan et al. 2007 emend. Kwon et al. 2014, Mangrovimonas yunxiaonensis Li et al. 2013, Antarcticimonas flava Yang et al. 2009 and Hoppeia youngheungensis Kwon et al. 2014.

    Science.gov (United States)

    Hameed, Asif; Shahina, Mariyam; Lai, Wei-An; Lin, Shih-Yao; Liu, You-Cheng; Hsu, Yi-Han; Young, Chiu-Chung

    2015-02-01

    A Gram-staining-negative, strictly aerobic, yellowish-orange, flexirubin-positive, rod-shaped, non-flagellated, non-spore-forming and non-gliding marine bacterium, designated strain CC-PY-50(T), was isolated from estuarine water off Pingtung, Taiwan. The strain produced zeaxanthin as a major carotenoid pigment, and showed highest pairwise 16S rRNA gene sequence similarity to Bizionia hallyeonensis T-y7(T) (93.9 %) followed by Corallibacter vietnamensis KMM 6217(T) (93.8 %), Geojedonia litorea YCS-16(T) (93.7 %) and other members of the family Flavobacteriaceae (5 % of total) fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH, C16 : 0 and C16 : 1ω6c and/or C16 : 1ω7c. The DNA G+C content was 37.1 mol% and menaquinone-6 (MK-6) was the sole respiratory quinone. Based on the phylogenetic evidence and several distinguishing phenotypic and chemotaxonomic features, strain CC-PY-50(T) is proposed to represent a novel genus and species of the family Flavobacteriaceae, for which the name Hanstruepera neustonica gen. nov., sp. nov. is proposed. The type strain of the type species Hanstruepera neustonica gen. nov., sp. nov. is CC-PY-50(T) ( = JCM 19743(T) = BCRC 80747(T)). Emended descriptions of the species Sediminibacter furfurosus, Mangrovimonas yunxiaonensis, Antarcticimonas flava and Hoppeia youngheungensis are also proposed. PMID:25351879

  13. Algoritmo Genético aplicado al problema de programación en procesos tecnológicos de maquinado con ambiente Flow Shop

    Directory of Open Access Journals (Sweden)

    José Eduardo Márquez Delgado

    2012-01-01

    Full Text Available Debido a las limitaciones de las técnicas de optimización convencionales, en el siguiente trabajo se presenta una metaheurística basada en un algoritmo genético (AG, para resolver problemas de programación de tipo flow shop, con el objetivo de minimizar el tiempo de finalización de todos los trabajos, más conocido como makespan. Este problema, considerado de difícil solución, es típico de la optimización combinatoria y se presenta en talleres con tecnología de maquinado, donde existen máquinas herramientas convencionales y se fabrican diferentes tipos de piezas que tienen en común una misma ruta tecnológica (orden del proceso. La solución propuesta se probó con problemas clásicos publicados por otros autores, obteniéndose resultados satisfactorios en cuanto a la calidad de las soluciones encontradas y el tiempo de cómputo empleado.

  14. Nasal Potential Difference in Cystic Fibrosis considering Severe CFTR Mutations

    Directory of Open Access Journals (Sweden)

    Ronny Tah Yen Ng

    2015-01-01

    Full Text Available The gold standard for diagnosing cystic fibrosis (CF is a sweat chloride value above 60 mEq/L. However, this historical and important tool has limitations; other techniques should be studied, including the nasal potential difference (NPD test. CFTR gene sequencing can identify CFTR mutations, but this method is time-consuming and too expensive to be used in all CF centers. The present study compared CF patients with two classes I-III CFTR mutations (10 patients (G1, CF patients with classes IV-VI CFTR mutations (five patients (G2, and 21 healthy subjects (G3. The CF patients and healthy subjects also underwent the NPD test. A statistical analysis was performed using the Mann-Whitney, Kruskal-Wallis, χ2, and Fisher’s exact tests, α=0.05. No differences were observed between the CF patients and healthy controls for the PDMax, Δamiloride, and Δchloride + free + amiloride markers from the NPD test. For the finger value, a difference between G2 and G3 was described. The Wilschanski index values were different between G1 and G3. In conclusion, our data showed that NPD is useful for CF diagnosis when classes I-III CFTR mutations are screened. However, if classes IV-VI are considered, the NPD test showed an overlap in values with healthy subjects.

  15. CFTR and Ca2+ signaling in cystic fibrosis

    Directory of Open Access Journals (Sweden)

    Fabrice eAntigny

    2011-10-01

    Full Text Available Among the diverse physiological functions exerted by calcium signaling in living cells, its role in the regulation of protein biogenesis and trafficking remains incompletely understood. In cystic fibrosis (CF disease the most common CFTR (Cystic Fibrosis Transmembrane conductance Regulator mutation, F508del-CFTR generates a misprocessed protein that is abnormally retained in the endoplasmic reticulum (ER compartment, rapidly degraded by the ubiquitine/proteasome pathway and hence absent at the plasma membrane of CF epithelial cells. Recent studies have demonstrated that intracellular calcium signals consequent to activation of apical G protein-coupled receptors (GPCRs by different agonists are increased in CF airway epithelia. Moreover, the regulation of various intracellular calcium storage compartments, such as ER is also abnormal in CF cells. Although the molecular mechanism to explain this increase remains puzzling in epithelial cells, the F508del-CFTR mutation is proposed to be the origin of abnormal Ca2+ influx linking the calcium signaling to CFTR pathobiology. This article reviews the relationships between CFTR and calcium signaling in the context of the genetic disease cystic fibrosis.

  16. A molecular mechanism for aberrantCFTR-dependent HCO3– transport in cystic fibrosis

    OpenAIRE

    Ko, Shigeru B. H.; Shcheynikov, Nikolay; Choi, Joo Young; Luo, Xiang; Ishibashi, Kenichi; Thomas, Philip J.; Kim, Joo Young; Kim, Kyung Hwan; Lee, Min Goo; Naruse, Satoru; Muallem, Shmuel

    2002-01-01

    Aberrant HCO3– transport is a hallmark of cystic fibrosis (CF) and is associated with aberrant Cl–-dependent HCO3– transport by the cystic fibrosis transmembrane conductance regulator (CFTR). We show here that HCO3– current by CFTR cannot account for CFTR-activated HCO3– transport and that CFTR does not activate AE1–AE4. In contrast, CFTR markedly activates Cl– and OH–/HCO3– transport by members of the SLC26 family DRA, SLC26A6 and pendrin. Most notably, the SLC26s are electrogenic transporte...

  17. Technology evaluation: AAV-CFTR vector, targeted genetics.

    Science.gov (United States)

    Tebbutt, S J

    1999-08-01

    Targeted Genetics is developing a gene therapy product, the AAV-CFTR vector system, for the treatment of cystic fibrosis (CF). This involves administration of an adeno-associated virus (AAV) vector containing CF transmembrane conductance regulator gene (CFTR) directly to the lungs of cystic fibrosis patients. The drug has Orphan Drug Status [325713]. Medeva acquired the worldwide commercial rights to the therapy in November 1998. Medeva expects to file a license application for the therapy, in the US and Europe, by 2002 [325713]. The therapy utilizing the normal CFTR gene entered phase II trials for sinusitis [197407] and phase II trials for CF in the first quarter of 1997. Lehman Brothers predicts filing in 2001/2002 [318119]. PMID:11713770

  18. CFTR: A New Horizon in the Pathomechanism and Treatment of Pancreatitis.

    Science.gov (United States)

    Hegyi, Péter; Wilschanski, Michael; Muallem, Shmuel; Lukacs, Gergely L; Sahin-Tóth, Miklós; Uc, Aliye; Gray, Michael A; Rakonczay, Zoltán; Maléth, József

    2016-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is an ion channel that conducts chloride and bicarbonate ions across epithelial cell membranes. Mutations in the CFTR gene diminish the ion channel function and lead to impaired epithelial fluid transport in multiple organs such as the lung and the pancreas resulting in cystic fibrosis. Heterozygous carriers of CFTR mutations do not develop cystic fibrosis but exhibit increased risk for pancreatitis and associated pancreatic damage characterized by elevated mucus levels, fibrosis, and cyst formation. Importantly, recent studies demonstrated that pancreatitis causing insults, such as alcohol, smoking, or bile acids, strongly inhibit CFTR function. Furthermore, human studies showed reduced levels of CFTR expression and function in all forms of pancreatitis. These findings indicate that impairment of CFTR is critical in the development of pancreatitis; therefore, correcting CFTR function could be the first specific therapy in pancreatitis. In this review, we summarize recent advances in the field and discuss new possibilities for the treatment of pancreatitis. PMID:26856995

  19. Mechanosensitive activation of CFTR by increased cell volume and hydrostatic pressure but not shear stress.

    Science.gov (United States)

    Vitzthum, Constanze; Clauss, Wolfgang G; Fronius, Martin

    2015-11-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl(-) channel that is essential for electrolyte and fluid homeostasis. Preliminary evidence indicates that CFTR is a mechanosensitive channel. In lung epithelia, CFTR is exposed to different mechanical forces such as shear stress (Ss) and membrane distention. The present study questioned whether Ss and/or stretch influence CFTR activity (wild type, ∆F508, G551D). Human CFTR (hCFTR) was heterologously expressed in Xenopus oocytes and the response to the mechanical stimulus and forskolin/IBMX (FI) was measured by two-electrode voltage-clamp experiments. Ss had no influence on hCFTR activity. Injection of an intracellular analogous solution to increase cell volume alone did not affect hCFTR activity. However, hCFTR activity was augmented by injection after pre-stimulation with FI. The response to injection was similar in channels carrying the common mutations ∆F508 and G551D compared to wild type hCFTR. Stretch-induced CFTR activation was further assessed in Ussing chamber measurements using Xenopus lung preparations. Under control conditions increased hydrostatic pressure (HP) decreased the measured ion current including activation of a Cl(-) secretion that was unmasked by the CFTR inhibitor GlyH-101. These data demonstrate activation of CFTR in vitro and in a native pulmonary epithelium in response to mechanical stress. Mechanosensitive regulation of CFTR is highly relevant for pulmonary physiology that relies on ion transport processes facilitated by pulmonary epithelial cells. PMID:26357939

  20. GEN 105 UOP course tutorial/tutorialoutlet

    OpenAIRE

    NARESH 1

    2015-01-01

    For more course tutorials visit www.tutorialoutlet.com     GEN 105 Assignment: Reading and Retention GEN 105 Assignment: Elevator Speech GEN 105 CheckPoint: Technological Tools GEN 105 CheckPoint: Distance Learning I GEN 105 CheckPoint: Distance Learning II GEN 105 CheckPoint: Communicating in Forums GEN 105 Week 2 Discussion Questions GEN 105 Week 4 Discussion Questions GEN 105 Week 6 Discussion Questions GEN 105 Week 8 Discussion...

  1. Gen quimérico que utiliza el gen o cDNA de la insulina, en especial para terapia génica de la diabetes

    OpenAIRE

    Bosch i Tubert, Fàtima; Valera Abril, Alfons

    2001-01-01

    Gen quimérico que utiliza el gen o CDNA de la insulina, en especial para terapia génica de la diabetes. El gen quimérico está dirigido por un promotor o fusión de promotores, los cuales preferentemente son regulables y están activados por el proceso diabético. Preferentemente, se obtiene por fusión del gen de la insulina humana al promotor de la PEPCK (p-enolpiruvato carboxiquinasa). Dicho promotor (fragmento - 460 pb a + 73 pb) está fusionado a la zona 5'' franqueante del gen de la insulina ...

  2. Parsers funcionales genéricos

    OpenAIRE

    Martínez López, Pablo E.; Nestares, Gustavo A.

    1996-01-01

    En este trabajo se propone el uso de técnicas avanzadas de programación funcional para ofrecer una solución genérica al problema de parsing. Este problema consiste en que, dada una lista de tokens que representan una estructura, debe obtenerse una representación elaborada de la misma. La solución propuesta es genérica en dos sentidos. Primero, la técnica de diseño utilizada consiste en dar una biblioteca de combinadores, los cuales pueden combinarse para escribir soluciones a problemas com...

  3. Partial correction of the CFTR-dependent ABPA mouse model with recombinant adeno-associated virus gene transfer of truncated CFTR gene.

    Science.gov (United States)

    Mueller, Christian; Torrez, Daniel; Braag, Sofia; Martino, Ashley; Clarke, Tracy; Campbell-Thompson, Martha; Flotte, Terence R

    2008-01-01

    Recently, we have developed a model of airway inflammation in a CFTR knockout mouse utilizing Aspergillus fumigatus crude protein extract (Af-cpe) to mimic allergic bronchopulmonary aspergillosis (ABPA) 1, an unusual IgE-mediated hypersensitivity syndrome seen in up to 15% of cystic fibrosis (CF) patients and rarely elsewhere. We hypothesized that replacement of CFTR via targeted gene delivery to airway epithelium would correct aberrant epithelial cytokine signaling and ameliorate the ABPA phenotype in CFTR-deficient (CFTR 489X - /-, FABP-hCFTR + / +) mice. CFTR knockout mice underwent intra-tracheal (IT) delivery of recombinant adeno-associated virus serotype 5 (rAAV5Delta-264CFTR) or rAAV5-GFP at 2.58 x 10(12) viral genomes/mouse. All mice were then sensitized with two serial injections (200 microg) of crude Af antigen via the intra-peritoneal (IP) route. Untreated mice were sensitized without virus exposure. Challenges were performed 2 weeks after final sensitization, using a 0.25% solution containing Aspergillus fumigatus crude protein extract delivered by inhalation on three consecutive days. The rAAV5Delta-264CFTR-treated mice had lower total serum IgE levels (172513 ng/ml +/- 1312) than rAAV5-GFP controls (26 892 ng/ml +/- 3715) (p = 0.037) and non-treated, sensitized controls (24 816 +/- 4219 ng/ml). Serum IgG1 levels also were lower in mice receiving the CFTR vector. Interestingly, splenocytes from rAAV5Delta-264CFTR-treated mice secreted less IL-13, INFg, TNFa, RANTES and GM-CSF after ConA stimulation. Gene therapy with rAAV5Delta-264CFTR attenuated the hyper-IgE response in this reproducible CF mouse model of ABPA, with systemic effects also evident in the cytokine response of stimulated splenocytes. PMID:18023072

  4. CFTR Mutations in Congenital Absence of Vas Deferens

    Directory of Open Access Journals (Sweden)

    Ramin Radpour

    2007-01-01

    Full Text Available A qualitative diagnosis of infertility requires attention to female and male physical abnormalities,endocrine anomalies and genetic conditions that interfere with reproduction. Many genes arelikely to be involved in the complex process of reproduction. Cystic fibrosis (CF incidence variesin different White people populations (a higher incidence of CF is observed in northern–westernEuropean populations than in southern European populations, and therefore the incidence ofcongenital bilateral absence of the vas deferens (CBAVD may also vary in different Whitepeople populations. As CF is mainly observed in White people, hardly any data are available ofCBAVD in non-White people, but frequent polymorphisms such as 5T are observed in mostpopulations. The spectrum and distribution of cystic fibrosis transmembrane conductanceregulator gene (CFTR mutations differs between CBAVD and CF patients, and even comparedwith control individuals. Combinations of particular alleles at several polymorphic loci yieldinsufficient functional CFTR. The combination of the 5T allele in one copy of the CFTR genewith a cystic fibrosis mutation in the other copy is the most common cause of CBAVD in Iran.Because of techniques such as intracytoplasmic sperm injection (ICSI, CBAVD patients are nowable to father children, however such couples have an increased risk of having a child with cysticfibrosis, and therefore genetic testing and counseling should be provided. Around 10% ofobstructive azoospermia is congenital and is due to mutations the CF gene. This paper reviews therelationship of mutations in the CFTR gene with CBAVD.

  5. Bioelectric characterization of epithelia from neonatal CFTR knockout ferrets

    NARCIS (Netherlands)

    J.T. Fisher (John); S.R. Tyler (Scott); Y. Zhang (Yulong); B.J. Lee (Ben); X. Liu (Xiaoming); X. Sun (Xinying); H. Sui (Hongshu); B. Liang (Bo); M. Luo (Ma); W. Xie (Weiliang); I. Yi (Iasson); W. Zhou (Weili); Y. Song (Yiqing); N. Keiser (Nicholas); K. Wang (Kai); H.R. de Jonge (Hugo); J.F. Engelhardt (John)

    2013-01-01

    textabstractCystic fibrosis (CF) is a life-shortening, recessive, multiorgan genetic disorder caused by the loss of CF transmembrane conductance regulator (CFTR) chloride channel function found in many types of epithelia. Animal models that recapitulate the human disease phenotype are critical to un

  6. Insulin-like growth factor 1 (IGF-1 enhances the protein expression of CFTR.

    Directory of Open Access Journals (Sweden)

    Ha Won Lee

    Full Text Available Low levels of insulin-like growth factor 1 (IGF-1 have been observed in the serum of cystic fibrosis (CF patients. However, the effects of low serum IGF-1 on the cystic fibrosis transmembrane conductance regulator (CFTR, whose defective function is the primary cause of cystic fibrosis, have not been studied. Here, we show in human cells that IGF-1 increases the steady-state levels of mature wildtype CFTR in a CFTR-associated ligand (CAL- and TC10-dependent manner; moreover, IGF-1 increases CFTR-mediated chloride transport. Using an acceptor photobleaching fluorescence resonance energy transfer (FRET assay, we have confirmed the binding of CAL and CFTR in the Golgi. We also show that CAL overexpression inhibits forskolin-induced increases in the cell-surface expression of CFTR. We found that IGF-1 activates TC10, and active TC10 alters the functional association between CAL and CFTR. Furthermore, IGF-1 and active TC10 can reverse the CAL-mediated reduction in the cell-surface expression of CFTR. IGF-1 does not increase the expression of ΔF508 CFTR, whose processing is arrested in the ER. This finding is consistent with our observation that IGF-1 alters the functional interaction of CAL and CFTR in the Golgi. However, when ΔF508 CFTR is rescued with low temperature or the corrector VRT-325 and proceeds to the Golgi, IGF-1 can increase the expression of the rescued ΔF508 CFTR. Our data support a model indicating that CAL-CFTR binding in the Golgi inhibits CFTR trafficking to the cell surface, leading CFTR to the degradation pathway instead. IGF-1-activated TC10 changes the interaction of CFTR and CAL, allowing CFTR to progress to the plasma membrane. These findings offer a potential strategy using a combinational treatment of IGF-1 and correctors to increase the post-Golgi expression of CFTR in cystic fibrosis patients bearing the ΔF508 mutation.

  7. Optimal correction of distinct CFTR folding mutants in rectal cystic fibrosis organoids.

    Science.gov (United States)

    Dekkers, Johanna F; Gogorza Gondra, Ricardo A; Kruisselbrink, Evelien; Vonk, Annelotte M; Janssens, Hettie M; de Winter-de Groot, Karin M; van der Ent, Cornelis K; Beekman, Jeffrey M

    2016-08-01

    Small-molecule therapies that restore defects in cystic fibrosis transmembrane conductance regulator (CFTR) gating (potentiators) or trafficking (correctors) are being developed for cystic fibrosis (CF) in a mutation-specific fashion. Options for pharmacological correction of CFTR-p.Phe508del (F508del) are being extensively studied but correction of other trafficking mutants that may also benefit from corrector treatment remains largely unknown.We studied correction of the folding mutants CFTR-p.Phe508del, -p.Ala455Glu (A455E) and -p.Asn1303Lys (N1303K) by VX-809 and 18 other correctors (C1-C18) using a functional CFTR assay in human intestinal CF organoids.Function of both CFTR-p.Phe508del and -p.Ala455Glu was enhanced by a variety of correctors but no residual or corrector-induced activity was associated with CFTR-p.Asn1303Lys. Importantly, VX-809-induced correction was most dominant for CFTR-p.Phe508del, while correction of CFTR-p.Ala455Glu was highest by a subgroup of compounds called bithiazoles (C4, C13, C14 and C17) and C5.These data support the development of mutation-specific correctors for optimal treatment of different CFTR trafficking mutants, and identify C5 and bithiazoles as the most promising compounds for correction of CFTR-p.Ala455Glu. PMID:27103391

  8. In vivo pharmacology and antidiarrheal efficacy of a thiazolidinone CFTR inhibitor in rodents.

    Science.gov (United States)

    Sonawane, N D; Muanprasat, Chatchai; Nagatani, Ray; Song, Yuanlin; Verkman, A S

    2005-01-01

    A small-molecule inhibitor of the cystic fibrosis transmembrane conductance regulator (CFTR), 3-[(3-trifluoromethyl)phenyl]-5-[(4-carboxyphenyl)methylene]-2-thioxo-4-thiazolidinone (CFTR(inh)-172), reduces enterotoxin-induced intestinal fluid secretion in rodents. Here, we study CFTR(inh)-172 pharmacology and antidiarrheal efficacy in rodents using (14)C-labeled CFTR(inh)-172, liquid chromatography/mass spectrometry, and a closed intestinal loop model of fluid secretion. CFTR(inh)-172 was cleared primarily by renal glomerular filtration without chemical modification. CFTR(inh)-172 accumulated in liver within 5 min after intravenous infusion in mice, and was concentrated fivefold in bile over blood. At 30-240 min, CFTR(inh)-172 was found mainly in liver, intestine, and kidney, with little detectable in the brain, heart, skeletal muscle, or lung. Pharmacokinetic analysis in rats following intravenous bolus infusion showed a distribution volume of 770 mL with redistribution and elimination half-times of 0.14 h and 10.3 h, respectively. CFTR(inh)-172 was stable in hepatic microsomes. Closed-loop studies in mice indicated that a single intraperitoneal injection of 20 microg CFTR(inh)-172 inhibited fluid accumulation at 6 h after cholera toxin by >90% in duodenum and jejunum, approximately 60% in ileum and accumulation of CFTR(inh)-172 account for its efficacy as an antidiarrheal. PMID:15761937

  9. How Phosphorylation and ATPase Activity Regulate Anion Flux though the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR).

    Science.gov (United States)

    Zwick, Matthias; Esposito, Cinzia; Hellstern, Manuel; Seelig, Anna

    2016-07-01

    The cystic fibrosis transmembrane conductance regulator (CFTR, ABCC7), mutations of which cause cystic fibrosis, belongs to the ATP-binding cassette (ABC) transporter family and works as a channel for small anions, such as chloride and bicarbonate. Anion channel activity is known to depend on phosphorylation by cAMP-dependent protein kinase A (PKA) and CFTR-ATPase activity. Whereas anion channel activity has been extensively investigated, phosphorylation and CFTR-ATPase activity are still poorly understood. Here, we show that the two processes can be measured in a label-free and non-invasive manner in real time in live cells, stably transfected with CFTR. This study reveals three key findings. (i) The major contribution (≥90%) to the total CFTR-related ATP hydrolysis rate is due to phosphorylation by PKA and the minor contribution (≤10%) to CFTR-ATPase activity. (ii) The mutant CFTR-E1371S that is still conductive, but defective in ATP hydrolysis, is not phosphorylated, suggesting that phosphorylation requires a functional nucleotide binding domain and occurs in the post-hydrolysis transition state. (iii) CFTR-ATPase activity is inversely related to CFTR anion flux. The present data are consistent with a model in which CFTR is in a closed conformation with two ATPs bound. The open conformation is induced by ATP hydrolysis and corresponds to the post-hydrolysis transition state that is stabilized by phosphorylation and binding of chloride channel potentiators. PMID:27226582

  10. Lumacaftor alone and combined with ivacaftor: preclinical and clinical trial experience of F508del CFTR correction.

    Science.gov (United States)

    Brewington, John J; McPhail, Gary L; Clancy, John P

    2016-01-01

    Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator protein (CFTR), leading to significant morbidity and mortality. CFTR is a chloride and bicarbonate channel at the epithelial cell membrane. The most common CFTR mutation is F508del, resulting in minimal CFTR at the plasma membrane. Current disease management is supportive, whereas an ultimate goal is to develop therapies to restore CFTR activity. We summarize experience with lumacaftor, a small molecule that increases F508del-CFTR levels at the plasma membrane. Lumacaftor in combination with ivacaftor, a modulator of CFTR gating defects, improves clinical outcome measures in patients homozygous for the F508del mutation. Lumacaftor represents a significant advancement in the treatment of biochemical abnormalities in CF. Further development of CFTR modulators will improve upon current therapies, although it remains unclear whether this approach will provide therapies for all CFTR mutations. PMID:26581802

  11. Low Genetic Diversity Among Garlic (Allium sativum L. Accessions Detected Using Random Amplified Polymorphic DNA (RAPD Escasa Diversidad Genética entre Accesiones de Ajo (Allium sativum L. Detectada Mediante ADN Polimórfico Amplificado al Azar (RAPD

    Directory of Open Access Journals (Sweden)

    Mario Paredes C

    2008-03-01

    Full Text Available Garlic (Allium sativum L. is a species of vegetative propagation, showing high morphological diversity. Besides, its clones have specific adaptations to different agroclimatic regions. The objective of this study was to determine the genetic diversity of 65 garlic clones collected in Chile and introduced from different countries, by using RAPD (Random Amplified Polymorphic DNA. Fourty random primers of 10 mers generated a total of 398 bands with an 87% of polymorphism. Each primer amplified between two and 20 bands. The size of the fragments obtained fluctuated between 3200 and 369 bp. The results showed that the clones analyzed had a genetic similarity rate of 94%. In addition, 70% of them were clustered in one major group. However, in spite of that situation several clones have different agronomic characteristicsEl ajo (Allium sativum L. es una especie de propagación vegetativa, que presenta una amplia variabilidad morfológica. Los clones de esta especie tienen una adaptación específica a diferentes regiones agroclimáticas. El objetivo de este estudio fue determinar la diversidad genética existente en 65 clones de ajos colectados en Chile e introducidos desde diferentes países, utilizando RAPD (ADN Polimórfico Amplificado al Azar. Para esta evaluación se utilizaron 40 partidores de 10-mers. Los partidores generaron entre dos y 20 bandas, observándose un alto número de patrones con bandas múltiples. Los fragmentos generados difieren en su tamaño entre 3.200 y 369 pb. Los partidores generaron 398 bandas, de las cuales un 87% fueron polimórficas. El análisis estadístico realizado detectó una similitud genética alta, de un 94% entre las accesiones evaluadas, donde aproximadamente un 70% de los clones formaron un grupo homogéneo. Sin embargo, este grupo incluye clones que presentan diferentes características agronómicas

  12. Estado emocional y creencias de salud en personas con agregación familiar al cáncer de mama que requieren consejo genético

    OpenAIRE

    Cabrera Torres, Esther

    2008-01-01

    Durante el año 2006, se produjeron 1.703.000 defunciones a causa del cáncer en Europa. El cáncer más frecuente fue el de mama, siendo el 13,5% de todos los casos de neoplasias. Sin embargo en España, los pacientes diagnosticados con cáncer de mama presentan mayor supervivencia que otros tipos tumorales; 94% al año, 84% a los 3 años y 78% a los 5 años. La mejora de la supervivencia es uno de los indicadores más importantes de la eficacia del sistema asistencial en la lucha contra el cáncer y ...

  13. Targeted Correction and Restored Function of the CFTR Gene in Cystic Fibrosis Induced Pluripotent Stem Cells

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    Ana M. Crane

    2015-04-01

    Full Text Available Recently developed reprogramming and genome editing technologies make possible the derivation of corrected patient-specific pluripotent stem cell sources—potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC lines. We then utilized zinc-finger nucleases (ZFNs, designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patient-derived lines via homology-directed repair (HDR. We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expressed the corrected CFTR gene; importantly, CFTR correction resulted in restored expression of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.

  14. Functional interaction between TRP4 and CFTR in mouse aorta endothelial cells

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    Droogmans Guy

    2001-05-01

    Full Text Available Abstract Background This study describes the functional interaction between the putative Ca2+ channel TRP4 and the cystic fibrosis transmembrane conductance regulator, CFTR, in mouse aorta endothelium (MAEC. Results MAEC cells express CFTR transcripts as shown by RT-PCR analysis. Application of a phosphorylating cocktail activated a Cl- current with characteristics similar to those of CFTR mediated currents in other cells types (slow activation by cAMP, absence of rectification, block by glibenclamide. The current is present in trp4 +/+ MAEC, but not in trp4 -/- cells, although the expression of CFTR seems unchanged in the trp4 deficient cells as judged from RT-PCR analysis. Conclusions It is concluded that TRP4 is necessary for CFTR activation in endothelium, possibly by providing a scaffold for the formation of functional CFTR channels.

  15. Lumacaftor-Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del CFTR

    DEFF Research Database (Denmark)

    Wainwright, Claire E; Elborn, J Stuart; Ramsey, Bonnie W;

    2015-01-01

    BACKGROUND: Cystic fibrosis is a life-limiting disease that is caused by defective or deficient cystic fibrosis transmembrane conductance regulator (CFTR) protein activity. Phe508del is the most common CFTR mutation. METHODS: We conducted two phase 3, randomized, double-blind, placebo......-controlled studies that were designed to assess the effects of lumacaftor (VX-809), a CFTR corrector, in combination with ivacaftor (VX-770), a CFTR potentiator, in patients 12 years of age or older who had cystic fibrosis and were homozygous for the Phe508del CFTR mutation. In both studies, patients were randomly...... homozygous for the Phe508del CFTR mutation. (Funded by Vertex Pharmaceuticals and others; TRAFFIC and TRANSPORT ClinicalTrials.gov numbers, NCT01807923 and NCT01807949.)....

  16. Proposal of Sphaerimonospora cavernae gen. nov., sp. nov. and transfer of Microbispora mesophila (Zhang et al., 1998) to Sphaerimonospora mesophila comb. nov. and Microbispora thailandensis (Duangmal et al., 2012) to Sphaerimonospora thailandensis comb. nov.

    Science.gov (United States)

    Mingma, Ratchanee; Duangmal, Kannika; Také, Akira; Inahashi, Yuki; O Mura, Satoshi; Takahashi, Yo Ko; Matsumoto, Atsuko

    2016-04-01

    The actinomycete strain N74T, isolated from cave soil, was studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain N74T formed a stable, distinct lineage cluster together with Microbispora thailandensis NN276T (99.3 % similarity) and Microbispora mesophila JCM 3151T (97.5 %). Strain N74T was observed to produce single spherical spores on aerial mycelium as reported for M. mesophila JCM 3151T and M. thailandensis NN276T but different from other known species of the genus Microbispora, which are characterized by pairs of spores on aerial hyphae. Multilocus sequence analyses based on concatenated partial gyrB, rpoB, atpD, recA and 16S rRNA gene sequences showed a clear distinction of strain N74T, M. mesophila JCM 3151T and M. thailandensis NN276T from other members of the genus Microbispora, although the chemotaxonomic characteristics of strain N74T were similar to the genus Microbispora; the cell wall contained meso-diaminopimelic acid and the whole-cell hydrolysate contained madurose as the diagnostic sugar. The major menaquinone was MK-9(H4). The fatty acid profile contained iso-C16 : 0. On the basis of morphological, chemotaxonomic and phylogenetic evidence, strain N74T is assigned to a novel species of a new genus, for which the name Sphaerimonospora cavernae gen. nov., sp. nov. is proposed. The type strain of Sphaerimonospora cavernae is N74T ( = BCC 77604T = NBRC 111481T). It is also proposed that M. mesophila and M. thailandensis be transferred to this genus as Sphaerimonospora mesophila comb. nov. (type strain JCM 3151T = NBRC 14179T = DSM 43048T) and Sphaerimonospora thailandensis comb. nov. (type strain NN276T = BCC 41490T = NBRC 107569T), respectively. PMID:26822211

  17. Adaptación al español del cuestionario Vecú et Sante Perçue de l'Adolescent (VSP-A: una medida genérica de calidad de vida para adolescentes

    Directory of Open Access Journals (Sweden)

    Serra-Sutton Vicky

    2002-01-01

    Full Text Available Fundamentos: En la última década se han desarrollado diversas medidas de calidad de vida relacionada con la salud para uso exclusivo en niños/as y adolescentes. No obstante, existen pocos instrumentos de estas características adaptados en España. El Vecú et Sante Perçue de l'adolescent (VSP-A es un instrumento genérico de calidad de vida relacionada con la salud para adolescentes de 11 a 17 años desarrollado en Francia. El objetivo de este estudio fue adaptar al español el VSP-A, como primera fase para la obtención del cuestionario. Métodos: Se adaptó la versión del VSP-A de 39 preguntas siguiendo la metodología de traducción directa e inversa incluyendo: 2 traducciones al español, puntuación del grado de dificultad (0 min-10 máx y clasificación de equivalencia semántica y cultural, 2 reuniones y discusión en paneles de adolescentes, así como reuniones de consenso del equipo de investigación. Finalmente se realizó una traducción inversa (retro-traducción al francés y se administró la versión final pre-test en la prueba piloto. Resultados: La mayoría de las preguntas se clasificaron como equivalentes (24 sobre 39. Tras las reuniones con adolescentes se modificaron algunas preguntas. Tras la retro-traducción, 3 preguntas necesitaron cambios menores. Conclusiones: La versión española del VSP-A parece semántica y culturalmente equivalente a la versión original en francés y adecuada para adolescentes en España. La sencillez de las preguntas, los comentarios de los adolescentes y la participación de los autores originales en el proceso de adaptación ha permitido obtener una versión pre-test adecuada. La siguiente fase del estudio es la comprobación de la fiabilidad y validez. Se espera que el VSP-A sea de utilidad para medir la calidad de vida relacionada con la salud en encuestas de salud o como instrumento de cribado en colegios o centros de atención primaria en nuestro medio.

  18. Impact of heterozygote CFTR Mutations in COPD patients with Chronic Bronchitis

    OpenAIRE

    Raju, S. Vamsee; Tate, Jody H; Peacock, Sandra KG; Fang, Ping; Oster, Robert A.; Dransfield, Mark T.; Steven M Rowe

    2014-01-01

    Background Cigarette smoking causes Chronic Obstructive Pulmonary Disease (COPD), the 3rd leading cause of death in the U.S. CFTR ion transport dysfunction has been implicated in COPD pathogenesis, and is associated with chronic bronchitis. However, susceptibility to smoke induced lung injury is variable and the underlying genetic contributors remain unclear. We hypothesized that presence of CFTR mutation heterozygosity may alter susceptibility to cigarette smoke induced CFTR dysfunction. Con...

  19. Stable ATP binding mediated by a partial NBD dimer of the CFTR chloride channel

    OpenAIRE

    Tsai, Ming-Feng; Li, Min; Hwang, Tzyh-Chang

    2010-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR), a member of the adenosine triphosphate (ATP) binding cassette (ABC) superfamily, is an ATP-gated chloride channel. Like other ABC proteins, CFTR encompasses two nucleotide binding domains (NBDs), NBD1 and NBD2, each accommodating an ATP binding site. It is generally accepted that CFTR’s opening–closing cycles, each completed within 1 s, are driven by rapid ATP binding and hydrolysis events in NBD2. Here, by recording CFTR currents in...

  20. Linfohistiocitosis hemofagocítica, el espectro desde la enfermedad genética al síndrome de activación macrofágica

    Directory of Open Access Journals (Sweden)

    Oscar Porras

    2011-06-01

    Full Text Available El compromiso de la regulación de la inflamación produce activación excesiva y expansión de macrófagos y linfocitos T que desencadenan una reacción inflamatoria severa, sin vías naturales de control. Los trastornos hemofagocíticos son la traducción clínica de este proceso inflamatorio. La linfohistiocitosis hemofagocítica se refiere a todas las variantes de esta patología, y el síndrome de activación macrofágica, a la variante asociada con enfermedad autoinmune. Los casos primarios se asocian con la forma familiar autosómica recesiva y los secundarios con inmunodeficiencias primarias, infección, malignidad y enfermedades autoinmunes. El principal distintivo de este grupo de patologías es la proliferación agresiva de macrófagos e histiocitos que fagocitan otras células sanguíneas. La reducción en la actividad de las células NK produce un aumento en la activación y expansión de linfocitos T, los cuales producen grandes cantidades de citoquinas. Las citoquinas inducen activación de macrófagos y células dendríticas, infiltración tisular y producción de interleuquinas, lo que genera una reacción inflamatoria severa, responsable del daño tisular y de las manifestaciones clínicas. El curso clínico se caracteriza principalmente por fiebre prolongada, hepatoesplenomegalia y citopenias. Los estudios de laboratorio muestran aumento de ferritina, triglicéridos e hipofibrinogenemia. La hemofagocitosis en médula ósea está presente en más del 80% de los casos al diagnóstico. El tratamiento está dirigido contra el linfocito T y los histiocitos hiperactivados, combinando quimioterapia con inmunosupresores y, en algunos casos, trasplante de células madre hematopoyéticas. Este tratamiento ha producido un cambio en la sobrevida de los pacientes. El protocolo de tratamiento HLH-2004 es una guía que estandariza el tratamiento, combinando etopósido, dexametazona y ciclosporina A. En Costa Rica se han reportado 60 casos en

  1. Osteoblast CFTR inactivation reduces differentiation and osteoprotegerin expression in a mouse model of cystic fibrosis-related bone disease.

    Directory of Open Access Journals (Sweden)

    Michael S Stalvey

    Full Text Available Low bone mass and increased fracture risk are recognized complications of cystic fibrosis (CF. CF-related bone disease (CFBD is characterized by uncoupled bone turnover--impaired osteoblastic bone formation and enhanced osteoclastic bone resorption. Intestinal malabsorption, vitamin D deficiency and inflammatory cytokines contribute to CFBD. However, epidemiological investigations and animal models also support a direct causal link between inactivation of skeletal cystic fibrosis transmembrane regulator (CFTR, the gene that when mutated causes CF, and CFBD. The objective of this study was to examine the direct actions of CFTR on bone. Expression analyses revealed that CFTR mRNA and protein were expressed in murine osteoblasts, but not in osteoclasts. Functional studies were then performed to investigate the direct actions of CFTR on osteoblasts using a CFTR knockout (Cftr-/- mouse model. In the murine calvarial organ culture assay, Cftr-/- calvariae displayed significantly less bone formation and osteoblast numbers than calvariae harvested from wildtype (Cftr+/+ littermates. CFTR inactivation also reduced alkaline phosphatase expression in cultured murine calvarial osteoblasts. Although CFTR was not expressed in murine osteoclasts, significantly more osteoclasts formed in Cftr-/- compared to Cftr+/+ bone marrow cultures. Indirect regulation of osteoclastogenesis by the osteoblast through RANK/RANKL/OPG signaling was next examined. Although no difference in receptor activator of NF-κB ligand (Rankl mRNA was detected, significantly less osteoprotegerin (Opg was expressed in Cftr-/- compared to Cftr+/+ osteoblasts. Together, the Rankl:Opg ratio was significantly higher in Cftr-/- murine calvarial osteoblasts contributing to a higher osteoclastogenesis potential. The combined findings of reduced osteoblast differentiation and lower Opg expression suggested a possible defect in canonical Wnt signaling. In fact, Wnt3a and PTH-stimulated canonical Wnt

  2. Cholic acid induces a Cftr dependent biliary secretion and liver growth response in mice.

    Directory of Open Access Journals (Sweden)

    Frank A J A Bodewes

    Full Text Available The cause of Cystic fibrosis liver disease (CFLD, is unknown. It is well recognized that hepatic exposure to hydrophobic bile salts is associated with the development of liver disease. For this reason, we hypothesize that, CFTR dependent variations, in the hepatic handling of hydrophobic bile salts, are related to the development CFLD. To test our hypothesis we studied, in Cftr-/- and control mice, bile production, bile composition and liver pathology, in normal feeding condition and during cholate exposure, either acute (intravenous or chronic (three weeks via the diet. In Cftr-/- and control mice the basal bile production was comparable. Intravenous taurocholate increased bile production to the same extent in Cftr-/- and control mice. However, chronic cholate exposure increased the bile flow significantly less in Cftr-/- mice than in controls, together with significantly higher biliary bile salt concentration in Cftr-/- mice. Prolonged cholate exposure, however, did not induce CFLD like pathology in Cftr-/- mice. Chronic cholate exposure did induce a significant increase in liver mass in controls that was absent in Cftr-/- mice. Chronic cholate administration induces a cystic fibrosis-specific hepatobiliary phenotype, including changes in bile composition. These changes could not be associated with CFLD like pathological changes in CF mouse livers. However, chronic cholate administration induces liver growth in controls that is absent in Cftr-/- mice. Our findings point to an impaired adaptive homeotrophic liver response to prolonged hydrophobic bile salt exposure in CF conditions.

  3. β2-Adrenergic receptor agonists activate CFTR in intestinal organoids and subjects with cystic fibrosis.

    Science.gov (United States)

    Vijftigschild, Lodewijk A W; Berkers, Gitte; Dekkers, Johanna F; Zomer-van Ommen, Domenique D; Matthes, Elizabeth; Kruisselbrink, Evelien; Vonk, Annelotte; Hensen, Chantal E; Heida-Michel, Sabine; Geerdink, Margot; Janssens, Hettie M; van de Graaf, Eduard A; Bronsveld, Inez; de Winter-de Groot, Karin M; Majoor, Christof J; Heijerman, Harry G M; de Jonge, Hugo R; Hanrahan, John W; van der Ent, Cornelis K; Beekman, Jeffrey M

    2016-09-01

    We hypothesized that people with cystic fibrosis (CF) who express CFTR (cystic fibrosis transmembrane conductance regulator) gene mutations associated with residual function may benefit from G-protein coupled receptor (GPCR)-targeting drugs that can activate and enhance CFTR function.We used intestinal organoids to screen a GPCR-modulating compound library and identified β2-adrenergic receptor agonists as the most potent inducers of CFTR function.β2-Agonist-induced organoid swelling correlated with the CFTR genotype, and could be induced in homozygous CFTR-F508del organoids and highly differentiated primary CF airway epithelial cells after rescue of CFTR trafficking by small molecules. The in vivo response to treatment with an oral or inhaled β2-agonist (salbutamol) in CF patients with residual CFTR function was evaluated in a pilot study. 10 subjects with a R117H or A455E mutation were included and showed changes in the nasal potential difference measurement after treatment with oral salbutamol, including a significant improvement of the baseline potential difference of the nasal mucosa (+6.35 mV, pCFTR activation when administered ex vivo to organoids.This proof-of-concept study suggests that organoids can be used to identify drugs that activate CFTR function in vivo and to select route of administration. PMID:27471203

  4. Determination of CFTR densities in erythrocyte plasma membranes using recognition imaging

    Science.gov (United States)

    Ebner, Andreas; Nikova, Dessy; Lange, Tobias; Häberle, Johannes; Falk, Sabine; Dübbers, Angelika; Bruns, Reimer; Hinterdorfer, Peter; Oberleithner, Hans; Schillers, Hermann

    2008-09-01

    CFTR (cystic fibrosis transmembrane conductance regulator) is a cAMP-regulated chloride (Cl-) channel that plays an important role in salt and fluid movement across epithelia. Cystic fibrosis (CF), the most common genetic disease among Caucasians, is caused by mutations in the gene encoding CFTR. The most predominant mutation, F508del, disturbs CFTR protein trafficking, resulting in a reduced number of CFTR in the plasma membrane. Recent studies indicate that CFTR is not only found in epithelia but also in human erythrocytes. Although considerable attempts have been made to quantify CFTR in cells, conclusions on numbers of CFTR molecules localized in the plasma membrane have been drawn indirectly. AFM has the power to provide the needed information, since both sub-molecular spatial resolution and direct protein recognition via antibody-antigen interaction can be observed. We performed a quantification study of the CFTR copies in erythrocyte membranes at the single molecule level, and compared the difference between healthy donors and CF patients. We detected that the number of CFTR molecules is reduced by 70% in erythrocytes of cystic fibrosis patients.

  5. Duplicated CFTR isoforms in eels diverged in regulatory structures and osmoregulatory functions.

    Science.gov (United States)

    Wong, Marty Kwok-Shing; Pipil, Supriya; Kato, Akira; Takei, Yoshio

    2016-09-01

    Two cystic fibrosis transmembrane conductance regulator (CFTR) isoforms, CFTRa and CFTRb, were cloned in Japanese eel and their structures and functions were studied in different osmoregulatory tissues in freshwater (FW) and seawater (SW) eels. Molecular phylogenetic results suggested that the CFTR duplication in eels occurred independently of the duplication event in salmonid. CFTRa was expressed in the intestine and kidney and downregulated in both tissues in SW eels, while CFTRb was specifically expressed in the gill and greatly upregulated in SW eels. Structurally, the CFTR isoforms are similar in most functional domains except the regulatory R domain, where the R domain of CFTRa is similar to that of human CFTR but the R domain of CFTRb is unique in having high intrinsic negative charges and fewer phosphorylation sites, suggesting divergence of isoforms in terms of gating properties and hormonal regulation. Immunohistochemical results showed that CFTR was localized on the apical regions of SW ionocytes, suggesting a Cl(-) secretory role as in other teleosts. In intestine and kidney, however, immunoreactive CFTR was mostly found in the cytosolic vesicles in FW eels, indicating that Cl(-) channel activity could be low at basal conditions, but could be rapidly increased by membrane insertion of the stored channels. Guanylin (GN), a known hormone that increases CFTR activity in mammalian intestine, failed to redistribute CFTR and to affect its expression in eel intestine. The results suggested that GN-independent CFTR regulation is present in eel intestine and kidney. PMID:27322796

  6. Targeting F508del-CFTR to develop rational new therapies for cystic fibrosis

    Institute of Scientific and Technical Information of China (English)

    Zhi-wei CAI; Jia LIU; Hong-yu LI; David N SHEPPARD

    2011-01-01

    The mutation F508del is the commonest cause of the genetic disease cystic fibrosis (CF). CF disrupts the function of many organs in the body, most notably the lungs, by perturbing salt and water transport across epithelial surfaces. F508del causes harm in two principal ways. First,the mutation prevents delivery of the cystic fibrosis transmembrane conductance regulator (CFTR) to its correct cellular location,the apical(lumen-facing) membrane of epithelial cells. Second, F508del perturbs the Cl- channel function of CFTR by disrupting channel gating. Here, we discuss the development of rational new therapies for CF that target F508del-CFTR.We highlight how structural studies provide new insight into the role of F508 in the regulation of channel gating by cycles of ATP binding and hydrolysis. We emphasize the use of high-throughput screening to identify lead compounds for therapy development.These compounds include CFTR correctors that restore the expression of F508del-CFTR at the apical membrane of epithelial cells and CFTR potentiators that rescue the F508del-CFTR gating defect. Initial results from clinical trials of CFTR correctors and potentiators augur well for the development of small molecule therapies that target the root cause of CF: mutations in CFTR.

  7. Determination of CFTR densities in erythrocyte plasma membranes using recognition imaging

    Energy Technology Data Exchange (ETDEWEB)

    Ebner, Andreas; Hinterdorfer, Peter [Institute for Biophysics, University of Linz, A-4040 Linz (Austria); Nikova, Dessy; Lange, Tobias; Bruns, Reimer; Oberleithner, Hans; Schillers, Hermann [Institute of Physiology II, University of Muenster, D-48149 Muenster (Germany); Haeberle, Johannes; Falk, Sabine; Duebbers, Angelika [Department of Pediatrics, University Hospitals of Muenster, D-48149 Muenster (Germany)], E-mail: schille@uni-muenster.de

    2008-09-24

    CFTR (cystic fibrosis transmembrane conductance regulator) is a cAMP-regulated chloride (Cl{sup -}) channel that plays an important role in salt and fluid movement across epithelia. Cystic fibrosis (CF), the most common genetic disease among Caucasians, is caused by mutations in the gene encoding CFTR. The most predominant mutation, F508del, disturbs CFTR protein trafficking, resulting in a reduced number of CFTR in the plasma membrane. Recent studies indicate that CFTR is not only found in epithelia but also in human erythrocytes. Although considerable attempts have been made to quantify CFTR in cells, conclusions on numbers of CFTR molecules localized in the plasma membrane have been drawn indirectly. AFM has the power to provide the needed information, since both sub-molecular spatial resolution and direct protein recognition via antibody-antigen interaction can be observed. We performed a quantification study of the CFTR copies in erythrocyte membranes at the single molecule level, and compared the difference between healthy donors and CF patients. We detected that the number of CFTR molecules is reduced by 70% in erythrocytes of cystic fibrosis patients.

  8. SNaPshot Assay for the Detection of the Most Common CFTR Mutations in Infertile Men

    OpenAIRE

    Predrag Noveski; Svetlana Madjunkova; Marija Mircevska; Toso Plaseski; Vanja Filipovski; Dijana Plaseska-Karanfilska

    2014-01-01

    Congenital bilateral absence of vas deferens (CBAVD) is the most common CFTR-related disorder (CFTR-RD) that explains about 1-2% of the male infertility cases. Controversial data have been published regarding the involvement of CFTR mutations in infertile men with non-obstructive azoospermia and oligozoospermia. Here, we describe single base extension (SNaPshot) assay for detection of 11 common CFTR mutations: F508del, G542X, N1303K, 621+1G->T, G551D, R553X, R1162X, W1282X, R117H, 2184insA an...

  9. SNaPshot assay for the detection of the most common CFTR mutations in infertile men.

    Directory of Open Access Journals (Sweden)

    Predrag Noveski

    Full Text Available Congenital bilateral absence of vas deferens (CBAVD is the most common CFTR-related disorder (CFTR-RD that explains about 1-2% of the male infertility cases. Controversial data have been published regarding the involvement of CFTR mutations in infertile men with non-obstructive azoospermia and oligozoospermia. Here, we describe single base extension (SNaPshot assay for detection of 11 common CFTR mutations: F508del, G542X, N1303K, 621+1G->T, G551D, R553X, R1162X, W1282X, R117H, 2184insA and 1717-1G->A and IVS8polyT variants. The assay was validated on 50 previously genotyped samples and was used to screen a total of 369 infertile men with different impairment of spermatogenesis and 136 fertile controls. Our results show that double heterozygosity of cystic fibrosis (CF and CFTR-related disorder (CFTR-RD mutations are found in a high percentage (22.7% of infertile men with obstructive azoospermia, but not in other studied groups of infertile men. The SNaPshot assay described here is an inexpensive, fast and robust method for primary screening of the most common CFTR mutations both in patients with classical CF and CFTR-RD. It can contribute to better understanding of the role of CFTR mutations in impaired spermatogenesis, ultimately leading to improved management of infertile men.

  10. RNA interference for CFTR attenuates lung fluid absorption at birth in rats

    Directory of Open Access Journals (Sweden)

    Folkesson Hans G

    2008-07-01

    Full Text Available Abstract Background Small interfering RNA (siRNA against αENaC (α-subunit of the epithelial Na channel and CFTR (cystic fibrosis transmembrane conductance regulator was used to explore ENaC and CTFR function in newborn rat lungs. Methods Twenty-four hours after trans-thoracic intrapulmonary (ttip injection of siRNA-generating plasmid DNA (pSi-0, pSi-4, or pSi-C2, we measured CFTR and ENaC expression, extravascular lung water, and mortality. Results αENaC and CFTR mRNA and protein decreased by ~80% and ~85%, respectively, following αENaC and CFTR silencing. Extravascular lung water and mortality increased after αENaC and CFTR-silencing. In pSi-C2-transfected isolated DLE cells there were attenuated CFTR mRNA and protein. In pSi-4-transfected DLE cells αENaC mRNA and protein were both reduced. Interestingly, CFTR-silencing also reduced αENaC mRNA and protein. αENaC silencing, on the other hand, only slightly reduced CFTR mRNA and protein. Conclusion Thus, ENaC and CFTR are both involved in the fluid secretion to absorption conversion around at birth.

  11. Tgf-β1 inhibits Cftr biogenesis and prevents functional rescue of ΔF508-Cftr in primary differentiated human bronchial epithelial cells.

    Directory of Open Access Journals (Sweden)

    Steven M Snodgrass

    Full Text Available CFTR is an integral transmembrane glycoprotein and a cAMP-activated Cl(- channel. Mutations in the CFTR gene lead to Cystic Fibrosis (CF-an autosomal recessive disease with majority of the morbidity and mortality resulting from airway infection, inflammation, and fibrosis. The most common disease-associated mutation in the CFTR gene-deletion of Phe508 (ΔF508 leads to a biosynthetic processing defect of CFTR. Correction of the defect and delivery of ΔF508-CFTR to the cell surface has been highly anticipated as a disease modifying therapy. Compared to promising results in cultured cell this approach was much less effective in CF patients in an early clinical trial. Although the cause of failure to rescue ΔF508-CFTR in the clinical trial has not been determined, presence of factor(s that interfere with the rescue in vivo could be considered. The cytokine TGF-β1 is frequently elevated in CF patients. TGF-β1 has pleiotropic effects in different disease models and genetic backgrounds and little is known about TGF-β1 effects on CFTR in human airway epithelial cells. Moreover, there are no published studies examining TGF-β1 effects on the functional rescue of ΔF508-CFTR. Here we found that TGF-β1 inhibits CFTR biogenesis by reducing mRNA levels and protein abundance in primary differentiated human bronchial epithelial (HBE cells from non-CF individuals. TGF-β1 inhibits CFTR biogenesis without compromising the epithelial phenotype or integrity of HBE cells. TGF-β1 also inhibits biogenesis and impairs the functional rescue of ΔF508-CFTR in HBE cells from patients homozygous for the ΔF508 mutation. Our data indicate that activation of TGF-β1 signaling may inhibit CFTR function in non-CF individuals and may interfere with therapies directed at correcting the processing defect of ΔF508-CFTR in CF patients.

  12. CFTR, PRSS1 und SPINK1 Varianten bei chronischer Pankreatitis

    OpenAIRE

    Rosendahl, Jonas

    2010-01-01

    Chronic pancreatitis (CP) is defined as a continuing or relapsing inflammatory disease of the pancreas, leading to endocrine and/or exocrine insufficiency and irreversible morphological changes. Currently, it is thought that the imbalance of pancreatic proteases and their inhibitors results in the development of chronic pancreatitis. Until now, genetic variations of four genes have been associated to chronic pancreatitis: PRSS1, PRSS2, SPINK1 and CFTR. Since direct DNA-sequencing has becom...

  13. CFTR gene mutations in isolated chronic obstructive pulmonary disease

    Energy Technology Data Exchange (ETDEWEB)

    Pignatti, P.F.; Bombien, C.; Marigo, C. [and others

    1994-09-01

    In order to identify a possible hereditary predisposition to the development of chronic obstructive pulmonary disease (COPD), we have looked for the presence of cystic fibrosis transmembrane regulator (CFTR) gene DNA sequence modifications in 28 unrelated patients with no signs of cystic fibrosis. The known mutations in Italian CF patients, as well as the most frequent worldwide CF mutations, were investigated. In addition, a denaturing gradient gel electrophoresis analysis of about half of the coding sequence of the gene in 56 chromosomes from the patients and in 102 chromosomes from control individuals affected by other pulmonary diseases and from normal controls was performed. Nine different CFTR gene mutations and polymorphisms were found in seven patients, a highly significant increase over controls. Two of the patients were compound heterozygotes. Two frequent CF mutations were detected: deletion F508 and R117H; two rare CF mutations: R1066C and 3667ins4; and five CF sequence variants: R75Q (which was also described as a disease-causing mutation in male sterility cases due to the absence of the vasa deferentia), G576A, 2736 A{r_arrow}G, L997F, and 3271+18C{r_arrow}T. Seven (78%) of the mutations are localized in transmembrane domains. Six (86%) of the patients with defined mutations and polymorphisms had bronchiectasis. These results indicate that CFTR gene mutations and sequence alterations may be involved in the etiopathogenesis of some cases of COPD.

  14. Mechanisms of CFTR functional variants that impair regulated bicarbonate permeation and increase risk for pancreatitis but not for cystic fibrosis.

    OpenAIRE

    Jessica LaRusch; Jinsei Jung; General, Ignacio J.; Lewis, Michele D; Hyun Woo Park; Brand, Randall E.; Andres Gelrud; Anderson, Michelle A.; Banks, Peter A; Darwin Conwell; Christopher Lawrence; Joseph Romagnuolo; John Baillie; Samer Alkaade; Gregory Cote

    2014-01-01

    CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev ) cause complete loss of CFTR function and result in cystic fibrosis (CF), a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize tha...

  15. Characterization of a critical role for CFTR chloride channels in cardioprotection against ischemia/reperfusion injury

    Institute of Scientific and Technical Information of China (English)

    Sunny Yang XIANG; Linda L YE; LI-lu Marie DUAN; Li-hui LIU; Zhi-dong GE; John A AUCHAMPACH; Garrett J GROSS; Dayue Darrel DUAN

    2011-01-01

    Aim: To further characterize the functional role of cystic fibrosis transmembrane conductance regulator (CFTR) in early and late (second window) ischemic preconditioning (IPC)- and postcondtioning (POC)-mediated cardioprotection against ischemia/reperfusion (I/R) injury.Methods: CFTR knockout (CFTR-/-) mice and age- and gender-matched wild-type (CFTR+/+) and heterozygous (CFTR+/-) mice were used.In in vivo studies, the animals were subjected to a 30-min coronary occlusion followed by a 40-min reperfusion. In ex vivo (isolate heart) studies, a 45-min global ischemia was applied. To evaluate apoptosis, the level of activated caspase 3 and TdT-mediated dUTP-X nick end labeling (TUNEL) were examined.Results: In the in vivo I/R models, early IPC significantly reduced the myocardial infarct size in wild-type (CFTR+/+) (from 40.4%±5.3% to 10.4%±2.0%, n=8, P<0.001) and heterozygous (CFTR+/-) littermates (from 39.4%±2.4% to 15.4%±5.1%, n=6, P<0.001) but failed to protect CFTR knockout (CFTR-/-) mice from I/R induced myocardial infarction (46.9%±6.2% vs 55.5%±7.8%, n=6, P>0.5). Similar results were observed in the in vivo late IPC experiments. Furthermore, in both in vivo and ex vivo I/R models, POC significantly reduced myocardial infarction in wild-type mice, but not in CFTR knockout mice. In ex vivo I/R models, targeted inactivation of CFTRgene abolished the protective effects of IPC against I/R-induced apoptosis.Conclusion: These results provide compelling evidence for a critical role for CFTR Cl- channels in IPC- and POC-mediated cardioprotection against I/R-induced myocardial injury.

  16. Mechanism-based corrector combination restores Delta F508-CFTR folding and function

    NARCIS (Netherlands)

    Okiyoneda, Tsukasa; Veit, Guido; Dekkers, Johanna F.; Bagdany, Miklos; Soya, Naoto; Xu, Haijin; Roldan, Ariel; Verkman, Alan S.; Kurth, Mark; Simon, Agnes; Hegedus, Tamas; Beekman, Jeffrey M.; Lukacs, Gergely L.

    2013-01-01

    The most common cystic fibrosis mutation, Delta F508 in nucleotide binding domain 1 (NBD1), impairs cystic fibrosis transmembrane conductance regulator (CFTR)-coupled domain folding, plasma membrane expression, function and stability. VX-809, a promising investigational corrector of Delta F508-CFTR

  17. Stimulation of Airway and Intestinal Mucosal Secretion by Natural Coumarin CFTR Activators

    OpenAIRE

    Hong eYang; Lina eXu; Yujie eSui; Xin eLiu; Chengyan eHe; Rouyu eFang; Jia eLiu; Feng eHao; Tong-Hui eMa

    2011-01-01

    Mutations of cystic fibrosis transmembrane conductance regulator (CFTR) cause lethal hereditary disease cystic fibrosis (CF) that involves extensive destruction and dysfunction of serous epithelium. Possible pharmacological therapy includes correction of defective intracellular processing and abnormal channel gating. In a previous study, we identified five natural coumarin potentiators of Δ508-CFTR including osthole, imperatorin, isopsoralen, praeruptorin A and scoparone. The present study wa...

  18. Advancing clinical development pathways for new CFTR modulators in cystic fibrosis.

    Science.gov (United States)

    Mayer-Hamblett, Nicole; Boyle, Michael; VanDevanter, Donald

    2016-05-01

    Cystic fibrosis (CF) is a life-shortening genetic disease affecting approximately 70 000 individuals worldwide. Until recently, drug development efforts have emphasised therapies treating downstream signs and symptoms resulting from the underlying CF biological defect: reduced function of the CF transmembrane conductance regulator (CFTR) protein. The current CF drug development landscape has expanded to include therapies that enhance CFTR function by either restoring wild-type CFTR protein expression or increasing (modulating) the function of mutant CFTR proteins in cells. To date, two systemic small-molecule CFTR modulators have been evaluated in pivotal clinical trials in individuals with CF and specific mutantCFTRgenotypes that have led to regulatory review and/or approval. Advances in the discovery of CFTR modulators as a promising new class of therapies have been impressive, yet work remains to develop highly effective, disease-modifying modulators for individuals of all CF genotypes. The objectives of this review are to outline the challenges and opportunities in drug development created by systemic genotype-specific CFTR modulators, highlight the advantages of sweat chloride as an established biomarker of CFTR activity to streamline early-phase development and summarise options for later phase clinical trial designs that respond to the adoption of approved genotype-specific modulators into standard of care. An optimal development framework will be needed to move the most promising therapies efficiently through the drug development pipeline and ultimately deliver efficacious and safe therapies to all individuals with CF. PMID:26903594

  19. Capacidad predictiva del modelo BCRAPro frente al profesional de enfermería en la selección de candidatos a estudio genético de cáncer de mama u ovario hereditario

    OpenAIRE

    Benito-Aracil, Llúcia; Yagüe Muñoz, Carmen; Iglesias Casals, Sílvia; Salinas Masdeu, Mònica; Teulé Vega, Àlex; Lázaro García, Conxi; Blanco Guillermo, Ignacio

    2010-01-01

    Objetivo: Comparar la capacidad predictiva del modelo predictivo BRCAPro y de los profesionales de enfermería con distintos niveles de formación/experiencia en la identificación de familias susceptibles de ser estudiadas genéticamente dada su historia personal y familiar de cáncer de mama. Método: Estudio descriptivo en el que 2 enfermeras con diferente grado de formación en consejo genético han estimado la probabilidad de ser portador de mutación en los genes BRCA1/BRCA2 de 157 familias. Se ...

  20. Evaluation of potential regulatory elements identified as DNase I hypersensitive sites in the CFTR gene

    DEFF Research Database (Denmark)

    Phylactides, M.; Rowntree, R.; Nuthall, H.;

    2002-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) gene shows a complex pattern of expression, with temporal and spatial regulation that is not accounted for by elements in the promoter. One approach to identifying the regulatory elements for CFTR is the mapping of DNase I...... hypersensitive sites (DHS) within the locus. We previously identified at least 12 clusters of DHS across the CFTR gene and here further evaluate DHS in introns 2,3,10,16,17a, 18, 20 and 21 to assess their functional importance in regulation of CFTR gene expression. Transient transfections of enhancer....../reporter constructs containing the DHS regions showed that those in introns 20 and 21 augmented the activity of the CFTR promoter. Structural analysis of the DNA sequence at the DHS suggested that only the one intron 21 might be caused by inherent DNA structures. Cell specificity of the DHS suggested a role for the...

  1. Involvement of CFTR in Uterine Bicarbonate Secretion and the Fertilizing Capacity of Sperm

    Institute of Scientific and Technical Information of China (English)

    WangXiao,Fei; ZhouChen-Xi; ShiQi-Xian; YuanYu-Ying; YuMei-Kuen; LouisChukwuemekaAjonuma

    2005-01-01

    Cystic fibrosis transmembrane conductance regulator (CFFR)is a cAMP-activated chloride channel expressed in a wide variety of epithelial cells,mutations of which are responsible for the hallmark defective chloride secretion observed in cystic fibrosis(CF).Although CFTR has been implicated in bicarbonate secretion,its ability to directly mediate bicarbonate secretion of any physiological significance has not been shown.We demonstrate here that endometriaI epithelial ceils possess a CFTR-mediated bicarbonate transport mechanism.Co-culture of sperm with endometrial ceils treated with antisense oligonucleotide against CFTR,or with bicarbonate secretion-defective CF epithelial cells,resulted in lower sperm capacitation and egg-fertilizing ability.These results are consistent with a critical role of CFTR in controlling uterine bicarbonate secretion and the fertilizing capacity of sperm,providing a link between defective CFTR and lower female fertility in CF.

  2. Manipulating proteostasis to repair the F508del-CFTR defect in cystic fibrosis.

    Science.gov (United States)

    Esposito, Speranza; Tosco, Antonella; Villella, Valeria R; Raia, Valeria; Kroemer, Guido; Maiuri, Luigi

    2016-12-01

    Cystic fibrosis (CF) is a lethal monogenic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene that entails the (diagnostic) increase in sweat electrolyte concentrations, progressive lung disease with chronic inflammation and recurrent bacterial infections, pancreatic insufficiency, and male infertility. Therapies aimed at restoring the CFTR defect have emerged. Thus, a small molecule which facilitates chloride channel opening, the potentiator Ivacaftor, has been approved for the treatment of CF patients bearing a particular class of rare CFTR mutations. However, small molecules that directly target the most common misfolded CFTR mutant, F508del, and improve its intracellular trafficking in vitro, have been less effective than expected when tested in CF patients, even in combination with Ivacaftor. Thus, new strategies are required to circumvent the F508del-CFTR defect. Airway and intestinal epithelial cells from CF patients bearing the F508del-CFTR mutation exhibit an impressive derangement of cellular proteostasis, with oxidative stress, overactivation of the tissue transglutaminase (TG2), and disabled autophagy. Proteostasis regulators such as cysteamine can rescue and stabilize a functional F508del-CFTR protein through suppressing TG2 activation and restoring autophagy in vivo in F508del-CFTR homozygous mice, in vitro in CF patient-derived cell lines, ex vivo in freshly collected primary patient's nasal cells, as well as in a pilot clinical trial involving homozygous F508del-CFTR patients. Here, we discuss how the therapeutic normalization of defective proteostasis can be harnessed for the treatment of CF patients with the F508del-CFTR mutation. PMID:26976279

  3. Thermal unfolding studies show the disease causing F508del mutation in CFTR thermodynamically destabilizes nucleotide-binding domain 1

    OpenAIRE

    Protasevich, Irina; Yang, Zhengrong; Wang, Chi; Atwell, Shane; Zhao, Xun; Emtage, Spencer; Wetmore, Diana; Hunt, John F.; Brouillette, Christie G

    2010-01-01

    Misfolding and degradation of CFTR is the cause of disease in patients with the most prevalent CFTR mutation, an in-frame deletion of phenylalanine (F508del), located in the first nucleotide-binding domain of human CFTR (hNBD1). Studies of (F508del)CFTR cellular folding suggest that both intra- and inter-domain folding is impaired. (F508del)CFTR is a temperature-sensitive mutant, that is, lowering growth temperature, improves both export, and plasma membrane residence times. Yet, paradoxicall...

  4. Targeting the Intracellular Environment in Cystic Fibrosis: Restoring Autophagy as a Novel Strategy to Circumvent the CFTR Defect.

    Science.gov (United States)

    Villella, Valeria Rachela; Esposito, Speranza; Bruscia, Emanuela M; Maiuri, Maria Chiara; Raia, Valeria; Kroemer, Guido; Maiuri, Luigi

    2013-01-01

    Cystic fibrosis (CF) patients harboring the most common deletion mutation of the CF transmembrane conductance regulator (CFTR), F508del, are poor responders to potentiators of CFTR channel activity which can be used to treat a small subset of CF patients who genetically carry plasma membrane (PM)-resident CFTR mutants. The misfolded F508del-CFTR protein is unstable in the PM even if rescued by pharmacological agents that prevent its intracellular retention and degradation. CF is a conformational disease in which defective CFTR induces an impressive derangement of general proteostasis resulting from disabled autophagy. In this review, we discuss how rescuing Beclin 1 (BECN1), a major player of autophagosome formation, either by means of direct gene transfer or indirectly by administration of proteostasis regulators, could stabilize F508del-CFTR at the PM. We focus on the relationship between the improvement of peripheral proteostasis and CFTR PM stability in F508del-CFTR homozygous bronchial epithelia or mouse lungs. Moreover, this article reviews recent pre-clinical evidence indicating that targeting the intracellular environment surrounding the misfolded mutant CFTR instead of protein itself could constitute an attractive therapeutic option to sensitize patients carrying the F508del-CFTR mutation to the beneficial action of CFTR potentiators on lung inflammation. PMID:23346057

  5. Targeting the intracellular environment in cystic fibrosis: restoring autophagy as a novel strategy to circumvent the CFTR defect

    Directory of Open Access Journals (Sweden)

    Valeria Rachela Villella

    2013-01-01

    Full Text Available Cystic fibrosis (CF patients harboring the most common deletion mutation of the cystic fibrosis transmembrane conductance regulator (CFTR, F508del, are poor responders to potentiators of CFTR channel activity which can be used to treat a small subset of CF patients who genetically carry plasma membrane-resident CFTR mutants. The misfolded F508del-CFTR protein is unstable in the plasma membrane even if rescued by pharmacological agents that prevent its intracellular retention and degradation. CF is a conformational disease in which defective CFTR induces an impressive derangement of general proteostasis resulting from disabled autophagy. In this review, we discuss how rescuing Beclin 1 (BECN1, a major player of autophagosome formation, either by means of direct gene transfer or indirectly by administration of proteostasis regulators, could stabilize F508del-CFTR at the plasma membrane. We focus on the relationship between the improvement of peripheral proteostasis and CFTR plasma membrane stability in F508del-CFTR homozygous bronchial epithelia or mouse lungs. Moreover, this article reviews recent preclinical evidence indicating that targeting the intracellular environment surrounding the misfolded mutant CFTR instead of protein itself could constitute an attractive therapeutic option to sensitize patients carrying the F508del-CFTR mutation to the beneficial action of CFTR potentiators on lung inflammation.

  6. A High-affinity Activator of G551D-CFTR Chloride Channel Identified By High Throughput Screening

    Institute of Scientific and Technical Information of China (English)

    ZHAO Lu; HE Cheng-yan; LIU Yan-li; ZHOU Hong-lan; ZHOU Jin-song; SHANG De-jing; YANG Hong

    2004-01-01

    A stably transfected CHO cell line coexpressing G551D-CFTR and iodide-sensitive yellow fluorescent protein mutant EYFP-H148Q-I152L was successfully established and used as assay model to identify small-molecule activators of G551D-CFTR chloride channel from 100000 diverse combinatorial compounds by high throughput screening on a customized Beckman robotic system. A bicyclooctane compound was identified to activate G551D-CFTR chloride channel with high-affinity(Kd=1.8 μmol/L). The activity of the bicyclooctane compound is G551D-CFTR-specific, reversible and non-toxic. The G551D-CFTR activator may be useful as a tool to study the mutant G551D-CFTR chloride channel structure and transport properties and as a candidate drug to cure cystic fibrosis caused by G551D-CFTR mutation.

  7. CFTR Deletion in Mouse Testis Induces VDAC1 Mediated Inflammatory Pathway Critical for Spermatogenesis

    Science.gov (United States)

    Huijuan, Liao; Jiang, Xie; Ming, Yang; Huaqin, Sun; Wenming, Xu

    2016-01-01

    Cystic fibrosis is the most common genetic disease among Caucasians and affects tissues including lung, pancreas and reproductive tracts. It has been shown that Endoplasmic Reticulum (ER) stress and heat shock response are two major deregulated functional modules related to CFTR dysfunction. To identify the impact of CFTR deletion during spermatogenesis, we examined the expression of spermiogenesis-related genes in the testis of CFTR mutant mice (CF mice). We confirmed expression changes of MSY2, a germ cell specific RNA binding protein, resulting from deletion of CFTR in testis. Furthermore, real time PCR and Western blot results showed that an inflammatory response was activated in CF mice testis, as reflected by the altered expression of cytokines. We demonstrate for the first time that expression of MSY2 is decreased in CF mice. Our results suggest that CFTR deletion in testis influences inflammatory responses and these features are likely to be due to the unique environment of the seminiferous tubule during the spermatogenesis process. The current study also suggests avenues to understand the pathophysiology of CFTR during spermatogenesis and provides targets for the possible treatment of CFTR-related infertility. PMID:27483469

  8. CFTR Deletion in Mouse Testis Induces VDAC1 Mediated Inflammatory Pathway Critical for Spermatogenesis.

    Science.gov (United States)

    Yan, Chen; Lang, Qin; Huijuan, Liao; Jiang, Xie; Ming, Yang; Huaqin, Sun; Wenming, Xu

    2016-01-01

    Cystic fibrosis is the most common genetic disease among Caucasians and affects tissues including lung, pancreas and reproductive tracts. It has been shown that Endoplasmic Reticulum (ER) stress and heat shock response are two major deregulated functional modules related to CFTR dysfunction. To identify the impact of CFTR deletion during spermatogenesis, we examined the expression of spermiogenesis-related genes in the testis of CFTR mutant mice (CF mice). We confirmed expression changes of MSY2, a germ cell specific RNA binding protein, resulting from deletion of CFTR in testis. Furthermore, real time PCR and Western blot results showed that an inflammatory response was activated in CF mice testis, as reflected by the altered expression of cytokines. We demonstrate for the first time that expression of MSY2 is decreased in CF mice. Our results suggest that CFTR deletion in testis influences inflammatory responses and these features are likely to be due to the unique environment of the seminiferous tubule during the spermatogenesis process. The current study also suggests avenues to understand the pathophysiology of CFTR during spermatogenesis and provides targets for the possible treatment of CFTR-related infertility. PMID:27483469

  9. Characterization of mitochondrial function in cells with impaired cystic fibrosis transmembrane conductance regulator (CFTR) function.

    Science.gov (United States)

    Atlante, Anna; Favia, Maria; Bobba, Antonella; Guerra, Lorenzo; Casavola, Valeria; Reshkin, Stephan Joel

    2016-06-01

    Evidence supporting the occurrence of oxidative stress in Cystic Fibrosis (CF) is well established and the literature suggests that oxidative stress is inseparably linked to mitochondrial dysfunction. Here, we have characterized mitochondrial function, in particular as it regards the steps of oxidative phosphorylation and ROS production, in airway cells either homozygous for the F508del-CFTR allele or stably expressing wt-CFTR. We find that oxygen consumption, ΔΨ generation, adenine nucleotide translocator-dependent ADP/ATP exchange and both mitochondrial Complex I and IV activities are impaired in CF cells, while both mitochondrial ROS production and membrane lipid peroxidation increase. Importantly, treatment of CF cells with the small molecules VX-809 and 4,6,4'-trimethylangelicin, which act as "correctors" for F508del CFTR by rescuing the F508del CFTR-dependent chloride secretion, while having no effect per sè on mitochondrial function in wt-CFTR cells, significantly improved all the above mitochondrial parameters towards values found in the airway cells expressing wt-CFTR. This novel study on mitochondrial bioenergetics provides a springboard for future research to further understand the molecular mechanisms responsible for the involvement of mitochondria in CF and identify the proteins primarily responsible for the F508del-CFTR-dependent mitochondrial impairment and thus reveal potential novel targets for CF therapy. PMID:27146408

  10. Evidence that CFTR is expressed in rat tracheal smooth muscle cells and contributes to bronchodilation

    Directory of Open Access Journals (Sweden)

    Mettey Yvette

    2006-08-01

    Full Text Available Abstract Background The airway functions are profoundly affected in many diseases including asthma, chronic obstructive pulmonary disease (COPD and cystic fibrosis (CF. CF the most common lethal autosomal recessive genetic disease is caused by mutations of the CFTR gene, which normally encodes a multifunctional and integral membrane protein, the CF transmembrane conductance regulator (CFTR expressed in airway epithelial cells. Methods To demonstrate that CFTR is also expressed in tracheal smooth muscle cells (TSMC, we used iodide efflux assay to analyse the chloride transports in organ culture of rat TSMC, immunofluorescence study to localize CFTR proteins and isometric contraction measurement on isolated tracheal rings to observe the implication of CFTR in the bronchodilation. Results We characterized three different pathways stimulated by the cAMP agonist forskolin and the isoflavone agent genistein, by the calcium ionophore A23187 and by hypo-osmotic challenge. The pharmacology of the cAMP-dependent iodide efflux was investigated in detail. We demonstrated in rat TSMC that it is remarkably similar to that of the epithelial CFTR, both for activation (using three benzo [c]quinolizinium derivatives and for inhibition (glibenclamide, DPC and CFTRinh-172. Using rat tracheal rings, we observed that the activation of CFTR by benzoquinolizinium derivatives in TSMC leads to CFTRinh-172-sensitive bronchodilation after constriction with carbachol. An immunolocalisation study confirmed expression of CFTR in tracheal myocytes. Conclusion Altogether, these observations revealed that CFTR in the airways of rat is expressed not only in the epithelial cells but also in tracheal smooth muscle cells leading to the hypothesis that this ionic channel could contribute to bronchodilation.

  11. Diversidad genética en pitahaya (Hylocereus undatus Haworth. Britton y Rose)

    OpenAIRE

    Juan Porfirio Legaria Solano; María Elisa Alvarado Cano; Ricardo Gaspar Hernández

    2005-01-01

    Se evaluó la variabilidad genética con marcadores RAPD (Polimorfismos en el ADN Amplificados al Azar) en 50 colectas de pitahaya (Hylocereus undatus Haworth, Britton and Rose) provenientes de nueve estados de México y una colecta de Colombia que se incluyó como testigo. Se detectó alta variabilidad genética (polimorfismo entre colectas de 92.5 %) en las poblaciones de pitahaya. Las huellas genéticas obtenidas permitieron la identificación individual de cada material genét...

  12. Caracterización de la diversidad genética en naranja y comparación del polimorfismo de microsatélites amplificados al azar (RAMs usando electroforesis de poliacrilamida y azarosa Characterization of the genetic diversity in orange, and comparison of polymorphism in randomly-amplifed microsatellites (RAMs, using polyacrylamide and agarose electrophoresis

    Directory of Open Access Journals (Sweden)

    Ana Cruz Morillo Coronado

    2009-10-01

    Full Text Available Se compararon las eficiencias de tres métodos de electroforesis en agarosa y poliacrilamida, usando la cámara pequeña de DNA Sequencing System y cámara grande OWL Sequi-Gen Sequencing Cell, en la detección del polimorfismo en 21 accesiones de naranja (Citrus sinensis con empleo del cebador CGA. El gel de poliacrilamida dio mejor resolución de los productos amplificados vía PCR producidos por RAMs. Este permitió una mejor detección de bandas de ADN polimórficas, lo que facilitó la identificación de la variabilidad genética. La electroforesis en agarosa puede ser más conveniente en otras aplicaciones, debido al bajo costo y fácil aplicación. El estudio de diversidad genética en naranja usando microsatélites RAMs diferenció 51 accesiones en siete grupos con 0.75 de similaridad y 0.25 de heterocigosidad, lo que revela bajo polimorfismo genético. La técnica RAMs permitió agrupar las accesiones en Comunes o Blancas, Navel y Pigmentadas o Sanguinas.We compared the efficiency of three methods of agarose and polyacrylamide electrophoresis (using the small tank of the DNA Sequencing System and the large OWL Sequi-Gen Sequencing Cell, for the detection of polymorphism in 21 accessions of orange (Citrus sinensis, using the primer CGA. The polyacrylamide gel gave better resolution of the PCR-amplified RAM products. This method allowed better detection of polymorphic DNA bands, facilitating the identification of genetic variability. The agarose electrophoresis may be more convenient in other applications, due to its low cost and easy implementation. The study of genetic diversity in orange using RAMs separated 51 accessions into seven groups with 0.75 similarity, and 0.25 heterozygosity, revealing low genetic polymorphism. The RAMs technique grouped the accessions into “Common or White”, “Navel” and “Pigmented or “Sanguine”.

  13. Optimizing nasal potential difference analysis for CFTR modulator development: assessment of ivacaftor in CF subjects with the G551D-CFTR mutation.

    Directory of Open Access Journals (Sweden)

    Steven M Rowe

    Full Text Available Nasal potential difference (NPD is used as a biomarker of the cystic fibrosis transmembrane conductance regulator (CFTR and epithelial sodium channel (ENaC activity. We evaluated methods to detect changes in chloride and sodium transport by NPD based on a secondary analysis of a Phase II CFTR-modulator study. Thirty-nine subjects with CF who also had the G551D-CFTR mutation were randomized to receive ivacaftor (Kalydeco™; also known as VX-770 in four doses or placebo twice daily for at least 14 days. All data were analyzed by a single investigator who was blinded to treatment assignment. We compared three analysis methods to determine the best approach to quantify changes in chloride and sodium transport: (1 the average of both nostrils; (2 the most-polarized nostril at each visit; and (3 the most-polarized nostril at screening carried forward. Parameters of ion transport included the PD change with zero chloride plus isoproterenol (CFTR activity, the basal PD, Ringer's PD, and change in PD with amiloride (measurements of ENaC activity, and the delta NPD (measuring CFTR and ENaC activity. The average and most-polarized nostril at each visit were most sensitive to changes in chloride and sodium transport, whereas the most-polarized nostril at screening carried forward was less discriminatory. Based on our findings, NPD studies should assess both nostrils rather than a single nostril. We also found that changes in CFTR activity were more readily detected than changes in ENaC activity, and that rigorous standardization was associated with relatively good within-subject reproducibility in placebo-treated subjects (± 2.8 mV. Therefore, we have confirmed an assay of reasonable reproducibility for detecting chloride-transport improvements in response to CFTR modulation.

  14. Activation Effect of Cathartic Natural Compound Rhein to CFTR Chloride Channel

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-activated chloride channel expressed in intestinal exocrine glands, which plays a key role in intestinal fluid secretion. A natural anthraquinone activator of CFTR Cl- channel, rhein, was identified by screening 217 single compounds from Chinese herbs via a cellbased halide-sensitive fluorescent assay. Rhein activates CFTR Cl- transportation in a dose-dependent manner in the presence of cAMP with a physiological concentration. This study provides a novel molecular pharmacological mechanism for the laxative drugs in Traditional Chinese Medicine such as aloe, cascara and senna.

  15. The "Goldilocks Effect" in Cystic Fibrosis: identification of a lung phenotype in the cftr knockout and heterozygous mouse

    Directory of Open Access Journals (Sweden)

    Bates Jason HT

    2004-07-01

    Full Text Available Abstract Background Cystic Fibrosis is a pleiotropic disease in humans with primary morbidity and mortality associated with a lung disease phenotype. However, knockout in the mouse of cftr, the gene whose mutant alleles are responsible for cystic fibrosis, has previously failed to produce a readily, quantifiable lung phenotype. Results Using measurements of pulmonary mechanics, a definitive lung phenotype was demonstrated in the cftr-/- mouse. Lungs showed decreased compliance and increased airway resistance in young animals as compared to cftr+/+ littermates. These changes were noted in animals less than 60 days old, prior to any long term inflammatory effects that might occur, and are consistent with structural differences in the cftr-/- lungs. Surprisingly, the cftr+/- animals exhibited a lung phenotype distinct from either the homozygous normal or knockout genotypes. The heterozygous mice showed increased lung compliance and decreased airway resistance when compared to either homozygous phenotype, suggesting a heterozygous advantage that might explain the high frequency of this mutation in certain populations. Conclusions In the mouse the gene dosage of cftr results in distinct differences in pulmonary mechanics of the adult. Distinct phenotypes were demonstrated in each genotype, cftr-/-, cftr +/-, and cftr+/+. These results are consistent with a developmental role for CFTR in the lung.

  16. The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Uses its C-Terminus to Regulate the A2B Adenosine Receptor.

    Science.gov (United States)

    Watson, Michael J; Lee, Shernita L; Marklew, Abigail J; Gilmore, Rodney C; Gentzsch, Martina; Sassano, Maria F; Gray, Michael A; Tarran, Robert

    2016-01-01

    CFTR is an apical membrane anion channel that regulates fluid homeostasis in many organs including the airways, colon, pancreas and sweat glands. In cystic fibrosis, CFTR dysfunction causes significant morbidity/mortality. Whilst CFTR's function as an ion channel has been well described, its ability to regulate other proteins is less understood. We have previously shown that plasma membrane CFTR increases the surface density of the adenosine 2B receptor (A2BR), but not of the β2 adrenergic receptor (β2AR), leading to an enhanced, adenosine-induced cAMP response in the presence of CFTR. In this study, we have found that the C-terminal PDZ-domain of both A2BR and CFTR were crucial for this interaction, and that replacing the C-terminus of A2BR with that of β2AR removed this CFTR-dependency. This observation extended to intact epithelia and disruption of the actin cytoskeleton prevented A2BR-induced but not β2AR-induced airway surface liquid (ASL) secretion. We also found that CFTR expression altered the organization of the actin cytoskeleton and PDZ-binding proteins in both HEK293T cells and in well-differentiated human bronchial epithelia. Furthermore, removal of CFTR's PDZ binding motif (ΔTRL) prevented actin rearrangement, suggesting that CFTR insertion in the plasma membrane results in local reorganization of actin, PDZ binding proteins and certain GPCRs. PMID:27278076

  17. Gen-Gen- und Gen-Umwelt-Interaktionsanalysen bei Kindern der multrizentrischen Allergie Studie

    OpenAIRE

    Deindl, Philipp

    2005-01-01

    Atopische Erkrankungen wie Asthma, atopische Dermatitis und allergische Rhinitis sind komplexe, multifaktorielle Erkrankungen. Diese Arbeit untersuchte Gen-Gen- und Gen-Umweltinteraktionen von insgesamt acht Polymorphismen in fünf Kandidatengenen. In einer großen deutschen Geburtskohorte (Multizentrische Allergie Studie, MAS 90) mit longitudinalen klinischen und serologischen Daten wurde der Effekt von Polymorphismen in Genen, die für Interleukin-13 (IL-13), Interleukin-4 (IL-4) und dessen...

  18. Potentiation of ΔF508- and G551D-CFTR-Mediated Cl- Current by Novel Hydroxypyrazolines.

    Science.gov (United States)

    Park, Jinhong; Khloya, Poonam; Seo, Yohan; Kumar, Satish; Lee, Ho K; Jeon, Dong-Kyu; Jo, Sungwoo; Sharma, Pawan K; Namkung, Wan

    2016-01-01

    The most common mutation of CFTR, affecting approximately 90% of CF patients, is a deletion of phenylalanine at position 508 (F508del, ΔF508). Misfolding of ΔF508-CFTR impairs both its trafficking to the plasma membrane and its chloride channel activity. To identify small molecules that can restore channel activity of ΔF508-CFTR, we synthesized and evaluated eighteen novel hydroxypyrazoline analogues as CFTR potentiators. To elucidate potentiation activities of hydroxypyrazolines for ΔF508-CFTR, CFTR activity was measured using a halide-sensitive YFP assay, Ussing chamber assay and patch-clamp technique. Compounds 7p, 7q and 7r exhibited excellent potentiation with EC50 value <10 μM. Among the compounds, 7q (a novel CFTR potentiator, CP7q) showed the highest potentiation activity with EC50 values of 0.88 ± 0.11 and 4.45 ± 0.31 μM for wild-type and ΔF508-CFTR, respectively. In addition, CP7q significantly potentiated chloride conductance of G551D-CFTR, a CFTR gating mutant; its maximal potentiation activity was 1.9 fold higher than the well-known CFTR potentiator genistein. Combination treatment with CP7q and VX-809, a corrector of ΔF508-CFTR, significantly enhanced functional rescue of ΔF508-CFTR compared with VX-809 alone. CP7q did not alter the cytosolic cAMP level and showed no cytotoxicity at the concentration showing maximum efficacy. The hydroxypyrazolines may be potential development candidates for drug therapy of cystic fibrosis. PMID:26863533

  19. Potentiation of ΔF508- and G551D-CFTR-Mediated Cl- Current by Novel Hydroxypyrazolines.

    Directory of Open Access Journals (Sweden)

    Jinhong Park

    Full Text Available The most common mutation of CFTR, affecting approximately 90% of CF patients, is a deletion of phenylalanine at position 508 (F508del, ΔF508. Misfolding of ΔF508-CFTR impairs both its trafficking to the plasma membrane and its chloride channel activity. To identify small molecules that can restore channel activity of ΔF508-CFTR, we synthesized and evaluated eighteen novel hydroxypyrazoline analogues as CFTR potentiators. To elucidate potentiation activities of hydroxypyrazolines for ΔF508-CFTR, CFTR activity was measured using a halide-sensitive YFP assay, Ussing chamber assay and patch-clamp technique. Compounds 7p, 7q and 7r exhibited excellent potentiation with EC50 value <10 μM. Among the compounds, 7q (a novel CFTR potentiator, CP7q showed the highest potentiation activity with EC50 values of 0.88 ± 0.11 and 4.45 ± 0.31 μM for wild-type and ΔF508-CFTR, respectively. In addition, CP7q significantly potentiated chloride conductance of G551D-CFTR, a CFTR gating mutant; its maximal potentiation activity was 1.9 fold higher than the well-known CFTR potentiator genistein. Combination treatment with CP7q and VX-809, a corrector of ΔF508-CFTR, significantly enhanced functional rescue of ΔF508-CFTR compared with VX-809 alone. CP7q did not alter the cytosolic cAMP level and showed no cytotoxicity at the concentration showing maximum efficacy. The hydroxypyrazolines may be potential development candidates for drug therapy of cystic fibrosis.

  20. Relationships among CFTR expression, HCO3- secretion, and host defense may inform gene- and cell-based cystic fibrosis therapies.

    Science.gov (United States)

    Shah, Viral S; Ernst, Sarah; Tang, Xiao Xiao; Karp, Philip H; Parker, Connor P; Ostedgaard, Lynda S; Welsh, Michael J

    2016-05-10

    Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel. Airway disease is the major source of morbidity and mortality. Successful implementation of gene- and cell-based therapies for CF airway disease requires knowledge of relationships among percentages of targeted cells, levels of CFTR expression, correction of electrolyte transport, and rescue of host defense defects. Previous studies suggested that, when ∼10-50% of airway epithelial cells expressed CFTR, they generated nearly wild-type levels of Cl(-) secretion; overexpressing CFTR offered no advantage compared with endogenous expression levels. However, recent discoveries focused attention on CFTR-mediated HCO3 (-) secretion and airway surface liquid (ASL) pH as critical for host defense and CF pathogenesis. Therefore, we generated porcine airway epithelia with varying ratios of CF and wild-type cells. Epithelia with a 50:50 mix secreted HCO3 (-) at half the rate of wild-type epithelia. Likewise, heterozygous epithelia (CFTR(+/-) or CFTR(+/∆F508)) expressed CFTR and secreted HCO3 (-) at ∼50% of wild-type values. ASL pH, antimicrobial activity, and viscosity showed similar relationships to the amount of CFTR. Overexpressing CFTR increased HCO3 (-) secretion to rates greater than wild type, but ASL pH did not exceed wild-type values. Thus, in contrast to Cl(-) secretion, the amount of CFTR is rate-limiting for HCO3 (-) secretion and for correcting host defense abnormalities. In addition, overexpressing CFTR might produce a greater benefit than expressing CFTR at wild-type levels when targeting small fractions of cells. These findings may also explain the risk of airway disease in CF carriers. PMID:27114540

  1. XIAO Pei-gen

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    <正>Academician of Chinese Academy of Engineering Editor-in-chief of Chinese Herbal Medicines(CHM)Honorary director of Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Beijing100193,China Tel/Fax:+86-10-62894462 E-mail:xiaopg@public.bta.net.cn Professor XIAO Pei-gen is the founder of the Institute of Medicinal Plant Development(IMPLAD),Chinese Academy of Medical Sciences(CAMS).He is also one of the founders and leading

  2. GEN 499 ASH course tutorial/tutorialoutlet

    OpenAIRE

    NARESH 34

    2015-01-01

    For more course tutorials visit www.tutorialoutlet.com       GEN 499 Week 1 DQ 1 Final Research Paper Topic and Plan (Ash) GEN 499 Week 1 DQ 2 Social Media (Ash) GEN 499 Week 2 DQ 1 Professional Resume and Cover Letter (Ash) GEN 499 Week 2 Assignment Critiquing Internet Sources (Ash) GEN 499 Week 3 DQ 1 Social Capital (Ash) GEN 499 Week 3 DQ 2 Federal Policy (Ash) GEN 499 Week 3 Assignment Annotated Bibliography (Ash) GEN 499 Week 4 DQ 1...

  3. Gen quimérico que comprende el gen o ADNc de la fructoquinasa para el desarrollo de aproximaciones terapéuticas a la intolerancia hereditaria a la fructosa

    OpenAIRE

    Bosch i Tubert, Fàtima; Feliu Albiñana, Juan Emilio; Pujol i Altarriba, Anna; Sánchez Gutiérrez, Julio César

    2003-01-01

    Gen quimérico para el desarrollo de aproximaciones terapéuticas a la intolerancia hereditaria a la fructosa. Comprende un promotor unido al gen o al ADNc de la fructoquinasa. Dicho gen quimérico se inserta en una célula hepática, permitiendo la expresión de fructoquinasa. También se refiere a un animal transgénico cuyo genoma comprende dicho gen quimérico y desarrollando, dicho animal transgénico, alteraciones asociadas a la intolerancia hereditaria a la fructosa tras la administración de fru...

  4. Side chain and backbone contributions of Phe508 to CFTR folding

    Energy Technology Data Exchange (ETDEWEB)

    Thibodeau, Patrick H.; Brautigam, Chad A.; Machius, Mischa; Thomas, Philip J. (U. of Texas-SMED)

    2010-12-07

    Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), an integral membrane protein, cause cystic fibrosis (CF). The most common CF-causing mutant, deletion of Phe508, fails to properly fold. To elucidate the role Phe508 plays in the folding of CFTR, missense mutations at this position were generated. Only one missense mutation had a pronounced effect on the stability and folding of the isolated domain in vitro. In contrast, many substitutions, including those of charged and bulky residues, disrupted folding of full-length CFTR in cells. Structures of two mutant nucleotide-binding domains (NBDs) reveal only local alterations of the surface near position 508. These results suggest that the peptide backbone plays a role in the proper folding of the domain, whereas the side chain plays a role in defining a surface of NBD1 that potentially interacts with other domains during the maturation of intact CFTR.

  5. La genómica de las plantas

    OpenAIRE

    Cánovas, Francisco M.

    2009-01-01

    La mejora vegetal es una actividad tan antigua como la propia agricultura. El estudio de las variedades genéticas de la vegetación permite conocer en profundidad sus funciones y mecanismos para localizar los beneficios alimenticios, textiles, cosméticos o combustibles que la naturaleza ha puesto al servicio del ser humano.

  6. Genética molecular del alcoholismo

    OpenAIRE

    Mauricio Rey-Buitrago

    2015-01-01

    El alcoholismo es una patología psiquiátrica compleja y de origen multifactorial en la que el factor genético explica alrededor del 50 % del fenómeno. Son numerosos los genes que se han asociado a esta enfermedad, pero su aporte individual es mínimo y contradictorio. Estos genes operan a través de características intermedias como la impulsividad y la sensibilidad al alcohol, lo que hace compleja la definición del fenotipo del alcoholismo. Los estudios de asociación de SNPs, de asociación a to...

  7. CFTR is required for maximal transepithelial liquid transport in pig alveolar epithelia

    OpenAIRE

    Li, Xiaopeng; Comellas, Alejandro P.; Karp, Philip H.; Ernst, Sarah E.; Moninger, Thomas O.; Gansemer, Nicholas D.; Taft, Peter J.; Pezzulo, Alejandro A; Michael V Rector; Rossen, Nathan; Stoltz, David A.; McCray, Paul B.; Welsh, Michael J.; Zabner, Joseph

    2012-01-01

    A balance between alveolar liquid absorption and secretion is critical for maintaining optimal alveolar subphase liquid height and facilitating gas exchange in the alveolar space. However, the role of cystic fibrosis transmembrane regulator protein (CFTR) in this homeostatic process has remained elusive. Using a newly developed porcine model of cystic fibrosis, in which CFTR is absent, we investigated ion transport properties and alveolar liquid transport in isolated type II alveolar epitheli...

  8. Physiological Adaptation of the Bacterium Lactococcus lactis in Response to the Production of Human CFTR*

    OpenAIRE

    A. Steen; Wiederhold, E.; T Gandhi; Breitling, R.; D. J. Slotboom

    2010-01-01

    Biochemical and biophysical characterization of CFTR (the cystic fibrosis transmembrane conductance regulator) is thwarted by difficulties to obtain sufficient quantities of correctly folded and functional protein. Here we have produced human CFTR in the prokaryotic expression host Lactococcus lactis. The full-length protein was detected in the membrane of the bacterium, but the yields were too low (< 0.1% of membrane proteins) for in vitro functional and structural characterization, and indu...

  9. Adeno-associated virus–targeted disruption of the CFTR gene in cloned ferrets

    OpenAIRE

    Sun, Xingshen; Yan, Ziying; Yi, Yaling; Li, Ziyi; Lei, Diana; Rogers, Christopher S.; Chen, Juan; Zhang, Yulong; Welsh, Michael J.; Leno, Gregory H.; Engelhardt, John F.

    2008-01-01

    Somatic cell gene targeting combined with nuclear transfer cloning presents tremendous potential for the creation of new, large-animal models of human diseases. Mouse disease models often fail to reproduce human phenotypes, underscoring the need for the generation and study of alternative disease models. Mice deficient for CFTR have been poor models for cystic fibrosis (CF), lacking many aspects of human CF lung disease. In this study, we describe the production of a CFTR gene–deficient model...

  10. Rare large homozygous CFTR gene deletion in an Iranian patient with cystic fibrosis

    OpenAIRE

    Farjadian, Shirin; Moghtaderi, Mozhgan; Zuntini, Roberta; Ferrari, Simona

    2014-01-01

    Cystic fibrosis, a common autosomal recessive genetic disorder among Caucasians, is caused by defects in the transmembrane conductance regulatory (CFTR) gene. The analysis of CFTR gene mutations is useful to better characterize the disease, and for preconceptional screening, prenatal and preimplantation genetic diagnosis. Here we report the results of a genetic analysis in a 16-year-old boy from southwestern Iran diagnosed as having cystic fibrosis in infancy based on gastrointestinal and pul...

  11. CFTR chloride channel as a molecular target of anthraquinone compounds in herbal laxatives

    Institute of Scientific and Technical Information of China (English)

    Hong YANG; Li-na XU; Cheng-yan HE; Xin LIU; Rou-yu FANG; Tong-hui MA

    2011-01-01

    Aim: To clarify whether CFTR is a molecular target of intestinal fluid secretion caused by the anthraquinone compounds from laxative herbal plants.Methods: A cell-based fluorescent assay to measure I- influx through CFTR chloride channel. A short-circuit current assay to measure transcellular Cl- current across single layer FRT cells and freshly isolated colon mucosa. A closed loop experiment to measure colon fluid secretion in vivo.Results: Anthraquinone compounds rhein, aloe-emodin and 1,8-dihydroxyanthraquinone (DHAN) stimulated l- influx through CFTR chloride channel in a dose-dependent manner in the presence of physiological concentration of cAMP. In the short-circuit current assay,the three compound enhanced Cl- currents in epithelia formed by CFTR-expressing FRT cells with EC5o values of 73±1.4, 56±1.7, and 50±0.5 μmol/L, respectively, and Rhein also enhanced Cl- current in freshly isolated rat colonic mucosa with a similar potency. These effects were completely reversed by the CFTR selective blocker CFTRinh-172. In in vivo closed loop experiments, rhein 2 mmol/L stimu-lated colonic fluid accumulation that was largely blocked by CFTRinh-172. The anthraquinone compounds did not elevate cAMP level in cultured FRT cells and rat colonic mucosa, suggesting a direct effect on CFTR activity.Conclusion: Natural anthraquinone compounds in vegetable laxative drugs are CFTR potsntiators that stimulated colonic chloride and fluid secretion. Thus CFTR chloride channel is a molecular target of vegetable laxative drugs.

  12. Phenotypic variability of R117H-CFTR expression within monozygotic twins.

    Science.gov (United States)

    Waller, Michael D; Simmonds, Nicholas J

    2016-08-01

    Whilst cystic fibrosis is a monogenic condition, variation in phenotype exists for the same CFTR genotype, which is influenced by multiple genetic and non-genetic (environmental) factors. The R117H-CFTR mutation has variability directly relating to in cis poly-thymidine alleles, producing a differing spectrum of disease. This paper provides evidence of extreme phenotype variability - including fertility status - in the context of male monogenetic twins, discussing mechanisms and highlighting the diagnostic and treatment challenges. PMID:27364092

  13. La gens Licinia y el Nordeste peninsular. Una aproximación al estudio de las formas de propiedad y de gestión de un rico patrimonio familiar

    Directory of Open Access Journals (Sweden)

    Berni, Piero

    2005-12-01

    Full Text Available In the last years, either progress in landscape archaeology as well as studies on the epigraphy of production areas, have contribute to increase our volume of information. However, such headway not always has come along with an improvement in the data interpretation, in other words, in the study of the complex forms of landownership and management of resources in Roman Spain. We believed it is necessary to integrate diverse types of existent documentation sources, and we propose in the present case to study a working model focus on the gens Licinia. The importance of this gens in the Northeast of the Peninsula and the outstanding volume of information preserved allow us to put forward a series of methodological and historical reflections we judge significant.En los últimos años, tanto el progreso de la arqueología del paisaje, como especialmente el de los estudios sobre la epigrafía de la producción, han contribuido a incrementar nuestro volumen de información, aunque ello no siempre ha ido acompañado de un progreso en la interpretación de los datos, es decir, en el estudio de las complejas formas de propiedad de la tierra y de la gestión de sus recursos en la Hispania romana. Creemos necesario, por tanto, integrar en este estudio los diversos tipos de fuentes documentales existentes y proponer un modelo de trabajo centrado en el caso de la gens Licinia, cuya importancia en el Nordeste peninsular por el notable volumen de información conservada nos permite plantear algunas reflexiones metodológicas e históricas que creemos significativas.

  14. Refining the continuum of CFTR-associated disorders in the era of newborn screening.

    Science.gov (United States)

    Levy, H; Nugent, M; Schneck, K; Stachiw-Hietpas, D; Laxova, A; Lakser, O; Rock, M; Dahmer, M K; Biller, J; Nasr, S Z; Baker, M; McColley, S A; Simpson, P; Farrell, P M

    2016-05-01

    Clinical heterogeneity in cystic fibrosis (CF) often causes diagnostic uncertainty in infants without symptoms and in older patients with milder phenotypes. We performed a cross-sectional evaluation of a comprehensive set of clinical and laboratory descriptors in a physician-defined cohort (N = 376; Children's Hospital of Wisconsin and the American Family Children's Hospital CF centers in Milwaukee and Madison, WI, USA) to determine the robustness of categorizing CF (N = 300), cystic fibrosis transmembrane conductance regulator (CFTR)-related disorder (N = 19), and CFTR-related (CRMS) metabolic syndrome (N = 57) according to current consensus guidelines. Outcome measures included patient demographics, clinical measures, sweat chloride levels, CFTR genotype, age at diagnosis, airway microbiology, pancreatic function, infection, and nutritional status. The CF cohort had a significantly higher median sweat chloride level (105 mmol/l) than CFTR-related disorder patients (43 mmol/l) and CFTR-related metabolic syndrome patients (35 mmol/l; p ≤ 0.001). Patient groups significantly differed in pancreatic sufficiency, immunoreactive trypsinogen levels, sweat chloride values, genotype, and positive Pseudomonas aeruginosa cultures (p ≤ 0.001). An automated classification algorithm using recursive partitioning demonstrated concordance between physician diagnoses and consensus guidelines. Our analysis suggests that integrating clinical information with sweat chloride levels, CFTR genotype, and pancreatic sufficiency provides a context for continued longitudinal monitoring of patients for personalized and effective treatment. PMID:26671754

  15. Synthesis and Characterization of A Small Molecule CFTR Chloride Channel Inhibitor

    Institute of Scientific and Technical Information of China (English)

    HE Cheng-yan; ZHANG Heng-jun; SU Zhong-min; ZHOU Jin-song; YANG Hong; MA Tong-hui

    2004-01-01

    A thiazolidinone CFTR inhibitor(CFTRinh-172) was synthesized by a three-step procedure with trifluromethylaniline as the starting material. The synthesized CFTR inhibitor was characterized structurally by means of 1H NMR and functionally in a CFTR-expressing cell line FRT/hCFTR/EYFP-H148Q by both fluorescent and electrophysiological methods. A large amount(100 g) of high-quality small molecule thiazolidinone CFTR chloride channel inhibitor, CFTRinh-172, can be produced with this simple three-step synthetic procedure. The structure of the final product 2-thioxo-3-(3-trifluromethylphenyl)-5-[4-carboxyphenyl-methylene]-4-thiazolidinone was confirmed by 1H NMR. The overall yield was 58% with a purity over 99% as analyzed by HPLC. The synthesized CFTRinh-172 specifically inhibited CFTR chloride channel function in a cell-based fluorescence assay(Kd≈1.5 μmol/L) and in a Ussing chamber-based short-circuit current assay(Kd≈0.2 μmol/L), indicating better quality than that of the commercial combinatorial compound. The synthesized inhibitor is nontoxic to cultured cells at a high concentration and to mouse at a high dose. The synthetic procedure developed here can be used to produce a large amount of the high-quality CFTRinh-172 suitable for antidiarrheal studies and for creation of cystic fibrosis models in large animals. The procedure can be used to synthesize radiolabled CFTRinh-172 for in vivo pharmacokinetics studies.

  16. A host defense mechanism involving CFTR-mediated bicarbonate secretion in bacterial prostatitis.

    Directory of Open Access Journals (Sweden)

    Chen Xie

    Full Text Available BACKGROUND: Prostatitis is associated with a characteristic increase in prostatic fluid pH; however, the underlying mechanism and its physiological significance have not been elucidated. METHODOLOGY/PRINCIPAL FINDINGS: In this study a primary culture of rat prostatic epithelial cells and a rat prostatitis model were used. Here we reported the involvement of CFTR, a cAMP-activated anion channel conducting both Cl(- and HCO(3(-, in mediating prostate HCO(3(- secretion and its possible role in bacterial killing. Upon Escherichia coli (E. coli-LPS challenge, the expression of CFTR and carbonic anhydrase II (CA II, along with several pro-inflammatory cytokines was up-regulated in the primary culture of rat prostate epithelial cells. Inhibiting CFTR function in vitro or in vivo resulted in reduced bacterial killing by prostate epithelial cells or the prostate. High HCO(3(- content (>50 mM, rather than alkaline pH, was found to be responsible for bacterial killing. The direct action of HCO(3(- on bacterial killing was confirmed by its ability to increase cAMP production and suppress bacterial initiation factors in E. coli. The relevance of the CFTR-mediated HCO(3(- secretion in humans was demonstrated by the upregulated expression of CFTR and CAII in human prostatitis tissues. CONCLUSIONS/SIGNIFICANCE: The CFTR and its mediated HCO(3(- secretion may be up-regulated in prostatitis as a host defense mechanism.

  17. Facilitating Structure-Function Studies of CFTR Modulator Sites with Efficiencies in Mutagenesis and Functional Screening.

    Science.gov (United States)

    Molinski, Steven V; Ahmadi, Saumel; Hung, Maurita; Bear, Christine E

    2015-12-01

    There are nearly 2000 mutations in the CFTR gene associated with cystic fibrosis disease, and to date, the only approved drug, Kalydeco, has been effective in rescuing the functional expression of a small subset of these mutant proteins with defects in channel activation. However, there is currently an urgent need to assess other mutations for possible rescue by Kalydeco, and further, definition of the binding site of such modulators on CFTR would enhance our understanding of the mechanism of action of such therapeutics. Here, we describe a simple and rapid one-step PCR-based site-directed mutagenesis method to generate mutations in the CFTR gene. This method was used to generate CFTR mutants bearing deletions (p.Gln2_Trp846del, p.Ser700_Asp835del, p.Ile1234_Arg1239del) and truncation with polyhistidine tag insertion (p.Glu1172-3Gly-6-His*), which either recapitulate a disease phenotype or render tools for modulator binding site identification, with subsequent evaluation of drug responses using a high-throughput (384-well) membrane potential-sensitive fluorescence assay of CFTR channel activity within a 1 wk time frame. This proof-of-concept study shows that these methods enable rapid and quantitative comparison of multiple CFTR mutants to emerging drugs, facilitating future large-scale efforts to stratify mutants according to their "theratype" or most promising targeted therapy. PMID:26385858

  18. Lack of CFTR in Skeletal Muscle Predisposes to Muscle Wasting and Diaphragm Muscle Pump Failure in Cystic Fibrosis Mice

    OpenAIRE

    Maziar Divangahi; Haouaria Balghi; Gawiyou Danialou; Comtois, Alain S.; Alexandre Demoule; Sheila Ernest; Christina Haston; Renaud Robert; Hanrahan, John W.; Danuta Radzioch; Petrof, Basil J

    2009-01-01

    Cystic fibrosis (CF) patients often have reduced mass and strength of skeletal muscles, including the diaphragm, the primary muscle of respiration. Here we show that lack of the CF transmembrane conductance regulator (CFTR) plays an intrinsic role in skeletal muscle atrophy and dysfunction. In normal murine and human skeletal muscle, CFTR is expressed and co-localized with sarcoplasmic reticulum-associated proteins. CFTR-deficient myotubes exhibit augmented levels of intracellular calcium aft...

  19. Defective CFTR expression and function are detectable in blood monocytes : development of a new blood test for cystic fibrosis

    OpenAIRE

    Sorio, C.; Buffelli, M.; C. Angiari; Ettorre, M; Johansson, J; M. Vezzalini; Viviani, L; Ricciardi, M.; G. Verzè; Assael, B M; P. Melotti

    2011-01-01

    Background Evaluation of cystic fibrosis transmembrane conductance regulator (CFTR) functional activity to assess new therapies and define diagnosis of cystic fibrosis (CF) is cumbersome. It is known that leukocytes express detectable levels of CFTR but the molecule has not been characterized in these cells. In this study we aim at setting up and validating a blood test to evaluate CFTR expression and function in leukocytes. Description Western blot, PCR, immunofluorescence and cell membrane ...

  20. The K+ channel opener 1-EBIO potentiates residual function of mutant CFTR in rectal biopsies from cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Eva K Roth

    Full Text Available BACKGROUND: The identification of strategies to improve mutant CFTR function remains a key priority in the development of new treatments for cystic fibrosis (CF. Previous studies demonstrated that the K⁺ channel opener 1-ethyl-2-benzimidazolone (1-EBIO potentiates CFTR-mediated Cl⁻ secretion in cultured cells and mouse colon. However, the effects of 1-EBIO on wild-type and mutant CFTR function in native human colonic tissues remain unknown. METHODS: We studied the effects of 1-EBIO on CFTR-mediated Cl⁻ secretion in rectal biopsies from 47 CF patients carrying a wide spectrum of CFTR mutations and 57 age-matched controls. Rectal tissues were mounted in perfused micro-Ussing chambers and the effects of 1-EBIO were compared in control tissues, CF tissues expressing residual CFTR function and CF tissues with no detectable Cl⁻ secretion. RESULTS: Studies in control tissues demonstrate that 1-EBIO activated CFTR-mediated Cl⁻ secretion in the absence of cAMP-mediated stimulation and potentiated cAMP-induced Cl⁻ secretion by 39.2±6.7% (P<0.001 via activation of basolateral Ca²⁺-activated and clotrimazole-sensitive KCNN4 K⁺ channels. In CF specimens, 1-EBIO potentiated cAMP-induced Cl⁻ secretion in tissues with residual CFTR function by 44.4±11.5% (P<0.001, but had no effect on tissues lacking CFTR-mediated Cl⁻ conductance. CONCLUSIONS: We conclude that 1-EBIO potentiates Cl⁻secretion in native CF tissues expressing CFTR mutants with residual Cl⁻ channel function by activation of basolateral KCNN4 K⁺ channels that increase the driving force for luminal Cl⁻ exit. This mechanism may augment effects of CFTR correctors and potentiators that increase the number and/or activity of mutant CFTR channels at the cell surface and suggests KCNN4 as a therapeutic target for CF.

  1. Development of allele-specific multiplex PCR to determine the length of poly-T in intron 8 of CFTR

    OpenAIRE

    Neng Chen; Prada, Anne E.

    2014-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation analysis has been implemented for Cystic Fibrosis (CF) carrier screening, and molecular diagnosis of CF and congenital bilateral absence of the vas deferens (CBAVD). Although poly-T allele analysis in intron 8 of CFTR is required when a patient is positive for R117H, it is not recommended for routine carrier screening. Therefore, commercial kits for CFTR mutation analysis were designed either to mask the poly-T allele re...

  2. Involvement of the Cdc42 pathway in CFTR post-translational turnover and in its plasma membrane stability in airway epithelial cells.

    Directory of Open Access Journals (Sweden)

    Romain Ferru-Clément

    Full Text Available Cystic fibrosis transmembrane conductance regulator (CFTR is a chloride channel that is expressed on the apical plasma membrane (PM of epithelial cells. The most common deleterious allele encodes a trafficking-defective mutant protein undergoing endoplasmic reticulum-associated degradation (ERAD and presenting lower PM stability. In this study, we investigated the involvement of the Cdc42 pathway in CFTR turnover and trafficking in a human bronchiolar epithelial cell line (CFBE41o- expressing wild-type CFTR. Cdc42 is a small GTPase of the Rho family that fulfils numerous cell functions, one of which is endocytosis and recycling process via actin cytoskeleton remodelling. When we treated cells with chemical inhibitors such as ML141 against Cdc42 and wiskostatin against the downstream effector N-WASP, we observed that CFTR channel activity was inhibited, in correlation with a decrease in CFTR amount at the cell surface and an increase in dynamin-dependent CFTR endocytosis. Anchoring of CFTR to the cortical cytoskeleton was then presumably impaired by actin disorganization. When we performed siRNA-mediated depletion of Cdc42, actin polymerization was not impacted, but we observed actin-independent consequences upon CFTR. Total and PM CFTR amounts were increased, resulting in greater activation of CFTR. Pulse-chase experiments showed that while CFTR degradation was slowed, CFTR maturation through the Golgi apparatus remained unaffected. In addition, we observed increased stability of CFTR in PM and reduction of its endocytosis. This study highlights the involvement of the Cdc42 pathway at several levels of CFTR biogenesis and trafficking: (i Cdc42 is implicated in the first steps of CFTR biosynthesis and processing; (ii it contributes to the stability of CFTR in PM via its anchoring to cortical actin; (iii it promotes CFTR endocytosis and presumably its sorting toward lysosomal degradation.

  3. Restoration of NBD1 thermal stability is necessary and sufficient to correct ΔF508 CFTR folding and assembly

    OpenAIRE

    He, Lihua; Aleksandrov, Andrei A.; An, Jianli; Cui, Liying; Yang, Zhengrong; Brouillette, Christie G; Riordan, John R.

    2014-01-01

    CFTR (ABCC7), unique among ABC exporters as an ion channel, regulates ion and fluid transport in epithelial tissues. Loss of function due to mutations in the cftr gene causes cystic fibrosis (CF). The most common CF-causing mutation, the deletion of F508 (ΔF508) from the first nucleotide binding domain (NBD1) of CFTR, results in misfolding of the protein and clearance by cellular quality control systems. The ΔF508 mutation has two major impacts on CFTR: reduced thermal stability of NBD1 and d...

  4. The mitochondrial complex I activity is reduced in cells with impaired cystic fibrosis transmembrane conductance regulator (CFTR function.

    Directory of Open Access Journals (Sweden)

    Angel G Valdivieso

    Full Text Available Cystic fibrosis (CF is a frequent and lethal autosomal recessive disease. It results from different possible mutations in the CFTR gene, which encodes the CFTR chloride channel. We have previously studied the differential expression of genes in CF and CF corrected cell lines, and found a reduced expression of MTND4 in CF cells. MTND4 is a mitochondrial gene encoding the MTND4 subunit of the mitochondrial Complex I (mCx-I. Since this subunit is essential for the assembly and activity of mCx-I, we have now studied whether the activity of this complex was also affected in CF cells. By using Blue Native-PAGE, the in-gel activity (IGA of the mCx-I was found reduced in CFDE and IB3-1 cells (CF cell lines compared with CFDE/6RepCFTR and S9 cells, respectively (CFDE and IB3-1 cells ectopically expressing wild-type CFTR. Moreover, colon carcinoma T84 and Caco-2 cells, which express wt-CFTR, either treated with CFTR inhibitors (glibenclamide, CFTR(inh-172 or GlyH101 or transfected with a CFTR-specific shRNAi, showed a significant reduction on the IGA of mCx-I. The reduction of the mCx-I activity caused by CFTR inhibition under physiological or pathological conditions may have a profound impact on mitochondrial functions of CF and non-CF cells.

  5. miR-16 rescues F508del-CFTR function in native cystic fibrosis epithelial cells.

    Science.gov (United States)

    Kumar, P; Bhattacharyya, S; Peters, K W; Glover, M L; Sen, A; Cox, R T; Kundu, S; Caohuy, H; Frizzell, R A; Pollard, H B; Biswas, R

    2015-11-01

    Cystic fibrosis (CF) is due to mutations in the CFTR gene, which prevents correct folding, trafficking and function of the mutant cystic fibrosis transmembrane conductance regulator (CFTR) protein. The dysfunctional effect of CFTR mutations, principally the F508del-CFTR mutant, is further manifested by hypersecretion of the pro-inflammatory chemokine interleukin-8 into the airway lumen, which further contributes to morbidity and mortality. We have hypothesized that microRNA (miR)-based therapeutics could rescue the dysfunctional consequences of mutant CFTR. Here we report that a miR-16 mimic can effectively rescue F508del-CFTR protein function in airway cell lines and primary cultures, of differentiated human bronchial epithelia from F508del homozygotes, which express mutant CFTR endogenously. We also identify two other miRs, miR-1 and miR-302a, which are also active. Although miR-16 is expressed at basal comparable levels in CF and control cells, miR-1 and miR-302a are undetectable. When miR mimics are expressed in CF lung or pancreatic cells, the expression of the F508del-CFTR protein is significantly increased. Importantly, miR-16 promotes functional rescue of the cyclic AMP-activated apical F508del-CFTR chloride channel in primary lung epithelial cells from CF patients. We interpret these findings to suggest that these miRs may constitute novel targets for CF therapy. PMID:26133785

  6. Gen-Umwelt-Interaktionen und Gen-Umwelt-Korrelationen bei psychiatrischen Erkrankungen

    Directory of Open Access Journals (Sweden)

    Winkler D

    2010-01-01

    Full Text Available Die seit Langem bestehende Frage, in welchem Ausmaß Anlage und Umwelt zu psychologischen Merkmalen und psychiatrischen Erkrankungen beitragen, wird durch Ergebnisse von Zwillingsstudien und in letzter Zeit durch Untersuchungen des Zusammenhangs zwischen molekulargenetischen Merkmalen und Umwelteinflüssen bereichert. Eine Gen-Umwelt-Interaktion liegt dann vor, wenn genetische Faktoren die Auswirkungen von Umweltbedingungen modulieren. Die Genetik kann weiters die Wahrscheinlichkeit der Exposition gegenüber bestimmten Umwelteinflüssen verändern, was als Gen- Umwelt-Korrelation bezeichnet wird. Beide Phänomene liegen aber häufig gleichzeitig vor, was eine besondere Herausforderung für die Konzeption von wissenschaftlichen Studien darstellt.

  7. Ubiquitination and Degradation of CFTR by the E3 Ubiquitin Ligase MARCH2 through Its Association with Adaptor Proteins CAL and STX6

    OpenAIRE

    Jie Cheng; William Guggino

    2013-01-01

    Golgi-localized cystic fibrosis transmembrane conductance regulator (CFTR)-associated ligand (CAL) and syntaxin 6 (STX6) regulate the abundance of mature, post-ER CFTR by forming a CAL/STX6/CFTR complex (CAL complex) that promotes CFTR degradation in lysosomes. However, the molecular mechanism underlying this degradation is unknown. Here we investigated the interaction of a Golgi-localized, membrane-associated RING-CH E3 ubiquitin ligase, MARCH2, with the CAL complex and the consequent bindin...

  8. Potentiators of Defective ΔF508-CFTR Gating that Do Not Interfere with Corrector Action.

    Science.gov (United States)

    Phuan, Puay-Wah; Veit, Guido; Tan, Joseph A; Finkbeiner, Walter E; Lukacs, Gergely L; Verkman, A S

    2015-10-01

    Combination drug therapies under development for cystic fibrosis caused by the ∆F508 mutation in cystic fibrosis transmembrane conductance regulator (CFTR) include a "corrector" to improve its cellular processing and a "potentiator" to improve its chloride channel function. Recently, it was reported that the approved potentiator N-(2,4-di-tert-butyl-5-hydroxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (Ivacaftor) reduces ∆F508-CFTR cellular stability and the efficacy of investigational correctors, including 3-(6-[([1-(2,2-difluoro-1,3-benzodioxol-5-yl)cyclopropyl]carbonyl) amino]-3-methyl-2-pyridinyl)-benzoic acid and 1-(2,2-difluoro-1,3-benzodioxol-5-yl)-N-(1-[(2R)-2,3-dihydroxypropyl]-6-fluoro-2-(2-hydroxy-1,1-dimethylethyl)-1H-indol-5-yl), which might contribute to the modest reported efficacy of combination therapy in clinical trials. Here, we report the identification and characterization of potentiators that do not interfere with ∆F508-CFTR stability or corrector action. High-throughput screening and structure-activity analysis identified several classes of potentiators that do not impair corrector action, including tetrahydrobenzothiophenes, thiooxoaminothiazoles, and pyrazole-pyrrole-isoxazoles. The most potent compounds have an EC(50) for ∆F508-CFTR potentiation down to 18 nM and do not reduce corrector efficacy in heterologous ∆F508-CFTR-expressing cells or primary cultures of ∆F508/∆F508 human bronchial epithelia. The ΔF508-CFTR potentiators also activated wild-type and G551D CFTR, albeit weakly. The efficacy of combination therapy for cystic fibrosis caused by the ∆F508 mutation may be improved by replacement of Ivacaftor with a potentiator that does not interfere with corrector action. PMID:26245207

  9. CFTR suppresses tumor progression through miR-193b targeting urokinase plasminogen activator (uPA) in prostate cancer.

    Science.gov (United States)

    Xie, C; Jiang, X H; Zhang, J T; Sun, T T; Dong, J D; Sanders, A J; Diao, R Y; Wang, Y; Fok, K L; Tsang, L L; Yu, M K; Zhang, X H; Chung, Y W; Ye, L; Zhao, M Y; Guo, J H; Xiao, Z J; Lan, H Y; Ng, C F; Lau, K M; Cai, Z M; Jiang, W G; Chan, H C

    2013-05-01

    Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is expressed in the epithelial cells of a wide range of organs/tissues from which most cancers are derived. Although accumulating reports have indicated the association of cancer incidence with genetic variations in CFTR gene, the exact role of CFTR in cancer development and the possible underlying mechanism have not been elucidated. Here, we report that CFTR expression is significantly decreased in both prostate cancer cell lines and human prostate cancer tissue samples. Overexpression of CFTR in prostate cancer cell lines suppresses tumor progression (cell growth, adhesion and migration), whereas knockdown of CFTR leads to enhanced malignancies both in vitro and in vivo. In addition, we demonstrate that CFTR knockdown-enhanced cell proliferation, cell invasion and migration are significantly reversed by antibodies against either urokinase plasminogen activator (uPA) or uPA receptor (uPAR), which are known to be involved in various malignant traits of cancer development. More interestingly, overexpression of CFTR suppresses uPA by upregulating the recently described tumor suppressor microRNA-193b (miR-193b), and overexpression of pre-miR-193b significantly reverses CFTR knockdown-enhanced malignant phenotype and abrogates elevated uPA activity in prostate cancer cell line. Finally, we show that CFTR gene transfer results in significant tumor repression in prostate cancer xenografts in vivo. Taken together, the present study has demonstrated a previously undefined tumor-suppressing role of CFTR and its involvement in regulation of miR-193b in prostate cancer development. PMID:22797075

  10. Funciones del gen relacionado a la latencia (gen LR) del herpes virus bovino tipo 1 (BoHV-1) y otras proteínas expresadas durante la latencia

    OpenAIRE

    S.E. Pérez

    2009-01-01

    El herpesvirus bovino tipo 1 (BoHV-1) causa diversos síndromes clínicos en el ganado. Al igual que otros alfa-herpesvirus, el ciclo de vida del BoHV-1 puede dividirse en tres etapas: infección aguda, latencia y reactivación. Durante la infección aguda, BoHV-1 expresa una cascada de genes altamente regulados. Sin embargo, durante la latencia, la expresión de genes virales se ve restringida al gen relacionado a la latencia (gen LR) y al ORF-E. El gen LR es poliadenilado y sufre empalmes alterna...

  11. A little CFTR goes a long way: CFTR-dependent sweat secretion from G551D and R117H-5T cystic fibrosis subjects taking ivacaftor.

    Directory of Open Access Journals (Sweden)

    Jessica E Char

    Full Text Available To determine if oral dosing with the CFTR-potentiator ivacaftor (VX-770, Kalydeco improves CFTR-dependent sweating in CF subjects carrying G551D or R117H-5T mutations, we optically measured sweat secretion from 32-143 individually identified glands in each of 8 CF subjects; 6 F508del/G551D, one G551D/R117H-5T, and one I507del/R117H-5T. Two subjects were tested only (- ivacaftor, 3 only (+ ivacaftor and 3 (+/- ivacaftor (1-5 tests per condition. The total number of gland measurements was 852 (- ivacaftor and 906 (+ ivacaftor. A healthy control was tested 4 times (51 glands. For each gland we measured both CFTR-independent (M-sweat and CFTR-dependent (C-sweat; C-sweat was stimulated with a β-adrenergic cocktail that elevated [cAMP]i while blocking muscarinic receptors. Absent ivacaftor, almost all CF glands produced M-sweat on all tests, but only 1/593 glands produced C-sweat (10 tests, 5 subjects. By contrast, 6/6 subjects (113/342 glands produced C-sweat in the (+ ivacaftor condition, but with large inter-subject differences; 3-74% of glands responded with C/M sweat ratios 0.04%-2.57% of the average WT ratio of 0.265. Sweat volume losses cause proportionally larger underestimates of CFTR function at lower sweat rates. The losses were reduced by measuring C/M ratios in 12 glands from each subject that had the highest M-sweat rates. Remaining losses were estimated from single channel data and used to correct the C/M ratios, giving estimates of CFTR function (+ ivacaftor  = 1.6%-7.7% of the WT average. These estimates are in accord with single channel data and transcript analysis, and suggest that significant clinical benefit can be produced by low levels of CFTR function.

  12. Expandiendo el espectro mutacional en pacientes chilenos con fibrosis quística Expanding the CFTR mutation spectrum in Chilean patients with cystic fibrosis

    Directory of Open Access Journals (Sweden)

    Marcos Vásquez D

    2012-06-01

    Full Text Available Introducción: La fibrosis quística (FQ es una enfermedad con herencia autosómica recesiva, que presenta una incidencia de 1 en 8.000 a 9.000 recién nacidos en Chile. A la fecha se han descrito más de 1.800 mutaciones diferentes en el gen CFTR. El diagnóstico molecular disponible consiste en el análisis de las 36 mutaciones presentes con mayor frecuencia en población caucásica, donde se describe una tasa de detección de un 85%. Sin embargo, en Chile el rendimiento corresponde a un 42%. Por esta razón, hemos iniciado un análisis sistemático en la región codificante del gen CFTR con elfin de identificar los restantes alelos en pacientes chilenos con FQ. Métodos: Análisis por secuenciación de los exones 6,7,14,19y 20, en 48pacientes chilenos del Programa Nacional de FQ. Se incluyeron pacientes con criterios clínicos y de laboratorio de FQ, y con sólo una mutación identificada en el panel de 36 mutaciones. Resultados: Se identificaron 3 mutaciones diferentes que no se analizan en el panel de diagnóstico molecular y que no habían sido reportadas en pacientes chilenos, totalizando 14 casos. Cuatro casos corresponden a una nueva mutación en el exón 14, que produce un corrimiento en el marco de lectura y un codón de término prematuro (c.2462_2463delGT/p.Ser821ArgfsX4. Ocho casos presentan la mutación c.3196C>T en el exón 20, mientras que en 2 casos se encontró la mutación c.3039delC en el exón 19. Ambas mutaciones han sido descritas previamente en otras poblaciones. Discusión: La identificación de estas mutaciones ha incrementado notablemente la tasa de detección obtenida en nuestros pacientes. Esto crea la necesidad de adaptar el análisis molecular inicial en pacientes chilenos con FQ, redundando en un diagnóstico de certeza en gran parte de los casos y permitiendo un adecuado asesoramiento genético para las familias.Introduction: Cystic Fibrosis (CF is an autosomal recessive disease and affects 1 in 8000

  13. THE CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR (CFTR) IS EXPRESSED IN MATURATION STAGE AMELOBLASTS, ODONTOBLASTS AND BONE CELLS

    OpenAIRE

    Bronckers, Antonius; Kalogeraki, Lida; Jorna, Huub J.N.; Wilke, Martina; Bervoets, Theodore J.; LYARUU, DONACIAN M.; Zandieh-Doulabi, Behrouz; DenBesten, Pamela; de Jonge, Hugo

    2009-01-01

    Patients with cystic fibrosis (CF) have mild defects in dental enamel. The gene mutated in these patients is CFTR, a Cl− channel involved in transepithelial salt- and water transport and bicarbonate secretion. We tested the hypothesis that Cftr channels are present and operating in the plasma membranes of mouse ameloblasts.

  14. Altered intestinal bile salt biotransformation in a cystic fibrosis (Cftr(-/-)) mouse model with hepato-biliary pathology

    NARCIS (Netherlands)

    Bodewes, Frank A. J. A.; van der Wulp, Mariette Y. M.; Beharry, Satti; Doktorova, Marcela; Havinga, Rick; Boverhof, Renze; Phillips, M. James; Durie, Peter R.; Verkade, Henkjan J.

    2015-01-01

    Background: Cftr(-/-tm1UC) mice develop progressive hepato-biliary pathology. We hypothesize that this liver pathology is related to alterations' in biliary bile hydrophobicity and bile salt metabolism in Cftr(-/-tm1Unc) mice. Methods: We determined bile production, biliary and fecal bile salt- and

  15. A Class of High-affinity Bicyclooctane G551D-CFTR Activators Identified by High Throughput Screening

    Institute of Scientific and Technical Information of China (English)

    HE Cheng-yan; ZHAO Lu; LIU Yan-li; XU Li-na; SHANG De-jing; YANG Hong

    2004-01-01

    The glycine-to-aspartic acid missense mutation at the codon 551(G551D) of the cystic fibrosis transmembrane conductance regulator(CFTR) is one of the five most frequent cystic fibrosis(CF) mutations associated with a severe CF phenotype. To explore the feasibility of pharmacological correction of disrupted activation of CFTR chloride channel caused by G551D mutation, we developed a halide-sensitive fluorescence miniassay for G551D-CFTR in Fisher rat thyroid(FRT) epithelial cells for the discovery of novel activators of G551D-CFTR. A class of bicyclooctane small molecule compounds that efficiently stimulate G551D-CFTR chloride channel activity was identified by high throughput screening via the FRT cell-based assay. This class of compounds selectively activates G551D-CFTR with a high affinity, whereas little effect of the compounds on wildtype CFTR can be seen. The discovery of a class of bicyclooctane G551D-CFTR activators will permit the analysis of structure-activity relationship of the compounds to identify ideal leads for in vivo therapeutic studies.

  16. The cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in maturation stage ameloblasts, odontoblasts and bone cells

    NARCIS (Netherlands)

    A. Bronckers; L. Kalogeraki; H.J.N. Jorna; M. Wilke; T.J. Bervoets; D.M. Lyaruu; B. Zandieh-Doulabi; P. Denbesten; H. de Jonge

    2010-01-01

    Patients with cystic fibrosis (CF) have mild defects in dental enamel. The gene mutated in these patients is CFTR, a Cl− channel involved in transepithelial salt and water transport and bicarbonate secretion. We tested the hypothesis that Cftr channels are present and operating in the plasma membran

  17. The primary folding defect and rescue of ΔF508 CFTR emerge during translation of the mutant domain

    NARCIS (Netherlands)

    Hoelen, H.M.; Kleizen, B.; Schmidt, A.; Richardson, J.; Charitou, P.; Braakman, L.J.; Thomas, P. J.

    2010-01-01

    In the vast majority of cystic fibrosis (CF) patients, deletion of residue F508 from CFTR is the cause of disease. F508 resides in the first nucleotide binding domain (NBD1) and its absence leads to CFTR misfolding and degradation. We show here that the primary folding defect arises during synthesis

  18. CFTR mediates bicarbonate-dependent activation of miR-125b in preimplantation embryo development

    Institute of Scientific and Technical Information of China (English)

    Yong Chao Lu; Alvin Chun Hang Ma; Anskar Yu Hung Leung; He Feng Huang; Hsiao Chang Chan; Hui Chen; Kin Lam Fok; Lai Ling Tsang; Mei Kuen Yu; Xiao Hu Zhang; Jing Chen; Xiaohua Jiang; Yiu Wa Chung

    2012-01-01

    Although HCO3-is known to be required for early embryo development,its exact role remains elusive.Here we report that HCO3-acts as an environmental cue in regulating miR-125b expression through CFTR-mediated influx during preimplantation embryo development.The results show that the effect of HCO3-on preimplantation embryo development can be suppressed by interfering the function of a HCO3--conducting channel,CFTR,by a specific inhibitor or gene knockout.Removal of extracellular HCO3-or inhibition of CFTR reduces miR-125b expression in 2 cell-stage mouse embryos.Knockdown of miR-125b mimics the effect of HCO3-removal and CFTR inhibition,while injection of miR-125b precursor reverses it.Downregulation of miR-125b upregulates p53 cascade in both human and mouse embryos.The activation of miR-125b is shown to be mediated by sAC/PKA-dependent nuclear shuttling of NF-KB.These results have revealed a critical role of CFTR in signal transduction linking the environmental HCO3-to activation of miR-125b during preimplantation embryo development and indicated the importance of ion channels in regulation of miRNAs.

  19. Self-reactive CFTR T cells in humans: implications for gene therapy.

    Science.gov (United States)

    Calcedo, Roberto; Griesenbach, Uta; Dorgan, Daniel J; Soussi, Samia; Boyd, A Christopher; Davies, Jane C; Higgins, Tracy E; Hyde, Stephen C; Gill, Deborah R; Innes, J Alastair; Porteous, David J; Alton, Eric W; Wilson, James M; Limberis, Maria P

    2013-09-01

    Cystic fibrosis (CF) is one of the most common autosomal recessive lethal disorders affecting white populations of northern European ancestry. To date there is no cure for CF. Life-long treatments for CF are being developed and include gene therapy and the use of small-molecule drugs designed to target specific cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations. Irrespective of the type of molecular therapy for CF, which may include gene replacement, exon skipping, nonsense suppression, or molecular correctors, because all of these modulate gene expression there is an inherent risk of activation of T cells against the wild-type version of CFTR. Here we report the validation of the human interferon-γ enzyme-linked immunospot assay and its application for the analysis of CFTR-specific T cell responses in patients with CF and in non-CF subjects. We found non-CF subjects with low levels of self-reactive CFTR-specific T cells in the United States and several patients with CF with low to high levels of self-reactive CFTR-specific T cells in both the United States and the United Kingdom. PMID:23790242

  20. Obligate coupling of CFTR pore opening to tight nucleotide-binding domain dimerization.

    Science.gov (United States)

    Mihályi, Csaba; Töröcsik, Beáta; Csanády, László

    2016-01-01

    In CFTR, the chloride channel mutated in cystic fibrosis (CF) patients, ATP-binding-induced dimerization of two cytosolic nucleotide binding domains (NBDs) opens the pore, and dimer disruption following ATP hydrolysis closes it. Spontaneous openings without ATP are rare in wild-type CFTR, but in certain CF mutants constitute the only gating mechanism, stimulated by ivacaftor, a clinically approved CFTR potentiator. The molecular motions underlying spontaneous gating are unclear. Here we correlate energetic coupling between residues across the dimer interface with spontaneous pore opening/closure in single CFTR channels. We show that spontaneous openings are also strictly coupled to NBD dimerization, which may therefore occur even without ATP. Coordinated NBD/pore movements are therefore intrinsic to CFTR: ATP alters the stability, but not the fundamental structural architecture, of open- and closed-pore conformations. This explains correlated effects of phosphorylation, mutations, and drugs on ATP-driven and spontaneous activity, providing insights for understanding CF mutation and drug mechanisms. PMID:27328319

  1. Pathophysiologic consequences following inhibition of a CFTR-dependent developmental cascade in the lung

    Directory of Open Access Journals (Sweden)

    Larson Janet E

    2005-02-01

    Full Text Available Abstract Background Examination of late gestation developmental genes in vivo may be limited by early embryonic lethality and compensatory mechanisms. This problem is particularly apparent in evaluating the developmental role of the cystic fibrosis transmembrane conductance regulator (CFTR gene in the cystic fibrosis (CF phenotype. A previously described transient in utero knockout (TIUKO technology was used to address the developmental role of CFTR in the rat lung. Results Rat fetuses transiently treated with antisense cftr in utero developed pathology that replicated aspects of the human CF phenotype. The TIUKO CF rat developed lung fibrosis, chronic inflammation, reactive airway disease, and the CF Antigen (MRP8/14, a marker for CF in human patients, was expressed. Conclusions The transient in utero antisense technology can be used to evaluate genes that exhibit either early lethality or compensating gene phenotypes. In the lung CFTR is part of a developmental cascade for normal secretory cell differentiation. Absence of CFTR results in a constitutive inflammatory process that is involved in some aspects of CF pathophysiology.

  2. Activation of CFTR-mediated CI-Transport by Capsaicinoids in Cell Culture Model

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xue-liang; HOU Ting-ting; GE Hong; SUN Juan-juan; YANG Hong; MA Tong-hui

    2009-01-01

    Previous studies reported that capsaicin potentiates ΔF508 mutant cystic fibrosis transmembrane conductance regulator(CFTR) channel gating defect by transfected cell-based assays.It has been postulated that orally ingested capsaicin may conceptually be used to develop a therapeutic strategy to treat gastrointestinal disorders in CF patients.We tried to reproduce and extend those pre-clinical data of previous studies.Cell-based fluorescence functional measurements in Fischer thyroid epithelial cells(FRT) expressing CFTR showed no effect of capsaicin on potentiating ΔF508-CFTR.while genistein showed a strongly positive activity.Studies show that capsaicin and dihydrocapsaicin activated cAMP-prestimulated wild-type CFTR in a dose-dependent manner with a maximal response of 70% of that activated by genistein,thus gave an apparent EC50 of (40.4±6.8)μmol/L and (150.2±7.4) μmol/L respectively.Preliminary study shows that the binding sites for capsaicin and dihydrocapsaicin may be probably partially overlapped with that for genistein because the maximal activation of wild-type CFTR with genistein is partially blocked by capsaicin and dihydrocapsaicin.

  3. First functional polymorphism in CFTR promoter that results in decreased transcriptional activity and Sp1/USF binding

    International Nuclear Information System (INIS)

    Growing evidences show that functionally relevant polymorphisms in various promoters alter both transcriptional activity and affinities of existing protein-DNA interactions, and thus influence disease progression in humans. We previously reported the -94G>T CFTR promoter variant in a female CF patient in whom any known disease-causing mutation has been detected. To investigate whether the -94G>T could be a regulatory variant, we have proceeded to in silico analyses and functional studies including EMSA and reporter gene assays. Our data indicate that the promoter variant decreases basal CFTR transcriptional activity in different epithelial cells and alters binding affinities of both Sp1 and USF nuclear proteins to the CFTR promoter. The present report provides evidence for the first functional polymorphism that negatively affects the CFTR transcriptional activity and demonstrates a cooperative role of Sp1 and USF transcription factors in transactivation of the CFTR gene promoter

  4. Conformationally rigid histone deacetylase inhibitors correct DF508-CFTR protein function

    DEFF Research Database (Denmark)

    Vickers, Chris J.; Olsen, Christian Adam; Hutt, Darren M.;

    2011-01-01

    Histone deacetylase (HDAC) inhibitors have shown partial efficacy toward correcting cystic fibrosis transmembrane conductance regulator (CFTR) protein function in ΔF508- CFTR models. While current treatment options for CF generally concentrate on disease symptoms such as management of inflammation...... and bacterial infection, therapy using HDAC inhibitors has the potential to treat and correct the underlying etiology associated with the disorder. Subsequently, we have synthesized conformationally well-defined cyclic tetrapeptide derivatives based on the natural product HDAC inhibitor Apicidin, in order...... to formulate a pharmacophore model to describe and enhance the bioactivity of these molecules. Through this study we have developed HDAC inhibitors which improve CFTR trafficking from the endoplasmic reticulum (ER) while ultimately increasing ion conductance across the plasma membrane of a lung epithelial cell...

  5. Lumacaftor/ivacaftor combination for cystic fibrosis patients homozygous for Phe508del-CFTR.

    Science.gov (United States)

    Zhang, W; Zhang, X; Zhang, Y H; Strokes, D C; Naren, A P

    2016-04-01

    Cystic fibrosis (CF) is a life-shortening inherited disease caused by the loss or dysfunction of the CF transmembrane conductance regulator (CFTR) channel activity resulting from mutations in the CFTR gene. Phe508del is the most prevalent mutation, with approximately 90% of all CF patients carrying it on at least one allele. Over the past two or three decades, significant progress has been made in understanding the pathogenesis of CF, and in the development of effective CF therapies. The approval of Orkambi® (lumacaftor/ivacaftor) marks another milestone in CF therapeutics development, which, with the advent of personalized medicine, could potentially revolutionize CF care and management. This article reviews the rationale, progress and future direction in the development of lumacaftor/ivacaftor combination to treat CF patients homozygous for the Phe508del-CFTR mutation. PMID:27252987

  6. Genética molecular del alcoholismo

    Directory of Open Access Journals (Sweden)

    Mauricio Rey-Buitrago

    2015-07-01

    Full Text Available El alcoholismo es una patología psiquiátrica compleja y de origen multifactorial en la que el factor genético explica alrededor del 50 % del fenómeno. Son numerosos los genes que se han asociado a esta enfermedad, pero su aporte individual es mínimo y contradictorio. Estos genes operan a través de características intermedias como la impulsividad y la sensibilidad al alcohol, lo que hace compleja la definición del fenotipo del alcoholismo. Los estudios de asociación de SNPs, de asociación a todo el genoma, de expresión y epigenéticos han identificado una amplia gama de variantes genéticas y epigenéticas, blancos para los estudios de susceptibilidad, diagnóstico y tratamiento farmacológico. Actualmente se comprenden mucho más estas relaciones y el desarrollo rápido de nuevas metodologías de estudio promete continuar este proceso, así como la generación de algoritmos de diagnóstico, prevención y tratamientos más acertados y confiables.

  7. Organismos modificados genéticamente en la alimentación humana

    OpenAIRE

    Barros Fernández, Paula

    2014-01-01

    El presente trabajo trata la controversia del tema de los organismos modificados genéticamente (OMG). Se mencionan los beneficios que aporta la ingeniería genética y también los principales riesgos y preocupaciones existentes en torno al consumo de los alimentos modificados genéticamente, reportando casos de estudios que así lo constatan.Se tratan temas como seguridad alimentaria, legislación y normativas de etiquetado de estos nuevos alimentos, señalando su relación con la salud. Además, se ...

  8. El papel del gen del transportador de serotonina en los trastornos de la conducta alimentaria

    OpenAIRE

    Sandra Hernández-Muñoz; Beatriz Camarena-Medellín

    2014-01-01

    Introducción: Al sistema serotoninérgico se lo ha implicado en la regulación del estado de ánimo y en la conducta alimentaria, por lo que el gen del transportador de serotonina (SLC6A4) es un buen candidato para el desarrollo de los trastornos de la conducta alimentaria (TCA). La mayoría de los estudios genéticos en los TCA se han centrado principalmente en un polimorfismo, el denominado 5-HTTLPR del gen SLC6A4. Objetivo: Realizar una revisión de los estudios de asociación entre el 5-HTTLPR y...

  9. Resveratrol increases F508del-CFTR dependent salivary secretion in cystic fibrosis mice

    Directory of Open Access Journals (Sweden)

    Barbara Dhooghe

    2015-07-01

    Full Text Available Cystic fibrosis (CF is a fatal genetic disease associated with widespread exocrine gland dysfunction. Studies have suggested activating effects of resveratrol, a naturally-occurring polyphenol compound with antioxidant and anti-inflammatory properties, on CF transmembrane conductance regulator (CFTR protein function. We assayed, in F508del-CFTR homozygous (CF and in wild-type mice, the effect of resveratrol on salivary secretion in basal conditions, in response to inhibition by atropine (basal β-adrenergic-dependent component and to stimulation by isoprenaline (CFTR-dependent component. Both components of the salivary secretion were smaller in CF mice than in controls. Two hours after intraperitoneal administration of resveratrol (50 mg/kg dissolved in DMSO, the compound was detected in salivary glands. As in both CF and in wild-type mice, DMSO alone increased the response to isoprenaline in males but not in females, the effect of resveratrol was only measured in females. In wild-type mice, isoprenaline increased secretion by more than half. In CF mice, resveratrol rescued the response to isoprenaline, eliciting a 2.5-fold increase of β-adrenergic-stimulated secretion. We conclude that the salivary secretion assay is suitable to test DMSO-soluble CFTR modulators in female mice. We show that resveratrol applied in vivo to mice reaches salivary glands and increases β-adrenergic secretion. Immunolabelling of CFTR in human bronchial epithelial cells suggests that the effect is associated with increased CFTR protein expression. Our data support the view that resveratrol is beneficial for treating CF. The salivary secretion assay has a potential application to test efficacy of novel CF therapies.

  10. A novel CFTR mutation found in a Chinese patient with cystic fibrosis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Background Cystic fibrosis (CF) is rare in Chinese. We investigated the mutations in the gene of cystic fibrosis transmembrane conductance regulator (CFTR) in a Chinese CF patient and reviewed the clinical features, gene mutations in Chinese CF cases. Methods Blood samples were collected from a previously reported CF girl and her parents. The 24 coding exons of CFTR of the proband were amplified and sequenced. Results A Chinese girl of 16 years old was diagnosed as CF at the age of 14. She had recurrent productive cough with bronchiectasis in bilateral upper lobes, parasinusitis and otitis media, but without pancreatic involvement. Her sweat chloride was (108.9 ±3.3) mmol/L. A heterozygous novel missense mutation of 699 C→A which results in the amino acid change of N189K was identified in exon 5. In addition, a heterozygous 3821-3823 delT mutation in exon 19 was found in CFTR. The mutation 699C→A was inherited from her father, and the 3821-3823delT mutation was from her mother. Twenty patients with CF in Chinese reported from 1974 to 2004 were also reviewed. DelF508 mutation was not found in the nine cases whose CFTR mutations were analyzed. Conclusions The CF proband carries two heterozygous mutations (699C→A and 3821-3823delT) in CFTR. 699C→A mutation is a novel mutation which is not reported previously. Review of reported Chinese cases suggests that the genotype of Chinese CF may be different from those of white cases. More studies are needed to understand the spectra of CFTR and clinical CF features in Chinese.

  11. Ribosomal Stalk Protein Silencing Partially Corrects the ΔF508-CFTR Functional Expression Defect.

    Science.gov (United States)

    Veit, Guido; Oliver, Kathryn; Apaja, Pirjo M; Perdomo, Doranda; Bidaud-Meynard, Aurélien; Lin, Sheng-Ting; Guo, Jingyu; Icyuz, Mert; Sorscher, Eric J; Hartman Iv, John L; Lukacs, Gergely L

    2016-05-01

    The most common cystic fibrosis (CF) causing mutation, deletion of phenylalanine 508 (ΔF508 or Phe508del), results in functional expression defect of the CF transmembrane conductance regulator (CFTR) at the apical plasma membrane (PM) of secretory epithelia, which is attributed to the degradation of the misfolded channel at the endoplasmic reticulum (ER). Deletion of phenylalanine 670 (ΔF670) in the yeast oligomycin resistance 1 gene (YOR1, an ABC transporter) of Saccharomyces cerevisiae phenocopies the ΔF508-CFTR folding and trafficking defects. Genome-wide phenotypic (phenomic) analysis of the Yor1-ΔF670 biogenesis identified several modifier genes of mRNA processing and translation, which conferred oligomycin resistance to yeast. Silencing of orthologues of these candidate genes enhanced the ΔF508-CFTR functional expression at the apical PM in human CF bronchial epithelia. Although knockdown of RPL12, a component of the ribosomal stalk, attenuated the translational elongation rate, it increased the folding efficiency as well as the conformational stability of the ΔF508-CFTR, manifesting in 3-fold augmented PM density and function of the mutant. Combination of RPL12 knockdown with the corrector drug, VX-809 (lumacaftor) restored the mutant function to ~50% of the wild-type channel in primary CFTRΔF508/ΔF508 human bronchial epithelia. These results and the observation that silencing of other ribosomal stalk proteins partially rescue the loss-of-function phenotype of ΔF508-CFTR suggest that the ribosomal stalk modulates the folding efficiency of the mutant and is a potential therapeutic target for correction of the ΔF508-CFTR folding defect. PMID:27168400

  12. Identification of CFTR Gene Mutations in Chinese Patients with Congenital Obstructive Azoospermia

    Institute of Scientific and Technical Information of China (English)

    曾国华; 吴开俊; 梅骅; 庄广伦

    2001-01-01

    Objective To analyze the frequency and hot spot of CFTR gene mutations in Chinese patients with congenital obstructive azoospermia Materials & Methods Mutations in CFTR exon 2,3,4,5,6a,8,10,11,12,13,15A 17b, 19A,20,21and 23 were detected. PCR-single strand conformation poly-morphism (SSCP) and direct sequencing were performed on 32 patients with congenital bilateral absence of the vas deferens (CBAVD), 17 patients with congenital unilateral absence of the vas deferens (CUAVD) and 50 normal Chinese.Results No CFTR gene mutations were detected in 50 normal Chinese. One CBAVD patient exhibited an abnormal band on SSCP for exon 10 of the CFTR gene and subsequent DNA sequencing showed a 3 bp deletion at position 1 653~ 1 655, which caused the deletion of a single amino acid, phenyalanine, in codon 508, i. e. , △F 508. A shift mutation was detected in another CBAVD patient in exon 2, a 1 bp deletion at position 225, 225 delC. One CUAVD patient exhibited an abnormal band on SSCP for exon 17 b of CFTR gene. Subsequent DNA sequencing showed a C-to-A transversion at position 3 295, which led to a predicted change of Leusine (codon 1 055,CUU) to Isoleucine (codon AUU), L1055I.Conclusion CFTR mutation could be detected in Chinese patients with congenital obstructive azoospermia. But no hot spots of mutations are discovered. 225 delC and L1055I are identified as two novel mutations, which are found only in Chinese.

  13. Impact of Cystic Fibrosis Transmembrane Regulator (CFTR gene mutations on male infertility

    Directory of Open Access Journals (Sweden)

    Jlenia Elia

    2014-09-01

    Full Text Available Objective. The aim of this study was to evaluate the prevalence of most common mutations and intron 8 5T (IVS8-5T polymorphism of CFTR gene in Italian: a azoospermic males; b non azoospermic subjects, male partners of infertile couples enrolled in assisted reproductive technology (ART programs. Material and methods. We studied 242 subjects attending our Andrology Unit (44 azoospermic subjects and 198 non azoospermic subjects, male partners of infertile couples enrolled in ART programs. Semen analysis, molecular analysis for CFTR gene mutations and genomic variant of IVS8-5T polymorphic tract, karyotype and chromosome Y microdeletions, hormonal profile (LH, FSH, Testosterone and seminal biochemical markers (fructose, citric acid and L-carnitine were carried out. Results. The prevalence of the common CFTR mutations and/or the IVS8-5T polymorphism was 12.9% (4/31 cases in secretory azoospermia, while in obstructive azoospermia was 84.6% (11/13 cases; in these, the most frequent mutations were the F508del, R117H and W1282X. Regarding the non azoospermic subjects, the prevalence of the CFTR and/or the IVS8-5T polymorphism was 11.1% (11/99 cases in severe dyspermia, 8.1% (6/74 cases in moderate dyspermia and finally 4.0% (1/25 cases in normospermic subjects. Conclusions. This study confirms the highly significant prevalence of CFTR mutations in males with bilateral absence of the vas deferens or ejaculatory ducts obstruction compared with subjects with secretory azoospermia. Moreover, the significant prevalence of mutations in severely dyspermic subjects may suggest the possible involvement of CFTR even in the spermatogenic process. This could explain the unsatisfactory recovery of sperm from testicular fine needle aspiration in patients affected by genital tract blockage.

  14. Ribosomal Stalk Protein Silencing Partially Corrects the ΔF508-CFTR Functional Expression Defect

    Science.gov (United States)

    Veit, Guido; Oliver, Kathryn; Apaja, Pirjo M.; Perdomo, Doranda; Bidaud-Meynard, Aurélien; Guo, Jingyu; Icyuz, Mert; Sorscher, Eric J.; Hartman IV, John L.; Lukacs, Gergely L.

    2016-01-01

    The most common cystic fibrosis (CF) causing mutation, deletion of phenylalanine 508 (ΔF508 or Phe508del), results in functional expression defect of the CF transmembrane conductance regulator (CFTR) at the apical plasma membrane (PM) of secretory epithelia, which is attributed to the degradation of the misfolded channel at the endoplasmic reticulum (ER). Deletion of phenylalanine 670 (ΔF670) in the yeast oligomycin resistance 1 gene (YOR1, an ABC transporter) of Saccharomyces cerevisiae phenocopies the ΔF508-CFTR folding and trafficking defects. Genome-wide phenotypic (phenomic) analysis of the Yor1-ΔF670 biogenesis identified several modifier genes of mRNA processing and translation, which conferred oligomycin resistance to yeast. Silencing of orthologues of these candidate genes enhanced the ΔF508-CFTR functional expression at the apical PM in human CF bronchial epithelia. Although knockdown of RPL12, a component of the ribosomal stalk, attenuated the translational elongation rate, it increased the folding efficiency as well as the conformational stability of the ΔF508-CFTR, manifesting in 3-fold augmented PM density and function of the mutant. Combination of RPL12 knockdown with the corrector drug, VX-809 (lumacaftor) restored the mutant function to ~50% of the wild-type channel in primary CFTRΔF508/ΔF508 human bronchial epithelia. These results and the observation that silencing of other ribosomal stalk proteins partially rescue the loss-of-function phenotype of ΔF508-CFTR suggest that the ribosomal stalk modulates the folding efficiency of the mutant and is a potential therapeutic target for correction of the ΔF508-CFTR folding defect. PMID:27168400

  15. Ribosomal Stalk Protein Silencing Partially Corrects the ΔF508-CFTR Functional Expression Defect.

    Directory of Open Access Journals (Sweden)

    Guido Veit

    2016-05-01

    Full Text Available The most common cystic fibrosis (CF causing mutation, deletion of phenylalanine 508 (ΔF508 or Phe508del, results in functional expression defect of the CF transmembrane conductance regulator (CFTR at the apical plasma membrane (PM of secretory epithelia, which is attributed to the degradation of the misfolded channel at the endoplasmic reticulum (ER. Deletion of phenylalanine 670 (ΔF670 in the yeast oligomycin resistance 1 gene (YOR1, an ABC transporter of Saccharomyces cerevisiae phenocopies the ΔF508-CFTR folding and trafficking defects. Genome-wide phenotypic (phenomic analysis of the Yor1-ΔF670 biogenesis identified several modifier genes of mRNA processing and translation, which conferred oligomycin resistance to yeast. Silencing of orthologues of these candidate genes enhanced the ΔF508-CFTR functional expression at the apical PM in human CF bronchial epithelia. Although knockdown of RPL12, a component of the ribosomal stalk, attenuated the translational elongation rate, it increased the folding efficiency as well as the conformational stability of the ΔF508-CFTR, manifesting in 3-fold augmented PM density and function of the mutant. Combination of RPL12 knockdown with the corrector drug, VX-809 (lumacaftor restored the mutant function to ~50% of the wild-type channel in primary CFTRΔF508/ΔF508 human bronchial epithelia. These results and the observation that silencing of other ribosomal stalk proteins partially rescue the loss-of-function phenotype of ΔF508-CFTR suggest that the ribosomal stalk modulates the folding efficiency of the mutant and is a potential therapeutic target for correction of the ΔF508-CFTR folding defect.

  16. Small-molecule CFTR activators increase tear secretion and prevent experimental dry eye disease.

    Science.gov (United States)

    Flores, Alyssa M; Casey, Scott D; Felix, Christian M; Phuan, Puay W; Verkman, A S; Levin, Marc H

    2016-05-01

    Dry eye disorders, including Sjögren's syndrome, constitute a common problem in the aging population, with limited effective therapeutic options available. The cAMP-activated Cl(-) channel cystic fibrosis transmembrane conductance regulator (CFTR) is a major prosecretory channel at the ocular surface. We investigated whether compounds that target CFTR can correct the abnormal tear film in dry eye. Small-molecule activators of human wild-type CFTR identified by high-throughput screening were evaluated in cell culture and in vivo assays, to select compounds that stimulate Cl(-)-driven fluid secretion across the ocular surface in mice. An aminophenyl-1,3,5-triazine, CFTRact-K089, fully activated CFTR in cell cultures with EC50 ∼250 nM and produced an ∼8.5 mV hyperpolarization in ocular surface potential difference. When delivered topically, CFTRact-K089 doubled basal tear volume for 4 h and had no effect in CF mice. CFTRact-K089 showed sustained tear film bioavailability without detectable systemic absorption. In a mouse model of aqueous-deficient dry eye produced by lacrimal ablation, topical administration of 0.1 nmol CFTRact-K089 3 times daily restored tear volume to basal levels, preventing corneal epithelial disruption when initiated at the time of surgery and reversing it when started after development of dry eye. Our results support the potential utility of CFTR-targeted activators as a novel prosecretory treatment for dry eye.-Flores, A. M., Casey, S. D., Felix, C. M., Phuan, P. W., Verkman, A. S., Levin, M. H. Small-molecule CFTR activators increase tear secretion and prevent experimental dry eye disease. PMID:26842854

  17. Estudio preliminar de la expresión del mensaje genético del transportador de serotonina en células mononucleares de sangre periférica en pacientes con dependencia al alcohol con y sin depresión mayor comórbida

    Directory of Open Access Journals (Sweden)

    Enrique Becerril-Villanueva

    2011-01-01

    Full Text Available La Encuesta Nacional de Adicciones 2008 reportó que en México existen 39 millones de personas que consumen alcohol y 4.8 millones presentan dependencia. A nivel mundial varios estudios indican que los pacientes con dependencia al alcohol (40 a 50% presentan comorbilidad con algún tipo de padecimiento psiquiátrico. La función serotoninérgica es un mediador clave en los estados de ánimo, la impulsividad y las conductas adictivas, entre ellas elconsumo de alcohol. Se ha reportado que el consumo excesivo de alcohol etílico disminuye los niveles de serotonina, aumenta la frecuencia de disparo de las neuronas serotoninérgicas en el núcleo del rafé y aumenta los niveles de serotonina en el núcleo accumbens. Las técnicas de biología molecular han permitido identificar factores de riesgo genético y se han seleccionado genes candidatos del sistema serotoninérgico, siendo uno de ellos el gen para el transportador de serotonina (5-HTT, el cual se ha demostrado que se encuentra asociado tanto a una mayor susceptibilidad para el establecimiento de la dependencia al alcohol como a la depresión mayor. Los receptores del sistema serotoninérgico como el 5-HTT, el 5-HT1 y el 5-HT2 se expresan tanto en las células del Sistema Nervioso como en las células del sistema inmunológico, lo que sugiere una similitud funcional de ambos sistemas. Es por ello que las células mononucleares de sangre periférica (PBMC han sido utilizadas como un modelo de estudio en los trastornos de dependencia al alcohol y en los psiquiátricos. El objetivo de este estudio fue evaluar los niveles de expresión del gen 5-HTT en células mononucleares de sangre periférica de pacientes con dependencia al alcohol con y sin depresión mayor comórbida. En el Servicio de Consulta Externa del Centro de Ayuda a Alcohólicos y Familiares (CAAF y en el Centro de Alcohólicos y Drogadictos «Carrasco» se evaluaron 70 pacientes durante el periodo comprendido entre febrero y

  18. Functional interaction between CFTR and the sodium-phosphate co-transport type 2a in Xenopus laevis oocytes.

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    Naziha Bakouh

    Full Text Available BACKGROUND: A growing number of proteins, including ion transporters, have been shown to interact with Cystic Fibrosis Transmembrane conductance Regulator (CFTR. CFTR is an epithelial chloride channel that is involved in Cystic Fibrosis (CF when mutated; thus a better knowledge of its functional interactome may help to understand the pathophysiology of this complex disease. In the present study, we investigated if CFTR and the sodium-phosphate co-transporter type 2a (NPT2a functionally interact after heterologous expression of both proteins in Xenopus laevis oocytes. METHODOLOGY/FINDINGS: NPT2a was expressed alone or in combination with CFTR in X. laevis oocytes. Using the two-electrode voltage-clamp technique, the inorganic phosphate-induced current (IPi was measured and taken as an index of NPT2a activity. The maximal IPi for NPT2a substrates was reduced when CFTR was co-expressed with NPT2a, suggesting a decrease in its expression at the oolemna. This was consistent with Western blot analysis showing reduced NPT2a plasma membrane expression in oocytes co-expressing both proteins, whereas NPT2a protein level in total cell lysate was the same in NPT2a- and NPT2a+CFTR-oocytes. In NPT2a+CFTR- but not in NPT2a-oocytes, IPi and NPT2a surface expression were increased upon PKA stimulation, whereas stimulation of Exchange Protein directly Activated by cAMP (EPAC had no effect. When NPT2a-oocytes were injected with NEG2, a short amino-acid sequence from the CFTR regulatory domain that regulates PKA-dependent CFTR trafficking to the plasma membrane, IPi values and NPT2a membrane expression were diminished, and could be enhanced by PKA stimulation, thereby mimicking the effects of CFTR co-expression. CONCLUSION/PERSPECTIVES: We conclude that when both CFTR and NPT2a are expressed in X. laevis oocytes, CFTR confers to NPT2a a cAMPi-dependent trafficking to the membrane. This functional interaction raises the hypothesis that CFTR may play a role in

  19. Mechanisms of CFTR functional variants that impair regulated bicarbonate permeation and increase risk for pancreatitis but not for cystic fibrosis.

    Directory of Open Access Journals (Sweden)

    Jessica LaRusch

    2014-07-01

    Full Text Available CFTR is a dynamically regulated anion channel. Intracellular WNK1-SPAK activation causes CFTR to change permeability and conductance characteristics from a chloride-preferring to bicarbonate-preferring channel through unknown mechanisms. Two severe CFTR mutations (CFTRsev cause complete loss of CFTR function and result in cystic fibrosis (CF, a severe genetic disorder affecting sweat glands, nasal sinuses, lungs, pancreas, liver, intestines, and male reproductive system. We hypothesize that those CFTR mutations that disrupt the WNK1-SPAK activation mechanisms cause a selective, bicarbonate defect in channel function (CFTRBD affecting organs that utilize CFTR for bicarbonate secretion (e.g. the pancreas, nasal sinus, vas deferens but do not cause typical CF. To understand the structural and functional requirements of the CFTR bicarbonate-preferring channel, we (a screened 984 well-phenotyped pancreatitis cases for candidate CFTRBD mutations from among 81 previously described CFTR variants; (b conducted electrophysiology studies on clones of variants found in pancreatitis but not CF; (c computationally constructed a new, complete structural model of CFTR for molecular dynamics simulation of wild-type and mutant variants; and (d tested the newly defined CFTRBD variants for disease in non-pancreas organs utilizing CFTR for bicarbonate secretion. Nine variants (CFTR R74Q, R75Q, R117H, R170H, L967S, L997F, D1152H, S1235R, and D1270N not associated with typical CF were associated with pancreatitis (OR 1.5, p = 0.002. Clones expressed in HEK 293T cells had normal chloride but not bicarbonate permeability and conductance with WNK1-SPAK activation. Molecular dynamics simulations suggest physical restriction of the CFTR channel and altered dynamic channel regulation. Comparing pancreatitis patients and controls, CFTRBD increased risk for rhinosinusitis (OR 2.3, p<0.005 and male infertility (OR 395, p<<0.0001. WNK1-SPAK pathway-activated increases in

  20. GenBank blastx search result: AK287488 [KOME

    Lifescience Database Archive (English)

    Full Text Available uctance regulator ATP-binding cassette sub-family C member (CFTR) and CTTNBP2 (CTTNBP2) genes, complete cds. PRI 6e-63 0 ... ...AK287488 J043029O04 DQ354388.1 DQ354388 Homo sapiens isolate cftr14683_A cystic fibrosis transmembrane cond

  1. GenBank blastx search result: AK287488 [KOME

    Lifescience Database Archive (English)

    Full Text Available uctance regulator ATP-binding cassette sub-family C member (CFTR) and CTTNBP2 (CTTNBP2) genes, complete cds. PRI 6e-63 0 ... ...AK287488 J043029O04 DQ354391.1 DQ354391 Homo sapiens isolate cftr01814_B cystic fibrosis transmembrane cond

  2. GenBank blastx search result: AK287488 [KOME

    Lifescience Database Archive (English)

    Full Text Available uctance regulator ATP-binding cassette sub-family C member (CFTR) and CTTNBP2 (CTTNBP2) genes, complete cds. PRI 6e-63 0 ... ...AK287488 J043029O04 DQ354389.1 DQ354389 Homo sapiens isolate cftr14683_B cystic fibrosis transmembrane cond

  3. GenBank blastx search result: AK287488 [KOME

    Lifescience Database Archive (English)

    Full Text Available uctance regulator ATP-binding cassette sub-family C member (CFTR) and CTTNBP2 (CTTNBP2) genes, complete cds. PRI 6e-63 0 ... ...AK287488 J043029O04 DQ354390.1 DQ354390 Homo sapiens isolate cftr01814_A cystic fibrosis transmembrane cond

  4. Equivalencia clínica entre el rociador nasal de propionato de fluticasona genérico y comercial en pacientes con rinitis alérgica: Clinical equival ence between generic and branded fluticasone propionate nasal spray in patients with alergic rhinitis

    Directory of Open Access Journals (Sweden)

    Vikki Brandi

    2009-10-01

    Full Text Available Objetivo. Establecer la equivalencia clínica de un rociador nasal de propionato de fluticasona (RNF genérico comparado con dos formulas farmacéuticas comerciales del mismo producto (Flonase® y Flixonase® durante la estación de polinización del cedro de montaña (Juniperus ashei en Texas, EEUU. Materiales y métodos. Estudio multicéntrico, aleatorizado, doble ciego, controlado con placebo, en grupos paralelos diseñado para investigar la seguridad y eficacia de RNF (200 mcg una vez al día, Flonase® (200 mcg una vez al día y Flixonase® (200 mcg una vez al día, comparados con placebo, administrados por 13 a 15 días. Los pacientes registraron diariamente, en la mañana y en la tarde, sus síntomas nasales totales (SNT. La variable de desenlace primaria fue la suma de SNT en la mañana y tarde + 1. Las variables de desenlace secundarias fueron los SNT AM + 1 y SNT PM + 1, y la evaluación de seguridad. Resultados. No se observó diferencia estadísticamente significativa en ningún día de estudio, ni en todo el periodo de tratamiento, ni al punto de final entre SNT promedio tanto de Flonase® como Flixonase® y RNF. La equivalencia clínica entre RNF y Flonase® (cociente= 0,98; intervalo de confianza [IC] al 90%, 0,91 a 1,06, y entre RNF y Flixonase® (cociente= 1,02; IC 90%, 0,94 a 1,10 fue demostrada tanto para la variable de desenlace primaria como para las otras variables de eficacia. Conclusiones. Estos resultados respaldan la equivalencia clínica entre RNF 200 mcg una vez al día tanto con Flonase® como Flixonase® en el tratamiento de rinitis alérgica estacional.

  5. The human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopy

    KAUST Repository

    Jourdain, P.

    2013-12-11

    The transmembrane water movements during cellular processes and their relationship to ionic channel activity remain largely unknown. As an example, in epithelial cells it was proposed that the movement of water could be directly linked to cystic fibrosis transmembrane conductance regulator (CFTR) protein activity through a cAMP-stimulated aqueous pore, or be dependent on aquaporin. Here, we used digital holographic microscopy (DHM) an interferometric technique to quantify in situ the transmembrane water fluxes during the activity of the epithelial chloride channel, CFTR, measured by patch-clamp and iodide efflux techniques. We showed that the water transport measured by DHM is fully inhibited by the selective CFTR blocker CFTRinh172 and is absent in cells lacking CFTR. Of note, in cells expressing the mutated version of CFTR (F508del-CFTR), which mimics the most common genetic alteration encountered in cystic fibrosis, we also show that the water movement is profoundly altered but restored by pharmacological manipulation of F508del-CFTR-defective trafficking. Importantly, whereas activation of this endogenous water channel required a cAMP-dependent stimulation of CFTR, activation of CFTR or F508del-CFTR by two cAMP-independent CFTR activators, genistein and MPB91, failed to trigger water movements. Finally, using a specific small-interfering RNA against the endogenous aquaporin AQP3, the water transport accompanying CFTR activity decreased. We conclude that water fluxes accompanying CFTR activity are linked to AQP3 but not to a cAMP-stimulated aqueous pore in the CFTR protein.

  6. Effective writing tasks and feedback for the iGen

    OpenAIRE

    Buyse, Kris

    2010-01-01

    Teaching foreign language writing often lacks adjustments to the requirements of today’s students of the “Internet Generation" (iGen): usually teachers set a —not extremely inspiring— theme, a deadline and then return a discouraging, manually underlined and/or annotated text without systematic labeling; at the end the annotated document is stored both by the teacher and the student in a bottomless box. Whereas earlier student generations were educated and trained to process conscientiously al...

  7. Polymorphism Trp64Arg of beta 3 adrenoreceptor gene: allelic frequencies and influence on insulin resistance in a multicenter study of Castilla-León Polimorfismo TRP64ARG del gen receptor beta 3: frecuencia alélica e influencia en la resistencia a la insulina en un estudio multicéntrico de Castilla y León

    Directory of Open Access Journals (Sweden)

    D. A. de Luis

    2010-04-01

    Full Text Available Background and objective: The genetic variant (Trp64Arg is a missense mutation located within the beta3 adrenoreceptor (Beta3AR. The aim of our study was to investigate the influence of Trp64Arg polymorphism in the Beta3AR gene on insulin resistance in obese patients and the allelic distribution of this polymorphismin a geographic area of Spain. Design: A population of 264 obese patients was analyzed. A bioimpedance, blood pressure, an assessment of nutritional intake, and biochemical parameters were measured. The beta 3 adrenoreceptor gene polymorphism(Trp64Arg was genotyped. Results: Two hundred and twenty six patients (77 males/149 females (85.6% had the genotype Trp64/Trp64 (wild type group with and average age of 41.12 ± 13.1 years and 38 patients (16 males/22 females Trp64/Arg64 (14.4% (mutant type group with an average age of 40.5 ± 12.7 years. High frequencies of Arg64 allele were observed in Salamanca and Valladolid. In the mutant type group, HOMA (3.75 ± 2.77 vs 5.27 ± 5.4; p Introducción y objetivos: La variante genética (Trp64Arg es una mutación localizada en el adrenoreceptor Beta 3 (Beta3AR. El objetivo de nuestro trabajo es evaluar la influencia de el polimorfismo Trp64Arg del gen de Beta3AR sobre la resistencia a la insulina en pacientes obesos, así como la distribución alélica de este polimorfismo en un área geográfica de España. Diseño: Una muestra de 264 pacientes obesos fue analizada. Se realizó una bioimpedancia, evaluación nutricional y análisis bioquímico. Se genotiparon a los pacientes en función delpolimorfismos Tr64Arg del gen adrenoreceptor-beta 3. Resultados: Un total de 227 pacientes (77 varones/149 mujeres (85,6% presentaron el genotipo Trp64/Trp64 (grupo genotipo salvaje, con una media de edad de 41,12 ± 13,1 años y un total de 38 pacientes (16 varones/22 mujeres Trp64/Arg64 (14,4% (grupo genotipo mutante con una edad media de 40,5 ± 12,7 años. Se detectó una alta frecuencia alélica (Arg64

  8. Study of CFTR delivery and stabilization at the plasma membrane

    OpenAIRE

    Moraes, Bruno José Rother Rocha de

    2010-01-01

    Tese de mestrado, Bioquímica (Bioquímica Médica), Universidade de Lisboa, Faculdade de Ciências, 2010 A fibrose quística (FQ) é a principal doença genética, autossómica recessiva, a afectar a população Caucasiana, embora se estenda igualmente a outras etnias. Apesar de caracterizada no pâncreas pela primeira vez em 1938, a FQ foi sucessivamente associada a defeitos em tecidos epiteliais exócrinos de outros órgãos, nomeadamente dos pulmões, onde se registam os efeitos mais nefastos, os quai...

  9. Distributed Generation Market Demand Model (dGen): Documentation

    Energy Technology Data Exchange (ETDEWEB)

    Sigrin, Benjamin [National Renewable Energy Lab. (NREL), Golden, CO (United States); Gleason, Michael [National Renewable Energy Lab. (NREL), Golden, CO (United States); Preus, Robert [National Renewable Energy Lab. (NREL), Golden, CO (United States); Baring-Gould, Ian [National Renewable Energy Lab. (NREL), Golden, CO (United States); Margolis, Robert [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2016-02-01

    The Distributed Generation Market Demand model (dGen) is a geospatially rich, bottom-up, market-penetration model that simulates the potential adoption of distributed energy resources (DERs) for residential, commercial, and industrial entities in the continental United States through 2050. The National Renewable Energy Laboratory (NREL) developed dGen to analyze the key factors that will affect future market demand for distributed solar, wind, storage, and other DER technologies in the United States. The new model builds off, extends, and replaces NREL's SolarDS model (Denholm et al. 2009a), which simulates the market penetration of distributed PV only. Unlike the SolarDS model, dGen can model various DER technologies under one platform--it currently can simulate the adoption of distributed solar (the dSolar module) and distributed wind (the dWind module) and link with the ReEDS capacity expansion model (Appendix C). The underlying algorithms and datasets in dGen, which improve the representation of customer decision making as well as the spatial resolution of analyses (Figure ES-1), also are improvements over SolarDS.

  10. Variabilidade genética de caracteres forrageiros em Paspalum

    Directory of Open Access Journals (Sweden)

    Emerson André Pereira

    2012-10-01

    Full Text Available O objetivo deste trabalho foi determinar a variabilidade genética e a expressão de caracteres de interesse forrageiro em espécies de Paspalum. Os experimentos foram conduzidos em diferentes locais e anos de cultivo, em delineamento de blocos ao acaso, com três repetições. Foram avaliados cinco acessos de P. nicorae e dois de P. guenoarum, além da cultivar Pensacola (P. notatum, utilizada como testemunha. Foram quantificados os seguintes caracteres: relação folha/colmo, índice de colheita e massa de matéria seca total, de folhas e de colmo. Tanto os efeitos principais (genótipos, anos e locais de cultivo quanto a interação entre os fatores tiveram influência significativa sobre os caracteres avaliados. Os acessos avaliados apresentam variabilidade genética em caracteres de interesse forrageiro, bem como desempenho variável de acordo com o local e o ano de cultivo. A produção de matéria seca total e de folhas são os caracteres que mais contribuem para a detecção da variabilidade genética observada, independentemente do ano de avaliação.

  11. Analysis of Y chromosome microdeletions and CFTR gene mutations as genetic markers of infertility in Serbian men

    Directory of Open Access Journals (Sweden)

    Dinić Jelena

    2007-01-01

    Full Text Available Background/Aim. Impaired fertility of a male partner is the main cause of infertility in up to one half of all infertile couples. At the genetic level, male infertility can be caused by chromosome aberrations or gene mutations. The presence and types of Y chromosome microdeletions and cystic fybrosis transmembrane conductance regulator (CFTR gene mutations as genetic cause of male infertility was tested in Serbian men. The aim of this study was to analyze CFTR gene mutations and Y chromosome microdelations as potential causes of male infertility in Serbian patients, as well as to test the hypothesis that CFTR mutations in infertile men are predominantly located in the several last exons of the gene. Methods. This study has encompassed 33 men with oligo- or azoospermia. The screening for Y chromosome microdeletions in the azoospermia factor (AZF region was performed by multiplex PCR analysis. The screening of the CFTR gene was performed by denaturing gradient gel electrophoresis (DGGE method. Results. Deletions on Y chromosome were detected in four patients, predominantly in AZFc region (four of total six deletions. Mutations in the CFTR gene were detected on eight out of 66 analyzed chromosomes of infertile men. The most common mutation was F508del (six of total eight mutations. Conclusion. This study confirmed that both Y chromosome microdeletions and CFTR gene mutations played important role in etiology of male infertility in Serbian infertile men. Genetic testing for Y chromosome microdeletions and CFTR gene mutations has been introduced in routine diagnostics and offered to couples undergoing assisted reproduction techniques. Considering that both the type of Y chromosome microdeletion and the type of CFTR mutation have a prognostic value, it is recommended that AZF and CFTR genotyping should not only be performed in patients with reduced sperm quality before undergoing assisted reproduction, but also for the purpose of preimplantation and

  12. Pseudomonas aeruginosa Reduces VX-809 Stimulated F508del-CFTR Chloride Secretion by Airway Epithelial Cells.

    Directory of Open Access Journals (Sweden)

    Bruce A Stanton

    Full Text Available P. aeruginosa is an opportunistic pathogen that chronically infects the lungs of 85% of adult patients with Cystic Fibrosis (CF. Previously, we demonstrated that P. aeruginosa reduced wt-CFTR Cl secretion by airway epithelial cells. Recently, a new investigational drug VX-809 has been shown to increase F508del-CFTR Cl secretion in human bronchial epithelial (HBE cells, and, in combination with VX-770, to increase FEV1 (forced expiratory volume in 1 second by an average of 3-5% in CF patients homozygous for the F508del-CFTR mutation. We propose that P. aeruginosa infection of CF lungs reduces VX-809 + VX-770- stimulated F508del-CFTR Cl secretion, and thereby reduces the clinical efficacy of VX-809 + VX-770.F508del-CFBE cells and primary cultures of CF-HBE cells (F508del/F508del were exposed to VX-809 alone or a combination of VX-809 + VX-770 for 48 hours and the effect of P. aeruginosa on F508del-CFTR Cl secretion was measured in Ussing chambers. The effect of VX-809 on F508del-CFTR abundance was measured by cell surface biotinylation and western blot analysis. PAO1, PA14, PAK and 6 clinical isolates of P. aeruginosa (3 mucoid and 3 non-mucoid significantly reduced drug stimulated F508del-CFTR Cl secretion, and plasma membrane F508del-CFTR.The observation that P. aeruginosa reduces VX-809 and VX-809 + VX-770 stimulated F508del CFTR Cl secretion may explain, in part, why VX-809 + VX-770 has modest efficacy in clinical trials.

  13. Three charged amino acids in extracellular loop 1 are involved in maintaining the outer pore architecture of CFTR.

    Science.gov (United States)

    Cui, Guiying; Rahman, Kazi S; Infield, Daniel T; Kuang, Christopher; Prince, Chengyu Z; McCarty, Nael A

    2014-08-01

    The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) bears six extracellular loops (ECL1-6); ECL1 is the site of several mutations associated with CF. Mutation R117H has been reported to reduce current amplitude, whereas D110H, E116K, and R117C/L/P may impair channel stability. We hypothesized that these amino acids might not be directly involved in ion conduction and permeation but may contribute to stabilizing the outer vestibule architecture in CFTR. We used cRNA injected oocytes combined with electrophysiological techniques to test this hypothesis. Mutants bearing cysteine at these sites were not functionally modified by extracellular MTS reagents and were blocked by GlyH-101 similarly to WT-CFTR. These results suggest that these three residues do not contribute directly to permeation in CFTR. In contrast, mutants D110R-, E116R-, and R117A-CFTR exhibited instability of the open state and significantly shortened burst duration compared with WT-CFTR and failed to be locked into the open state by AMP-PNP (adenosine 5'-(β,γ-imido) triphosphate); charge-retaining mutants showed mainly the full open state with comparably longer open burst duration. These interactions suggest that these ECL1 residues might be involved in maintaining the outer pore architecture of CFTR. A CFTR homology model suggested that E116 interacts with R104 in both the closed and open states, D110 interacts with K892 in the fully closed state, and R117 interacts with E1126 in the open state. These interactions were confirmed experimentally. The results suggest that D110, E116, and R117 may contribute to stabilizing the architecture of the outer pore of CFTR by interactions with other charged residues. PMID:25024266

  14. Estructura y diversidad genética en vacas Holstein de Antioquia usando un polimorfismo del gen bGH

    Directory of Open Access Journals (Sweden)

    Juan Rincon F.

    2013-03-01

    Full Text Available Objetivo. Determinar las frecuencias alélicas y genotípicas del polimorfismo del intrón 3 del gen bGH y estimar algunos parámetros de estructura poblacional en ganado Holstein. Materiales y métodos. El estudio se realizó con 1366 vacas Holstein en 120 hatos de 11 municipios del departamento de Antioquia. Se extrajo DNA por el método de Salting out y la genotipificación se realizó usando la técnica de PCR-RFLPs. La diversidad genética se determinó mediante la comparación de las heterocigosidades, El equilibrio de Hardy-Weinberg (HW y la diferenciación genética entre las poblaciones se realizó usando el software Arlequín 2.0 Las frecuencias alélicas y genotípicas se evaluaron mediante el paquete estadístico SAS®. Resultados. Las frecuencias genotípicas encontradas fueron 0.764 (+/+, 0.223 (+/- y 0.013 (-/- y las frecuencias alélicas 0.876 (+ y 0.124 (-. No se encontraron desviaciones del Equilibrio de Hardy Weinberg en ninguna de las subpoblaciones. La diversidad genética determinada mediante la comparación de las heterocigosidades fue relativamente baja entre poblaciones pero al interior de estas no. El valor de FST de toda la población fue de 0.0068 y significativo (p<0.05, algunos FST pareados también lo fueron, tomando valores desde 0.0 a 0.13. Los estadísticos FIT y FIS no fueron significativos. Conclusiones. El gen bGH es un candidato interesante para evaluar características de importancia económica ya que no parece haber sido sometido a selección directa, presenta una variabilidad media en las poblaciones, observándose diferenciación genética significativa entre distintos municipios, producto de los diferentes sistemas de producción y acceso a las biotecnologías.

  15. FutureGen Project Report

    Energy Technology Data Exchange (ETDEWEB)

    Cabe, Jim; Elliott, Mike

    2010-09-30

    This report summarizes the comprehensive siting, permitting, engineering, design, and costing activities completed by the FutureGen Industrial Alliance, the Department of Energy, and associated supporting subcontractors to develop a first of a kind near zero emissions integrated gasification combined cycle power plant and carbon capture and storage project (IGCC-CCS). With the goal to design, build, and reliably operate the first IGCC-CCS facility, FutureGen would have been the lowest emitting pulverized coal power plant in the world, while providing a timely and relevant basis for coal combustion power plants deploying carbon capture in the future. The content of this report summarizes key findings and results of applicable project evaluations; modeling, design, and engineering assessments; cost estimate reports; and schedule and risk mitigation from initiation of the FutureGen project through final flow sheet analyses including capital and operating reports completed under DOE award DE-FE0000587. This project report necessarily builds upon previously completed siting, design, and development work executed under DOE award DE-FC26- 06NT4207 which included the siting process; environmental permitting, compliance, and mitigation under the National Environmental Policy Act; and development of conceptual and design basis documentation for the FutureGen plant. For completeness, the report includes as attachments the siting and design basis documents, as well as the source documentation for the following: • Site evaluation and selection process and environmental characterization • Underground Injection Control (UIC) Permit Application including well design and subsurface modeling • FutureGen IGCC-CCS Design Basis Document • Process evaluations and technology selection via Illinois Clean Coal Review Board Technical Report • Process flow diagrams and heat/material balance for slurry-fed gasifier configuration • Process flow diagrams and heat/material balance

  16. Algoritmos genéticos locales

    OpenAIRE

    García-Martínez, Carlos; Lozano, Manuel

    2007-01-01

    Los Algoritmos Genéticos Locales son procedimientos que iterativamente re nan soluciones dadas. Su diferencia con procedimientos de mejora iterativa clásicos reside en el uso de operadores genéticos para realizar el re namiento. En este estudio presentamos un nuevo Algoritmo Genético Local Binario basado en un Algoritmo Genético Estacionario. Hemos comparado el Algoritmo Genético Local Binario con otros procedimientos de mejora iterativa de la literatura. Los res...

  17. CFTR delivery to 25% of surface epithelial cells restores normal rates of mucus transport to human cystic fibrosis airway epithelium.

    Directory of Open Access Journals (Sweden)

    Liqun Zhang

    2009-07-01

    Full Text Available Dysfunction of CFTR in cystic fibrosis (CF airway epithelium perturbs the normal regulation of ion transport, leading to a reduced volume of airway surface liquid (ASL, mucus dehydration, decreased mucus transport, and mucus plugging of the airways. CFTR is normally expressed in ciliated epithelial cells of the surface and submucosal gland ductal epithelium and submucosal gland acinar cells. Critical questions for the development of gene transfer strategies for CF airway disease are what airway regions require CFTR function and how many epithelial cells require CFTR expression to restore normal ASL volume regulation and mucus transport to CF airway epithelium? An in vitro model of human CF ciliated surface airway epithelium (CF HAE was used to test whether a human parainfluenza virus (PIV vector engineered to express CFTR (PIVCFTR could deliver sufficient CFTR to CF HAE to restore mucus transport, thus correcting the CF phenotype. PIVCFTR delivered CFTR to >60% of airway surface epithelial cells and expressed CFTR protein in CF HAE approximately 100-fold over endogenous levels in non-CF HAE. This efficiency of CFTR delivery fully corrected the basic bioelectric defects of Cl(- and Na(+ epithelial ion transport and restored ASL volume regulation and mucus transport to levels approaching those of non-CF HAE. To determine the numbers of CF HAE surface epithelial cells required to express CFTR for restoration of mucus transport to normal levels, different amounts of PIVCFTR were used to express CFTR in 3%-65% of the surface epithelial cells of CF HAE and correlated to increasing ASL volumes and mucus transport rates. These data demonstrate for the first time, to our knowledge, that restoration of normal mucus transport rates in CF HAE was achieved after CFTR delivery to 25% of surface epithelial cells. In vivo experimentation in appropriate models will be required to determine what level of mucus transport will afford clinical benefit to CF patients

  18. Dexamethasone regulates CFTR expression in Calu-3 cells with the involvement of chaperones HSP70 and HSP90.

    Directory of Open Access Journals (Sweden)

    Luiz Felipe M Prota

    Full Text Available BACKGROUND: Dexamethasone is widely used for pulmonary exacerbation in patients with cystic fibrosis, however, not much is known about the effects of glucocorticoids on the wild-type cystic fibrosis channel transmembrane regulator (CFTR. Our aim was to determine the effects of dexamethasone treatment on wild-type CFTR expression. METHODS AND RESULTS: Dose-response (1 nM to 10 µM and time course (3 to 48 h curves were generated for dexamethasone for mRNA expression in Calu-3 cells using a real-time PCR. Within 24 h, dexamethasone (10 nM showed a 0.3-fold decrease in CFTR mRNA expression, and a 3.2-fold increase in αENaC mRNA expression compared with control groups. Dexamethasone (10 nM induced a 1.97-fold increase in the total protein of wild-type CFTR, confirmed by inhibition by mifepristone. To access surface protein expression, biotinylation followed by Western blotting showed that dexamethasone treatment led to a 2.35-fold increase in the amount of CFTR in the cell surface compared with the untreated control groups. Once protein translation was inhibited with cycloheximide, dexamethasone could not increase the amount of CFTR protein. Protein stability was assessed by inhibition of protein synthesis with cycloheximide (50 µg/ml at different times in cells treated with dexamethasone and in untreated cells. Dexamethasone did not alter the degradation of wild-type CFTR. Assessment of the B band of CFTR within 15 min of metabolic pulse labeling showed a 1.5-fold increase in CFTR protein after treatment with dexamethasone for 24 h. Chaperone 90 (HSP90 binding to CFTR increased 1.55-fold after treatment with dexamethasone for 24 h, whereas chaperone 70 (HSP70 binding decreased 0.30 fold in an immunoprecipitation assay. CONCLUSION: Mature wild-type CFTR protein is regulated by dexamethasone post transcription, involving cotranslational mechanisms with HSP90 and HSP70, which enhances maturation and expression of wild-type CFTR.

  19. Cytoplasmic pathway followed by chloride ions to enter the CFTR channel pore.

    Science.gov (United States)

    El Hiani, Yassine; Negoda, Alexander; Linsdell, Paul

    2016-05-01

    Most ATP-binding cassette (ABC) proteins function as ATP-dependent membrane pumps. One exception is the cystic fibrosis transmembrane conductance regulator (CFTR), an ABC protein that functions as a Cl(-) ion channel. As such, the CFTR protein must form a continuous pathway for the movement of Cl(-) ions from the cytoplasm to the extracellular solution when in its open channel state. Extensive functional investigations have characterized most parts of this Cl(-) permeation pathway. However, one region remains unexplored-the pathway connecting the cytoplasm to the membrane-spanning pore. We used patch clamp recording and extensive substituted cysteine accessibility mutagenesis to identify amino acid side-chains in cytoplasmic regions of CFTR that lie close to the pathway taken by Cl(-) ions as they pass from the cytoplasm through this pathway. Our results suggest that Cl(-) ions enter the permeation pathway via a single lateral tunnel formed by the cytoplasmic parts of the protein, and then follow a fairly direct central pathway towards the membrane-spanning parts of the protein. However, this pathway is not lined continuously by any particular part of the protein; instead, the contributions of different cytoplasmic regions of the protein appear to change as the permeation pathway approaches the membrane, which appears to reflect the ways in which different cytoplasmic regions of the protein are oriented towards its central axis. Our results allow us to define for the first time the complete Cl(-) permeation pathway in CFTR, from the cytoplasm to the extracellular solution. PMID:26659082

  20. Mutation Analysis of CFTR Gene in 70 Iranian Cystic Fibrosis Patients

    Directory of Open Access Journals (Sweden)

    Reza Alibakhshi Mahdi Zamani

    2006-03-01

    Full Text Available Cystic fibrosis (CF is the most common inherited disorder in Caucasian populations, with over 1400 cystic fibrosis transmembrane conductance regulator (CFTR mutations. The type of mutations and their distributions varies widely between different countries and/or ethnic groups. Seventy Iranian cystic fibrosis patients were screened for the CFTR gene mutation using ARMS/PCR (amplification refractory mutation system for the following mutations: ∆F508, N1303K, G542X, 1717-1G>A, R553X, W1282X, G551D, 621+1G>T, ∆I507 and R560T. Single strand conformation polymorphism (SSCP analysis of exons 3, 7, 10, 11 and 17b, including both the exon/intron junctions, of the CFTR gene was performed in patients in whom no mutation could be identified on one or both CFTR genes. As a result of this screening, only three mutations were found: ∆F508 mutation was found in 25 (17.8% alleles, N1303K in six (4.3% alleles and G542X in five (3.6% alleles. Thus, a total of 3 mutations cover 25.7% of CF alleles. These finding will be used for planning future screening and appropriate genetic counseling programs in Iranian CF patients.

  1. Comprehensive and accurate mutation scanning of the CFTR gene by two-dimensional DNA electrophoresis

    NARCIS (Netherlands)

    Wu, Y; Hofstra, RMW; Scheffer, H; Uitterlinden, AG; Mullaart, E; Buys, CHCM; Vijg, J

    1996-01-01

    The large number of possible disease causing mutations in the 27 exons of the cystic fibrosis transmembrane conductance regulator (CFTR) gene has severely limited direct diagnosis of cystic fibrosis (CF) patients and carriers by mutation detection. Here we show that in principle testing for mutation

  2. Cholic acid induces a Cftr dependent biliary secretion and liver growth response in mice

    NARCIS (Netherlands)

    Bodewes, Frank A. J. A.; Bijvelds, Marcel J.; de Vries, Willemien; Baller, Juul F. W.; Gouw, Annette S. H.; de Jonge, Hugo R.; Verkade, Henkjan J.

    2015-01-01

    The cause of Cystic fibrosis liver disease (CFLD), is unknown. It is well recognized that hepatic exposure to hydrophobic bile salts is associated with the development of liver disease. For this reason, we hypothesize that, CFTR dependent variations, in the hepatic handling of hydrophobic bile salts

  3. Critical Role of Cystic Fibrosis Transmembrane Conductance Regulation(CFTR)in Female Reproduction

    Institute of Scientific and Technical Information of China (English)

    Hsiao Chang CHAN

    2003-01-01

    @@ Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-activated Cl- channel, mutations of which are responsible for defective Cl- and/or HCO-3 secretions seen in cystic fibrosis (CF), a common lethal genetic disease affecting most exocrine glands/organs, including the lungs, intestine, pancreas and reproductive tracts of both sexes.

  4. Cholic acid induces a Cftr dependent biliary secretion and liver growth response in mice

    NARCIS (Netherlands)

    F.A.J.A. Bodewes (Frank); M.J.C. Bijvelds (Marcel); De Vries, W. (Willemien); Baller, J.F.W. (Juul F. W.); A.S.H. Gouw (Annette); H.R. de Jonge (Hugo); H.J. Verkade (Henkjan)

    2015-01-01

    textabstractThe cause of Cystic fibrosis liver disease (CFLD), is unknown. It is well recognized that hepatic exposure to hydrophobic bile salts is associated with the development of liver disease. For this reason, we hypothesize that, CFTR dependent variations, in the hepatic handling of hydrophobi

  5. Analysis of mutations in the cystic fibrosis transmembrane regulator (CFTR gene in patients with obstructive azoospermia

    Directory of Open Access Journals (Sweden)

    Andrea L.F. Bernardino

    2003-01-01

    Full Text Available Congenital bilateral absence of the vas deferens (CBAVD accounts for 1%-2% of sterility in men. A high incidence of mutations, as well as the involvement of the 5T variant of the T tract length in intron 8 of the cystic fibrosis conductance regulator (CFTR gene, have been previously described in males with CBAVD. Herein we report the screening for mutations and for the 5T variant of the CFTR gene in 17 patients with CBAVD and three others with non-CABVD obstructive azoospermia. In the CBAVD group, three patients (15% were compound heterozygotes for mutations, and five patients (25% had a mutation in one allele and the 5T variant in the other; the 5T variant was also present in two other patients, one of them being homozygous. The most frequent mutation was DF508, present on five chromosomes (12.5%. A novel missense mutation (A399D was detected in a Japanese CBVAD patient. Our results yield further evidence for a strong association between male obstructive azoospermia caused by CBAVD and mutation/5T variant in the CFTR gene. The search for CFTR mutations in such patients is thus recommended for genetic counseling of couples who undergo assisted fertilization due to CBAVD.

  6. Personalized medicine in cystic fibrosis: genistein supplementation as a treatment option for patients with a rare S1045Y-CFTR mutation.

    Science.gov (United States)

    Arora, Kavisha; Yarlagadda, Sunitha; Zhang, Weiqiang; Moon, ChangSuk; Bouquet, Erin; Srinivasan, Saumini; Li, Chunying; Stokes, Dennis C; Naren, Anjaparavanda P

    2016-08-01

    Cystic fibrosis (CF) is a life-shortening disease caused by the mutations that generate nonfunctional CF transmembrane conductance regulator (CFTR) protein. A rare serine-to-tyrosine (S1045Y) CFTR mutation was earlier reported to result in CF-associated fatality. We identified an African-American patient with the S1045Y mutation in CFTR, as well as a stop-codon mutation, who has a mild CF phenotype. The underlying mechanism of CF caused by S1045Y-CFTR has not been elucidated. In this study, we determined that S1045Y-CFTR exhibits twofold attenuated function compared with wild-type (WT)-CFTR. We report that serine-to-tyrosine mutation leads to increased tyrosine phosphorylation of S1045Y-CFTR, followed by recruitment and binding of E3-ubiquitin ligase c-cbl, resulting in enhanced ubiquitination and passage of S1045Y-CFTR in the endosome/lysosome degradative compartments. We demonstrate that inhibition of tyrosine phosphorylation partially rescues S1045Y-CFTR surface expression and function. Based on our findings, it could be suggested that consuming genistein (a tyrosine phosphorylation inhibitor) would likely ameliorate CF symptoms in individuals with S1045Y-CFTR, providing a unique personalized therapy for this rare CF mutation. PMID:27261451

  7. Important role of platelets in modulating endotoxin-induced lung inflammation in CFTR-deficient mice.

    Directory of Open Access Journals (Sweden)

    Caiqi Zhao

    Full Text Available Mutation of CFTR (cystic fibrosis transmembrane conductance regulator leads to cystic fibrosis (CF. Patients with CF develop abnormalities of blood platelets and recurrent lung inflammation. However, whether CFTR-mutated platelets play a role in the development of lung inflammation is elusive. Therefore, we intratracheally challenged wildtype and F508del (a common type of CFTR mutation mice with LPS to observe changes of F508del platelets in the peripheral blood and indexes of lung inflammation (BAL neutrophils and protein levels. Furthermore, we investigated whether or not and how F508del platelets modulate the LPS-induced acute lung inflammation by targeting anti-platelet aggregation, depletion of neutrophils, reconstitution of bone marrow or neutrophils, blockade of P-selectin glycoprotein ligand-1 (PSGL-1, platelet activating factor (PAF, and correction of mutated CFTR trafficking. We found that LPS-challenged F508del mice developed severe thrombocytopenia and had higher levels of plasma TXB2 coincided with neutrophilic lung inflammation relative to wildtype control. Inhibition of F508del platelet aggregation or depletion of F508del neutrophils diminished the LPS-induced lung inflammation in the F508del mice. Moreover, wildtype mice reconstituted with either F508del bone marrow or neutrophils developed worse thrombocytopenia. Blocking PSGL-1, platelet activating factor (PAF, or rectifying trafficking of mutated CFTR in F508del mice diminished and alveolar neutrophil transmigration in the LPS-challenged F508del mice. These findings suggest that F508del platelets and their interaction with neutrophils are requisite for the development of LPS-induced lung inflammation and injury. As such, targeting platelets might be an emerging strategy for dampening recurrent lung inflammation in cystic fibrosis patients.

  8. CFTR gene transfer to lung epithelium--on the trail of a target cell.

    Science.gov (United States)

    O'Dea, S; Harrison, D J

    2002-05-01

    Cystic fibrosis (CF) is a lethal inherited disease that afflicts up to 1 in 2,500 people in the western world. Since 1989, when mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene were identified as responsible for the disease, intense effort has been applied to the development of replacement gene therapy strategies to cure CF. Problems with basic gene delivery techniques along with limited knowledge of the pathogenesis of CF have hindered progress so far. However, recent insights into the expression patterns and functions of CFTR in developing and adult lungs are now advancing our understanding of this disease. It is becoming apparent that progress in gene delivery to cure CF may be best served by identification of a target cell(s) around which gene transfer strategies can be specifically tailored to most closely reproduce the effects of normal CFTR expression. In fact, accurate restoration of endogenous expression patterns may be crucial, not only for disease reversal, but also to avoid potentially deleterious effects of inappropriate expression. This approach is in turn confounded however, by ill-defined stem and progenitor cell pathways within the lung epithelium. Nonetheless, studies to date suggest that these pathways are relatively plastic and may respond differently during homeostasis compared with repair following injury. It may therefore be feasible to target the lung epithelium in a non-cell specific manner and allow endogenous differentiation pathways to subsequently establish correct CFTR distribution patterns. In this review, emerging information on CFTR expression and function in developing and adult lungs is discussed in the context of putative stem cell populations and their potential for current gene delivery approaches. PMID:12109214

  9. Stimulation of airway and intestinal mucosal secretion by natural coumarin CFTR activators

    Directory of Open Access Journals (Sweden)

    Hong eYang

    2011-09-01

    Full Text Available Mutations of cystic fibrosis transmembrane conductance regulator (CFTR cause lethal hereditary disease cystic fibrosis (CF that involves extensive destruction and dysfunction of serous epithelium. Possible pharmacological therapy includes correction of defective intracellular processing and abnormal channel gating. In a previous study, we identified five natural coumarin potentiators of Δ508-CFTR including osthole, imperatorin, isopsoralen, praeruptorin A and scoparone. The present study was designed to determine the activity of these coumarine compounds on CFTR activity in animal tissues as a primary evaluation of their therapeutic potential. In the present study, we analyzed the affinity of these coumarin potentiators in activating wild-type CFTR and found that they are all potent activators. Osthole showed the highest affinity with Kd values <50 nmol/L as determined by Ussing chamber short-circuit current assay. Stimulation of rat colonic mucosal secretion by osthole was tested by the Ussing chamber short-circuit current assay. Osthole reached maximal activation of colonic Cl- secretion at 5 mol/L. Stimulation of mouse tracheal mucosal secretion was analyzed by optical measurement of single gland secretion. Fluid secretion rate of tracheal single submucosal gland stimulated by osthole at 10mol/L was 3-fold more rapid than that in negative control. In both cases the stimulated secretions were fully abolished by CFTRinh-172. In conclusion, the effective stimulation of Cl– and fluid secretion in colonic and tracheal mucosa by osthole suggested the therapeutic potential of natural coumarine compounds for the treatment of cystic fibrosis and other CFTR-related diseases.

  10. Cell cycle checkpoints in Caenorhabditis elegans: the 14-3-3 gene par-5 is required for germline development and DNA damage response / Checkpoints del ciclo celular en Caenorhabditis elegans: el gen 14-3-3, par-5, es necesario para el desarrollo y respuesta al daño genómico de la línea germinal

    OpenAIRE

    Aristizábal Corrales, David

    2012-01-01

    [spa] Las proteínas 14-3-3 han sido ampliamente estudiadas desde levadura hasta mamíferos y han sido asociadas con múltiples roles en procesos como ciclo celular, apoptosis y la respuesta al estrés. Así mismo estas proteínas se han visto involucradas en enfermedades neurodegenerativas y cáncer. De hecho, las proteínas 14-3-3 han sido propuestas como posibles agentes terapéuticos en el tratamiento contra el cáncer. En mamíferos existen 7 genes que codifican para proteínas 14-3-3, mientras en C...

  11. GenLab, Laboratorio Virtual de Genética

    Directory of Open Access Journals (Sweden)

    García Sergio

    2000-12-01

    Full Text Available

    GenLab es el nombre que tiene el software diseñado por nosotros, en el cual se modela el proceso meiótico y la fecundación en organismos diploides. El objetivo de esta aplicación es ilustrar el resultado de un cruce determinado, tratando de ser lo más ajustados a la realidad. La modelación de la reproducción sexual se realiza internamente y el GenLab se limita a presentar los resultados según el número de descendencia seleccionado para un cruce específico, esto significa que se puede escoger una gran cantidad de características para los parentales y se puede estudiar la frecuencia de estos en la descendencia. El modelo cuenta con base de datos donde están almacenados algunos de los locus de Drosophila melanogaster junto con su ubicación en centimorgans 1. EI propósito de este modelo es servir como herramienta pedagógica  y didáctica tanto en universidades como en colegios, facilitando el aprendizaje de algunos principios básicos de la genética, por lo cual puede ser usado si se cuenta con una conexión a Internet y un navegador visitando http://biologia.unal.edu.co/fidel.

  12. Taxonomic dissection of the genus Micrococcus: Kocuria gen. nov., Nesterenkonia gen. nov., Kytococcus gen. nov., Dermacoccus gen. nov., and Micrococcus Cohn 1872 gen. emend.

    Science.gov (United States)

    Stackebrandt, E; Koch, C; Gvozdiak, O; Schumann, P

    1995-10-01

    The results of a phylogenetic and chemotaxonomic analysis of the genus Micrococcus indicated that it is significantly heterogeneous. Except for Micrococcus lylae, no species groups phylogenetically with the type species of the genus, Micrococcus luteus. The other members of the genus form three separate phylogenetic lines which on the basis of chemotaxonomic properties can be assigned to four genera. These genera are the genus Kocuria gen. nov. for Micrococcus roseus, Micrococcus varians, and Micrococcus kristinae, described as Kocuria rosea comb. nov., Kocuria varians comb. nov., and Kocuria kristinae comb. nov., respectively; the genus Nesterenkonia gen. nov. for Micrococcus halobius, described as Nesterenkonia halobia comb. nov.; the genus Nesterenkonia gen. nov. for Micrococcus halobius, described as Nesterenkonia halobia comb. nov.; the genus Dermacoccus gen. nov. for Micrococcus nishinomiyaensis, described as Dermacoccus nishinomiyaensis comb. nov.; and the genus Kytocossus gen. nov. for Micrococcus sedentarius, described as Kytococcus sedentarius comb. nov. M. luteus and M. lylae, which are closely related phylogenetically but differ in some chemotaxonomic properties, are the only species that remain in the genus Micrococcus Cohn 1872. An emended description of the genus Micrococcus is given [corrected]. PMID:7547287

  13. The calpain, caspase 12, caspase 3 cascade leading to apoptosis is altered in F508del-CFTR expressing cells.

    Directory of Open Access Journals (Sweden)

    Mathieu Kerbiriou

    Full Text Available In cystic fibrosis (CF, the most frequent mutant variant of the cystic fibrosis transmembrane conductance regulator (CFTR, F508del-CFTR protein, is misfolded and retained in the endoplasmic reticulum (ER. We previously showed that the unfolded protein response (UPR may be triggered in CF. Since prolonged UPR activation leads to apoptosis via the calcium-calpain-caspase-12-caspase-3 cascade and because apoptosis is altered in CF, our aim was to compare the ER stress-induced apoptosis pathway between wild type (Wt and F508del-CFTR expressing cells. Here we show that the calcium-calpain-caspase-12-caspase-3 cascade is altered in F508del-CFTR expressing cells. We propose that this alteration is involved in the altered apoptosis triggering observed in CF.

  14. Analysis of CFTR Gene Mutations in Children with Cystic Fibrosis, First Report from North-East of Iran

    OpenAIRE

    Atieh Mehdizadeh Hakkak; Mohammad Keramatipour; Saeid Talebi; Azam Brook; Jalil Tavakol Afshari; Amin Raazi; Hamid Reza Kianifar

    2013-01-01

     Objective(s):  More than 1500 registered mutations in cystic fibrosis transmembrane regulator (CFTR) gene are responsible for dysfunction of an ion channel protein and a wide spectrum of clinical manifestations in patients with cystic fibrosis (CF). This study was performed to investigate the frequency of a number of well-known CFTR mutations in North Eastern Iranian CF patients. Material and Methods: A total number of 56 documented CF patients participated in this study. Peripheral blood...

  15. The Novel CFTR Mutation A457P in a Male with a Delayed Diagnosis of Cystic Fibrosis

    OpenAIRE

    Zuckerman, Jonathan B.; Yarmus, Lonny B.; Sosnay, Patrick R.; Cole, Kate H.

    2011-01-01

    Cystic fibrosis (CF) is an autosomal recessive disease that may be caused by more than 1000 different mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We describe the case of a CF patient who was initially diagnosed at 16 years of age after presenting with mild respiratory compromise and pancreatic sufficiency. When genetic testing was first performed using a CF mutation panel, only a single F508del CFTR allele was identified. We subsequently performed testing...

  16. Regulation of CFTR Expression and Arginine Vasopressin Activity Are Dependent on Polycystin-1 in Kidney-Derived Cells

    Directory of Open Access Journals (Sweden)

    Carolina Monteiro de Lemos Barbosa

    2016-01-01

    Full Text Available Background: Autosomal dominant polycystic kidney disease (ADPKD is characterized by the development of multiple, progressive, fluid-filled renal cysts that distort the renal parenchyma, leading to end-stage renal failure, mainly after the fifth decade of life. ADPKD is caused by a mutation in the PKD1 or PKD2 genes that encode polycystin-1 (PC-1 and polycystin-2 (PC-2, respectively. PC-1 is an important regulator of several signaling pathways and PC-2 is a nonselective calcium channel. The CFTR chloride channel is responsible for driving net fluid secretion into the cysts, promoting cyst growth. Arginine vasopressin hormone (AVP, in turn, is capable of increasing cystic intracellular cAMP, contributing to cell proliferation, transepithelial fluid secretion, and therefore to disease progression. The aim of this study was to assess if AVP can modulate CFTR and whether PC-1 plays a role in this potential modulation. Methods: M1 cells, derived from mouse cortical collecting duct, were used in the current work. The cells were treated with 10-7 M AVP hormone and divided into two main groups: transfected cells superexpressing PC-1 (Transf and cells not transfected (Ctrl. CFTR expression was assessed by immunodetection, CFTR mRNA levels were quantified by quantitative reverse transcription-polymerase chain reaction, and CFTR net ion transport was measured using the Ussing chamber technique. Results: AVP treatment increased the levels of CFTR protein and mRNA. CFTR short-circuit currents were also increased. However, when PC-1 was overexpressed in M1 cells, no increase in any of these parameters was detected. Conclusions: CFTR chloride channel expression is increased by AVP in M1 cells and PC-1 is capable of regulating this modulation.

  17. Increased plasma membrane cholesterol in cystic fibrosis cells correlates with CFTR genotype and depends on de novo cholesterol synthesis

    OpenAIRE

    Sonawane Nitin D; Previs Stephen F; Jiang Dechen; Ruddy Jennifer; Manson Mary E; West Richard H; Fang Danjun; Burgess James D; Kelley Thomas J

    2010-01-01

    Abstract Background Previous observations demonstrate that Cftr-null cells and tissues exhibit alterations in cholesterol processing including perinuclear cholesterol accumulation, increased de novo synthesis, and an increase in plasma membrane cholesterol accessibility compared to wild type controls. The hypothesis of this study is that membrane cholesterol accessibility correlates with CFTR genotype and is in part influenced by de novo cholesterol synthesis. Methods Electrochemical detectio...

  18. Analysis of Y chromosome microdeletions and CFTR gene mutations as genetic markers of infertility in Serbian men

    OpenAIRE

    Dinić Jelena; Kušić Jelena; Nikolić Аleksandra; Divac Aleksandra; Ristanović Momčilo; Radojković Dragica

    2007-01-01

    Background/Aim. Impaired fertility of a male partner is the main cause of infertility in up to one half of all infertile couples. At the genetic level, male infertility can be caused by chromosome aberrations or gene mutations. The presence and types of Y chromosome microdeletions and cystic fybrosis transmembrane conductance regulator (CFTR) gene mutations as genetic cause of male infertility was tested in Serbian men. The aim of this study was to analyze CFTR gene mutations and Y chromosome...

  19. Lack of CFTR in skeletal muscle predisposes to muscle wasting and diaphragm muscle pump failure in cystic fibrosis mice.

    Directory of Open Access Journals (Sweden)

    Maziar Divangahi

    2009-07-01

    Full Text Available Cystic fibrosis (CF patients often have reduced mass and strength of skeletal muscles, including the diaphragm, the primary muscle of respiration. Here we show that lack of the CF transmembrane conductance regulator (CFTR plays an intrinsic role in skeletal muscle atrophy and dysfunction. In normal murine and human skeletal muscle, CFTR is expressed and co-localized with sarcoplasmic reticulum-associated proteins. CFTR-deficient myotubes exhibit augmented levels of intracellular calcium after KCl-induced depolarization, and exposure to an inflammatory milieu induces excessive NF-kB translocation and cytokine/chemokine gene upregulation. To determine the effects of an inflammatory environment in vivo, sustained pulmonary infection with Pseudomonas aeruginosa was produced, and under these conditions diaphragmatic force-generating capacity is selectively reduced in Cftr(-/- mice. This is associated with exaggerated pro-inflammatory cytokine expression as well as upregulation of the E3 ubiquitin ligases (MuRF1 and atrogin-1 involved in muscle atrophy. We conclude that an intrinsic alteration of function is linked to the absence of CFTR from skeletal muscle, leading to dysregulated calcium homeostasis, augmented inflammatory/atrophic gene expression signatures, and increased diaphragmatic weakness during pulmonary infection. These findings reveal a previously unrecognized role for CFTR in skeletal muscle function that may have major implications for the pathogenesis of cachexia and respiratory muscle pump failure in CF patients.

  20. EPAC1 activation by cAMP stabilizes CFTR at the membrane by promoting its interaction with NHERF1.

    Science.gov (United States)

    Lobo, Miguel J; Amaral, Margarida D; Zaccolo, Manuela; Farinha, Carlos M

    2016-07-01

    Cyclic AMP (cAMP) activates protein kinase A (PKA) but also the guanine nucleotide exchange factor 'exchange protein directly activated by cAMP' (EPAC1; also known as RAPGEF3). Although phosphorylation by PKA is known to regulate CFTR channel gating - the protein defective in cystic fibrosis - the contribution of EPAC1 to CFTR regulation remains largely undefined. Here, we demonstrate that in human airway epithelial cells, cAMP signaling through EPAC1 promotes CFTR stabilization at the plasma membrane by attenuating its endocytosis, independently of PKA activation. EPAC1 and CFTR colocalize and interact through protein adaptor NHERF1 (also known as SLC9A3R1). This interaction is promoted by EPAC1 activation, triggering its translocation to the plasma membrane and binding to NHERF1. Our findings identify a new CFTR-interacting protein and demonstrate that cAMP activates CFTR through two different but complementary pathways - the well-known PKA-dependent channel gating pathway and a new mechanism regulating endocytosis that involves EPAC1. The latter might constitute a novel therapeutic target for treatment of cystic fibrosis. PMID:27206858

  1. Le goût de Genève

    OpenAIRE

    Lévy, Bertrand

    2006-01-01

    Le livre contient une anthologie de textes littéraires sur Genève, avec 5 sections : L'esprit de Genève, voir Genève, goûter Genève, vivre à Genève, annexes (renseignements pratiques et bibliographie)

  2. ATP and AMP Mutually Influence Their Interaction with the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) at Separate Binding Sites*

    OpenAIRE

    Randak, Christoph O.; Dong, Qian; Ver Heul, Amanda R.; Elcock, Adrian H.; Welsh, Michael J.

    2013-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel in the ATP-binding cassette (ABC) transporter protein family. In the presence of ATP and physiologically relevant concentrations of AMP, CFTR exhibits adenylate kinase activity (ATP + AMP ⇆ 2 ADP). Previous studies suggested that the interaction of nucleotide triphosphate with CFTR at ATP-binding site 2 is required for this activity. Two other ABC proteins, Rad50 and a structural maintenance of chromosome protein, ...

  3. Acute administration of ivacaftor to people with cystic fibrosis and a G551D-CFTR mutation reveals smooth muscle abnormalities

    Science.gov (United States)

    Adam, Ryan J.; Hisert, Katherine B.; Dodd, Jonathan D.; Grogan, Brenda; Launspach, Janice L.; Barnes, Janel K.; Gallagher, Charles G.; Sieren, Jered P.; Gross, Thomas J.; Fischer, Anthony J.; Cavanaugh, Joseph E.; Hoffman, Eric A.; Singh, Pradeep K.; Welsh, Michael J.; McKone, Edward F.; Stoltz, David A.

    2016-01-01

    Background Airflow obstruction is common in cystic fibrosis (CF), yet the underlying pathogenesis remains incompletely understood. People with CF often exhibit airway hyperresponsiveness, CF transmembrane conductance regulator (CFTR) is present in airway smooth muscle (ASM), and ASM from newborn CF pigs has increased contractile tone, suggesting that loss of CFTR causes a primary defect in ASM function. We hypothesized that restoring CFTR activity would decrease smooth muscle tone in people with CF. Methods To increase or potentiate CFTR function, we administered ivacaftor to 12 adults with CF with the G551D-CFTR mutation; ivacaftor stimulates G551D-CFTR function. We studied people before and immediately after initiation of ivacaftor (48 hours) to minimize secondary consequences of CFTR restoration. We tested smooth muscle function by investigating spirometry, airway distensibility, and vascular tone. Results Ivacaftor rapidly restored CFTR function, indicated by reduced sweat chloride concentration. Airflow obstruction and air trapping also improved. Airway distensibility increased in airways less than 4.5 mm but not in larger-sized airways. To assess smooth muscle function in a tissue outside the lung, we measured vascular pulse wave velocity (PWV) and augmentation index, which both decreased following CFTR potentiation. Finally, change in distensibility of <4.5-mm airways correlated with changes in PWV. Conclusions Acute CFTR potentiation provided a unique opportunity to investigate CFTR-dependent mechanisms of CF pathogenesis. The rapid effects of ivacaftor on airway distensibility and vascular tone suggest that CFTR dysfunction may directly cause increased smooth muscle tone in people with CF and that ivacaftor may relax smooth muscle. Funding This work was funded in part from an unrestricted grant from the Vertex Investigator-Initiated Studies Program. PMID:27158673

  4. Differential expression of CFTR gene in the mouse intestinal tissues%CFTR 基因在小鼠肠道组织中的差异表达

    Institute of Scientific and Technical Information of China (English)

    王月影; 韩莹倩; 查光明; 汪新建; 李和平

    2014-01-01

    Object This experiment was conducted to study the relationship between CFTR gene expression in the intestinal tissues and secretory diarrhea.Methods Twenty-four Kunming mice were selected, half male and half female, and were randomly divided into 3 groups ( n=8 in each group):control group with intraperitoneal injection of 0.2 mL nor-mal saline, and the experimental group of mice by intraperitoneal injection of lipopolysaccharide(LPS) (6 mg/kg· bw). The mental state and intestinal morphology of the mice at 1 h and 8 h after LPS injection were observed to assess whether the secretory diarrhea model was successfully established.The expression of CFTR gene segments of intestine tissue was de-tected by fluorescence quantitative PCR.Results LPS induced secretory diarrhea.CFTR gene was expressed in the mouse duodenum, jejunum, ileum and colon tissues with different expression abundance.It was highest in the colon, but the difference was not significant between intestinal segments.Compared with the control group, LPS up-regulated the tran-scription level of CFTR gene in the duodenum, jejunum and ileum, and down-regulated the transcription of CFTR gene in the colon.Conclusions The results of our study suggest that the changes of the transcriptional level of CFTR gene are closely related with the diarrhea induced by LPS and the effects in different intestinal segments on the diarrhea is different. The jejunum plays a crucial role and the colon plays a least role in the Cl-secretion.%目的:研究肠道组织CFTR基因表达与分泌性腹泻发生的关系。方法选取KM小鼠24只,雌雄各半,随机分为3组(每组8只):对照组经小鼠腹腔注射0.2 mL生理盐水,实验组小鼠经腹腔注射LPS[6 mg/(kg· bw)]分别作用1 h、8 h,于注射后通过小鼠精神状态、肠道组织形态学判定分泌性腹泻模型的建立,利用荧光定量PCR法检测各段肠道组织CFTR基因的表达。结果 LPS成功诱导小鼠发生了分泌

  5. Algoritmos genéticos incrementales

    OpenAIRE

    Nesmachnow, Sergio; Dominioni, Federico; Musso, Pablo

    2005-01-01

    Este artículo presenta el estudio, diseño e implementación de un modelo particular de algoritmos genéticos paralelos, los algoritmos genéticos incrementales. La principal ventaja del motor de algoritmos genéticos implementado es su potencialidad de ejecución en entornos no dedicados, detectando la carga generada por los usuarios del entorno y consumiendo de un modo “inteligente” los recursos computacionales disponibles. Se presenta la descripción teórica del modelo de algoritmos genéticos inc...

  6. Comportamento de dois genótipos de milho cultivados em sistema de aléias preestabelecido com diferentes leguminosas arbóreas Behaviour of two maize genotypes grown in alley cropping system pre-established with diferents leguminous trees

    Directory of Open Access Journals (Sweden)

    Andréia Araújo Lima Leite

    2008-12-01

    Full Text Available O cultivo em aléias tem sido recomendado como alternativa para a substituição da agricultura de corte e queima, no trópico úmido, devido à grande capacidade de produção de matéria orgânica e de reciclagem de nutrientes, mas algumas dúvidas quanto à sustentabilidade e à competição interespecífica são persistentes. O objetivo no trabalho foi avaliar a viabilidade da cultura do milho em um sistema de cultivo em aléias de leguminosas arbóreas. O delineamento experimental utilizado foi em blocos casualisados, com quatro repetições dos tratamentos: aléias de sombreiro (Clitoria fairchildiana, ingá (Inga edulis, guandu (Cajanus cajan e leucena (Leucaena leucocephala e uma testemunha sem aléias. Foram avaliadas a remobilização de carbono e nitrogênio, massa de grãos, massa de mil grãos e competição interespecífica entre as cultivares de milho e as leguminosas. A produção de grãos foi maior nas parcelas com C. fairchildiana e L. leucocephala. A produtividade do híbrido de milho foi superior à da variedade em todos os tratamentos. A produtividade e a massa de mil grãos de milho não são negativamente afetadas pela distância da linha da leguminosa arbórea. Esse estudo conclui que o sistema de aléias com leguminosas arbóreas é uma alternativa importante ao manejo sustentável dos agroecossistemas no tropico úmido. Além disso, nessa região a produtividade em grãos na cultura do milho é favorecida no sistema de aléias preeestabelecidas com as leguminosas arbóreas sombreiro, ingá e leucena e pela utilização de genótipos eficientes no aproveitamento do nitrogênio, cujo sincronismo entre a liberação e a absorção do N aplicado por meio das leguminosas deve ser aprimorado.Alley cropping has been recommended as alternative land use to slash-and-burn agriculture in humid tropics. However, interespecific competition between cash crop and hedgerow can reduce this potential. This study aimed to evaluate the

  7. CFTR depletion results in changes in fatty acid composition and promotes lipogenesis in intestinal Caco 2/15 cells.

    Directory of Open Access Journals (Sweden)

    Geneviève Mailhot

    Full Text Available BACKGROUND: Abnormal fatty acid composition (FA in plasma and tissue lipids frequently occurs in homozygous and even in heterozygous carriers of cystic fibrosis transmembrane conductance regulator (CFTR mutations. The mechanism(s underlying these abnormalities remained, however, poorly understood despite the potentially CFTR contributing role. METHODOLOGY/PRINCIPAL FINDINGS: The aim of the present study was to investigate the impact of CFTR depletion on FA uptake, composition and metabolism using the intestinal Caco-2/15 cell line. shRNA-mediated cftr gene silencing induced qualitative and quantitative modifications in FA composition in differentiated enterocytes as determined by gas-liquid chromatography. With the cftr gene disruption, there was a 1,5 fold increase in the total FA amount, largely attributable to monounsaturated and saturated FA compared to controls. The activity of delta-7 desaturase, estimated by the 16:1(n-7/16:0, was significantly higher in knockdown cells and consistent with the striking elevation of the n-7 FA family. When incubated with [14C]-oleic acid, CFTR-depleted cells were capable of quick incorporation and export to the medium concomitantly with the high protein expression of L-FABP known to promote intracellular FA trafficking. Accordingly, lipoprotein vehicles (CM, VLDL, LDL and HDL, isolated from CFTR knockdown cells, exhibited higher levels of radiolabeled FA. Moreover, in the presence of [14C]-acetate, knockdown cells exhibited enhanced secretion of newly synthesized phospholipids, triglycerides, cholesteryl esters and free FA, thereby suggesting a stimulation of the lipogenic pathway. Conformably, gene expression of SREBP-1c, a key lipogenic transcription factor, was increased while protein expression of the phosphorylated and inactive form of acetylCoA carboxylase was reduced, confirming lipogenesis induction. Finally, CFTR-depleted cells exhibited lower gene expression of transcription factors (PPARalpha

  8. Inflammation and CFTR: Might Neutrophils be the Key in Cystic Fibrosis?

    OpenAIRE

    Witko-Sarsat, V; Sermet-Gaudelus, I; Lenoir, G.; Descamps-Latscha, B

    1999-01-01

    The aim of this hypothesis is to provide new insights into the still unclear mechanisms governing airway inflammation in cystic fibrosis. Although the genetic basis of cystic fibrosis as well as the molecular structure of cystic fibrosis transmembrane regulator (CFTR), the mutated protein which causes the disease, have been well defined, a clear relationship between the genetic defect and the pulmonary pathophysiology, especially chronic infections and neutrophil-dominated airway inflammation...

  9. Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in allergic bronchopulmonary aspergillosis.

    OpenAIRE

    Miller, P. W.; Hamosh, A.; Macek, M.; Greenberger, P. A.; MacLean, J; Walden, S M; Slavin, R G; Cutting, G R

    1996-01-01

    The etiology of allergic bronchopulmonary aspergillosis (ABPA) is not well understood. A clinical phenotype resembling the pulmonary disease seen in cystic fibrosis (CF) patients can occur in some individuals with ABPA. Reports of familial occurrence of ABPA and increased incidence in CF patients suggest a possible genetic basis for the disease. To test this possibility, the entire coding region of the cystic fibrosis transmembrane regulator (CFTR) gene was analyzed in 11 individuals who met ...

  10. Impact of Cystic Fibrosis Transmembrane Regulator (CFTR) gene mutations on male infertility

    OpenAIRE

    Jlenia Elia; Rossella Mazzilli; Michele Delfino; Maria Piane; Cristina Bozzao; Vincenzo Spinosa; Luciana Chessa; Fernando Mazzilli

    2014-01-01

    Objective. The aim of this study was to evaluate the prevalence of most common mutations and intron 8 5T (IVS8-5T) polymorphism of CFTR gene in Italian: a) azoospermic males; b) non azoospermic subjects, male partners of infertile couples enrolled in assisted reproductive technology (ART) programs. Material and methods. We studied 242 subjects attending our Andrology Unit (44 azoospermic subjects and 198 non azoospermic subjects, male partners of infertile couples enrolled in ART programs). S...

  11. Phosphorylation of CFTR and other membrane proteins: a role in biogenesis and traffic?

    OpenAIRE

    Romeiras, Francisco Maria de Sousa de Macedo Malta, 1986-

    2009-01-01

    Tese de mestrado, Bioquímica (Bioquímica Médica), Universidade de Lisboa, Faculdade de Ciências, 2009 Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder among Caucasians, is caused by mutations in the gene that encodes for the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). With more than 1600 mutations reported, a single mutation– the deletion of phenylalanine residue at position 508 – accounts for more than 70% of chromosomes worldwide. The presence of ...

  12. Regulation of CFTR Cl− channel gating by ATP binding and hydrolysis

    OpenAIRE

    Ikuma, Mutsuhiro; Welsh, Michael J.

    2000-01-01

    Opening and closing of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel is regulated by the interaction of ATP with its two cytoplasmic nucleotide-binding domains (NBD). Although ATP hydrolysis by the NBDs is required for normal gating, the influence of ATP binding versus hydrolysis on specific steps in the gating cycle remains uncertain. Earlier work showed that the absence of Mg2+ prevents hydrolysis. We found that even in the absence of Mg2+, ATP could support cha...

  13. Atencion integral al paciente con poliquistosis renal genética: Perfil clínico y experiencia vital subjetiva Integral care of patient with genetic renal polycystosis: Clinical profile and subject vital experience

    Directory of Open Access Journals (Sweden)

    M Calderó Urrea

    2007-09-01

    Full Text Available La poliquistosis renal autosómica dominante afecta al 5%-10% de personas en programa de hemodiálisis o de trasplante renal. Este estudio tiene un doble objetivo: conocer el perfil clínico del paciente con poliquistosis renal y comprender cómo esta enfermedad influye en las diferentes etapas vitales. El diseño de investigación es cuantitativo y cualitativo para dar respuesta a cada uno de los objetivos propuestos. La perspectiva cuantitativa abarca un estudio descriptivo retrospectivo de revisión de las historias clínicas de los pacientes atendidos en nuestro centro con este diagnóstico entre los años 2000 y 2005 (N=161. Para abarcar aspectos cualitativos de la enfermedad se han realizado dos entrevistas en profundidad, semiestructuradas, a personas cuyo diagnóstico ha actuado como condicionante en la toma de decisiones personales y familiares. El perfil de paciente con poliquistosis renal es el de una persona de 58 años de media con poliquistosis hepática, así como antecedentes familiares de la enfermedad y diversas patologías asociadas, siendo la más frecuente la hipertensión arterial. Las entrevistas en profundidad denotan la presencia de dolor crónico, con una alteración de la vida cotidiana y de la dinámica familiar. La incerteza en cuanto a su evolución y tratamiento produce ansiedad y un desgaste emocional progresivo.Autosomic dominant renal polycystosis affects 5%-10% of people on haemodialysis or with kidney transplants. This study has a two-fold purpose: to determine the clinical profile of the patient with renal polycystosis and to understand how this dise ase affects different vital stages. The research design is quantitative and qualitative in order to meet each of the proposed goals. The quantitative perspective encompasses a retrospective descriptive review of the case histories of the patients treated at our centre with this diagnosis between the years 2000 and 2005 (N=161. To cover qualitative aspects of the

  14. Frecuencias alélicas y fenotípicas de los marcadores genéticos enzimáticos: Fosfoglucomutasa 1 (FGM 1, Esterasa D (EsD y Fosfatasa ácida eritrocitaria (FAE en población residente en la ciudad de Medellín, 1996-1997

    Directory of Open Access Journals (Sweden)

    María Victoria Hurtado Ángel

    2000-01-01

    Full Text Available Este estudio descriptivo de las enzimas fosfoglucomutasa 1 (fgm 1, esterasa D (EsD y Fosfatasa ácida eritrocitaria (FAE como marcadores genéticos polimórficos, se realizó en 145 muestras de sangre, tomadas a igual númerode donantes de los tres principales bancos de sangre de la ciudad, por el método de electroforesis convencional, con sistema de refrigeración, en gel de agarosa. La probabilidad de identificación para cada uno de los marcadores fue buena; la mejor fue la subtipificación de la FGM1, seguida de la FAE por ser las más polimórficas. Igual resultado se obtuvo para el poder de discriminación. Las frecuencias alélicas y fenotípicas se encontraron de manera representativa en la población, en la que estaban presentes 21 de los 22 fenotipos posibles. Los resultados obtenidos muestran que los tres marcadores enzimáticos estudiados se encuentran en equilibrio de Hardy-Weinberg, lo cual se comprobó aplicando el método estadístico 2 y la corrección de Bonferroni para la FGM 1 subtipo; por ello son de gran utilidad para cálculos de probabilidad en casos forenses con población residente en Antioquia, pues se encontró que dada la procedencia de los donantes y sus padres se puede hacer inferencia a todo el departamento.

  15. Justicia en salud y genética

    Directory of Open Access Journals (Sweden)

    Maria Graciela De Ortuzar

    2014-06-01

    Full Text Available Las expectativas puestas en el conocimiento genético exceden el ámbito de la medicina tradiciona, debido a que la intervención directa en la lotería natural demandaría el replanteamiento de conceptos centrales de justicia en salud: necesidades médicas, enfermedad, normalidad, e igualdad de oportunidades en el acceso a la salud. El punto en debate es sí el replanteo de dichos conceptos conlleva un cambio radical en las teorías de justicia (libertariana y/o liberal, mostrando su obsolescencia, o sí simplemente se requiere ampliar dichos conceptos claves por fallas estructurales en las mismas teorías. Como hipótesis general considero que los supuestos cuestionamientos, lejos de socavar las bases de las teorías de justicia, sólo ponen en evidencia sus viejos problemas estructurales. Por razones expositivas, dividiré la presentación tres partes. En la Primera parte, analizo la teoría libertariana, estudiando las contradicciones del modelo a través del impacto de la información genética en el seguro privado de salud. En la Segunda Parte, desarrollo la propuesta alternativa liberal rawlsianadanielsiana del modelo de seguro público, evaluando las implicaciones de la genética a partir de la crítica de su concepto biológico de enfermedad y su restricción al acceso a la salud por necesidades naturales. En la Tercera parte presento un modelo integral de necesidades y capacidades básicas, comprendiendo la prevención, el tratamiento y el mejoramiento moralmente permisible (genético y no genético.Mi aporte principal consiste en la elaboración de este modelo normativo integral de necesidades y capacidades para la regulación conjunta de la información y terapia genética con los restantes problemas de salud.

  16. Increased susceptibility of Cftr-/- mice to LPS-induced lung remodeling.

    Science.gov (United States)

    Bruscia, Emanuela M; Zhang, Ping-Xia; Barone, Christina; Scholte, Bob J; Homer, Robert; Krause, Diane S; Egan, Marie E

    2016-04-15

    Cystic fibrosis (CF) is caused by homozygous mutations of the CF transmembrane conductance regulator (CFTR) Cl(-) channel, which result in chronic pulmonary infection and inflammation, the major cause of morbidity and mortality. Although these processes are clearly related to each other, each is likely to contribute to the pathology differently. Understanding the contribution of each of these processes to the overall pathology has been difficult, because they are usually so intimately connected. Various CF mouse models have demonstrated abnormal immune responses compared with wild-type (WT) littermates when challenged with live bacteria or bacterial products acutely. However, these studies have not investigated the consequences of persistent inflammation on lung tissue in CF mice, which may better model the lung pathology in patients. We characterized the lung pathology and immune response of Cftr(-/-) (CF) and Cftr(+/+) (WT) mice to chronic administration of Pseudomonas aeruginosa lipopolysaccharide (LPS). We show that, after long-term repeated LPS exposure, CF mice develop an abnormal and persistent immune response, which is associated with more robust structural changes in the lung than those observed in WT mice. Although CF mice and their WT littermates develop lung pathology after chronic exposure to LPS, the inflammation and damage resolve in WT mice. However, CF mice do not recover efficiently, and, as a consequence of their chronic inflammation, CF mice are more susceptible to morphological changes and lung remodeling. This study shows that chronic inflammation alone contributes significantly to aspects of CF lung pathology. PMID:26851259

  17. The power stroke driven by ATP binding in CFTR as studied by molecular dynamics simulations.

    Science.gov (United States)

    Furukawa-Hagiya, Tomoka; Furuta, Tadaomi; Chiba, Shuntaro; Sohma, Yoshiro; Sakurai, Minoru

    2013-01-10

    Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel belonging to the ATP binding cassette (ABC) protein superfamily. Currently, it remains unclear how ATP binding causes the opening of the channel gate at the molecular level. To clarify this mechanism, we first constructed an atomic model of the inward-facing CFTR using the X-ray structures of other ABC proteins. Molecular dynamics (MD) simulations were then performed to explore the structure and dynamics of the inward-facing CFTR in a membrane environment. In the MgATP-bound state, two nucleotide-binding domains (NBDs) formed a head-to-tail type of dimer, in which the ATP molecules were sandwiched between the Walker A and signature motifs. Alternatively, one of the final MD structures in the apo state was similar to that of a "closed-apo" conformation found in the X-ray analysis of ATP-free MsbA. Principal component analysis for the MD trajectory indicated that NBD dimerization causes significant structural and dynamical changes in the transmembrane domains (TMDs), which is likely indicative of the formation of a chloride ion access path. This study suggests that the free energy gain from ATP binding acts as a driving force not only for NBD dimerization but also for NBD-TMD concerted motions. PMID:23214920

  18. Detection of phospho-sites generated by protein kinase CK2 in CFTR: mechanistic aspects of Thr1471 phosphorylation.

    Directory of Open Access Journals (Sweden)

    Andrea Venerando

    Full Text Available By mass spectrometry analysis of mouse Cystic Fibrosis Transmembrane-conductance Regulator (mCFTR expressed in yeast we have detected 21 phosphopeptides accounting for 22 potential phospho-residues, 12 of which could be unambiguously assigned. Most are conserved in human CFTR (hCFTR and the majority cluster in the Regulatory Domain, lying within consensus sequences for PKA, as identified in previous mammalian studies. This validates our yeast expression model. A number of phospho-residues were novel and human conserved, notably mouse Ser670, Ser723, Ser737, and Thr1467, that all lie in acidic sequences, compatible with their phosphorylation by protein kinase CK2. Thr1467 is localized in the C-terminal tail, embedded in a functionally important and very acidic sequence (EETEEE which displays an optimal consensus for protein kinase CK2. Herein, we show that Thr1467, homologous to human Thr1471 is readily phosphorylated by CK2. Indeed a 42 amino acid peptide encompassing the C-terminal segment of human CFTR is readily phosphorylated at Thr1471 with favorable kinetics (Km 1.7 µM by CK2 holoenzyme, but neither by its isolated catalytic subunit nor by other acidophilic Ser/Thr kinases (CK1, PLK2/3, GCK/FAM20C. Our finding that by treating CFTR expressing BHK cells with the very specific CK2 inhibitor CX4945, newly synthesized wild type CFTR (and even more its Phe508del mutant accumulates more abundantly than in the absence of CK2 inhibitor, supports the conclusion that phosphorylation of CFTR by CK2 correlates with decreased stability of the protein.

  19. Environmental management in PowerGen

    International Nuclear Information System (INIS)

    PowerGen has developed a company wide Environmental Compliance Policy. This incorporates an environmental management system and sets down Minimum Management Standards. The Policy is supported by an environmental audit programme. PowerGen has participated in the development of standardised Environmental Management Systems within BS7750 and the European Community Eco - Management and Audit Scheme. 1 fig

  20. Genética del asma Genetics of asthma

    Directory of Open Access Journals (Sweden)

    S. de Arriba Méndez

    2010-01-01

    Full Text Available El asma tiene una clara agregación familiar, siendo más frecuente el desarrollo de asma en un niño si sus padres son asmáticos; asimismo, es mayor la concordancia en los gemelos idénticos y son numerosos los estudios epidemiológicos que han destacado que los antecedentes familiares constituyen un importante factor de riesgo para desarrollar asma. Diferentes estudios con gemelos han concluido que la herencia del asma está en torno al 60%, destacando por tanto la importancia de dichos factores genéticos...

  1. Frigitilla gen. nov., a new genus of Amazonian Mutillidae (Hymenoptera).

    Science.gov (United States)

    Bartholomay, Pedro R; Williams, Kevin A; Luz, David R; Morato, Elder F

    2015-01-01

    Mutilla frigidula Cresson, 1902 was transferred to Tobantilla by Williams et al. (2011), based on morphological similarities with females of that genus. Discovery of the male of this species indicated significant morphological differences from Tobantilla. We therefore erect the genus, Frigitilla gen. nov., for Mutilla frigidula. Herein, we describe the male of F. frigidula (Cresson, 1902), comb. nov., associate it with its host (Trypoxylon spp.), and discuss its relations to other mutillid genera. The impact of collecting method and specimen age on the integumental coloration of specimens is discussed, emphasizing the need to diagnose species on consistent structural features rather than differences in color pattern. PMID:26249052

  2. A novel treatment of cystic fibrosis acting on-target: cysteamine plus epigallocatechin gallate for the autophagy-dependent rescue of class II-mutated CFTR.

    Science.gov (United States)

    Tosco, A; De Gregorio, F; Esposito, S; De Stefano, D; Sana, I; Ferrari, E; Sepe, A; Salvadori, L; Buonpensiero, P; Di Pasqua, A; Grassia, R; Leone, C A; Guido, S; De Rosa, G; Lusa, S; Bona, G; Stoll, G; Maiuri, M C; Mehta, A; Kroemer, G; Maiuri, L; Raia, V

    2016-08-01

    We previously reported that the combination of two safe proteostasis regulators, cysteamine and epigallocatechin gallate (EGCG), can be used to improve deficient expression of the cystic fibrosis transmembrane conductance regulator (CFTR) in patients homozygous for the CFTR Phe508del mutation. Here we provide the proof-of-concept that this combination treatment restored CFTR function and reduced lung inflammation (Ptreatment in vitro. We assessed individual responses to cysteamine plus EGCG in a single-centre, open-label phase-2 trial. The combination treatment decreased sweat chloride from baseline, increased both CFTR protein and function in nasal cells, restored autophagy in such cells, decreased CXCL8 and TNF-α in the sputum, and tended to improve respiratory function. These positive effects were particularly strong in patients carrying Phe508del CFTR mutations in homozygosity or heterozygosity. However, a fraction of patients bearing other CFTR mutations failed to respond to therapy. Importantly, the same patients whose primary nasal brushed cells did not respond to cysteamine plus EGCG in vitro also exhibited deficient therapeutic responses in vivo. Altogether, these results suggest that the combination treatment of cysteamine plus EGCG acts 'on-target' because it can only rescue CFTR function when autophagy is functional (in mice) and improves CFTR function when a rescuable protein is expressed (in mice and men). These results should spur the further clinical development of the combination treatment. PMID:27035618

  3. ホスファチジン酸による CFTR の細胞内輸送制御機構および CFTR 機能破綻により生じる掻痒病態発症機構の解明

    OpenAIRE

    橋本, 泰明; ハシモト, ヤスアキ; Hashimoto, Yasuaki

    2008-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) in which mutation isthe cause of cystic fibrosis (CF) is a polytopic integral membrane protein that mediatestransepithelial chloride transport across epithelial cells in airways, pancreas, intestines andsweat glands. In addition, CFTR also regulates other various molecules (ion channels andreceptors) such as ENaC by forming multimolecular complex called “Transportsome”.Therefore, CFTR molecule is an extremely important molecule fo...

  4. MARCH2 regulates autophagy by promoting CFTR ubiquitination and degradation and PIK3CA-AKT-MTOR signaling.

    Science.gov (United States)

    Xia, Dan; Qu, Liujing; Li, Ge; Hongdu, Beiqi; Xu, Chentong; Lin, Xin; Lou, Yaxin; He, Qihua; Ma, Dalong; Chen, Yingyu

    2016-09-01

    MARCH2 (membrane-associated RING-CH protein 2), an E3 ubiquitin ligase, is mainly associated with the vesicle trafficking. In the present study, for the first time, we demonstrated that MARCH2 negatively regulates autophagy. Our data indicated that overexpression of MARCH2 impaired autophagy, as evidenced by attenuated levels of LC3B-II and impaired degradation of endogenous and exogenous autophagic substrates. By contrast, loss of MARCH2 expression had the opposite effects. In vivo experiments demonstrate that MARCH2 knockout mediated autophagy results in an inhibition of tumorigenicity. Further investigation revealed that the induction of autophagy by MARCH2 deficiency was mediated through the PIK3CA-AKT-MTOR signaling pathway. Additionally, we found that MARCH2 interacts with CFTR (cystic fibrosis transmembrane conductance regulator), promotes the ubiquitination and degradation of CFTR, and inhibits CFTR-mediated autophagy in tumor cells. The functional PDZ domain of MARCH2 is required for the association with CFTR. Thus, our study identified a novel negative regulator of autophagy and suggested that the physical and functional connection between the MARCH2 and CFTR in different conditions will be elucidated in the further experiments. PMID:27308891

  5. Genève Reconnaissante

    CERN Multimedia

    2001-01-01

    Robert Cailliau (centre), with Geneva's Mayor Alain Vaissade (left) and Jean Erhardt, Secretary General of the Administrative Council of Geneva (right). Geneva recognised the contribution of two CERN people to the reputation of the city last Tuesday when Mayor Alain Vaissade presented the Genève Reconaissante Medal to Tim Berners-Lee and Robert Cailliau. Berners-Lee, who was not able to be present in person, invented the World Wide Web at CERN just over a decade ago, while Cailliau was his first collaborator. Quoting Cailliau, Vaissade said that whilst there is no doubt that something like the Web would have appeared sooner or later, the fact that it happened at CERN, in Geneva, was no accident. Both the Laboratory and the city are places where people from around the world meet and work in harmony.

  6. Marcadores genéticos relacionados con el déficit cognitivo en el envejecimiento

    OpenAIRE

    Solé-Padullés, Cristina; Clemente, Imma C.; Bartrés Faz, David

    2004-01-01

    En el presente trabajo se revisan estudios realizados en relación a los marcadores genéticos de vulnerabilidad al deterioro cognitivo en el envejecimiento. En concreto, se estudia el papel de varios polimorfismos genéticos en las entidades de Alteración de la Memoria Asociada a la Edad (AAMI, del inglés Age Associated Memory Impairment) y Alteración Cognitiva Leve (MCI, del inglés Mild Cognitive Impairment), por ser condiciones de alto riesgo para el desarrollo ...

  7. Enfermedades de base genética Genetically based diseases

    Directory of Open Access Journals (Sweden)

    D. González-Lamuño

    2008-01-01

    Full Text Available La genética constituye uno de los mayores avances científicos del siglo XX, que comienza con el redescubrimiento de las leyes de Mendel y termina con la elaboración del primer "borrador" de la secuencia completa del genoma humano. La genética utiliza diferentes estrategias de investigación, como los estudios de gemelos y de adopción, que investigan la influencia de los factores genéticos y ambientales, y las estrategias para identificar genes específicos (genética molecular. Además del importante grado de discapacidad que generan, el impacto social de las enfermedades hereditarias es enorme, por su carácter potencialmente recurrente en una misma familia y por el elevado coste socio-sanitario derivado de la enorme carga de cuidados que requiere. El diagnóstico de las enfermedades hereditarias presenta características diferenciadoras muy significativas ya que el resultado de un diagnóstico genético tiene no sólo efectos sobre el paciente sino también sobre todos los individuos emparentados. Por tanto, la unidad de estudio en el diagnóstico genético es la familia y todo proceso de diagnóstico implica una investigación familiar. También conviene tener en cuenta que los protocolos de diagnóstico se desarrollan de forma paralela a la investigación básica y generalmente están poco estandarizados. Los resultados obtenidos en los estudios genéticos y el tipo de información que se facilita al paciente y a su familia deben ser matizados dentro del proceso del "consejo genético".Genetics is one of the greatest scientific advances of the XX century, which begins with the rediscovery of Mendel’s laws and culminates in the elaboration of the first "draft" of the complete sequence of the human genome. Genetics employs different research strategies, such as the study of twins and adoption, investigating the influence of genetic and environmental factors, and strategies for identifying specific genes (molecular genetics. Besides the

  8. Warum Englisch allein als Wissenschaftssprache nicht genügt

    OpenAIRE

    Baschera, Marco

    2010-01-01

    Einst war Latein Lingua franca der intellektuellen Welt. Gegenwärtig wird darüber debattiert, ob sich Englisch dafür eignet. Doch Latein war eine künstliche Gelehrtensprache, während Englisch heute einem unkontrollierbaren Sprachwandel unterworfen ist. Kulturell unterschiedene Denkweisen vermag es nicht zu spiegeln.

  9. Del pasado al futuro de las razas bovinas de carne autóctonas. Análisis genealógico y de marcadores SNP para la implementación de la selección genómica

    OpenAIRE

    Cañas Álvarez, Jhon Jacobo

    2015-01-01

    Las técnicas actuales de genotipado masivo de marcadores SNP han proporcionado una herramienta muy útil, tanto para determinar la diversidad como para la mejora genética animal. Sin embargo, en las razas españolas de vacuno de carne su aplicabilidad no ha sido tan evidente hasta ahora. Es por esta razón, que las preguntas más importantes que se plantearon en esta tesis doctoral fueron: 1) determinar la cantidad de variación genética que hay en las principales razas autóctonas de ganado de car...

  10. Thermal unfolding studies show the disease causing F508del mutation in CFTR thermodynamically destabilizes nucleotide-binding domain 1

    Science.gov (United States)

    Protasevich, Irina; Yang, Zhengrong; Wang, Chi; Atwell, Shane; Zhao, Xun; Emtage, Spencer; Wetmore, Diana; Hunt, John F; Brouillette, Christie G

    2010-01-01

    Misfolding and degradation of CFTR is the cause of disease in patients with the most prevalent CFTR mutation, an in-frame deletion of phenylalanine (F508del), located in the first nucleotide-binding domain of human CFTR (hNBD1). Studies of (F508del)CFTR cellular folding suggest that both intra- and inter-domain folding is impaired. (F508del)CFTR is a temperature-sensitive mutant, that is, lowering growth temperature, improves both export, and plasma membrane residence times. Yet, paradoxically, F508del does not alter the fold of isolated hNBD1 nor did it seem to perturb its unfolding transition in previous isothermal chemical denaturation studies. We therefore studied the in vitro thermal unfolding of matched hNBD1 constructs ±F508del to shed light on the defective folding mechanism and the basis for the thermal instability of (F508del)CFTR. Using primarily differential scanning calorimetry (DSC) and circular dichroism, we show for all hNBD1 pairs studied, that F508del lowers the unfolding transition temperature (Tm) by 6–7°C and that unfolding occurs via a kinetically-controlled, irreversible transition in isolated monomers. A thermal unfolding mechanism is derived from nonlinear least squares fitting of comprehensive DSC data sets. All data are consistent with a simple three-state thermal unfolding mechanism for hNBD1 ± F508del: N(±MgATP) ⇄ IT(±MgATP) → AT → (AT)n. The equilibrium unfolding to intermediate, IT, is followed by the rate-determining, irreversible formation of a partially folded, aggregation-prone, monomeric state, AT, for which aggregation to (AT)n and further unfolding occur with no detectable heat change. Fitted parameters indicate that F508del thermodynamically destabilizes the native state, N, and accelerates the formation of AT. PMID:20687133

  11. Correction of chloride transport and mislocalization of CFTR protein by vardenafil in the gastrointestinal tract of cystic fibrosis mice.

    Directory of Open Access Journals (Sweden)

    Barbara Dhooghe

    Full Text Available Although lung disease is the major cause of mortality in cystic fibrosis (CF, gastrointestinal (GI manifestations are the first hallmarks in 15-20% of affected newborns presenting with meconium ileus, and remain major causes of morbidity throughout life. We have previously shown that cGMP-dependent phosphodiesterase type 5 (PDE5 inhibitors rescue defective CF Transmembrane conductance Regulator (CFTR-dependent chloride transport across the mouse CF nasal mucosa. Using F508del-CF mice, we examined the transrectal potential difference 1 hour after intraperitoneal injection of the PDE5 inhibitor vardenafil or saline to assess the amiloride-sensitive sodium transport and the chloride gradient and forskolin-dependent chloride transport across the GI tract. In the same conditions, we performed immunohistostaining studies in distal colon to investigate CFTR expression and localization. F508del-CF mice displayed increased sodium transport and reduced chloride transport compared to their wild-type littermates. Vardenafil, applied at a human therapeutic dose (0.14 mg/kg used to treat erectile dysfunction, increased chloride transport in F508del-CF mice. No effect on sodium transport was detected. In crypt colonocytes of wild-type mice, the immunofluorescence CFTR signal was mostly detected in the apical cell compartment. In F508del-CF mice, a 25% reduced signal was observed, located mostly in the subapical region. Vardenafil increased the peak of intensity of the fluorescence CFTR signal in F508del-CF mice and displaced it towards the apical cell compartment. Our findings point out the intestinal mucosa as a valuable tissue to study CFTR transport function and localization and to evaluate efficacy of therapeutic strategies in CF. From our data we conclude that vardenafil mediates potentiation of the CFTR chloride channel and corrects mislocalization of the mutant protein. The study provides compelling support for targeting the cGMP signaling pathway in CF

  12. Increased plasma membrane cholesterol in cystic fibrosis cells correlates with CFTR genotype and depends on de novo cholesterol synthesis

    Directory of Open Access Journals (Sweden)

    Sonawane Nitin D

    2010-05-01

    Full Text Available Abstract Background Previous observations demonstrate that Cftr-null cells and tissues exhibit alterations in cholesterol processing including perinuclear cholesterol accumulation, increased de novo synthesis, and an increase in plasma membrane cholesterol accessibility compared to wild type controls. The hypothesis of this study is that membrane cholesterol accessibility correlates with CFTR genotype and is in part influenced by de novo cholesterol synthesis. Methods Electrochemical detection of cholesterol at the plasma membrane is achieved with capillary microelectrodes with a modified platinum coil that accepts covalent attachment of cholesterol oxidase. Modified electrodes absent cholesterol oxidase serves as a baseline control. Cholesterol synthesis is determined by deuterium incorporation into lipids over time. Incorporation into cholesterol specifically is determined by mass spectrometry analysis. All mice used in the study are on a C57Bl/6 background and are between 6 and 8 weeks of age. Results Membrane cholesterol measurements are elevated in both R117H and ΔF508 mouse nasal epithelium compared to age-matched sibling wt controls demonstrating a genotype correlation to membrane cholesterol detection. Expression of wt CFTR in CF epithelial cells reverts membrane cholesterol to WT levels further demonstrating the impact of CFTR on these processes. In wt epithelial cell, the addition of the CFTR inhibitors, Gly H101 or CFTRinh-172, for 24 h surprisingly results in an initial drop in membrane cholesterol measurement followed by a rebound at 72 h suggesting a feedback mechanism may be driving the increase in membrane cholesterol. De novo cholesterol synthesis contributes to membrane cholesterol accessibility. Conclusions The data in this study suggest that CFTR influences cholesterol trafficking to the plasma membrane, which when depleted, leads to an increase in de novo cholesterol synthesis to restore membrane content.

  13. The Mitochondrial Complex I Activity Is Reduced in Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    OpenAIRE

    Valdivieso, Angel G.; Clauzure, Mariángeles; Marín, María C.; Taminelli, Guillermo L.; Massip Copiz, María M.; Sánchez, Francisco; Schulman, Gustavo; Teiber, María L.; Santa-Coloma, Tomás A.

    2012-01-01

    Cystic fibrosis (CF) is a frequent and lethal autosomal recessive disease. It results from different possible mutations in the CFTR gene, which encodes the CFTR chloride channel. We have previously studied the differential expression of genes in CF and CF corrected cell lines, and found a reduced expression of MTND4 in CF cells. MTND4 is a mitochondrial gene encoding the MTND4 subunit of the mitochondrial Complex I (mCx-I). Since this subunit is essential for the assembly and activity of mCx-...

  14. Distribution of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR Mutations in a Cohort of Patients Residing in Palestine.

    Directory of Open Access Journals (Sweden)

    Issa Siryani

    Full Text Available Cystic fibrosis (CF is an autosomal recessive inherited life-threatening disorder that causes severe damage to the lungs and the digestive system. In Palestine, mutations in the Cystic Fibrosis Transmembrane Conductance Regulator gene (CFTR that contributes to the clinical presentation of CF are ill defined. A cohort of thirty three clinically diagnosed CF patients from twenty one different Palestinian families residing in the central and southern part of Palestine were incorporated in this study. Sweat chloride testing was performed using the Sweat Chek Conductivity Analyzer (ELITECH Group, France to confirm the clinical diagnosis of CF. In addition, nucleic acid from the patients' blood samples was extracted and the CFTR mutation profiles were assessed by direct sequencing of the CFTR 27 exons and the intron-exon boundaries. For patient's DNA samples where no homozygous or two heterozygous CFTR mutations were identified by exon sequencing, DNA samples were tested for deletions or duplications using SALSA MLPA probemix P091-D1 CFTR assay. Sweat chloride testing confirmed the clinical diagnosis of CF in those patients. All patients had NaCl conductivity >60 mmol/l. In addition, nine different CFTR mutations were identified in all 21 different families evaluated. These mutations were c.1393-1G>A, F508del, W1282X, G85E, c.313delA, N1303K, deletion exons 17a-17b-18, deletion exons 17a-17b and Q1100P. c.1393-1G>A was shown to be the most frequent occurring mutation among tested families. We have profiled the underling mutations in the CFTR gene of a cohort of 21 different families affected by CF. Unlike other studies from the Arab countries where F508del was reported to be the most common mutation, in southern/central Palestine, the c.1393-1G>A appeared to be the most common. Further studies are needed per sample size and geographic distribution to account for other possible CFTR genetic alterations and their frequencies. Genotype

  15. Three charged amino acids in extracellular loop 1 are involved in maintaining the outer pore architecture of CFTR

    OpenAIRE

    Cui, Guiying; Rahman, Kazi S.; Infield, Daniel T.; Kuang, Christopher; Prince, Chengyu Z.; McCarty, Nael A.

    2014-01-01

    The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) bears six extracellular loops (ECL1–6); ECL1 is the site of several mutations associated with CF. Mutation R117H has been reported to reduce current amplitude, whereas D110H, E116K, and R117C/L/P may impair channel stability. We hypothesized that these amino acids might not be directly involved in ion conduction and permeation but may contribute to stabilizing the outer vestibule architecture in CFTR. We used cRNA injected oo...

  16. Estudi comparatiu de l'estructura del gen "Adh" a vàries espècies de "Drosophila"

    OpenAIRE

    Marfany i Nadal, Gemma

    1991-01-01

    [cat] S'ha caracterizat l'estructura de la regió genòmica del gen "Adh" a quatre espècies del gènere "Drosophila", pertanyents al subgrup "obscura": "D. ambigua", "D. subobscura", "D. madeirensis" i "D. guanche" amb els objectius d'analitzar l'organització i l'evolució d'aquesta regió genòmica dins d'aquest gènere i clarificar les relacions filogenètiques d'aquestes espècies. Per assolir aquestes fites, es van construir llibreries genòmiques de cada espècie que es van crivellar amb una sonda ...

  17. Preserving Accuracy in GenBank

    DEFF Research Database (Denmark)

    Bidartondo, M.I.; Bruns, T. D.; Blackwell, M.;

    2008-01-01

    GenBank, the public repository for nucleotide and protein sequences, is a critical resource for molecular biology, evolutionary biology, and ecology. While some attention has been drawn to sequence errors (1), common annotation errors also reduce the value of this database. In fact, for organisms...... such as fungi, which are notoriously difficult to identify, up to 20% of DNA sequence records may have erroneous lineage designations in GenBank (2). Gene function annotation in protein sequence databases is similarly error-prone (3, 4). Because identity and function of new sequences are often...... unsustainable over the long term as authors eventually leave the field. Although it is possible to link third-party databases to GenBank records, this is a short-term solution that has little guarantee of permanence. Similarly, the current third-party annotation option in GenBank (TPA) complicates rather than...

  18. CFTR and cystic fibrosis%CFTR与囊性纤维化

    Institute of Scientific and Technical Information of China (English)

    王瑞; 李学军

    2006-01-01

    囊性纤维化跨膜传导调节因子(CFTR)是一种cAMP激活的ATP门控性氯离子通道,表达于气道,消化道和生殖道上皮细胞的顶部质膜中.囊性纤维化(CF)是白人中最常见的遗传性疾病之一,由CFTR基因突变造成.对CFTR基因的破译使人们进一步了解CF的发病机制,并为该疾病的诊断提供了新的线索.

  19. Development of ARMS PCR tests for detection of common CFTR gene mutations

    OpenAIRE

    Livshits L. A.; Pampukha V. M.; Soloviov O. O.

    2010-01-01

    Aim. To develop diagnostic assays, based on the amplification refractory mutation system (ARMS) principle, for detection of common mutations in the CFTR gene using two approaches: standard PCR with further gel-electrophoresis and Real-Time PCR with SYBR Green. Materials. For this study we have chosen the following mutations: dF508, W1282X, R117H, 621 + 1G > T, 2143delT with the frequencies in Ukraine: dF508 – 43.3 %; 2143delT – 1.38 %; W1282X – 1.1 %; R117H, 621 + 1G > T – < 0.6 %. For the de...

  20. The zebrafish Kupffer's vesicle as a model system for the molecular mechanisms by which the lack of Polycystin-2 leads to stimulation of CFTR

    Directory of Open Access Journals (Sweden)

    Mónica Roxo-Rosa

    2015-11-01

    Full Text Available In autosomal dominant polycystic kidney disease (ADPKD, cyst inflation and continuous enlargement are associated with marked transepithelial ion and fluid secretion into the cyst lumen via cystic fibrosis transmembrane conductance regulator (CFTR. Indeed, the inhibition or degradation of CFTR prevents the fluid accumulation within cysts. The in vivo mechanisms by which the lack of Polycystin-2 leads to CFTR stimulation are an outstanding challenge in ADPKD research and may bring important biomarkers for the disease. However, hampering their study, the available ADPKD in vitro cellular models lack the three-dimensional architecture of renal cysts and the ADPKD mouse models offer limited access for live-imaging experiments in embryonic kidneys. Here, we tested the zebrafish Kupffer's vesicle (KV as an alternative model-organ. KV is a fluid-filled vesicular organ, lined by epithelial cells that express both CFTR and Polycystin-2 endogenously, being each of them easily knocked-down. Our data on the intracellular distribution of Polycystin-2 support its involvement in the KV fluid-flow induced Ca2+-signalling. Mirroring kidney cysts, the KV lumen inflation is dependent on CFTR activity and, as we clearly show, the knockdown of Polycystin-2 results in larger KV lumens through overstimulation of CFTR. In conclusion, we propose the zebrafish KV as a model organ to study the renal cyst inflation. Favouring its use, KV volume can be easily determined by in vivo imaging offering a live readout for screening compounds and genes that may prevent cyst enlargement through CFTR inhibition.

  1. Development of rAAV2-CFTR: History of the First rAAV Vector Product to be Used in Humans.

    Science.gov (United States)

    Loring, Heather S; ElMallah, Mai K; Flotte, Terence R

    2016-04-01

    The first human gene therapy trials using recombinant adeno-associated virus (rAAV) vectors were performed in cystic fibrosis (CF) patients. Over 100 CF patients were enrolled in 5 separate trials of rAAV2-CFTR administration via nasal, endobronchial, maxillary sinus, and aerosol delivery. Recombinant AAV vectors were designed to deliver the CF transmembrane regulator (CFTR) gene and correct the basic CFTR defect by restoring chloride transport and reverting the upregulation of proinflammatory cytokines. However, vector DNA expression was limited in duration because of the low incidence of integration and natural airway epithelium turnover. In addition, repeated administration of AAV-CFTR vector resulted in a humoral immune response that prevented effective gene transfer from subsequent doses of vector. AAV serotype 2 was used in human trials before the comparison with other serotypes and determination that serotypes 1 and 5 not only possess higher tropism for the airway epithelium, but also are capable of bypassing the binding and trafficking processes-both were important hindrances to the effectiveness of rAAV2. Although rAAV-CFTR gene therapy does not appear likely to supplant newer small-molecule CFTR modulators in the near future, early work with rAAV-CFTR provided an important foundation for later use of rAAV in humans. PMID:26895204

  2. Measurements of CFTR-mediated Cl- secretion in human rectal biopsies constitute a robust biomarker for Cystic Fibrosis diagnosis and prognosis.

    Directory of Open Access Journals (Sweden)

    Marisa Sousa

    Full Text Available BACKGROUND: Cystic Fibrosis (CF is caused by ∼1,900 mutations in the CF transmembrane conductance regulator (CFTR gene encoding for a cAMP-regulated chloride (Cl(- channel expressed in several epithelia. Clinical features are dominated by respiratory symptoms, but there is variable organ involvement thus causing diagnostic dilemmas, especially for non-classic cases. METHODOLOGY/PRINCIPAL FINDINGS: To further establish measurement of CFTR function as a sensitive and robust biomarker for diagnosis and prognosis of CF, we herein assessed cholinergic and cAMP-CFTR-mediated Cl(- secretion in 524 freshly excised rectal biopsies from 118 individuals, including patients with confirmed CF clinical diagnosis (n=51, individuals with clinical CF suspicion (n=49 and age-matched non-CF controls (n=18. Conclusive measurements were obtained for 96% of cases. Patients with "Classic CF", presenting earlier onset of symptoms, pancreatic insufficiency, severe lung disease and low Shwachman-Kulczycki scores were found to lack CFTR-mediated Cl(- secretion (<5%. Individuals with milder CF disease presented residual CFTR-mediated Cl(- secretion (10-57% and non-CF controls show CFTR-mediated Cl(- secretion ≥ 30-35% and data evidenced good correlations with various clinical parameters. Finally, comparison of these values with those in "CF suspicion" individuals allowed to confirm CF in 16/49 individuals (33% and exclude it in 28/49 (57%. Statistical discriminant analyses showed that colonic measurements of CFTR-mediated Cl(- secretion are the best discriminator among Classic/Non-Classic CF and non-CF groups. CONCLUSIONS/SIGNIFICANCE: Determination of CFTR-mediated Cl(- secretion in rectal biopsies is demonstrated here to be a sensitive, reproducible and robust predictive biomarker for the diagnosis and prognosis of CF. The method also has very high potential for (pre-clinical trials of CFTR-modulator therapies.

  3. Targeted Integration of a Super-Exon into the CFTR Locus Leads to Functional Correction of a Cystic Fibrosis Cell Line Model.

    Science.gov (United States)

    Bednarski, Christien; Tomczak, Katja; Vom Hövel, Beate; Weber, Wolf-Michael; Cathomen, Toni

    2016-01-01

    In vitro disease models have enabled insights into the pathophysiology of human disease as well as the functional evaluation of new therapies, such as novel genome engineering strategies. In the context of cystic fibrosis (CF), various cellular disease models have been established in recent years, including organoids based on induced pluripotent stem cell technology that allowed for functional readouts of CFTR activity. Yet, many of these in vitro CF models require complex and expensive culturing protocols that are difficult to implement and may not be amenable for high throughput screens. Here, we show that a simple cellular CF disease model based on the bronchial epithelial ΔF508 cell line CFBE41o- can be used to validate functional CFTR correction. We used an engineered nuclease to target the integration of a super-exon, encompassing the sequences of CFTR exons 11 to 27, into exon 11 and re-activated endogenous CFTR expression by treating CFBE41o- cells with a demethylating agent. We demonstrate that the integration of this super-exon resulted in expression of a corrected mRNA from the endogenous CFTR promoter and used short-circuit current measurements in Ussing chambers to corroborate restored ion transport of the repaired CFTR channels. In conclusion, this study proves that the targeted integration of a large super-exon in CFTR exon 11 leads to functional correction of CFTR, suggesting that this strategy can be used to functionally correct all CFTR mutations located downstream of the 5' end of exon 11. PMID:27526025

  4. Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

    OpenAIRE

    Wang, Chi; Protasevich, Irina; Yang, Zhengrong; Seehausen, Derek; Skalak, Timothy; Zhao, Xun; Atwell, Shane; Spencer Emtage, J; Wetmore, Diana R.; Brouillette, Christie G; Hunt, John F.

    2010-01-01

    The lethal genetic disease cystic fibrosis is caused predominantly by in-frame deletion of phenylalanine 508 in the cystic fibrosis transmembrane conductance regulator (CFTR). F508 is located in the first nucleotide-binding domain (NBD1) of CFTR, which functions as an ATP-gated chloride channel on the cell surface. The F508del mutation blocks CFTR export to the surface due to aberrant retention in the endoplasmic reticulum. While it was assumed that F508del interferes with NBD1 folding, bioph...

  5. Novel residues lining the CFTR chloride channel pore identified by functional modification of introduced cysteines.

    Science.gov (United States)

    Fatehi, Mohammad; Linsdell, Paul

    2009-04-01

    Substituted cysteine accessibility mutagenesis (SCAM) has been used widely to identify pore-lining amino acid side chains in ion channel proteins. However, functional effects on permeation and gating can be difficult to separate, leading to uncertainty concerning the location of reactive cysteine side chains. We have combined SCAM with investigation of the charge-dependent effects of methanethiosulfonate (MTS) reagents on the functional permeation properties of cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels. We find that cysteines substituted for seven out of 21 continuous amino acids in the eleventh and twelfth transmembrane (TM) regions can be modified by external application of positively charged [2-(trimethylammonium)ethyl] MTS bromide (MTSET) and negatively charged sodium [2-sulfonatoethyl] MTS (MTSES). Modification of these cysteines leads to changes in the open channel current-voltage relationship at both the macroscopic and single-channel current levels that reflect specific, charge-dependent effects on the rate of Cl(-) permeation through the channel from the external solution. This approach therefore identifies amino acid side chains that lie within the permeation pathway. Cysteine mutagenesis of pore-lining residues also affects intrapore anion binding and anion selectivity, giving more information regarding the roles of these residues. Our results demonstrate a straightforward method of screening for pore-lining amino acids in ion channels. We suggest that TM11 contributes to the CFTR pore and that the extracellular loop between TMs 11 and 12 lies close to the outer mouth of the pore. PMID:19381710

  6. Identification of Herbal Compound lmperatorin with Adverse Effects on ANO1 and CFTR Chloride Channels

    Institute of Scientific and Technical Information of China (English)

    HAO Feng; YI Fei; ZHANG Di; NING Yan; SU Wei-heng; FENG Xue-chao; YANG Hong; MA Tong-hui

    2011-01-01

    Calcium-activated chloride channels(CaCCs) are the crucial regulators of transepithelial fluid secretion,smooth muscle contraction and sensory transduction. Recently, compelling evidence has indicated that TMEM 16A(ANO 1 or anoctamin-i ) is a bona fide calcium-acvtivated chloride channel. A few small molecule CaCCs regulators are available for functional and therapeutic studies. We screened 126 natural compounds from Chinese herbs. Screening was performed with an iodide influx assay in Fischer rat thyroid epithelial cells to coexpress ANOI and an iodide-sensitive fluorescent indicator(EYFP-HI48Q/I152L). lmperatorin, a coumarin compound, was identifled to inhibit ANOl-mediated chloride transport activated by multiple calcium-elevating agonists. The inhibitory effect is dose-dependent with IC50 ~14.63 μmol/L. Interestingly, imperatorin activated CFTR chloride channel with EC50 ~35.52 μmol/L. The adverse effects of imperatorin on CaCC and CFTR chloride channels will make it useful in pharmacological dissection of chloride transport in airway and intestinal epithelium. Further studies are required to evaluate the therapeutic effects of imperatorin on hypertension, asthma and certain tumors.

  7. Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in allergic bronchopulmonary aspergillosis

    Energy Technology Data Exchange (ETDEWEB)

    Miller, P.W.; Hamosh, A.; Macek, M. Jr. [John Hopkins Univ. School of Medicine, Baltimore, MD (United States)] [and others

    1996-07-01

    The etiology of allergic bronchopulmonary aspergillosis (ABPA) is not well understood. A clinical phenotype resembling the pulmonary disease seen in cystic fibrosis (CF) patients can occur in some individuals with ABPA. Reports of familial occurrence of ABPA and increased incidence in CF patients suggest a possible genetic basis for the disease. To test this possibility, the entire coding region of the cystic fibrosis transmembrane regulator (CFTR) gene was analyzed in 11 individuals who met strict criteria for the diagnosis of ABPA and had normal sweat electrolytes ({le}40 mmol/liter). One patient carried two CF mutations ({Delta}F508/R347H), and five were found to carry one CF mutation (four {Delta}F508; one R117H). The frequency of the {Delta}F508 mutation in patients with ABPA was significantly higher than in 53 Caucasian patients with chronic bronchitis (P < .0003) and the general population (P < .003). These results suggest that CFTR plays an etiologic role in a subset of ABPA patients. 54 refs., 2 tabs.

  8. Unleashing Gen Y: Marketing Mars to Millennials

    Science.gov (United States)

    Leahy, Bart D.; Hidalgo, Loretta; Kloberdanz, Cassie

    2007-01-01

    Space advocates need to engage Generation Y (born 1977-1999).This outreach is necessary to recruit the next generation of scientists and engineers to explore Mars. Space advocates in the non-profit, private, and government sectors need to use a combination of technical communication, marketing, and politics, to develop messages that resonate with Gen Y. Until now, space messages have been generated by and for college-educated white males; Gen Y is much more diverse, including as much as one third minorities. Young women, too, need to be reached. My research has shown that messages emphasizing technology, fun, humor, and opportunity are the best means of reaching the Gen Y audience of 60 million (US population is 300 million). The important things space advocates must avoid are talking down to this generation, making false promises, or expecting them to "wait their turn" before they can participate. This is the MTV generation! We need to find ways of engaging Gen Y now to build a future where human beings can live and work on the planet Mars. In addition to the messages themselves, advocates need to keep up with Gen Y' s social networking and use of iPods, cell phones, and the Internet. NASA and space advocacy groups can use these tools for "viral marketing," where young people share targeted space-related information via cell phones or the Internet because they like it. Overall, Gen Y is a socially dynamic and media-savvy group; advocates' space messages need to be sincere, creative, and placed in locations where Gen Y lives. Mars messages must be memorable!

  9. Cystic fibrosis and the role of gastrointestinal outcome measures in the new era of therapeutic CFTR modulation

    NARCIS (Netherlands)

    Bodewes, Frank A J A; Verkade, Henkjan J; Taminiau, Jan A J M; Borowitz, Drucy; Wilschanski, Michael

    2015-01-01

    With the development of new drugs that directly affect CFTR protein function, clinical trials are being designed or initiated for a growing number of patients with cystic fibrosis. The currently available and accepted clinical endpoints, FEV1 and BMI, have limitations. The aim of this report is to d

  10. Heterogeneity of phenotype in two cystic fibrosis patients homozygous for the CFTR exon 11 mutation G551D.

    OpenAIRE

    Parad, R B

    1996-01-01

    In the heterozygous state, the cystic fibrosis transmembrane conductance regulator (CFTR) exon 11 mutation G551D has been described as "severe," causing pancreatic insufficiency. Two cystic fibrosis (CF) patients homozygous for this mutation showed a mild rather than severe pancreatic phenotype and a variable pulmonary phenotype.

  11. Ursodeoxycholate modulates bile flow and bile salt pool independently from the cystic fibrosis transmembrane regulator (Cftr) in mice

    NARCIS (Netherlands)

    Bodewes, Frank A. J. A.; Wouthuyzen-Bakker, Marjan; Bijvelds, Marcel J.; Havinga, Rick; de Jonge, Hugo R.; Verkade, Henkjan J.

    2012-01-01

    Bodewes FAJA, Wouthuyzen-Bakker M, Bijvelds MJ, Havinga R, de Jonge HR, Verkade HJ. Ursodeoxycholate modulates bile flow and bile salt pool independently from the cystic fibrosis transmembrane regulator (Cftr) in mice. Am J Physiol Gastrointest Liver Physiol 302: G1035-G1042, 2012. First published F

  12. Rescue of Murine F508del CFTR Activity in Native Intestine by Low Temperature and Proteasome Inhibitors

    NARCIS (Netherlands)

    M. Wilke (Martina); A.G. Bot (Alice); H. Jorna (Huub); B.J. Scholte (Bob); H.R. de Jonge (Hugo)

    2012-01-01

    textabstractMost patients with Cystic Fibrosis (CF) carry at least one allele with the F508del mutation, resulting in a CFTR chloride channel protein with a processing, gating and stability defect, but with substantial residual activity when correctly sorted to the apical membranes of epithelial cel

  13. Development of allele-specific multiplex PCR to determine the length of poly-T in intron 8 of CFTR

    Directory of Open Access Journals (Sweden)

    Neng Chen

    2014-07-01

    Full Text Available Cystic fibrosis transmembrane conductance regulator (CFTR gene mutation analysis has been implemented for Cystic Fibrosis (CF carrier screening, and molecular diagnosis of CF and congenital bilateral absence of the vas deferens (CBAVD. Although poly-T allele analysis in intron 8 of CFTR is required when a patient is positive for R117H, it is not recommended for routine carrier screening. Therefore, commercial kits for CFTR mutation analysis were designed either to mask the poly-T allele results, unless a patient is R117H positive, or to have the poly-T analysis as a standalone reflex test using the same commercial platform. There are other standalone assays developed to detect poly-T alleles, such as heteroduplex analysis, High Resolution Melting (HRM curve analysis, allele-specific PCR (AS-PCR and Sanger sequencing. In this report, we developed a simple and easy-to-implement multiplex AS-PCR assay using unlabeled standard length primers, which can be used as a reflex or standalone test for CFTR poly-T track analysis. Out of 115 human gDNA samples tested, results from our new AS-PCR matched to the previous known poly-T results or results from Sanger sequencing.

  14. Cystic fibrosis transmembrane conductance regulator (CFTR allelic variants relate to shifts in faecal microbiota of cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Serena Schippa

    Full Text Available INTRODUCTION: In this study we investigated the effects of the Cystic Fibrosis Transmembrane conductance Regulator (CFTR gene variants on the composition of faecal microbiota, in patients affected by Cystic Fibrosis (CF. CFTR mutations (F508del is the most common lead to a decreased secretion of chloride/water, and to mucus sticky secretions, in pancreas, respiratory and gastrointestinal tracts. Intestinal manifestations are underestimated in CF, leading to ileum meconium at birth, or small bowel bacterial overgrowth in adult age. METHODS: Thirty-six CF patients, fasting and under no-antibiotic treatment, were CFTR genotyped on both alleles. Faecal samples were subjected to molecular microbial profiling through Temporal Temperature Gradient Electrophoresis and species-specific PCR. Ecological parameters and multivariate algorithms were employed to find out if CFTR variants could be related to the microbiota structure. RESULTS: Patients were classified by two different criteria: 1 presence/absence of F508del mutation; 2 disease severity in heterozygous and homozygous F508del patients. We found that homozygous-F508del and severe CF patients exhibited an enhanced dysbiotic faecal microbiota composition, even within the CF cohort itself, with higher biodiversity and evenness. We also found, by species-specific PCR, that potentially harmful species (Escherichia coli and Eubacterium biforme were abundant in homozygous-F508del and severe CF patients, while beneficial species (Faecalibacterium prausnitzii, Bifidobacterium spp., and Eubacterium limosum were reduced. CONCLUSIONS: This is the first report that establishes a link among CFTR variants and shifts in faecal microbiota, opening the way to studies that perceive CF as a 'systemic disease', linking the lung and the gut in a joined axis.

  15. Aislamiento y caracterización de ABG1, un gen esencial del hongo patógeno oportunista Candida Albicans.

    OpenAIRE

    Veses Jiménez, Verónica

    2005-01-01

    RESUMEN Mediante el inmunorrastreo de una genoteca de expresión de Candida albicans con un anticuerpo policlonal específico de la forma micelial de dicho hongo (PAb anti-gt), se ha aislado un nuevo gen de este microorganismo. La represión del gen aislado ocasionó un severo defecto en la morfogénesis de C. albicans, caracterizado por la formación de cadenas de levaduras, de tamaño decreciente en dirección al extremo apical de la cadena, por lo que el gen fue denominado ABG1 (del inglés, alt...

  16. Defective fluid secretion from submucosal glands of nasal turbinates from CFTR-/- and CFTR (ΔF508/ΔF508 pigs.

    Directory of Open Access Journals (Sweden)

    Hyung-Ju Cho

    Full Text Available BACKGROUND: Cystic fibrosis (CF, caused by reduced CFTR function, includes severe sinonasal disease which may predispose to lung disease. Newly developed CF pigs provide models to study the onset of CF pathophysiology. We asked if glands from pig nasal turbinates have secretory responses similar to those of tracheal glands and if CF nasal glands show reduced fluid secretion. METHODOLOGY/PRINCIPAL FINDINGS: Unexpectedly, we found that nasal glands differed from tracheal glands in five ways, being smaller, more numerous (density per airway surface area, more sensitive to carbachol, more sensitive to forskolin, and nonresponsive to Substance P (a potent agonist for pig tracheal glands. Nasal gland fluid secretion from newborn piglets (12 CF and 12 controls in response to agonists was measured using digital imaging of mucus bubbles formed under oil. Secretion rates were significantly reduced in all conditions tested. Fluid secretory rates (Controls vs. CF, in pl/min/gland were as follows: 3 µM forskolin: 9.2±2.2 vs. 0.6±0.3; 1 µM carbachol: 143.5±35.5 vs. 52.2±10.3; 3 µM forskolin + 0.1 µM carbachol: 25.8±5.8 vs. CF 4.5±0.9. We also compared CF(ΔF508/ΔF508 with CFTR(-/- piglets and found significantly greater forskolin-stimulated secretion rates in the ΔF508 vs. the null piglets (1.4±0.8, n = 4 vs. 0.2±0.1, n = 7. An unexpected age effect was also discovered: the ratio of secretion to 3 µM forskolin vs. 1 µM carbachol was ∼4 times greater in adult than in neonatal nasal glands. CONCLUSIONS/SIGNIFICANCE: These findings reveal differences between nasal and tracheal glands, show defective fluid secretion in nasal glands of CF pigs, reveal some spared function in the ΔF508 vs. null piglets, and show unexpected age-dependent differences. Reduced nasal gland fluid secretion may predispose to sinonasal and lung infections.

  17. VARIABILIDAD DEL GEN NUCLEAR G3PDH EN JATROPHA CURCAS L.

    Directory of Open Access Journals (Sweden)

    Sonia Castro Guzmán

    2015-11-01

    Full Text Available Jatrophacurcas L.es una especie nativa de América tropical; en nuestro país se ha venido utilizando principalmente como medicinal y alimenticia desde la época prehispánica. Actualmente el aceite extraído de sus semillas ha adquirido importancia internacional ya que puede ser transformado a biodiesel. El conocimiento que existe sobre la variabilidad genética de la especie es escaso. Un gen utilizado con éxito para el estudio de patrones de variación y origen de la yuca (ManihotesculentaL. y del caco (Theobromacacao L. es el gen nuclearG3pdh(Gliceraldehído3 fosfato deshidrogenasa involucrado en la fotosíntesis. Con base en ello, en este trabajo se exploró la variabilidad del gen G3pdh en individuos deJ. curcasprovenientes de 15 poblaciones de los estados de Veracruz, Campeche, Yucatán y Quintana Roo. Para obtener el gen G3pdh completo de alrededor de 1000 pares de bases, se amplificó utilizando losprimersdiseñados porStrandet al. (1997 y los dosprimersinternos reverse diseñados porOlsenySchaal(1999 para obtener segmentos más cortos, de 600 y 800 pares de bases aproximadamente. Por primera vez se amplificaron aproximadamente 500pby los resultados demuestran que el gen G3pdh es útil para analizar la variabilidad deJ. curcas, y con un importante potencial para evaluar la distribución y evolución de sus poblaciones en México, conocer las relaciones ancestro descendiente a nivel poblacional y explicar las causas de la distribución de los distintoshaplotipos.

  18. Mapas genéticos em plantas

    OpenAIRE

    Carneiro Monalisa Sampaio; Vieira Maria Lucia Carneiro

    2002-01-01

    Ao lado dos projetos de seqüenciamento e das análises do cariótipo pelas técnicas de hibridização in situ, o desenvolvimento de mapas genéticos fundamentados em marcadores de DNA tem propiciado consideráveis avanços à genômica de plantas. Esta revisão aborda as premissas básicas utilizadas para o mapeamento genético e suas principais aplicações, especialmente para o melhoramento vegetal. Fundamentos teóricos sobre segregação, recombinação e ligação são considerados e relacionados à construção...

  19. Clusters of Cl- channels in CFTR-expressing Sf9 cells switch spontaneously between slow and fast gating modes

    DEFF Research Database (Denmark)

    Larsen, Erik Hviid; Price, E. M.; Gabriel, S. E.;

    1996-01-01

    The Sf9 insect Spodoptora frugiperda cell line was used for heterologous expression of the cloned human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA, or the cloned ß-galactosidase gene, using the baculovirus Autographa califonica as the infection vector. Using application of the...... patch-clamp technique, evidence for functional expression of CFTR was obtained according to the following three criteria. Firstly, whole-cell currents recorded 2 days after infection with CFTR revealed a statistically significant increase of membrane conductance, ˜25 times above that of mock......-infected control cells, with the reversal potential of the major current component being governed by the chloride equilibrium potential (E Cl). Secondly, in contrast to uninfected cells and cells infected with ß-galactosidase, the membrane conductance to chloride of CFTR-injected cells was stimulated by cytosolic...

  20. Purinergic regulation of CFTR and Ca2+ -activated Cl- channels and K+ channels in human pancreatic duct epithelium

    DEFF Research Database (Denmark)

    Wang, Jing; Haanes, Kristian A; Novak, Ivana

    2013-01-01

    dependent on intracellular Ca(2+). Apically applied ATP/UTP stimulated CF transmembrane conductance regulator (CFTR) and Ca(2+)-activated Cl(-) (CaCC) channels, which were inhibited by CFTRinh-172 and niflumic acid, respectively. The basolaterally applied ATP stimulated CFTR. In CFPAC-1 cells, which have...... mutated CFTR, basolateral ATP and UTP had negligible effects. In addition to Cl(-) transport in Capan-1 cells, the effects of 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DC-EBIO) and clotrimazole indicated functional expression of the intermediate conductance K(+) channels (IK, KCa3.1). The...... receptors both Cl(-) channels (TMEM16A/ANO1 and CFTR) and K(+) channels (IK). The K(+) channels provide the driving force for Cl(-)-channel-dependent secretion, and luminal ATP provided locally or secreted from acini may potentiate secretory processes. Future strategies in augmenting pancreatic duct...

  1. Thermodynamic study of the native and phosphorylated regulatory domain of the CFTR

    Energy Technology Data Exchange (ETDEWEB)

    Marasini, Carlotta, E-mail: marasini@ge.ibf.cnr.it [Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Via De Marini 6, 16149 Genova (Italy); Galeno, Lauretta; Moran, Oscar [Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Via De Marini 6, 16149 Genova (Italy)

    2012-07-06

    Highlights: Black-Right-Pointing-Pointer CFTR mutations produce cystic fibrosis. Black-Right-Pointing-Pointer Chloride transport depends on the regulatory domain phosphorylation. Black-Right-Pointing-Pointer Regulatory domain is intrinsically disordered. Black-Right-Pointing-Pointer Secondary structure and protein stability change upon phosphorylation. -- Abstract: The regulatory domain (RD) of the cystic fibrosis transmembrane conductance regulator (CFTR), the defective protein in cystic fibrosis, is the region of the channel that regulates the CFTR activity with multiple phosphorylation sites. This domain is an intrinsically disordered protein, characterized by lack of stable or unique tertiary structure. The disordered character of a protein is directly correlated with its function. The flexibility of RD may be important for its regulatory role: the continuous conformational change may be necessary for the progressive phosphorylation, and thus activation, of the channel. However, the lack of a defined and stable structure results in a considerable limitation when trying to in build a unique molecular model for the RD. Moreover, several evidences indicate significant structural differences between the native, non-phosphorylated state, and the multiple phosphorylated state of the protein. The aim of our work is to provide data to describe the conformations and the thermodynamic properties in these two functional states of RD. We have done the circular dichroism (CD) spectra in samples with a different degree of phosphorylation, from the non-phosphorylated state to a bona fide completely phosphorylated state. Analysis of CD spectra showed that the random coil and {beta}-sheets secondary structure decreased with the polypeptide phosphorylation, at expenses of an increase of {alpha}-helix. This observation lead to interpret phosphorylation as a mechanism favoring a more structured state. We also studied the thermal denaturation curves of the protein in the two

  2. Thermodynamic study of the native and phosphorylated regulatory domain of the CFTR

    International Nuclear Information System (INIS)

    Highlights: ► CFTR mutations produce cystic fibrosis. ► Chloride transport depends on the regulatory domain phosphorylation. ► Regulatory domain is intrinsically disordered. ► Secondary structure and protein stability change upon phosphorylation. -- Abstract: The regulatory domain (RD) of the cystic fibrosis transmembrane conductance regulator (CFTR), the defective protein in cystic fibrosis, is the region of the channel that regulates the CFTR activity with multiple phosphorylation sites. This domain is an intrinsically disordered protein, characterized by lack of stable or unique tertiary structure. The disordered character of a protein is directly correlated with its function. The flexibility of RD may be important for its regulatory role: the continuous conformational change may be necessary for the progressive phosphorylation, and thus activation, of the channel. However, the lack of a defined and stable structure results in a considerable limitation when trying to in build a unique molecular model for the RD. Moreover, several evidences indicate significant structural differences between the native, non-phosphorylated state, and the multiple phosphorylated state of the protein. The aim of our work is to provide data to describe the conformations and the thermodynamic properties in these two functional states of RD. We have done the circular dichroism (CD) spectra in samples with a different degree of phosphorylation, from the non-phosphorylated state to a bona fide completely phosphorylated state. Analysis of CD spectra showed that the random coil and β-sheets secondary structure decreased with the polypeptide phosphorylation, at expenses of an increase of α-helix. This observation lead to interpret phosphorylation as a mechanism favoring a more structured state. We also studied the thermal denaturation curves of the protein in the two conditions, monitoring the changes of the mean residue ellipticity measured at 222 nm as a function of temperature

  3. Measurements of CFTR-Mediated Cl− Secretion in Human Rectal Biopsies Constitute a Robust Biomarker for Cystic Fibrosis Diagnosis and Prognosis

    OpenAIRE

    Sousa, Marisa; Servidoni, Maria F; Vinagre, Adriana M.; Ramalho, Anabela S; Bonadia, Luciana C.; Felício, Verónica; Ribeiro, Maria A..; Uliyakina, Inna; Marson A, Fernando; Kmit, Arthur; Cardoso, Silvia R.; Ribeiro, José D; Carmen S. Bertuzzo; Sousa, Lisete; Kunzelmann, Karl

    2012-01-01

    BACKGROUND: Cystic Fibrosis (CF) is caused by ∼1,900 mutations in the CF transmembrane conductance regulator (CFTR) gene encoding for a cAMP-regulated chloride (Cl(-)) channel expressed in several epithelia. Clinical features are dominated by respiratory symptoms, but there is variable organ involvement thus causing diagnostic dilemmas, especially for non-classic cases. METHODOLOGY/PRINCIPAL FINDINGS: To further establish measurement of CFTR function as a sensitive and robust biomarker fo...

  4. Increased efficacy of VX-809 in different cellular systems results from an early stabilization effect of F508del-CFTR.

    Science.gov (United States)

    Farinha, Carlos M; Sousa, Marisa; Canato, Sara; Schmidt, André; Uliyakina, Inna; Amaral, Margarida D

    2015-08-01

    Cystic fibrosis (CF), the most common recessive autosomal disease among Caucasians, is caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR) protein. The most common mutation, F508del, leads to CFTR impaired plasma membrane trafficking. Therapies modulating CFTR basic defect are emerging, such as VX-809, a corrector of F508del-CFTR traffic which just succeeded in a Phase III clinical trial. We recently showed that VX-809 is additive to two other correctors (VRT-325 and compound 4a). Here, we aimed to determine whether the differential rescuing by these compounds results from cell-specific factors or rather from distinct effects at the early biogenesis and/or processing. The rescuing efficiencies of the above three correctors were first compared in different cellular models (primary respiratory cells, cystic fibrosis bronchial epithelial and baby hamster kidney [BHK] cell lines) by functional approaches: micro-Ussing chamber and iodide efflux. Next, biochemical methods (metabolic labeling, pulse-chase and immunoprecipitation) were used to determine their impact on CFTR biogenesis / processing. Functional analyses revealed that VX-809 has the greatest rescuing efficacy and that the relative efficiencies of the three compounds are essentially maintained in all three cellular models tested. Nevertheless, biochemical data show that VX-809 significantly stabilizes F508del-CFTR immature form, an effect that is not observed for C3 nor C4. VX-809 and C3 also significantly increase accumulation of immature CFTR. Our data suggest that VX-809 increases the stability of F508del-CFTR immature form at an early phase of its biogenesis, thus explaining its increased efficacy when inducing its rescue. PMID:26171232

  5. Restoration of R117H CFTR folding and function in human airway cells through combination treatment with VX-809 and VX-770.

    Science.gov (United States)

    Gentzsch, Martina; Ren, Hong Y; Houck, Scott A; Quinney, Nancy L; Cholon, Deborah M; Sopha, Pattarawut; Chaudhry, Imron G; Das, Jhuma; Dokholyan, Nikolay V; Randell, Scott H; Cyr, Douglas M

    2016-09-01

    Cystic fibrosis (CF) is a lethal recessive genetic disease caused primarily by the F508del mutation in the CF transmembrane conductance regulator (CFTR). The potentiator VX-770 was the first CFTR modulator approved by the FDA for treatment of CF patients with the gating mutation G551D. Orkambi is a drug containing VX-770 and corrector VX809 and is approved for treatment of CF patients homozygous for F508del, which has folding and gating defects. At least 30% of CF patients are heterozygous for the F508del mutation with the other allele encoding for one of many different rare CFTR mutations. Treatment of heterozygous F508del patients with VX-809 and VX-770 has had limited success, so it is important to identify heterozygous patients that respond to CFTR modulator therapy. R117H is a more prevalent rare mutation found in over 2,000 CF patients. In this study we investigated the effectiveness of VX-809/VX-770 therapy on restoring CFTR function in human bronchial epithelial (HBE) cells from R117H/F508del CF patients. We found that VX-809 stimulated more CFTR activity in R117H/F508del HBEs than in F508del/F508del HBEs. R117H expressed exclusively in immortalized HBEs exhibited a folding defect, was retained in the ER, and degraded prematurely. VX-809 corrected the R117H folding defect and restored channel function. Because R117 is involved in ion conductance, VX-770 acted additively with VX-809 to restore CFTR function in chronically treated R117H/F508del cells. Although treatment of R117H patients with VX-770 has been approved, our studies indicate that Orkambi may be more beneficial for rescue of CFTR function in these patients. PMID:27402691

  6. Involvement of F1296 and N1303 of CFTR in induced-fit conformational change in response to ATP binding at NBD2

    OpenAIRE

    Szollosi, A; P.; Vergani; Csanady, L.

    2010-01-01

    The chloride ion channel cystic fibrosis transmembrane conductance regulator (CFTR) displays a typical adenosine trisphosphate (ATP)-binding cassette (ABC) protein architecture comprising two transmembrane domains, two intracellular nucleotide-binding domains (NBDs), and a unique intracellular regulatory domain. Once phosphorylated in the regulatory domain, CFTR channels can open and close when supplied with cytosolic ATP. Despite the general agreement that formation of a head-to-tail NBD dim...

  7. Genómica funcional de la elongación transcripcional

    OpenAIRE

    Rodríguez Gil, Alfonso

    2008-01-01

    Los principales objetivos de esta Tesis Doctoral son: Desarrollo de un nuevo método para el estudio de la distribución intragénica de la RNA polimerasa II a escala genómica. Aplicación del método desarrollado al estudio de mutantes afectados en la elongación transcripcional. Identificación de nuevos factores que afectan a la elongación transcripcional

  8. Kontrolle der Expression des UNUSUAL FLORAL ORGANS (UFO) Gens in Arabidopsis thaliana

    OpenAIRE

    Hobe, Martin

    2004-01-01

    Die vorliegende Arbeit befaßt sich mit der Kontrolle des Expressionsmusters des UNUSUAL FLORAL ORGANS (UFO) Gens von Arabidopsis thaliana. UFO wird im Sproß- und Blütenmeristemen aller Entwicklungsstadien der Pflanze exprimiert. In Blütenmeristemen agiert UFO als Kofaktor von LEAFY (LFY) bei der Aktivierung der Organidentitätsgene des zweiten und dritten Wirtels. UFO stellt also einen generellen Faktor der Musterbildung in Meristemen dar. Um regulatorische Gene, die die Expression von UFO bee...

  9. GenBank blastx search result: AK062648 [KOME

    Lifescience Database Archive (English)

    Full Text Available smembrane helix receptor pseudogene, the 3' end of NR5A1 gen...e for nuclear receptor (subfamily 5, group A) member 1, a novel gene and four CpG islands, complete sequence.|PRI PRI 4e-74 +2 ... ...EK6 gene for never in mitosis gene a (NIMA)-related kinase 6, a pseudogene similar to part of neuronal pentraxin II, a seven tran...AK062648 001-105-E04 AL137846.24 Human DNA sequence from clone RP11-101K10 on chrom...osome 9 Contains the PSMB7 gene for proteasome (prosome, macropain) beta type 7 subunit, the 3' end of the N

  10. GenBank blastx search result: AK112000 [KOME

    Lifescience Database Archive (English)

    Full Text Available smembrane helix receptor pseudogene, the 3' end of NR5A1 gen...e for nuclear receptor (subfamily 5, group A) member 1, a novel gene and four CpG islands, complete sequence.|PRI PRI 2e-12 +3 ... ...EK6 gene for never in mitosis gene a (NIMA)-related kinase 6, a pseudogene similar to part of neuronal pentraxin II, a seven tran...AK112000 001-035-B07 AL137846.24 Human DNA sequence from clone RP11-101K10 on chrom...osome 9 Contains the PSMB7 gene for proteasome (prosome, macropain) beta type 7 subunit, the 3' end of the N

  11. GenBank blastx search result: AK058440 [KOME

    Lifescience Database Archive (English)

    Full Text Available smembrane helix receptor pseudogene, the 3' end of NR5A1 gen...e for nuclear receptor (subfamily 5, group A) member 1, a novel gene and four CpG islands, complete sequence.|PRI PRI 2e-59 +1 ... ...EK6 gene for never in mitosis gene a (NIMA)-related kinase 6, a pseudogene similar to part of neuronal pentraxin II, a seven tran...AK058440 001-015-F11 AL137846.24 Human DNA sequence from clone RP11-101K10 on chrom...osome 9 Contains the PSMB7 gene for proteasome (prosome, macropain) beta type 7 subunit, the 3' end of the N

  12. GenBank blastx search result: AK104361 [KOME

    Lifescience Database Archive (English)

    Full Text Available smembrane helix receptor pseudogene, the 3' end of NR5A1 gen...e for nuclear receptor (subfamily 5, group A) member 1, a novel gene and four CpG islands, complete sequence.|PRI PRI 2e-15 +3 ... ...EK6 gene for never in mitosis gene a (NIMA)-related kinase 6, a pseudogene similar to part of neuronal pentraxin II, a seven tran...AK104361 001-035-C04 AL137846.24 Human DNA sequence from clone RP11-101K10 on chrom...osome 9 Contains the PSMB7 gene for proteasome (prosome, macropain) beta type 7 subunit, the 3' end of the N

  13. Genetic and bibliographic information: Abcc9 [GenLibi

    Lifescience Database Archive (English)

    Full Text Available Abcc9 ATP-binding cassette, sub-family C (CFTR/MRP), member 9 rat Hypertension ... (MeSH) Cardiovasc ... lar Diseases (C14) > Vascular Diseases (C14.907) > Hypertension ... (C14.907.489) 04A0394477; 05A0803372 ...

  14. Thermal unfolding studies show the disease causing F508del mutation in CFTR thermodynamically destabilizes nucleotide-binding domain 1.

    Science.gov (United States)

    Protasevich, Irina; Yang, Zhengrong; Wang, Chi; Atwell, Shane; Zhao, Xun; Emtage, Spencer; Wetmore, Diana; Hunt, John F; Brouillette, Christie G

    2010-10-01

    Misfolding and degradation of CFTR is the cause of disease in patients with the most prevalent CFTR mutation, an in-frame deletion of phenylalanine (F508del), located in the first nucleotide-binding domain of human CFTR (hNBD1). Studies of (F508del)CFTR cellular folding suggest that both intra- and inter-domain folding is impaired. (F508del)CFTR is a temperature-sensitive mutant, that is, lowering growth temperature, improves both export, and plasma membrane residence times. Yet, paradoxically, F508del does not alter the fold of isolated hNBD1 nor did it seem to perturb its unfolding transition in previous isothermal chemical denaturation studies. We therefore studied the in vitro thermal unfolding of matched hNBD1 constructs ±F508del to shed light on the defective folding mechanism and the basis for the thermal instability of (F508del)CFTR. Using primarily differential scanning calorimetry (DSC) and circular dichroism, we show for all hNBD1 pairs studied, that F508del lowers the unfolding transition temperature (T(m)) by 6-7°C and that unfolding occurs via a kinetically-controlled, irreversible transition in isolated monomers. A thermal unfolding mechanism is derived from nonlinear least squares fitting of comprehensive DSC data sets. All data are consistent with a simple three-state thermal unfolding mechanism for hNBD1 ± F508del: N(±MgATP) I(T)(±MgATP) → A(T) → (A(T))(n). The equilibrium unfolding to intermediate, I(T), is followed by the rate-determining, irreversible formation of a partially folded, aggregation-prone, monomeric state, A(T), for which aggregation to (A(T))(n) and further unfolding occur with no detectable heat change. Fitted parameters indicate that F508del thermodynamically destabilizes the native state, N, and accelerates the formation of A(T). PMID:20687133

  15. Asociación del gen bola-drb3.2 con el virus de la leucosis bovina (vlb) en ganado criollo hartón del valle

    OpenAIRE

    Posso Terranova, Andrés Mauricio; Muñoz Florez, Jaime Eduardo; Giovambattista, Guillermo; Alvarez Franco, Luz Angela

    2014-01-01

    En cien muestras de ganado criollo hartón del Valle (HV) del Banco de ADN del Laboratorio de genética animal de la Universidad Nacional de Colombia sede Palmira se determinó la presencia del VLB siguiendo la metodología PCR anidado descrita por Beier et al. (2001) y se genotipificaron los animales para el gen DRB3.2* utilizando la metodología de PCR-SBT (Sequence Based Typings) descrita por Takeshima et al. (2009). Se encontró el porcentaje de presencia del virus. Para el gen BoLA-DRB3.2* se ...

  16. Análisis de la variabiblidad genética de Quercus Suber L. mediante marcadores moleculares y su aplicación a la conservación de recursos genéticos

    OpenAIRE

    Jiménez Sancho, María del Pilar

    2000-01-01

    En la presente tesis se realiza el estudio de la variabilidad genética de Quercus suber L. A través de dos tipos de marcadores moleculares: isoenzimas y ADN de cloroplastos. La variación enzimática se estima a partir de 22 poblaciones, mediante 9 loci polimórficos. A partir de las frecuencias alélicas se obtienen diversos parámetros de diversidad genética entre y dentro de poblaciones, que ofrecen valores tipicos de las especies del genero. Se observa una gran diversificación intrapoblacional...

  17. Evolución genómica por diseño molecular de levaduras industriales

    OpenAIRE

    SANI, DANIELE

    2013-01-01

    En esta Tesis Doctoral se propone una alternativa a la coyuntura actual de rechazo social frente al uso de OMGs en la industria agroalimentaria, mediante la demostración y el desarrollo de un nuevo concepto sobre el uso de las técnicas de biología molecular en la obtención de levaduras modificadas genéticamente, el concepto de Evolución Genómica mediante Diseño Molecular. La idea básica de este nuevo concepto es simple y se basa en imitar a la propia naturaleza en su const...

  18. Anomalías y displasias dentarias de origen genético-hereditario Inherited dental abnormalities and dysplasias

    OpenAIRE

    J. Martín-González; B. Sánchez-Domínguez; M.L. Tarilonte-Delgado; L. Castellanos-Cosano; J.M. Llamas-Carreras; F.J. López-Frías; J.J. Segura-Egea

    2012-01-01

    Las alteraciones del desarrollo embriológico de la dentición provocan anomalías y displasias dentarias. Los factores etiopatogénicos implicados en las alteraciones del desarrollo dentario son básicamente dos: genéticos y ambientales. Según la fase del desarrollo en que afecten al órgano del esmalte y a los tejidos dentarios, aparecerán diferentes anomalías y/o displasias dentales. El control genético del desarrollo dentario se lleva a cabo mediante dos procesos: a) control de la histogénesis ...

  19. Los algoritmos genéticos y el método de generación y prueba

    Directory of Open Access Journals (Sweden)

    Luis Roberto Ojeda Ch.

    2011-01-01

    Full Text Available Este trabajo pretende mostrar dos paradigmas de solución de problemas en inteligencia artificial. Los algoritmos genéticos han tomado una posición destacada en los últimos tiempos y el método de generación y prueba, cuyo perfil se aproxima al fundamento de los algoritmos genéticos constituye la forma de diseño de DENDRAL, un producto ampliamente reconocido en IA. Se desea mostrar que ambos paradigmas se ubican naturalmente dentro de las expectativas de la inteligencia artificial.

  20. Los algoritmos genéticos y el método de generación y prueba

    OpenAIRE

    Luis Roberto Ojeda Ch.

    2011-01-01

    Este trabajo pretende mostrar dos paradigmas de solución de problemas en inteligencia artificial. Los algoritmos genéticos han tomado una posición destacada en los últimos tiempos y el método de generación y prueba, cuyo perfil se aproxima al fundamento de los algoritmos genéticos constituye la forma de diseño de DENDRAL, un producto ampliamente reconocido en IA. Se desea mostrar que ambos paradigmas se ubican naturalmente dentro de las expectativas de la inteligencia artificial.

  1. Frecuencias alélicas y fenotípicas de los marcadores genéticos enzimáticos: Fosfoglucomutasa 1 (FGM 1), Esterasa D (EsD) y Fosfatasa ácida eritrocitaria (FAE) en población residente en la ciudad de Medellín, 1996-1997

    OpenAIRE

    María Victoria Hurtado Ángel; Nora María Giraldo Ramírez; Silvia García Jaramillo; Ofelia Calle Avendaño

    2000-01-01

    Este estudio descriptivo de las enzimas fosfoglucomutasa 1 (fgm 1), esterasa D (EsD) y Fosfatasa ácida eritrocitaria (FAE) como marcadores genéticos polimórficos, se realizó en 145 muestras de sangre, tomadas a igual númerode donantes de los tres principales bancos de sangre de la ciudad, por el método de electroforesis convencional, con sistema de refrigeración, en gel de agarosa. La probabilidad de identificación para cada uno de los marcadores fue buena; la mejor fue la subtipificación de ...

  2. EFFECTS OF MONOCARBOXYLIC ACID DERIVATIVES ON CARDIAC VENTRICULAR CFTR Cl-CHANNELS IN GUINEA PIG%单羧酸类Cl-通道阻断剂对心室肌CFTR Cl-通道的影响

    Institute of Scientific and Technical Information of China (English)

    周士胜; 臧益民

    1999-01-01

    本文采用全细胞膜片箝与细胞内灌注技术,观察了单羧酸类 Cl-通道阻断剂对豚鼠心室肌囊性纤维变性膜透性调节蛋白(CFTR)Cl-电流的影响,细胞外9-AC以可逆方式增强异丙肾上腺素(ISO)激发的CFTR Cl-的外向电流成分,5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB)和二苯胺羧酸(DPC)对ISO发的CFTR Cl-电流的作用呈现先增强后抑制的双相效应.细胞内NPPB表现为增强ISO激发作用.结果表明,单羧酸类Cl-通道阻断剂在细胞上有不同的作用位点,该类药物作用效果的差异可能与此有关.

  3. Endocytic Sorting of CFTR variants Monitored by Single Cell Fluorescence Ratio Image Analysis (FRIA) in Living Cells

    Science.gov (United States)

    Barriere, H.; Apaja, P.; Okiyoneda, T.; Lukacs, G. L.

    2016-01-01

    Summary The wild-type CFTR channel undergoes constitutive internalization and recycling at the plasma membrane. This process is initiated by the recognition of the Tyr- and di-Leu-based endocytic motifs of CFTR by the AP-2 adaptor complex, leading to the formation of clathrin-coated vesicles and the channel delivery to sorting/recycling endosomes. Accumulating evidence suggests that conformationally defective mutant CFTRs (e.g. rescued ΔF508 and glycosylation-deficient channel) are unstable at the plasma membrane and undergo augmented ubiquitination in post-Golgi compartments. Ubiquitination conceivably accounts for the metabolic instability at cell surface by provoking accelerated internalization, as well as rerouting the channel from recycling towards lysosomal degradation. We developed an in vivo fluorescence ratio imaging assay (FRIA) that in concert with genetic manipulation can be utilized to establish the post-endocytic fate and sorting determinants of mutant CFTRs. PMID:21594793

  4. Restoration of NBD1 thermal stability is necessary and sufficient to correct ∆F508 CFTR folding and assembly.

    Science.gov (United States)

    He, Lihua; Aleksandrov, Andrei A; An, Jianli; Cui, Liying; Yang, Zhengrong; Brouillette, Christie G; Riordan, John R

    2015-01-16

    Cystic fibrosis transmembrane conductance regulator (CFTR) (ABCC7), unique among ABC exporters as an ion channel, regulates ion and fluid transport in epithelial tissues. Loss of function due to mutations in the cftr gene causes cystic fibrosis. The most common cystic-fibrosis-causing mutation, the deletion of F508 (ΔF508) from the first nucleotide binding domain (NBD1) of CFTR, results in misfolding of the protein and clearance by cellular quality control systems. The ΔF508 mutation has two major impacts on CFTR: reduced thermal stability of NBD1 and disruption of its interface with membrane-spanning domains (MSDs). It is unknown if these two defects are independent and need to be targeted separately. To address this question, we varied the extent of stabilization of NBD1 using different second-site mutations and NBD1 binding small molecules with or without NBD1/MSD interface mutation. Combinations of different NBD1 changes had additive corrective effects on ∆F508 maturation that correlated with their ability to increase NBD1 thermostability. These effects were much larger than those caused by interface modification alone and accounted for most of the correction achieved by modifying both the domain and the interface. Thus, NBD1 stabilization plays a dominant role in overcoming the ΔF508 defect. Furthermore, the dual target approach resulted in a locked-open ion channel that was constitutively active in the absence of the normally obligatory dependence on phosphorylation by protein kinase A. Thus, simultaneous targeting of both the domain and the interface, as well as being non-essential for correction of biogenesis, may disrupt normal regulation of channel function. PMID:25083918

  5. Restoration of NBD1 thermal stability is necessary and sufficient to correct ΔF508 CFTR folding and assembly

    Science.gov (United States)

    He, Lihua; Aleksandrov, Andrei A; An, Jianli; Cui, Liying; Yang, Zhengrong; Brouillette, Christie G.; Riordan, John R

    2015-01-01

    CFTR (ABCC7), unique among ABC exporters as an ion channel, regulates ion and fluid transport in epithelial tissues. Loss of function due to mutations in the cftr gene causes cystic fibrosis (CF). The most common CF-causing mutation, the deletion of F508 (ΔF508) from the first nucleotide binding domain (NBD1) of CFTR, results in misfolding of the protein and clearance by cellular quality control systems. The ΔF508 mutation has two major impacts on CFTR: reduced thermal stability of NBD1 and disruption of its interface with membrane-spanning domains (MSDs). It is unknown if these two defects are independent and need to be targeted separately. To address this question we varied the extent of stabilization of NBD1 using different second site mutations and NBD1 binding small molecules with or without NBD1/MSD interface mutation. Combinations of different NBD1 changes had additive corrective effects on ΔF508 maturation that correlated with their ability to increase NBD1 thermostability. These effects were much larger than those caused by interface modification alone and accounted for most of the correction achieved by modifying both the domain and the interface. Thus, NBD1 stabilization plays a dominant role in overcoming the ΔF508 defect. Furthermore, the dual target approach resulted in a locked-open ion channel that was constitutively active in the absence of the normally obligatory dependence on phosphorylation by protein kinase A. Thus, simultaneous targeting of both the domain and the interface, as well as being non-essential for correction of biogenesis, may disrupt normal regulation of channel function. PMID:25083918

  6. Analysis of the CFTR gene in Venezuelan cystic fibrosis patients, identification of six novel cystic fibrosis-causing genetic variants

    OpenAIRE

    Sánchez K; de Mendonca E; Matute X; Chaustre I; Villalón M; Takiff H

    2016-01-01

    Karen Sánchez,1 Elizabeth de Mendonca,1 Xiorama Matute,2 Ismenia Chaustre,2 Marlene Villalón,3 Howard Takiff4 1Unit of Genetic and Forensic Studies, Venezuelan Institute for Scientific Research (IVIC), 2Hospital JM de los Ríos, 3Hospital José Ignacio Baldo, Algodonal, National Reference Unit, 4Laboratory of Molecular Genetics, Venezuelan Institute for Scientific Research (IVIC), Caracas, Venezuela. Abstract: The mutations in the CFTR gene found in patients with cy...

  7. Analysis of the CFTR gene in Venezuelan cystic fibrosis patients, identification of six novel cystic fibrosis-causing genetic variants

    OpenAIRE

    Sánchez, Karen

    2016-01-01

    Karen Sánchez,1 Elizabeth de Mendonca,1 Xiorama Matute,2 Ismenia Chaustre,2 Marlene Villalón,3 Howard Takiff4 1Unit of Genetic and Forensic Studies, Venezuelan Institute for Scientific Research (IVIC), 2Hospital JM de los Ríos, 3Hospital José Ignacio Baldo, Algodonal, National Reference Unit, 4Laboratory of Molecular Genetics, Venezuelan Institute for Scientific Research (IVIC), Caracas, Venezuela. Abstract: The mutations in the CFTR gene found in ...

  8. Frequency of CFTR, SPINK1, and Cathepsin B Gene Mutation in North Indian Population: Connections between Genetics and Clinical Data

    OpenAIRE

    Shweta Singh; Gourdas Choudhuri; Sarita Agarwal

    2014-01-01

    Objectives. Genetic mutations and polymorphisms have been correlated with chronic pancreatitis (CP). This study aims to investigate the association of genetic variants of cystic fibrosis transmembrane conductance regulator (CFTR) and serine protease inhibitor Kazal type 1 (SPINK-1) genes and Cathepsin B gene polymorphisms with CP and to associate genetic backgrounds with clinical phenotypes. Methods. 150 CP patients and 150 normal controls were enrolled consecutively. We analyzed SPINK-1 N34S...

  9. Detection of Five Common CFTR Mutations by Rapid-Cycle Real-Time Amplification Refractory Mutation System PCR

    OpenAIRE

    Dempsey, Eugene; Barton, Davis; Ryan, Fergus X.

    2004-01-01

    Cystic fibrosis is the most common autosomal recessive disease in Caucasian populations and has a carrier frequency of 1 in 25 (1 ). The gene involved codes for the cystic fibrosis transmembrane conductance regulator (CFTR), a membrane-associated protein involved in ion transport across the plasma membrane of epithelial cells. To date more than 1000 mutations have been described in this gene, and most are rare (2 ). By focusing on five common mutations it is possible to detect the diseasecaus...

  10. The Novel CFTR Mutation A457P in a Male with a Delayed Diagnosis of Cystic Fibrosis

    Directory of Open Access Journals (Sweden)

    Kate H. Cole

    2011-01-01

    Full Text Available Cystic fibrosis (CF is an autosomal recessive disease that may be caused by more than 1000 different mutations in the cystic fibrosis transmembrane conductance regulator (CFTR gene. We describe the case of a CF patient who was initially diagnosed at 16 years of age after presenting with mild respiratory compromise and pancreatic sufficiency. When genetic testing was first performed using a CF mutation panel, only a single F508del CFTR allele was identified. We subsequently performed testing, which revealed a previously unreported mutation: A457P (p.Ala457Pro, c.1369G>C. The patient's clinical course through adulthood is described, and genotype-phenotype correlation is discussed. The A457P mutation appears to confer a relatively mild phenotype, as is usually observed with CFTR class IV–VI defects. With the advent of more comprehensive and widely available genetic testing techniques, identification of CF genotypes in patients with milder disease variants may help stratify patients for targeted therapy and prevent late complications of the disease.

  11. Genética de poplaciones de Atta sexdens rubropilosa (Hymenoptera: Formicidae) POPULATION GENETICS OF Atta sexdens rubropilosa (HYMENOPTERA: FORMICIDAE)

    OpenAIRE

    LIRIANA BELIZÁRIO CANTAGALLI; CLAUDETE APARECIDA MANGOLIN; MARIA CLAUDIA COLLA RUVOLO TAKASUSUKI

    2013-01-01

    La variabilidad genética de las hormigas Atta sexdens rubropilosa colectadas en cinco lugares distintos de Brasil fueron evaluados por la técnica PCR-RAPD. Un total de 15 primers produjeron 148 fragmentos, de los cuales 123 fueron polimórficos, lo que corresponden al 83,11 %. La estimación de la diversidad genética por el índice de Shannon fue 0,3836 y el desviación estándar fue de ± 0,2335. Estos valores demuestran una alta diversidad genética. El valor de G ST fue 0,2372 y ΦPT = 0,184 l...

  12. Diagnostico genético prenatal y aborto. Dos cuestiones de eugenesia y discriminación

    OpenAIRE

    FABIOLA VILLELA CORTÉS; Jorge E. Linares Salgado

    2012-01-01

    Los avances en genética seguidos de las nuevas tecnologías en la detección temprana de afecciones genéticas conllevan dilemas bioéticos sobre el uso adecuado de estas técnicas, la información que se le da a la mujer embarazada y la decisión que ella tomará al recibirla. Detectar a tiempo anomalías genéticas permite, en algunas ocasiones, el inicio de un tratamiento adecuado que permita que el niño por nacer no desarrolle una enfermedad discapacitante, como el caso de la fenilcetonuria, o una ...

  13. Structure of wild type and mutant F508del CFTR: A small-angle X-ray scattering study of the protein-detergent complexes.

    Science.gov (United States)

    Pollock, Naomi L; Satriano, Letizia; Zegarra-Moran, Olga; Ford, Robert C; Moran, Oscar

    2016-04-01

    CFTR is an anionic channel expressed in epithelia whose mutations cause cystic fibrosis. Wild (WT) and mutated (F508del) types were over-expressed in yeast, solubilised in the detergent LPG-14 and purified. The detergent-CFTR complexes were studied by SAXS techniques using a solvent of variable density. The final result of the study is the numerical value of a set of parameters: molecular mass, volume and radius of gyration, average electron density and second moment of the electron density fluctuations inside the particles. It is also shown that in the complex the centres of gravity of CFTR and of the detergent are displaced relative to each other. The analysis of these parameters led to the determination of the size and shape of the volumes occupied by protein and by detergent in the complex. WT-CFTR to be an elongated molecule (maximum diameter ∼12.4nm) which spans a flat detergent micelle. The distance distribution function, P(r) confirms that the WT-CFTR is elongated and with an inhomogeneous electronic density. The F508del-CFTR molecule is also elongated (maximum diameter ∼13.2nm), but the associated detergent micelle hides a larger surface, plausibly related to an increased exposure of hydrophobic portions of the mutated protein. The corresponding P(r) is consistent with the presence of well defined domains, probably linked by flexible regions. These differences suggest that the full-length mutant F508del-CFTR has a detectably different conformation, in contrast to the minor differences observed for the isolated F508-containing domain. We interpret the data in terms of an incomplete post-translational assembly of the protein domains. PMID:26850167

  14. Structures and stability of metal-doped GenM (n = 9, 10 clusters

    Directory of Open Access Journals (Sweden)

    Wei Qin

    2015-06-01

    Full Text Available The lowest-energy structures of neutral and cationic GenM (n = 9, 10; M = Si, Li, Mg, Al, Fe, Mn, Pb, Au, Ag, Yb, Pm and Dy clusters were studied by genetic algorithm (GA and first-principles calculations. The calculation results show that doping of the metal atoms and Si into Ge9 and Ge10 clusters is energetically favorable. Most of the metal-doped Ge cluster structures can be viewed as adding or substituting metal atom on the surface of the corresponding ground-state Gen clusters. However, the neutral and cationic FeGe9,10,MnGe9,10 and Ge10Al are cage-like with the metal atom encapsulated inside. Such cage-like transition metal doped Gen clusters are shown to have higher adsorption energy and thermal stability. Our calculation results suggest that Ge9,10Fe and Ge9Si would be used as building blocks in cluster-assembled nanomaterials because of their high stabilities.

  15. Avances genéticos y moleculares en el estudio de trastornos mentales

    Directory of Open Access Journals (Sweden)

    Daisy Natalia Salamanca-Ortíz

    2014-04-01

    Full Text Available Actualmente, con el aumento de la disponibilidad de técnicas para la realización de estudios genéticos, han aparecido nuevas áreas del conocimiento como la epigenética y la farmacogenética. Estas nuevas áreas han permitido esclarecer las bases genéticas implicadas en la aparición de las enfermedades que afectan al ser humano. Dada su aplicación global, la psiquiatría no ha sido ajena al alcance de estas nuevas disciplinas, lo cual se refleja en el gran número de estudios realizados a nivel mundial que han relacionado la presencia de variantes genéticas en los individuos con la aparición de trastornos mentales específicos. De la misma forma, otros estudios han evidenciado que la efectividad de los tratamientos farmacológicos empleados en psiquiatría se correlaciona con polimorfismos en las enzimas encargadas de metabolizar dichos medicamentos. Por lo tanto, es importante que los psiquiatras conozcan los aspectos básicos de estas relaciones para obtener mejores resultados en el diagnóstico y tratamiento de los principales trastornos mentales.

  16. Structures and stability of metal-doped GenM (n = 9, 10) clusters

    International Nuclear Information System (INIS)

    The lowest-energy structures of neutral and cationic GenM (n = 9, 10; M = Si, Li, Mg, Al, Fe, Mn, Pb, Au, Ag, Yb, Pm and Dy) clusters were studied by genetic algorithm (GA) and first-principles calculations. The calculation results show that doping of the metal atoms and Si into Ge9 and Ge10 clusters is energetically favorable. Most of the metal-doped Ge cluster structures can be viewed as adding or substituting metal atom on the surface of the corresponding ground-state Gen clusters. However, the neutral and cationic FeGe9,10,MnGe9,10 and Ge10Al are cage-like with the metal atom encapsulated inside. Such cage-like transition metal doped Gen clusters are shown to have higher adsorption energy and thermal stability. Our calculation results suggest that Ge9,10Fe and Ge9Si would be used as building blocks in cluster-assembled nanomaterials because of their high stabilities

  17. Safety Assurance in NextGen

    Science.gov (United States)

    HarrisonFleming, Cody; Spencer, Melissa; Leveson, Nancy; Wilkinson, Chris

    2012-01-01

    The generation of minimum operational, safety, performance, and interoperability requirements is an important aspect of safely integrating new NextGen components into the Communication Navigation Surveillance and Air Traffic Management (CNS/ATM) system. These requirements are used as part of the implementation and approval processes. In addition, they provide guidance to determine the levels of design assurance and performance that are needed for each element of the new NextGen procedures, including aircraft, operator, and Air Navigation and Service Provider. Using the enhanced Airborne Traffic Situational Awareness for InTrail Procedure (ATSA-ITP) as an example, this report describes some limitations of the current process used for generating safety requirements and levels of required design assurance. An alternative process is described, as well as the argument for why the alternative can generate more comprehensive requirements and greater safety assurance than the current approach.

  18. Genève grandeur cultures.

    Directory of Open Access Journals (Sweden)

    Vincent Kaufmann

    2005-04-01

    Full Text Available Equinoxe , revue genevoise de sciences humaines, consacre son dernier numéro à l’émergence et au développement des contre-cultures à Genève depuis la fin des années 1960. Celui-ci se présente comme « une réflexion critique sur trois décennies de pratiques culturelles et sur la manière dont celles-ci sont parvenues non sans heurts à se faire une place dans la cité ». L’intérêt de la démarche réside dans le fait qu’à Genève, contrairement à Berlin, Paris, Barcelone ...

  19. Gen. Keen discusses Americas with JFSSPP

    OpenAIRE

    2011-01-01

    Army Lt. Gen. P. K. Keen, he Militay Deputy Commander for USSOUTHCOM, met with Foreign Area Officers (FAOs) attending a Latin American in-residence course at NPS for the School of International Graduate Studies (SIGS) Joint Foreign Area OFficer Skill Sustainment Pilot Program (JFSSPP). Keen spoke for nearly an hour to more than 35 participants in the course, which ran from February 21 though March 4. JFSSPP students are non-matriculated students who come to NPS for topical short courses rela...

  20. Spatial positioning of CFTR's pore-lining residues affirms an asymmetrical contribution of transmembrane segments to the anion permeation pathway.

    Science.gov (United States)

    Gao, Xiaolong; Hwang, Tzyh-Chang

    2016-05-01

    The structural composition of CFTR's anion permeation pathway has been proposed to consist of a short narrow region, flanked by two wide inner and outer vestibules, based on systematic cysteine scanning studies using thiol-reactive probes of various sizes. Although these studies identified several of the transmembrane segments (TMs) as pore lining, the exact spatial relationship between pore-lining elements remains under debate. Here, we introduce cysteine pairs in several key pore-lining positions in TM1, 6, and 12 and use Cd(2+) as a probe to gauge the spatial relationship of these residues within the pore. We find that inhibition of single cysteine CFTR mutants, such as 102C in TM1 or 341C in TM6, by intracellular Cd(2+) is readily reversible upon removal of the metal ion. However, the inhibitory effect of Cd(2+) on the double mutant 102C/341C requires the chelating agent dithiothreitol (DTT) for rapid reversal, indicating that 102C and 341C are close enough to the internal edge of the narrow region to coordinate one Cd(2+) ion between them. We observe similar effects of extracellular Cd(2+) on TM1/TM6 cysteine pairs 106C/337C, 107C/337C, and 107C/338C, corroborating the idea that these paired residues are physically close to each other at the external edge of the narrow region. Although these data paint a picture of relatively symmetrical contributions to CFTR's pore by TM1 and TM6, introducing cysteine pairs between TM6 and TM12 (348C/1141C, 348C/1144C, and 348C/1145C) or between TM1 and TM12 (95C/1141C) yields results that contest the long-held principle of twofold pseudo-symmetry in the assembly of ABC transporters' TMs. Collectively, these findings not only advance our current understanding of the architecture of CFTR's pore, but could serve as a guide for refining computational models of CFTR by imposing physical constraints among pore-lining residues. PMID:27114613

  1. Parámetros y tendencias genéticas para características de crecimiento predestete en la población mexicana de Simmental

    OpenAIRE

    Javier Rosales; Elzo, Mauricio A.; Moisés Montaño; Vicente Eliezer Vega

    2004-01-01

    Se utilizaron registros de pesos al nacimiento (18,383) y al destete (9,023) de la población mexicana de ganado Simmental, para estimar varianzas genéticas aditivas directas y maternas, heredabilidades, correlaciones y tendencias genéticas. La información se obtuvo en 434 hatos de socios de la Asociación Mexicana Simmental-Simbrah, entre 1980 y 1999. El grupo contemporáneo se definió como hatoaño- estación. Se utilizó un modelo animal de dos características con efectos aditivos directos y mat...

  2. Mucopolisacaridosis II: nueva mutación patogénica en gen IDS

    Directory of Open Access Journals (Sweden)

    Eugenia Pérez-Elizondo

    2014-12-01

    Full Text Available La mucopolisacaridosis tipo II es una enfermedad lisosomal producida por la deficiencia de la enzima iduronato 2 sulfatasa. Es una condición infrecuente de herencia recesiva ligada al X, que puede producir importante discapacidad progresiva. El análisis molecular es una técnica útil en la confirmación diagnóstica, que además permite detección de portadores asintomáticos, brindando la oportunidad de asesoría genética. Se presenta el caso de un paciente con mucopolisacaridosis tipo II, en quien se documentó una nueva mutación patogénica en el Gen IDS.

  3. SFRs and GEN IV: ASN actions

    International Nuclear Information System (INIS)

    ASN carries out several actions regarding the 4th generation of reactors (GEN IV) and the sodium fast reactors (SFRs). First of all, ASN considers that GEN IV reactors have to be safer than the EPR reactor currently under construction in France (at Flamanville). Regarding GEN IV in general, the “generation IV international Forum” (GIF) has identified six technologies of reactors for which the possibility of an industrial development could be considered (SFR, GFR, HTR/VHTR, LFR, MSR, SCWR) ; these six technologies include fast reactors. Based on documents sent by French nuclear operators at its request, ASN will organize a technical expertise, with the technical safety organization IRSN and ASN Advisory Committees, in order to have an overview, in terms of safety, R&D needs and possibility of transmutation of these six technologies. The conclusions of the technical expertise are expected by the end of 2013. Concerning SFR in particular, the French operator CEA submitted to ASN in June 2012 a file presenting the general safety orientations of the prototype reactor ASTRID (Advanced Sodium Technological Reactor for Industrial Demonstration). ASN has organized, with IRSN and ASN Advisory Committees, a technical expertise to analyse this file. The conclusions of this technical expertise are expected by mid-2013. Besides, a specific analysis is going on about the transmutation. The conclusions of the analysis could be known in 2013, subject to the transmission of documents by the end of 2012

  4. Amniotic Mesenchymal Stem Cells: A New Source for Hepatocyte-Like Cells and Induction of CFTR Expression by Coculture with Cystic Fibrosis Airway Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Valentina Paracchini

    2012-01-01

    Full Text Available Cystic fibrosis (CF is a monogenic disease caused by mutations in the CF transmembrane conductance regulator (CFTR gene, with lung and liver manifestations. Because of pitfalls of gene therapy, novel approaches for reconstitution of the airway epithelium and CFTR expression should be explored. In the present study, human amniotic mesenchymal stem cells (hAMSCs were isolated from term placentas and characterized for expression of phenotypic and pluripotency markers, and for differentiation potential towards mesoderm (osteogenic and adipogenic lineages. Moreover, hAMSCs were induced to differentiate into hepatocyte-like cells, as demonstrated by mixed function oxidase activity and expression of albumin, alpha1-antitrypsin, and CK19. We also investigated the CFTR expression in hAMSCs upon isolation and in coculture with CF airway epithelial cells. Freshly isolated hAMSCs displayed low levels of CFTR mRNA, which even decreased with culture passages. Following staining with the vital dye CM-DiI, hAMSCs were mixed with CFBE41o- respiratory epithelial cells and seeded onto permeable filters. Flow cytometry demonstrated that 33–50% of hAMSCs acquired a detectable CFTR expression on the apical membrane, a result confirmed by confocal microscopy. Our data show that amniotic MSCs have the potential to differentiate into epithelial cells of organs relevant in CF pathogenesis and may contribute to partial correction of the CF phenotype.

  5. Comparative ex vivo, in vitro and in silico analyses of a CFTR splicing mutation: Importance of functional studies to establish disease liability of mutations.

    Science.gov (United States)

    Ramalho, Anabela S; Clarke, Luka A; Sousa, Marisa; Felicio, Verónica; Barreto, Celeste; Lopes, Carlos; Amaral, Margarida D

    2016-01-01

    The Cystic Fibrosis p.Ile1234Val missense mutation actually creates a new dual splicing site possibly used either as a new acceptor or donor. Here, we aimed to test the accuracy of in silico predictions by comparing them with in vitro and ex vivo functional analyses of this mutation for an accurate CF diagnosis/prognosis. To this end, we applied a new in vitro strategy using a CFTR mini-gene which includes the complete CFTR coding sequence plus intron 22 (short version) which allows the assessment of alternatively spliced mRNA levels as well as the properties of the resulting abnormal CFTR protein regarding processing, intracellular localization and function. Our data demonstrate that p.Ile1234Val leads to usage of the alternative splicing donor (but not acceptor) resulting in alternative CFTR transcripts lacking 18 nts of exon 22 which produce a truncated CFTR protein with residual Cl- channel function. These results recapitulate data from native tissues of a CF patient. In conclusion, the existing in silico prediction models have limited application and ex vivo functional assessment of mutation effects should be made. Alternatively the in vitro strategy adopted here can be applied to assess the disease liability of mutations for an accurate CF diagnosis/prognosis. PMID:25735457

  6. Aïllament, identificació i caracterització de nematodes entomopatògens en sòls de cultius del Baix Llobregat i les muntanyes de Prades i possible utilització per al control de plagues

    OpenAIRE

    Suárez Cano, Pedro

    2015-01-01

    Durant els últims anys s'estan realitzant nombrosos estudis sobre l'ús de nematodes com a mètode de control biològic. Mitjançant el present treball s'ha pretès realitzar un aïllament de nematodes entomopatògens en les zones de Prades i del Baix Llobregat i un cop aïllats i identificats realitzar diferents proves d'eficàcia d'aquests aïllats com mètodes de control de diferents plagues. Per dur a terme aquests assajos s'han realitzat diferents sistemes pel manteniment i producció de les pob...

  7. Generic classification of the Archiborborinae (Diptera: Sphaeroceridae), with a revision of Antrops Enderlein, Coloantrops gen. nov., Maculantrops gen. nov., Photoantrops gen. nov., and Poecilantrops gen. nov.

    Science.gov (United States)

    Kits, Joel H; Marshall, Stephen A

    2013-01-01

    The Archiborborinae comprise a diverse clade of flies in the family Sphaeroceridae. We here revise the generic classification, redefining the genus Antrops Enderlein and naming 5 new genera: Boreantrops gen. nov., Coloantrops gen. nov., Maculantrops gen. nov., Photoantrops gen. nov., and Poecilantrops gen. nov. The genus Archiborborus, until recently a paraphyletic assemblage including most of the described species in the subfamily, is treated as a junior synonym of Antrops (syn. nov.) We revise the genera Antrops (53 species, including 40 sp. nov.: Antrops anovariegatus, Antrops aurantifemur, Antrops baeza, Antrops bellavista, Antrops biflavus, Antrops bucki, Antrops carpishensis, Antrops cochabamba, Antrops cochinoca, Antrops coniobaptos, Antrops coroico, Antrops cotopaxi, Antrops didactylos, Antrops diversipennis, Antrops eurus, Antrops fulgiceps, Antrops fuliginosus, Antrops guandera, Antrops guaramacalensis, Antrops inca, Antrops juninensis, Antrops mucarensis, Antrops niger, Antrops papallacta, Antrops pecki, Antrops podocarpus, Antrops quadrilobus, Antrops siberia, Antrops sierrazulensis, Antrops tachira, Antrops tequendama, Antrops tetrastichus, Antrops tumbrensis, Antrops unduavi, Antrops variegatus, Antrops versabilis, Antrops vittatus, Antrops yungas, and Antrops zongo and the following comb. nov.: Antrops annulatus (Richards), Antrops chaetosus (Richards), Antrops femoralis (Blanchard), Antrops hirtus (Bigot), Antrops maculipennis (Duda), Antrops maximus (Richards), Antrops microphthalmus (Richards), Antrops quadrinotus (Bigot), Antrops setosus (Duda), Antrops simplicimanus (Richards), Antrops nitidicollis (Becker), and Antrops orbitalis (Duda)), Coloantrops (1 species: Coloantrops daedalus, sp. nov.), Maculantrops (2 species, Maculantrops hirtipes (Macquart) comb. nov. and Maculantrops altiplanus, sp. nov.), Photoantrops (1 species: Pho-toantrops echinus sp. nov.), and Poecilantrops (10 species: Poecilantrops baorucensis, Poecilantrops boraceiensis

  8. Market share scenarios for Gen-DIII and gen-IV reactors in Europe

    International Nuclear Information System (INIS)

    Nuclear energy is back on the agenda worldwide in order to meet growing energy demand and especially the growth in electricity demand. Many objectives direct to an increased use of nuclear energy, i.e. minimising energy costs, reducing climate change effects and others. In the light of the potential renewed growth of nuclear energy, the public demands a clear view on what nuclear energy may contribute towards meeting these objectives and especially how nuclear energy may address some socio-political obstructions with respect to economics, radioactive waste, safety and proliferation of fissile materials. To address these questions, the future nuclear reactor park mix in Europe has been analysed applying an integrated dynamic process modelling technique. Various market share scenarios for nuclear energy are derived including sub-variants with regard to the intra-nuclear options. In the analyses, it is assumed that different types of new reactors may be built, taking into account the introduction date of considered Gen-Ill (i.e. EPR) and Gen-IV (i.e. SCWR, HTR, FR) reactors, and the economic evaluation of the complete fuel cycle. The assessment was undertaken using the DANESS code (Dynamic Analysis of Nuclear Energy System Strategies). The analyses show that given the considered realistic nuclear energy demand and given a limited number of available Gen-III and Gen-IV reactor types, the future European nuclear park will exist of combinations of Gen-III and Gen-IV reactors. This mix will always consist of a set of reactor types each having its specific strengths. The analyses also highlight the triggers influencing the choice between different nuclear energy deployment scenarios. (authors)

  9. In vitro and in vivo Functional Characterization of Gutless Recombinant SV40-derived CFTR Vectors

    OpenAIRE

    Mueller, Christian; Strayer, Marlene S.; Sirninger, Jeffery; Braag, Sofia; Branco, Francisco; Louboutin, Jean-Pierre; Flotte, Terence R.; Strayer, David S.

    2009-01-01

    In cystic fibrosis (CF) respiratory failure caused by progressive airway obstruction and tissue damage is primarily a result of the aberrant inflammatory responses to lung infections with Pseudomonas aeruginosa. Despite considerable improvement in patient survival, conventional therapies are mainly supportive. Recent progress towards gene therapy for CF has been encouraging; however, several factors such as immune response and transduced cell turnover remain as potential limitations to CF gen...

  10. An unexpected effect of TNF-α on F508del-CFTR maturation and function [v1; ref status: indexed, http://f1000r.es/5jf

    Directory of Open Access Journals (Sweden)

    Sara Bitam

    2015-07-01

    Full Text Available Cystic fibrosis (CF is a multifactorial disease caused by mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR, which encodes a cAMP-dependent Cl- channel. The most frequent mutation, F508del, leads to the synthesis of a prematurely degraded, otherwise partially functional protein. CFTR is expressed in many epithelia, with major consequences in the airways of patients with CF, characterized by both fluid transport abnormalities and persistent inflammatory responses. The relationship between the acute phase of inflammation and the expression of wild type (WT CFTR or F508del-CFTR is poorly understood. The aim of the present study was to investigate this effect. The results show that 10 min exposure to TNF-alpha (0.5-50ng/ml of F508del-CFTR-transfected HeLa cells and human bronchial cells expressing F508del-CFTR in primary culture (HBE leads to the maturation of F508del-CFTR and induces CFTR chloride currents. The enhanced CFTR expression and function upon TNFα is sustained, in HBE cells, for at least 24 h. The underlying mechanism of action involves a protein kinase C (PKC signaling pathway, and occurs through insertion of vesicles containing F508del-CFTR to the plasma membrane, with TNFα behaving as a corrector molecule. In conclusion, a novel and unexpected action of TNFα has been discovered and points to the importance of systematic studies on the roles of inflammatory mediators in the maturation of abnormally folded proteins in general and in the context of CF in particular.

  11. An unexpected effect of TNF-α on F508del-CFTR maturation and function [v2; ref status: indexed, http://f1000r.es/5tv

    Directory of Open Access Journals (Sweden)

    Sara Bitam

    2015-09-01

    Full Text Available Cystic fibrosis (CF is a multifactorial disease caused by mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR, which encodes a cAMP-dependent Cl- channel. The most frequent mutation, F508del, leads to the synthesis of a prematurely degraded, otherwise partially functional protein. CFTR is expressed in many epithelia, with major consequences in the airways of patients with CF, characterized by both fluid transport abnormalities and persistent inflammatory responses. The relationship between the acute phase of inflammation and the expression of wild type (WT CFTR or F508del-CFTR is poorly understood. The aim of the present study was to investigate this effect. The results show that 10 min exposure to TNF-alpha (0.5-50ng/ml of F508del-CFTR-transfected HeLa cells and human bronchial cells expressing F508del-CFTR in primary culture (HBE leads to the maturation of F508del-CFTR and induces CFTR chloride currents. The enhanced CFTR expression and function upon TNFα is sustained, in HBE cells, for at least 24 h. The underlying mechanism of action involves a protein kinase C (PKC signaling pathway, and occurs through insertion of vesicles containing F508del-CFTR to the plasma membrane, with TNFα behaving as a corrector molecule. In conclusion, a novel and unexpected action of TNFα has been discovered and points to the importance of systematic studies on the roles of inflammatory mediators in the maturation of abnormally folded proteins in general and in the context of CF in particular.

  12. Apratylenchus vietnamensis gen. n., sp. n. and A. binhi gen. n., sp. n., sedentary Pratylenchidae (Nematoda:Tylenchida) from coffee in Vietnam, with proposal of Apratylenchinae subfam. n.

    NARCIS (Netherlands)

    Trinh, P.Q.; Waeyenberge, L.; Nguyen, C.N.; Baldwin, J.G.; Karssen, G.; Moens, M.

    2009-01-01

    Apratylenchinae subfam. n. is proposed within the Pratylenchidae to include Apratylenchus gen. n. and two new species, A. vietnamensis gen. n., sp. n. and A. binhi gen. n., sp. n., from coffee plantations in Vietnam. Apratylenchus gen. n. is distinguished by the presence of transverse rows of cuticu

  13. Türk ankilozan spondilitli hastalarda HLA-B27 ve MEFV gen mutasyonları arasındaki ilişki

    OpenAIRE

    SARIKAYA PEKACAR, Filiz

    2015-01-01

    Ankilozan spondilit (AS), patogenezinde bir çok genetik, immunolojik ve çevresel faktörün birlikte rol oynadığı  inflamatuvar bir hastalıktır. Genetik faktörler arasında en büyük pay HLA -B27’nindir. Genom  çapında ilişkilendirme çalışmaları sonucu AS ile ilgili bir çok gen bulunmuş MHC dışı genlerin hastalığın  gelişiminde rol oynadığı düşünülmüştür.
    Bu tez çalışmasında HLA-B27, MEFV gen mutasyonları, IL12B, IL23R ve ERAP1 gen polimorfiz...

  14. Aconselhamento genético Genetic counseling

    Directory of Open Access Journals (Sweden)

    João Monteiro de Pina-Neto

    2008-08-01

    Full Text Available OBJETIVO: Esta revisão sobre aconselhamento genético (AG teve o objetivo de mostrar os conceitos atuais e os princípios filosóficos e éticos aceitos na grande maioria dos países e recomendados pela Organização Mundial da Saúde, as fases do processo, seus resultados e o impacto psicológico de uma doença genética em uma família. FONTES DOS DADOS: Os conceitos apresentados são baseados em uma síntese histórica da literatura sobre AG desde a década de 1930 até o momento atual, sendo que os artigos citados representam os principais trabalhos publicados e que hoje fundamentam a teoria e a prática do AG. SÍNTESE DOS DADOS: O AG modernamente é definido como um processo de comunicação que trata dos problemas humanos relacionados à ocorrência de uma doença genética em uma família. É fundamental que os profissionais da saúde conheçam os aspectos psicológicos desencadeados pela doença genética e como estes aspectos podem ser manejados. Vivemos ainda na genética humana e médica uma fase de predomínio dos aspectos técnicos e científicos e de pouca ênfase no estudo das reações emocionais e dos processos de adaptação das pessoas a estas doenças, o que leva ao baixo entendimento dos clientes sobre os fatos ocorridos, com conseqüências negativas sobre a vida familiar e para a sociedade. CONCLUSÕES: Conclui-se pela necessidade de que as famílias com doenças genéticas sejam encaminhadas para AG e que os profissionais desta área invistam mais na humanização do atendimento, desenvolvendo mais as técnicas do AG psicológico não-diretivo.OBJECTIVE: The objective of this review of genetic counseling (GC is to describe the current concepts and philosophical and ethical principles accepted by the great majority of countries and recommended by the World Health Organization, the stages of the process, its results and the psychological impact that a genetic disease has on a family. SOURCES: The concepts presented are

  15. Marcadores genéticos del bovino criollo : Su germoplasma como recurso genético

    OpenAIRE

    Quinteros, Indalecio Rodolfo

    1994-01-01

    En la presente comunicación se propone continuar en la investigación del germoplasma del ganado bovino por los Marcadores Genéticos Eritrocitarios y Bioquímicos en correspondencia a poblaciones de distintas regiones de Argentina y América, como contribución a otros tipos de estudios

  16. TidGen Power System Commercialization Project

    Energy Technology Data Exchange (ETDEWEB)

    Sauer, Christopher R. [President & CEO; McEntee, Jarlath [VP Engineering & CTO

    2013-12-30

    ORPC Maine, LLC, a wholly-owned subsidiary of Ocean Renewable Power Company, LLC (collectively ORPC), submits this Final Technical Report for the TidGen® Power System Commercialization Project (Project), partially funded by the U.S. Department of Energy (DE-EE0003647). The Project was built and operated in compliance with the Federal Energy Regulatory Commission (FERC) pilot project license (P-12711) and other permits and approvals needed for the Project. This report documents the methodologies, activities and results of the various phases of the Project, including design, engineering, procurement, assembly, installation, operation, licensing, environmental monitoring, retrieval, maintenance and repair. The Project represents a significant achievement for the renewable energy portfolio of the U.S. in general, and for the U.S. marine hydrokinetic (MHK) industry in particular. The stated Project goal was to advance, demonstrate and accelerate deployment and commercialization of ORPC’s tidal-current based hydrokinetic power generation system, including the energy extraction and conversion technology, associated power electronics, and interconnection equipment capable of reliably delivering electricity to the domestic power grid. ORPC achieved this goal by designing, building and operating the TidGen® Power System in 2012 and becoming the first federally licensed hydrokinetic tidal energy project to deliver electricity to a power grid under a power purchase agreement in North America. Located in Cobscook Bay between Eastport and Lubec, Maine, the TidGen® Power System was connected to the Bangor Hydro Electric utility grid at an on-shore station in North Lubec on September 13, 2012. ORPC obtained a FERC pilot project license for the Project on February 12, 2012 and the first Maine Department of Environmental Protection General Permit issued for a tidal energy project on January 31, 2012. In addition, ORPC entered into a 20-year agreement with Bangor Hydro Electric

  17. Genética de la conservación: la aplicación de los conceptos de la evolución a la conservación de la diversidad biológica

    OpenAIRE

    Caballero Rúa, Armando

    2013-01-01

    La comprensión de las fuerzas de cambio evolutivo que actúan sobre las poblaciones, en conjunción con las técnicas más modernas de análisis genético, lleva aplicándose en los últimos veinte años al servicio de la conservación de la biodiversidad. Esta nueva aplicación de la genética ha recibido el nombre de genética de la conservación.

  18. Análisis de la asociación entre sexo y diferencias en pigmentación humana según el genotipo del gen MC1R

    OpenAIRE

    Hernando, Bárbara; Ibarrola-Villava, Maider; Llorca-Cardeñosa, Marta J.; Ribas, Gloria; Martínez-Cadenas, Conrado

    2016-01-01

    Introducción: La pigmentación cutánea basal y la respuesta al sol por bronceamiento son rasgos hereditarios influidos por varios genes, entre los que el gen mc1r es uno de los más importantes. Mutaciones en este gen afectan a los niveles y tipos de me-lanina dando lugar a patrones alterados de pigmentación. Recientemente, se ha iden-tificado una asociación entre el genotipo del gen oca2, el color de ojos y el sexo, su-giriendo que existe un factor relacionado con el sexo que contribuye a las ...

  19. Aconselhamento Genético Genetic Counseling

    Directory of Open Access Journals (Sweden)

    Décio Brunoni

    2002-01-01

    Full Text Available O Aconselhamento Genético (AG deve ser desenvolvido nas unidades de saúde como um atendimento médico multiprofissional e interdisciplinar. A maioria da população brasileira não tem acesso a esses serviços. Os grupos, centros e serviços de AG existentes no Brasil podem, em curto espaço de tempo, preparar profissionais para atender a rede do Sistema Único de Saúde (SUS. O Ministério da Saúde tem os instrumentos necessários para viabilizar esta estratégia. Medidas que visem incrementar o ensino da genética médica nos cursos de graduação e em curso de especialização/aperfeiçoamento podem ser efetivadas. O envolvimento da população, de pacientes e famílias no conhecimento dos fatores genéticos que agravam a saúde é desejável e pode ser alcançado por ações de genética comunitária como tem sido recomendado pela Organização Mundial da Saúde (OMS.Genetic counseling (GC is a medical multiprofessional and interdisciplinar health care process. Most of the Brazilian people don't have access to these services. In Brazil there are many GC groups, centers and services in condition to train health care professionals to the Federal Brazilian Public System of Health (Sistema Único de Saúde - SUS. Ministry of Health could easily created an Office of Genetic Services to organize and launch trainning programs. It is also necessary improve genetic educational programs for the health professions as well as educate the public in genetic disorders and birth defects. Support of parent/patient organizations is also desirable. The implementation of such community genetics programs has been recommended by the World Health Organization (WHO.

  20. Gen IV Materials Handbook Functionalities and Operation

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Weiju [ORNL

    2009-12-01

    This document is prepared for navigation and operation of the Gen IV Materials Handbook, with architecture description and new user access initiation instructions. Development rationale and history of the Handbook is summarized. The major development aspects, architecture, and design principles of the Handbook are briefly introduced to provide an overview of its past evolution and future prospects. Detailed instructions are given with examples for navigating the constructed Handbook components and using the main functionalities. Procedures are provided in a step-by-step fashion for Data Upload Managers to upload reports and data files, as well as for new users to initiate Handbook access.

  1. Genética, performance física humana e doping genético: o senso comum versus a realidade científica

    Directory of Open Access Journals (Sweden)

    Rodrigo Gonçalves Dias

    2011-02-01

    Full Text Available Atletas de elite são reconhecidos como fenômenos esportivos e o potencial para atingir níveis superiores de performance no esporte está parcialmente sob o controle de genes. A excelência atlética é essencialmente multifatorial e determinada por complexas interações entre fatores ambientais e genéticos. Existem aproximadamente 10 milhões de variantes genéticas dispersas por todo o genoma humano e uma parcela destas variantes têm demonstrado influenciar a responsividade ao treinamento físico. Os fenótipos de performance física humana parecem ser altamente poligênicos e alguns estudos têm comprovado a existência de raras combinações genotípicas em atletas. No entanto, os mecanismos pelos quais genes se interagem para amplificar a performance física são desconhecidos. O conhecimento sobre os genes que influenciam a treinabilidade somado ao potencial uso indevido dos avanços da terapia gênica, como a possível introdução de genes em células de atletas, fez surgir o termo doping genético, um novo e censurado método de amplificação da performance física, além dos limites fisiológicos. Aumentos na hipertrofia muscular esquelética e nos níveis de hematócrito estão sendo conseguidos através da manipulação da expressão de genes específicos, mas a grande parte das impressionáveis alterações foi obtida em experimentação com animais de laboratório. A compreensão dos resultados científicos envolvendo genética, performance física humana e doping genético é uma difícil tarefa. Com o propósito de evitar a contínua má interpretação e propagação de conceitos errôneos, esta revisão, intencionalmente, vem discutir as evidências científicas produzidas até o momento sobre o tema, permitindo a compreensão do atual "estado da arte"

  2. Disruption of Interleukin-1β Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    OpenAIRE

    Clauzure, Mariángeles; Valdivieso, Angel G.; Massip Copiz, María M.; Schulman, Gustavo; Teiber, María Luz; Santa-Coloma, Tomás A.

    2014-01-01

    Patients with cystic fibrosis (CF) have elevated concentration of cytokines in sputum and a general inflammatory condition. In addition, CF cells in culture produce diverse cytokines in excess, including IL-1β. We have previously shown that IL-1β, at low doses (∼30 pM), can stimulate the expression of CFTR in T84 colon carcinoma cells, through NF-κB signaling. However, at higher doses (>2.5 ng/ml, ∼150 pM), IL-1β inhibit CFTR mRNA expression. On the other hand, by using differential display, ...

  3. Correlaciones fenotípicas, ambientales y genéticas en berenjena

    Directory of Open Access Journals (Sweden)

    Hermes Araméndiz Tatis

    2009-10-01

    Full Text Available En el Centro de Investigaciones Turipaná de la Corporación Colombiana de Investigaciones (Corpoica (Cereté, Córdoba, Colombia -8° 31' N y 75° 58' O, a 13 m.s.n.m se estudiaron las correlaciones, ambientales y genéticas entre seis caracteres de 24 cultivares de berenjena (Solanum melongena. Para el efecto se utilizó un diseño de bloques completamente al azar con tres repeticiones y unidades experimentales de 10 m². Los resultados mostraron que las correlaciones fueron de mayor o igual magnitud que las fenotípicas, mientras que las ambientales fueron de escaso valor. El número de frutos y el rendimiento estuvieron genéticamente correlacionados (r = 0.56, P < 0.01, la longitud y la resistencia del fruto mostraron correlación genética negativa (r = -0.68, P < 0.01 y entre el rendimiento y peso de fruto la correlación fue muy baja (r = 0.04. El número de frutos y su peso de frutos se correlacionaron de manera negativa (r =-0.63, P < 0.01. El número de frutos por planta puede ser utilizado como criterio de selección para la obtención de cultivares de berenjena de altos rendimientos.

  4. Correlaciones fenotípicas, ambientales y genéticas en berenjena

    Directory of Open Access Journals (Sweden)

    Cardona Ayala Carlos Enrique

    2009-12-01

    Full Text Available En el Centro de Investigaciones Turipaná de la Corporación Colombiana de Investigaciones (Corpoica (Cereté, Córdoba, Colombia –8° 31' N y 75° 58' O, a 13 m.s.n.m se estudiaron las correlaciones, ambientales y genéticas entre seis caracteres de 24 cultivares de berenjena (Solanum melongena. Para el efecto se utilizó un diseño de bloques completamente al azar con tres repeticiones y unidades experimentales de 10 m2. Los resultados mostraron que las correlaciones fueron de mayor o igual magnitud que las fenotípicas, mientras que las ambientales fueron de escaso valor. El número de frutos y el rendimiento estuvieron genéticamente correlacionados (r = 0.56, P < 0.01, la longitud y la resistencia del fruto mostraron correlación genética negativa (r = -0.68, P < 0.01 y entre el rendimiento y peso de fruto la correlación fue muy baja (r = 0.04. El número de frutos y su peso de frutos se correlacionaron de manera negativa (r =-0.63, P < 0.01. El número de frutos por planta puede ser utilizado como criterio de selección para la obtención de cultivares de berenjena de altos rendimientos.

  5. El papel del gen del transportador de serotonina en los trastornos de la conducta alimentaria

    Directory of Open Access Journals (Sweden)

    Sandra Hernández-Muñoz

    2014-10-01

    Full Text Available Introducción: Al sistema serotoninérgico se lo ha implicado en la regulación del estado de ánimo y en la conducta alimentaria, por lo que el gen del transportador de serotonina (SLC6A4 es un buen candidato para el desarrollo de los trastornos de la conducta alimentaria (TCA. La mayoría de los estudios genéticos en los TCA se han centrado principalmente en un polimorfismo, el denominado 5-HTTLPR del gen SLC6A4. Objetivo: Realizar una revisión de los estudios de asociación entre el 5-HTTLPR y los TCA, como anorexia nerviosa, bulimia nerviosa y trastornos alimentarios no especificados. Método: Se realizó una búsqueda en MEDLINE, ISI y PubMed de las palabras clave «transportador de serotonina» y «TCA». Conclusiones: Según la revisión de 37 artículos originales, la variante S del 5-HTTLPR es un factor de riesgo de anorexia nerviosa. Además, se encontró asociación entre el alelo S y el índice de masa corporal, impulsividad, ansiedad, depresión y el tiempo de evolución en TCA. Sin embargo, en bulimia nerviosa no se reporta asociación con las variantes del 5-HTTLPR.

  6. Diversidade genética em palmeiras através de isoenzimas e RAPD

    Directory of Open Access Journals (Sweden)

    SAWAZAKI H. E.

    1998-01-01

    Full Text Available Estudou-se, mediante polimorfismo enzimático em gel de poliacrilamida e polimorfismo de DNA com base na amplificação de segmentos de DNA ao acaso, denominado RAPD, a variabilidade genética em algumas espécies e ecótipos de palmeiras dos gêneros Euterpe, Bactris, Elaeis e Syagrus. Os extratos de folhas de mudas dessas palmeiras foram analisados para as isoenzimas de malato desidrogenase (MDH, leucinoaminopeptidase (LAP, glutamato oxaloacetato transaminase (GOT, fosfoglucose isomerase (PGI, fosfoglucose mutase (PGM, fosfatase ácida (ACP, peroxidase (PRX, esterase (EST e para os marcadores RAPD, utilizando os "primers" dos kits A e B da Operon Technologies. Verificou-se grande variabilidade genética interespecífica, comprovada pelos dendrogramas UPGMA, com reconhecimento de híbridos. Foram observadas várias bandas além das referidas pela literatura em gel de amido. Os resultados dos marcadores RAPD comprovaram os das isoenzimas com maior eficácia, pois possibilitaram facilmente a análise de grande número de marcadores genéticos.

  7. The analysis of some CFTR gene mutations in a small group of cf patients from southern part of Romania

    Directory of Open Access Journals (Sweden)

    Lucian GAVRILA

    2009-05-01

    Full Text Available Cystic fibrosis is the most common hereditary disease in European descendant populations, with prevalencedepending on ethnic groups studied. In contrast to other European countries, there is little information regarding the frequency ofCFTR mutations for the Southern part of Romania. The aim of this study was to test the presence of nine CFTR mutations in CFpatients from the Southern part of Romania, using complementary analysis methods. We investigated a group of unrelated CFpatients (n=19 and, when possible, their voluntary parents (n=15. We observed that the most frequently worldwide CF mutation,delta F508, was present in 17 of our patients (89.5% in homozygous (n=7 or heterozygous (n=10 condition and absent in 2 cases(10.5%. This mutation was also detected in ten parents, seven of them (100% have homozygous children and three (37.5%have heterozygous children for delta F508 mutation. None of the G542X, S549N, G551D, R553X, R560T, S1255X, W1282X andN1303K mutations have been detected in the samples from patients or parents. Our results are partially similar with those reportedin neighbouring countries where the delta F508 is the most common mutation detected and the frequency of R560T, S549N, G551D andS1255X mutations is near zero. The enlargement of this study could give a better result regarding the spectrum of CFTR mutationsin Romanian patients with CF.

  8. Engenharia Genética: busca da mercadoria perfeita?

    OpenAIRE

    José Neivaldo de Souza

    2014-01-01

    Resenha:Engenharia Genética: busca da mercadoria perfeita? -SANDEL, Michael J. Contra a Perfeição: ética na era da engenharia genética. 1ª.ed. Tradução por Ana Carolina Mesquita. Rio de janeiro: Civilização Brasileira, 2013. 160p. 

  9. Hoja informativa de pruebas genéticas

    Science.gov (United States)

    Trata de pruebas genéticas para riesgos heredados de cáncer; incluye tipos de pruebas, para quién son, explicación y confidencialidad de los resultados. Ofrece información de pruebas genéticas a domicilio o pruebas para el consumidor.

  10. Towards an International Culture: Gen Y Students and SNS?

    Science.gov (United States)

    Lichy, Jessica

    2012-01-01

    This article reports the findings of a small-scale investigation into the Internet user behaviour of generation Y (Gen Y) students, with particular reference to social networking sites. The study adds to the literature on cross-cultural Internet user behaviour with specific reference to Gen Y and social networking. It compares how a cohort of…

  11. Ética y genética

    OpenAIRE

    2002-01-01

    Este documento contiene una transcripción de las ponencias presentadas en la Mesa Redonda sobre Ética y Genética celebrada en Oviedo, el día 10 de octubre de 2001, y organizada por el Real Patronato sobre Discapacidad en el marco del XXI Congreso Nacional de Genética Humana.

  12. ALS Association

    Science.gov (United States)

    ... toward a world without ALS! Walk to Defeat ALS® Walk to Defeat ALS® draws people of all ... We need your help. I Will Advocate National ALS Registry The National ALS Registry is a congressionally ...

  13. Thaumetopoea pityocampa i nematodes entomopatògens : un mètode alternatiu de control biològic de la plaga

    OpenAIRE

    Franquet Barrera, Laura; García del Pino, Fernando

    2009-01-01

    L'objectiu principal d'aquesta investigació és determinar la viabilitat dels nematodes entomopatògens per controlar les plagues de Thaumetopoea pityocampa, tenint en compte que presenten tot un seguit de característiques que els fan adequats per al control de diferents plagues d'insectes.

  14. Caracterización genética de la raza chato murciano

    OpenAIRE

    Herrero Medrano, Juan Manuel

    2012-01-01

    El Chato Murciano es una raza local porcina autóctona del sureste de España y en peligro de extinción. Esta raza, al igual que otras muchas razas locales, está amenazada por los efectos de la endogamia y el cruce con otras razas porcinas. En el presente trabajo se utilizaron conjuntamente tres marcadores genéticos, concretamente un panel de 34 microsatélites, un panel de alta densidad de SNPs y un fragmento de ADN mitocondrial que se analizaron en la práctica totalidad de la...

  15. Sèrie Tipologies Comercials. Comerç Just. Part 2. Orígens

    OpenAIRE

    Juan Vigaray, María Dolores de; González Gascón, Elena

    2014-01-01

    En aquest vídeo veurem quina ha estat l'expansió del Comerç Just des dels seus orígens després de la II Guerra Mundial. Des del primer exemple, el 1946 a Estats Units, quan l'organització Ten Thousand Villages va començar a vendre treballs de costura produïts per dones pobres de Puerto Rico, fins al llançament del seu segell actual, juntament amb la marca FTO (Fair Trade Organizations), el 2002.

  16. Vizier: un sistema de recomendación genérico y multidimensional

    OpenAIRE

    Carrillo Ramos, Angela Cristina

    2012-01-01

    Los Sistemas de Recomendación, al ser herramientas que puedan ser utilizadas para identificar fácilmente productos de interés, han surgido para apoyar, aumentar y automatizar el proceso natural y diario de compartir recomendaciones, y de esta manera, reducir un espacio de alternativas. Este artículo tiene como objetivo presentar Vizier, un framework construido bajo un enfoque genérico, que proporciona servicios a aplicaciones de recuperación de información para que éstas a su vez, se encuentr...

  17. Interrupciones de embarazo por causa genética Interruptions of pregnancy through genetic

    OpenAIRE

    Isabel Figueroa Calderón; Daymi Saavedra Moredo; Yudith de la Torres Sieres; Mayra Sánchez Lueiro

    2012-01-01

    Introducción: las malformaciones congénitas incrementan notablemente las tasas de mortalidad infantil y comprometen la calidad de vida del infante. Objetivo: cuantificar las Interrupciones de causa genética y conocer las malformaciones congénitas más frecuentes en la población. Métodos: se realizó un estudio observacional descriptivo en el Hospital Docente Provincial "Ana Betancourt de Mora" de Camagüey, en el período comprendido del 1ro. de enero de 2004 al 31 de diciembre de 2011. El univer...

  18. La figura del autor en el arte genético y transgénico

    OpenAIRE

    Matewecki, Natalia

    2006-01-01

    Las prácticas artísticas contemporáneas redefinen constantemente el estatuto del arte tradicional al tensionar, por ejemplo, los límites entre el arte y la ciencia. El arte genético y el arte transgénico plantean un trabajo multidisciplinario en el que participan artistas y científicos a la vez, dando lugar así a la construcción de nuevas figuras de autor como las de artista-científico, artista-investigador, artista-técnico o científico-artista. La figura de artista se configura, según Dan...

  19. Optimización global con algoritmos genéticos

    OpenAIRE

    Carretero López, Félix

    2010-01-01

    El objetivo principal de este TFC es dar a conocer al lector el mundo de los métodos de optimización global mediante los algoritmos genéticos. Para alcanzar este objetivo empezaremos por exponer las bases teóricas en las que se fundamentan y a continuación las intentaremos corroborar empíricamente mediante un conjunto de pruebas simples. A partir de los resultados y con todo lo aprendido intentaremos resolver un problema real más complejo. En concreto intentaremos equilibrar la carga de las b...

  20. Université de Genève

    CERN Multimedia

    2008-01-01

    Ecole de physique - Département de physique nucléaire et corspusculaire 24, quai Ernest-Ansermet 1211 GENÈVE 4 Tél: (022) 379 62 73 - Fax: (022) 379 69 92 Lundi 1er décembre 2008 PARTICLE PHYSICS SEMINAR at 17.00 hrs – Stückelberg Auditorium Superconducting Interfaces between Insulating Oxide Prof. Jean-Marc TRISCONE / Université de Genève At interfaces between complex oxides, electronic systems with unusual properties can be generated. A striking example is the interface between LaAlO3 and SrTiO3, two good insulating perovskite oxides, which was found in 2004 to be conducting with a high mobility. We recently discovered that the ground state of this system is a superconducting condensate, with a critical temperature of about 200 mK. The characteristics observed for the superconducting transitions are consistent with a two-dimensional superconducting sheet as thin as a few nanometers. Recent field effect experiments revealed the sensitivity of the normal and superconducting states to the carrier d...

  1. Université de Genève

    CERN Multimedia

    2008-01-01

    Ecole de physique - Département de physique nucléaire et corspusculaire 24, quai Ernest-Ansermet - 1211 GENÈVE 4 Tél: (022) 379 62 73 - Fax: (022) 379 69 92 Lundi 1er décembre 2008 PARTICLE PHYSICS SEMINAR at 17.00 hrs – Stückelberg Auditorium Superconducting Interfaces between Insulating Oxide Prof. Jean-Marc TRISCONE / Université de Genève At interfaces between complex oxides, electronic systems with unusual properties can be generated. A striking example is the interface between LaAlO3 and SrTiO3, two good insulating perovskite oxides, which was found in 2004 to be conducting with a high mobility. We recently discovered that the ground state of this system is a superconducting condensate, with a critical temperature of about 200 mK. The characteristics observed for the superconducting transitions are consistent with a two-dimensional superconducting sheet as thin as a few nanometers. Recent field effect experiments revealed the sensitivity of the normal and superconducting states to the carrier ...

  2. Development of ARMS PCR tests for detection of common CFTR gene mutations

    Directory of Open Access Journals (Sweden)

    Livshits L. A.

    2010-09-01

    Full Text Available Aim. To develop diagnostic assays, based on the amplification refractory mutation system (ARMS principle, for detection of common mutations in the CFTR gene using two approaches: standard PCR with further gel-electrophoresis and Real-Time PCR with SYBR Green. Materials. For this study we have chosen the following mutations: dF508, W1282X, R117H, 621 + 1G > T, 2143delT with the frequencies in Ukraine: dF508 – 43.3 %; 2143delT – 1.38 %; W1282X – 1.1 %; R117H, 621 + 1G > T – T, 2143delT mutations. To validate the developed assays we have analyzed control DNA samples with the following mutations: W1282X (n = 3, R117H (n = 2, 621 + 1G > T (n = 1, 2143delT (n = = 1. For validation of the dF508 assay we have analyzed 100 heterozygous carriers and 50 homozygous carriers. We have analyzed 48 patients with cystic fibrosis, in which only one mutation was previously detected in combination with unknown mutant variant, using the developed ARMS assay for the 2143delT mutation, and detected 4 heterozygous carriers. No differences were observed in comparison with the standard protocols. Conclusions. It was shown that ARMS is a reliable, rapid and inexpensive method, and the developed assays can be applied in the standard PCR protocol with further gel-electrophoresis as well as using Real-Time PCR with SYBR Green for the molecular genetic diagnostics of cystic fibrosis.

  3. Lamotheoxyuris ackerti n. gen., n. comb. (Nematoda: Heteroxynematidae parasite of Neotoma spp. (Rodentia: Muridae Lamotheoxyuris ackerti n. gen., n. comb. (Nematoda: Heteroxynematidae parásito de Neotoma spp. (Rodentia: Muridae

    Directory of Open Access Journals (Sweden)

    JORGE FALCÓN-ORDAZ

    2010-06-01

    Full Text Available On the basis of the revision of the type material of Aspiculuris ackerti Kruidenier & Mehra, 1959, and new specimens collected from Neotoma nelsoni Goldman, 1905 (Rodentia: Cricetidae, in Veracruz, Mexico, we herein to which A. ackerti is transferred as Lamotheoxyuris ackerti This new genus differs from all other genera included in 1 mouth surrounded by six lips; 2 extension of lateral alae describe a new genus (Lamotheoxyuris n. gen., (Kruidener & Mehra, 1959 n. gen., n. comb. Heteroxynematinae by the following main traits: reduced; and 3 lack of caudal alae.Con base en la revisión del material tipo de Aspiculuris ackerti Kruidenier y Mehra, 1959 y de nuevos ejemplares recolectados en Neotoma nelsoni Goldman, 1905 (Rodentia: Cricetidae, en Veracruz, México, se describe un nuevo género (Lamotheoxyuris n. gen., al que A. ackerti es transferido como Lamotheoxyuris ackerti (Kruidener y Mehra, 1959 n. gen., n. comb. Este nuevo género se distingue de todos los demás géneros incluidos en Heteroxynematinae por las siguientes características: 1 presencia de seis labios rodeando la boca; 2 extensión reducida del ala lateral; y 3 carencia de ala caudal.

  4. Telefoninterviews als Datenerhebungsstrategie: Erfahrungen und Empfehlungen

    OpenAIRE

    Burke, Lisa A.; Miller, Monica K.

    2001-01-01

    In dem vorliegenden Beitrag werden konkrete Vorschläge zur Nutzung von Telefoninterviews als Datenerhebungsstrategie gemacht. Unsere Überlegungen sind an all jene adressiert, die telefonische Interviews durchführen (wollen), wobei sie vor allem für diejenigen Forscher(innen) hilfreich sein dürften, die über keine oder nur geringe Erfahrungen mit dieser Erhebungsmethode verfügen. Es werden zahlreiche praktische Empfehlungen gegeben, die üblicherweise in der traditionellen Forschungsliteratur n...

  5. Polimorfismo genético relacionado con la probabilidad de desarrollar asma ocupacional en trabajadores expuestos a isocianatos

    Directory of Open Access Journals (Sweden)

    Gaetano Pepe Betancourt

    2014-03-01

    Full Text Available Introducción: El desarrollo tecnológico ha traído como consecuencia el uso de sustancias químicas potencialmente perjudiciales para la salud de los trabajadores. Particularmente el uso de isocianatos ha resultado en una mayor morbilidad de patología respiratoria, especialmente el asma. Considerando que no todos los trabajadores expuestos desarrollan la enfermedad se ha propuesto un modelo de interacción gen-medioambiental, el cual trata de explicar la predisposición genética que tienen algunos individuos a desarrollar asma ocupacional y otros no. Objetivo: Conocer la evidencia científica relacionada con el polimorfismo genético y la susceptibilidad que tienen los trabajadores expuestos a isocianatos a desarrollar asma ocupacional. Metodología: Se realizó una revisión sistemática mediante una búsqueda bibliográfica utilizando las bases de datos PubMedline, así como en los repositorios Dialnet y ELSEVIER. Se extrajeron los artículos relacionados al objetivo de esta revisión, no se aplicaron filtros de temporalidad, utilizándose los siguientes descriptores: MeSH Major Topic, MeSH Terms. El periodo de búsqueda fue desde el 20 de noviembre de 2013 y finalizó el 15 de diciembre de 2013. El nivel de evidencia se estableció de acuerdo a los criterios GRADE. Resultados: Se analizaron a texto completo 42 artículos, la evidencia científica se sustentó en 11 estudios de casos-controles. Dada la complejidad del polimorfismo genético asociado con la expresión fenotípica de la enfermedad, como limitación de los estudios, los autores coinciden que el tamaño muestral no es suficientemente grande, sin embargo después de ajustar los factores de confusión los artículos encontrados tuvieron un nivel de evidencia B de GRADE. Conclusión: La genética tiene una influencia significativa en el asma ocupacional inducida por isocianatos. El peso de la susceptibilidad genética y de la interacción gen-medioambiente aún no se han

  6. Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium.

    Science.gov (United States)

    McNeer, Nicole Ali; Anandalingam, Kavitha; Fields, Rachel J; Caputo, Christina; Kopic, Sascha; Gupta, Anisha; Quijano, Elias; Polikoff, Lee; Kong, Yong; Bahal, Raman; Geibel, John P; Glazer, Peter M; Saltzman, W Mark; Egan, Marie E

    2015-01-01

    Cystic fibrosis (CF) is a lethal genetic disorder most commonly caused by the F508del mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. It is not readily amenable to gene therapy because of its systemic nature and challenges including in vivo gene delivery and transient gene expression. Here we use triplex-forming peptide nucleic acids and donor DNA in biodegradable polymer nanoparticles to correct F508del. We confirm modification with sequencing and a functional chloride efflux assay. In vitro correction of chloride efflux occurs in up to 25% of human cells. Deep-sequencing reveals negligible off-target effects in partially homologous sites. Intranasal delivery of nanoparticles in CF mice produces changes in the nasal epithelium potential difference assay, consistent with corrected CFTR function. Also, gene correction is detected in the nasal and lung tissue. This work represents facile genome engineering in vivo with oligonucleotides using a nanoparticle system to achieve clinically relevant levels of gene editing without off-target effects. PMID:25914116

  7. Türkiye'de kültür ve doğal çipura (Sparus aurata l.) populasyonları arasındaki filogenetik farklılıkların mitokondriyal gen bölgelerinin dna dizi analizleri ile tanımlanması

    OpenAIRE

    DOĞANKAYA, Levent

    2010-01-01

      Bu çalışmada, kıyılarımızda bulunan doğal çipura (Sparus aurata)  populasyonları ile balık çiftliklerinde üretilen çipura populasyonlarının filogenetik ilişkilerinin incelenmesi ve bu stoklar arasındaki farklılıkların belirlenmesi amaçlanmıştır.

    Araştırma için Adana, Mersin, Antalya, Bodrum ve İzmir olmak üzere beş istasyon seçilmiştir. Her istasyonda doğal ve kültür balık stoklarından temin edilen örneklerin karaciğerlerinde...

  8. Structures of a minimal human CFTR first nucleotide-binding domain as a monomer, head-to-tail homodimer, and pathogenic mutant

    Energy Technology Data Exchange (ETDEWEB)

    Atwell, Shane; Brouillette, Christie G.; Conners, Kris; Emtage, Spencer; Gheyi, Tarun; Guggino, William B.; Hendle, Jorg; Hunt, John F.; Lewis, Hal A.; Lu, Frances; Protasevich, Irina I.; Rodgers, Logan A.; Romero, Rich; Wasserman, Stephen R.; Weber, Patricia C.; Wetmore, Diana; Zhang, Feiyu F.; Zhao, Xun (Cystic); (UAB); (JHU); (Columbia); (Lilly)

    2010-04-26

    Upon removal of the regulatory insert (RI), the first nucleotide binding domain (NBD1) of human cystic fibrosis transmembrane conductance regulator (CFTR) can be heterologously expressed and purified in a form that remains stable without solubilizing mutations, stabilizing agents or the regulatory extension (RE). This protein, NBD1 387-646({Delta}405-436), crystallizes as a homodimer with a head-to-tail association equivalent to the active conformation observed for NBDs from symmetric ATP transporters. The 1.7-{angstrom} resolution X-ray structure shows how ATP occupies the signature LSGGQ half-site in CFTR NBD1. The {Delta}F508 version of this protein also crystallizes as a homodimer and differs from the wild-type structure only in the vicinity of the disease-causing F508 deletion. A slightly longer construct crystallizes as a monomer. Comparisons of the homodimer structure with this and previously published monomeric structures show that the main effect of ATP binding at the signature site is to order the residues immediately preceding the signature sequence, residues 542-547, in a conformation compatible with nucleotide binding. These residues likely interact with a transmembrane domain intracellular loop in the full-length CFTR channel. The experiments described here show that removing the RI from NBD1 converts it into a well-behaved protein amenable to biophysical studies yielding deeper insights into CFTR function.

  9. The CFTR polymorphisms poly-T, TG-repeats and M470V in Chinese males with congenital bilateral absence of the vas deferens

    Institute of Scientific and Technical Information of China (English)

    Wu-Hua Ni; Lei Jiang; Qian-Jin Fei; Jian-Yuan Jin; Xu Yang; Xue-Feng Huang

    2012-01-01

    Congenital bilateral absence of the vas deferens (CBAVD) is a frequent cause of obstructive azoospermia,and mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene have also been frequently identified in patients with CBAVD.However,the distribution of the CFTR polymorphisms M470V,poly-T,TG-repeats and F508del mutation in the Chinese CBAVD population with presumed low cystic fibrosis (CF) frequency remains to be evaluated.Samples obtained from 109 Chinese infertile males with CBAVD and 104 normal controls were analyzed for the presence of CFTR (TG)m(T)n,M470V and F508del by PCR amplification followed by direct sequencing.Our study showed that the F508del mutation was not found in our patients.The 5T mutation was present with high frequency in Chinese CBAVD patients and IVS8-5T linked to either 12 or 13 TG repeats was highly prevalent among CBAVD patients (97.22% of 72 cases and 96.91% of 97 alleles with IVS8-5T).Moreover,a statistically significant relationship between TG12-5T-V470 haplotype and CBAVD was detected.This study indicated that the CFTR polymorphisms poly-T,TG-repeats and M470V might affect the process of CBAVD in the Chinese population.

  10. Contribution of CFTR to Alveolar Fluid Clearance by Lipoxin A4 via PI3K/Akt Pathway in LPS-Induced Acute Lung Injury

    Directory of Open Access Journals (Sweden)

    Yi Yang

    2013-01-01

    Full Text Available The lipoxins are the first proresolution mediators to be recognized and described as the endogenous “braking signals” for inflammation. We evaluated the anti-inflammatory and proresolution bioactions of lipoxin A4 in our lipopolysaccharide (LPS-induced lung injury model. We demonstrated that lipoxin A4 significantly improved histology of rat lungs and inhibited IL-6 and TNF-α in LPS-induced lung injury. In addition, lipoxin A4 increased alveolar fluid clearance (AFC and the effect of lipoxin A4 on AFC was abolished by CFTRinh-172 (a specific inhibitor of CFTR. Moreover, lipoxin A4 could increase cystic fibrosis transmembrane conductance regulator (CFTR protein expression in vitro and in vivo. In rat primary alveolar type II (ATII cells, LPS decreased CFTR protein expression via activation of PI3K/Akt, and lipoxin A4 suppressed LPS-stimulated phosphorylation of Akt. These results showed that lipoxin A4 enhanced CFTR protein expression and increased AFC via PI3K/Akt pathway. Thus, lipoxin A4 may provide a potential therapeutic approach for acute lung injury.

  11. Lack of association between UGT1A7, UGT1A9, ARP, SPINK1 and CFTR gene polymorphisms and pancreatic cancer in Italian patients

    Institute of Scientific and Technical Information of China (English)

    Ada Piepoli; Annamaria Gentile; Maria Rosa Valvano; Daniela Barana; Cristina Oliani; Rosa Cotugno; Michele Quitadamo; Angelo Andriulli; Francesco Perri

    2006-01-01

    AIM: To investigate simultaneously UGT1A7, UGT1A9,ARP, SPINK and CFTR genes to verify whether genetic polymorphisms predispose to the development of pancreatic cancer (PC).METHODS: Genomic DNA of 61 pancreatic cancer patients and 105 healthy controls (HC) were analyzed.UGT1,47 genotyping was determined by PCR-RFLP analysis. Specific PCR and sequencing were used to analyze genetic variants of UGT1A9, ARP, SPINK1 and CFTR genes.RESULTS: Four different alleles (*1: WT;*2: N129Kand R131K;*3: N129K, R131K, and W208R;and *4:W208R) in UGT1A7 and three different alleles (*1: WT;*4: Y242X;and *5: D256N) in UGT1A9 were detected.All UGT1A polymorphisms were observed at similar frequency in PC patients and HC. Seven different alleles in ARP were found in PC patients and HC at similar frequency. The SPINK1 mutations N34S and P55Soccurred in five PC patients with a prevalence (4.1%) not significantly different from that observed (2.0%) in HC.The only CFTR ΔF508 mutation was recognized in three PC patients with a prevalence (4.9%) similar to HC.CONCLUSION: UGT1A7, UGT1A9, ARP, SPINK1 and CFTR gene polymorphisms are not associated with PC in Italian patients.

  12. Progesterone Downregulates Oestrogen-Induced Expression of CFTR and SLC26A6 Proteins and mRNA in Rats’ Uteri

    Directory of Open Access Journals (Sweden)

    K. Gholami

    2012-01-01

    Full Text Available Under progesterone (P dominance, fluid loss assists uterine closure which is associated with pH reduction. We hypothesize that P inhibits uterine fluid secretion and HCO3- transport. Aim. to investigate the expression of Cystic Fibrosis Transmembrane Regulator (CFTR and Cl−/HCO3- exchanger (SLC26A6 under P effect. Method. Uteri from ovariectomized steroid replaced and intact rats at different stages of oestrous cycle were analyzed for changes in protein and mRNA expressions. Results. P inhibits CFTR and SLC26A6 proteins and mRNA expression while oestrogen (E causes vice versa. E treatment followed by P causes a reduction in these transporters’ mRNA and protein. Similar changes occur throughout the oestrous cycle; that is, CFTR mRNA expression was high at proestrus while SLC26A6 mRNA and protein expressions were increased at proestrus and estrus. At diestrus, however, the expression of these transporters’ protein and mRNA was reduced. Conclusion. Inhibition of CFTR and SLC26A6 expressions may explain the reduced fluid volume and pH under P-mediated effect.

  13. Estimación de componentes de varianza genética con cruzas autofecundadas de progenitores no endogámicos.

    OpenAIRE

    Jaime Sahagún Castellanos

    2003-01-01

    En el fitomejoramiento genético, la estimación de componentes de la varianza genética frecuentemente requiere hacer cruzas, tarea que se complica por la reducida producción de semilla para evaluación. Para incrementar la cantidad de semilla se ha sugerido el uso de cruzas autofecundadas (CA), pero la teoría de este enfoque se ha restringido al caso, muy limitado, de progenitores homocigóticos con un modelo de dos alelos. Aquí se pretende, para poblaciones panmícticas con multialelismo: 1) Der...

  14. Búsqueda de factores genéticos de susceptibilidad a la artrosis: fenotipos extremos, variantes raras, microsatélites y metaanálisis de genes candidatos

    OpenAIRE

    Rodríguez Fontenla, María Cristina

    2014-01-01

    diversas alteraciones en los tejidos de la articulación. Se trata de una enfermedad compleja de naturaleza multifactorial en la que participan factores sistémicos, biomecánicos y genéticos. El estudio de los factores genéticos implicados en la susceptibilidad a la OA es un campo complejo que ha progresado en los últimos años. En la actualidad se puede hablar de un total de nueve genes y/o loci asociados con OA en población europea al nivel requerido en los GWAS (Genome Wide Associ...

  15. Evaluación del polimorfismo del gen leptina en bovinos en el sistema doble proposito en Chiapas, México.

    OpenAIRE

    Ortiz Salazar, Jorge Alberto

    2011-01-01

    La leptina es una hormona proteica de 16 KDa, compuesta de 146 aminoácidos y es sintetizada principalmente por el tejido adiposo. En el eje hipotálamo-hipófisisgonadal, la leptina juega un papel muy importante en la regulación de la reproducción de los mamíferos. La mutación del gen leptina TT está asociado con la calidad de la carne y leche en bovinos. El objetivo de este estudio fue estimar las frecuencias genotípicas y alélicas del polimorfismo (SNP, Single Nucleotide Polymorphism) del gen...

  16. Diversidad y estructura genética de accesiones de palma de aceite (Elaeis guineesis Jacq.) provenientes de Camerún

    OpenAIRE

    Jose Estiben Pacheco Diaz; Diana Marcela Arias Moreno; Zaida Zarely Ojeda Perez; Hernán Mauricio Romero Angulo

    2014-01-01

    Título en español: Diversidad y estructura genética de accesiones de palma de aceite (Elaeis guineesis Jacq.) provenientes de CamerúnTítulo en ingles: Diversity and genetic structure of oil palm accession (Elaeis guineesis Jacq.)  from CameroonTítulo corto: Diversidad genética de accesiones de palma de aceiteResumen: La palma de aceite Elaeis guineesis Jacq. posee gran importancia debido al aceite que se extrae de sus frutos, del cual se obtienen derivados refinados de gran valor comercial co...

  17. Clonación artificial de un controlador on-line, basado en lógica difusa y algoritmos genéticos

    OpenAIRE

    Javier Ballesteros; Alonso Guevara

    2006-01-01

    Los Algoritmos Genéticos son procedimientos adaptativos para la búsqueda de soluciones en espacios complejos, inspirados en la evolución biológica, con patrones de operaciones basados en el principio darwiniano de reproducción y supervivencia de los individuos que mejor se adaptan al entorno en que viven. En este artículo se presenta un estudio sobre los Algoritmos Genéticos y la Lógica Difusa, para desarrollar una metodología propuesta y replicar la caja negra de un controlador, utilizando p...

  18. The GenDev Curriculum Development Workshop.

    Science.gov (United States)

    D'cunha, J

    1997-01-01

    This article describes the second Curriculum Development Workshop held in May 1997 at the Asian Institute of Technology (AIT) in Bangkok, Thailand. The workshop aimed to review critically and restructure the Gender and Development Studies (GenDev) curriculum and to assess AIT's role in training gender experts for the region. Participants included 22 people from 16 countries in Asia, Europe, and the US who were teaching graduate students about gender issues and who were activists with nongovernmental organizations working on gender issues. It was determined that the following were required courses: Culture, Knowledge and Gender Relations; Gender, Technology, and Development; Principles of Gender Research and Methodology in Science and Technology; and Gender Analysis and Field Methods. Other suggested core courses included: Gender and Natural Resource Management; Enterprise Management, Technology, and Gender; Gender and Agrarian Reform; Urbanization: A Gender Perspective; Gender-Responsive Development Planning; and Gender and Economic Change: Past and Present Concerns. Participants distinguished between GenDev courses offered to anyone attending AIT and training courses designed to produce gender experts in the region. The aim of training courses for AIT graduate students was to sensitize potential managers, technologists, and others on gender issues and to create awareness of the importance of including gender perspectives within decision-making, policy formation, and implementation. Training courses to produce gender experts should be directed to those with a prior background in gender studies and include gender analysis in field methods. Participants agreed that there should be an independent and autonomous field of gender and development studies. Participants made six recommendations for such a field of study. PMID:12179927

  19. Aplicación de la Genética a la Cría de las Plantas.

    Directory of Open Access Journals (Sweden)

    Hutchinson J. B.

    1949-09-01

    Full Text Available La influencia de la ciencia de la genética en el arte de la cría de plantas ha sido mucho menos profunda de cuanto esperaban los primeros genetistas. Al dilucidar el mecanismo hereditario, fueron utilizados únicamente factores que originaban grandes diferencias, sin caer en la cuenta inmediata de que ellos son de pequeña importancia naturalmente o bajo la selección artificial. El progreso en el control genético de las pequeñas diferencias ha sido difícil y lento, considerando todavía muchos genetistas que los "genes" de menor categoría no valen la pena de ser investigados. Algunos principios genéticos han sido aplicados con éxito por los criadores, pero, a menudo, los intrincados principios no son totalmente comprendidos y la aplicaci6n se ha hecho con frecuencia sin apoyo experimental, basando las deducciones en observaciones generales sobre el material de cría. En relación con los métodos de cría es notable la frecuencia con que una práctica de fundamental importancia se justifica por una declaración que principia así: "Mi impresión es . .. ". Será tarea del genetista substituir la intuición del criador, por la evidencia objetiva. El objeto de este escrito consiste en revisar la aplicación de la teoría genética al cultivo del algodón en particular y bosquejar los problemas en los cuales el genetista podrá ayudar al criador.

  20. Sobre el significado del descubrimiento del gen FOXP2

    OpenAIRE

    Víctor Manuel LONGA MARTÍNEZ

    2006-01-01

    El reciente descubrimiento del gen FOXP2 ha ofrecido la primera evidencia clara de la base genética del lenguaje, mostrando una correlación inequívoca desde la perspectiva genética entre una versión mutada de F0XP2 y los trastornos lingüísticos de diferente tipo sufridos por una familia inglesa, conocida como KE. El objetivo central del presente trabajo es discutir diferentes aspectos relacionados con tal descubrimiento; especialmente, la discusión del significado de FOXP2 con ...

  1. Estudio de bloques constructivos en algoritmos genéticos

    OpenAIRE

    Murias Rodríguez, Angel

    2007-01-01

    Estudio experimental sobre el efecto de las modificaciones en los bloques constructivos. Para conseguirlo, primero se ha diseñado un algoritmo genético genérico con posibilidad de adaptarlo a un problema específico (función de objetivo a optimizar) e incorporar fácilmente diferentes características que pueden tener estos algoritmos. Como referencia de comparación se ha diseñado y implementado un algoritmo genético estándar y los algoritmos con las modificaciones. Se han probado los algorit...

  2. Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis.

    Science.gov (United States)

    Wang, Chi; Protasevich, Irina; Yang, Zhengrong; Seehausen, Derek; Skalak, Timothy; Zhao, Xun; Atwell, Shane; Spencer Emtage, J; Wetmore, Diana R; Brouillette, Christie G; Hunt, John F

    2010-10-01

    The lethal genetic disease cystic fibrosis is caused predominantly by in-frame deletion of phenylalanine 508 in the cystic fibrosis transmembrane conductance regulator (CFTR). F508 is located in the first nucleotide-binding domain (NBD1) of CFTR, which functions as an ATP-gated chloride channel on the cell surface. The F508del mutation blocks CFTR export to the surface due to aberrant retention in the endoplasmic reticulum. While it was assumed that F508del interferes with NBD1 folding, biophysical studies of purified NBD1 have given conflicting results concerning the mutation's influence on domain folding and stability. We have conducted isothermal (this paper) and thermal (accompanying paper) denaturation studies of human NBD1 using a variety of biophysical techniques, including simultaneous circular dichroism, intrinsic fluorescence, and static light-scattering measurements. These studies show that, in the absence of ATP, NBD1 unfolds via two sequential conformational transitions. The first, which is strongly influenced by F508del, involves partial unfolding and leads to aggregation accompanied by an increase in tryptophan fluorescence. The second, which is not significantly influenced by F508del, involves full unfolding of NBD1. Mg-ATP binding delays the first transition, thereby offsetting the effect of F508del on domain stability. Evidence suggests that the initial partial unfolding transition is partially responsible for the poor in vitro solubility of human NBD1. Second-site mutations that increase the solubility of isolated F508del-NBD1 in vitro and suppress the trafficking defect of intact F508del-CFTR in vivo also stabilize the protein against this transition, supporting the hypothesize that it is responsible for the pathological trafficking of F508del-CFTR. PMID:20687163

  3. Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

    Science.gov (United States)

    Wang, Chi; Protasevich, Irina; Yang, Zhengrong; Seehausen, Derek; Skalak, Timothy; Zhao, Xun; Atwell, Shane; Spencer Emtage, J; Wetmore, Diana R; Brouillette, Christie G; Hunt, John F

    2010-01-01

    The lethal genetic disease cystic fibrosis is caused predominantly by in-frame deletion of phenylalanine 508 in the cystic fibrosis transmembrane conductance regulator (CFTR). F508 is located in the first nucleotide-binding domain (NBD1) of CFTR, which functions as an ATP-gated chloride channel on the cell surface. The F508del mutation blocks CFTR export to the surface due to aberrant retention in the endoplasmic reticulum. While it was assumed that F508del interferes with NBD1 folding, biophysical studies of purified NBD1 have given conflicting results concerning the mutation's influence on domain folding and stability. We have conducted isothermal (this paper) and thermal (accompanying paper) denaturation studies of human NBD1 using a variety of biophysical techniques, including simultaneous circular dichroism, intrinsic fluorescence, and static light-scattering measurements. These studies show that, in the absence of ATP, NBD1 unfolds via two sequential conformational transitions. The first, which is strongly influenced by F508del, involves partial unfolding and leads to aggregation accompanied by an increase in tryptophan fluorescence. The second, which is not significantly influenced by F508del, involves full unfolding of NBD1. Mg-ATP binding delays the first transition, thereby offsetting the effect of F508del on domain stability. Evidence suggests that the initial partial unfolding transition is partially responsible for the poor in vitro solubility of human NBD1. Second-site mutations that increase the solubility of isolated F508del-NBD1 in vitro and suppress the trafficking defect of intact F508del-CFTR in vivo also stabilize the protein against this transition, supporting the hypothesize that it is responsible for the pathological trafficking of F508del-CFTR. PMID:20687163

  4. 公丁香提取物抑制CFTR氯离子通道的发现与研究%The extract of clove inhibits CFTR chloride channel

    Institute of Scientific and Technical Information of China (English)

    栾剑; 张耀方; 杨红

    2015-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is an epithelial chloride chan‐nel .In recent years ,the blockers of CFTR become the new hot spot in the treatment of secretory di‐arrhea .The aim of this research is using high‐throughput screening techniques screened blockers of CFTR chloride channel from traditional Chinese medicine .In this study ,after 40000 fractions of Chi‐nese herbal medicine have been screened ,clove extract was found .In cell‐based fluorescence assays and voltage clamp experiments ,the best active fraction‐E06 significantly blocks CFTR chloride chan‐nel .Therefore ,clove extract screened from traditional Chinese medicine blocks CFTR chloride chan‐nel and provides a theoretical basis for the in‐depth study of anti‐diarrheal drugs .%囊性纤维化跨膜电导调节因子(CFTR)是一种上皮细胞顶膜中表达的氯离子通道,是近年来治疗分泌型腹泻的新热点。利用高通量筛选技术,自中国传统中药中筛选能够抑制CFTR氯离子通道的中药组分。结果显示,自500种中草药的40000种中药组分中筛选到公丁香。经细胞荧光实验和电压膜片钳实验验证公丁香最佳活性孔———E06对CFTR具有明显的抑制作用,IC50=103 mg/L 。本研究结果为深入探讨公丁香的抗泻药物研发提供理论依据。

  5. Intención de compra de medicamentos genéricos por parte de los usuarios de Asturias

    Directory of Open Access Journals (Sweden)

    González Hernando Santiago

    2003-01-01

    Full Text Available Fundamento: Conocer las percepciones de los consumidores acerca del riesgo asociado al uso de medicamentos genéricos y los factores que más influyen en la intención de solicitar un genérico al médico (prescriptor y/o al farmacéutico, a fin de determinar posibles barreras o frenos a la aceptación de los mismos y obtener información que apoye la toma de decisiones de los gestores sanitarios. Métodos: Estudio sobre utilización de medicamentos centrado en la disposición de los pacientes a solicitar una EFG. En esta investigación transversal cuantitativa se entrevistó personalmente a 542 individuos, a la salida de un centro de salud o de un establecimiento de farmacia en Asturias. En el cuestionario se incluía una escala de medición del riesgo percibido en la compra de un medicamento con 15 atributos agrupados en cinco dimensiones. Asimismo se recogió información sobre la intención de consumir medicamentos genéricos y sobre las características demográficas y socioeconómicas de los entrevistados. Para el análisis de los resultados se aplicaron un análisis factorial confirmatorio, regresión múltiple y análisis univariable. El tratamiento de los datos se efectuó con los programas estadísticos EQS y SPSS. Resultados: Percepción media del riesgo (escalas de 1 a 7: funcional: 2,75; físico: 2,68; financiero: 2,19; psicológico: 1,99; social: 1,42. Factores influyentes sobre la intención de solicitar genéricos al médico: riesgo psicológico (p=0,000. Sobre la solicitud al farmacéutico: riesgo psicológico (p=0,000 y riesgo social (p=0,020. Conclusiones: Los agentes interesados en el desarrollo en el mercado de las EFG deben mantener sus esfuerzos de comunicación hacia la equiparación de los aspectos funcionales y financieros entre especialidades del fabricante y especialidades genéricas, pero no deben dejar de lado aspectos psicológicos y sociales del comportamiento de compra del consumidor.

  6. Uso de algoritmos genéticos para detección de objetos en tiempo real

    OpenAIRE

    Martínez Gómez, Jesús; Gómez Martín, José Antonio; García Varea, Ismael; Matellán Olivera, Vicente

    2009-01-01

    El artículo presenta un sistema de Visión para la detección de objetos mediante el uso de algoritmos genéticos. La principal aportación del mismo consiste en la adaptación al tiempo real de algoritmos generalmente utilizados offline. Para conseguir esto se necesita una ejecución eficiente que reduzca al máximo la complejidad del algoritmo. El ámbito de aplicación del sistema es el campeonato de fútbol robótico RoboCup, en concreto la categoría Standard Platform, y debe permi...

  7. Clonación artificial de un controlador on-line, basado en lógica difusa y algoritmos genéticos

    Directory of Open Access Journals (Sweden)

    Javier Ballesteros

    2006-10-01

    Full Text Available Los Algoritmos Genéticos son procedimientos adaptativos para la búsqueda de soluciones en espacios complejos, inspirados en la evolución biológica, con patrones de operaciones basados en el principio darwiniano de reproducción y supervivencia de los individuos que mejor se adaptan al entorno en que viven. En este artículo se presenta un estudio sobre los Algoritmos Genéticos y la Lógica Difusa, para desarrollar una metodología propuesta y replicar la caja negra de un controlador, utilizando procedimientos de obtención del conjunto de reglas de inferencia, agrupamiento difuso y después aplicar el desarrollo del algoritmo genético simple con algunas alteraciones, buscando el objetivo del trabajo propuesto.

  8. Comportamento de dois genótipos de milho cultivados em sistema de aléias preestabelecido com diferentes leguminosas arbóreas Behaviour of two maize genotypes grown in alley cropping system pre-established with diferents leguminous trees

    OpenAIRE

    Andréia Araújo Lima Leite; Altamiro Souza de Lima Ferraz Junior; Emanoel Gomes de Moura; Alana das Chagas Ferreira Aguiar

    2008-01-01

    O cultivo em aléias tem sido recomendado como alternativa para a substituição da agricultura de corte e queima, no trópico úmido, devido à grande capacidade de produção de matéria orgânica e de reciclagem de nutrientes, mas algumas dúvidas quanto à sustentabilidade e à competição interespecífica são persistentes. O objetivo no trabalho foi avaliar a viabilidade da cultura do milho em um sistema de cultivo em aléias de leguminosas arbóreas. O delineamento experimental utilizado foi em blocos c...

  9. Desempenho de genótipos de batata-doce submetidos ao efeito da calagem em Rio Largo-Alagoas

    Directory of Open Access Journals (Sweden)

    Islan Diego Espindula de Carvalho

    2015-09-01

    Full Text Available O objetivo desse trabalho foi avaliar o desempenho de genótipos de batata-doce submetidos ao efeito da calagem em Rio Largo-AL. O delineamento experimental utilizado foi o de blocos casualizados no esquema fatorial (7 x 2,  com sete genótipos de batata-doce e dois tipos de correção do solo, em três repetições. As variáveis avaliadas foram: Número de Raízes Comerciais (NRC; Número Total de Raízes (NTR; Comprimento de Raízes Comerciais (CRC; Diâmetro de Raízes Comerciais (DRC; Rendimento de Raízes Comerciais (RRC; Rendimento Total de Raízes (RTR e Peso Médio de Raízes Comerciais (PMRC. Dentre os genótipos de batata-doce avaliados no experimento, o Clone-06 apresentou o melhor desempenho, superando todos os genótipos avaliados, inclusive as testemunhas, Rainha de Penedo e Sergipana, tanto em produtividade, com rendimento médio de 15,79 t.ha-1de raízes comerciais, quanto em qualidade, apresentando dimensões de raízes comerciais dentro dos padrões de comercialização como Extra A. O desempenho dos genótipos, bem como a grande maioria das variáveis de batata-doce não foram influenciadas pela correção do solo.

  10. GenBank blastx search result: AK243655 [KOME

    Lifescience Database Archive (English)

    Full Text Available putative transcriptional activator; similar to cAMP-receptor (CAP, CRP) protein: PIR Access...ion Number A26049; similar to fnr (nirR) gene product encoded by GenBank Accession Number J01608;

  11. GenBank blastx search result: AK062102 [KOME

    Lifescience Database Archive (English)

    Full Text Available transcriptional activator; similar to cAMP-receptor (CAP, CRP) protein: PIR Access...ion Number A26049; similar to fnr (nirR) gene product encoded by GenBank Accession Number J01608; aspartic

  12. GenBank blastx search result: AK242621 [KOME

    Lifescience Database Archive (English)

    Full Text Available putative transcriptional activator; similar to cAMP-receptor (CAP, CRP) protein: PIR Access...ion Number A26049; similar to fnr (nirR) gene product encoded by GenBank Accession Number J01608;

  13. Structure and Dynamics of NBD1 from CFTR Characterized Using Crystallography and Hydrogen/Deuterium Exchange Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, H.A.; Wang, C.; Zhao, X.; Hamuro, Y.; Conners, K.; Kearins, M.C.; Lu, F.; Sauder, J.M.; Molnar, K.S.; Coales, S.J.; Maloney, P.C.; Guggino, W.B.; Wetmore, D.R.; Weber, P.C.; Hunt, J.F. (SGX); (ExSAR); (Cystic); (JHU-MED); (Columbia)

    2012-04-30

    The {Delta}F508 mutation in nucleotide-binding domain 1 (NBD1) of the cystic fibrosis transmembrane conductance regulator (CFTR) is the predominant cause of cystic fibrosis. Previous biophysical studies on human F508 and {Delta}F508 domains showed only local structural changes restricted to residues 509-511 and only minor differences in folding rate and stability. These results were remarkable because {Delta}F508 was widely assumed to perturb domain folding based on the fact that it prevents trafficking of CFTR out of the endoplasmic reticulum. However, the previously reported crystal structures did not come from matched F508 and {Delta}F508 constructs, and the {Delta}F508 structure contained additional mutations that were required to obtain sufficient protein solubility. In this article, we present additional biophysical studies of NBD1 designed to address these ambiguities. Mass spectral measurements of backbone amide {sup 1}H/{sup 2}H exchange rates in matched F508 and {Delta}F508 constructs reveal that {Delta}F508 increases backbone dynamics at residues 509-511 and the adjacent protein segments but not elsewhere in NBD1. These measurements also confirm a high level of flexibility in the protein segments exhibiting variable conformations in the crystal structures. We additionally present crystal structures of a broader set of human NBD1 constructs, including one harboring the native F508 residue and others harboring the {Delta}F508 mutation in the presence of fewer and different solubilizing mutations. The only consistent conformational difference is observed at residues 509-511. The side chain of residue V510 in this loop is mostly buried in all non-{Delta}F508 structures but completely solvent exposed in all {Delta}F508 structures. These results reinforce the importance of the perturbation {Delta}F508 causes in the surface topography of NBD1 in a region likely to mediate contact with the transmembrane domains of CFTR. However, they also suggest that increased

  14. ANALISIS MOLEKULER GEN OMP LEPTOSPIRA UNTUK DIAGNOSIS SEROLOGIS SPESIFIK

    OpenAIRE

    Rosdiana Natzir; Zaraswati Dwyana

    2007-01-01

    Leptospira dapat menginfeksi ginjal, hati, paru-paru, otak dan mata, bahkan dapat menyebabkan kematian, hal ini sering terjadi, bila diagnose dan penanganannya tidak tepat. Dalam pengembangan untuk pencarian terget yang penting untuk membuat serodiagnose terhadap Leptospira, gen OMP merupakan target ideal, dan perlu pengetahuan mengenai gen OMP ini. Pada penelitian ini dilakukan ekstraksi DNA dengan menggunakan Wizard Genomic DNA Purification Kit dari Promega. Setelah itu dilakukan amplif...

  15. Diagnosis of cystic fibrosis in the kindred of an infant with CFTR-related metabolic syndrome: Importance of follow-up that includes monitoring sweat chloride concentrations over time

    OpenAIRE

    Williams, SN; Nussbaum, E.; Chin, TW; Do, PCM; Singh, KE; Randhawa, I

    2014-01-01

    Newly implemented newborn screening (NBS) programs in California have resulted in a large subset of patients in whom at least two cystic fibrosis transmembrane conductance regulator (CFTR) mutations are identified, but subsequent sweat chloride analysis reveals normal or indeterminate values. These patients are diagnosed with CFTR-Related Metabolic Syndrome (CRMS). However, the natural progression and management of these patients are not clearly understood and frequently after the age of 1-ye...

  16. Diversidad y diferenciación genética de la yuca ( Manihot esculenta Crantz con marcadores microsatélites en poblaciones de África y Latinoamérica

    Directory of Open Access Journals (Sweden)

    Fregene Martin

    2004-12-01

    Full Text Available Se estudió la diversidad y diferenciación genética de 224 accesiones de yuca tradicionalmente cultivadas en Uganda. Adicionalmente, se incluyeron estudios previos de diversidad, 20 materiales de Tanzania, 20 de Ghana, 22 de Nigeria, 20 de Guatemala y 12 accesiones representando la colección núcleo de Latinoamérica, mantenidas en CIAT. Nueve grupos basados en el país de origen fueron creados para estudiar la variación genética dentro y entre países. Usando secuencias simples repetidas (SSR o marcadores microsatélites, la variación en las frecuencias alélicas en 35 loci no ligados sirvió para estimar los parámetros de diversidad y diferenciación genética. Los resultados afirman una divergencia genética entre accesiones africanas y latinoamericanas, y una fuerte diferenciación de algunas accesiones de Guatemala con respecto a los
    otros países. Ellos también muestran una alta diversidad genética dentro de países y una moderada diferenciación entre ellos. En particular Uganda mantiene alta diversidad genética
    dentro Distritos aún después de una reciente epidemia de CMD (cassava mosaic disease pero baja diferenciación entre ellos. Se discuten las posibles fuerzas implicadas en la dinámica de la diversidad genética, la importancia de Guatemala en los programas de mejoramiento de yuca en la búsqueda de grupos con potencial heterótico, el bajo impacto causado por CMD en la constitución genética del cultivo en Uganda y la observación de una distribución continua de la diversidad genética.

  17. New sequestrate fungi from Guyana: Jimtrappea guyanensis gen. sp. nov., Castellanea pakaraimophila gen. sp. nov., and Costatisporus cyanescens gen. sp. nov. (Boletaceae, Boletales).

    Science.gov (United States)

    Smith, Matthew E; Amses, Kevin R; Elliott, Todd F; Obase, Keisuke; Aime, M Catherine; Henkel, Terry W

    2015-12-01

    Jimtrappea guyanensis gen. sp. nov., Castellanea pakaraimophila gen. sp. nov., and Costatisporus cyanescens gen. sp. nov. are described as new to science. These sequestrate, hypogeous fungi were collected in Guyana under closed canopy tropical forests in association with ectomycorrhizal (ECM) host tree genera Dicymbe (Fabaceae subfam. Caesalpinioideae), Aldina (Fabaceae subfam. Papilionoideae), and Pakaraimaea (Dipterocarpaceae). Molecular data place these fungi in Boletaceae (Boletales, Agaricomycetes, Basidiomycota) and inform their relationships to other known epigeous and sequestrate taxa within that family. Macro- and micromorphological characters, habitat, and multi-locus DNA sequence data are provided for each new taxon. Unique morphological features and a molecular phylogenetic analysis of 185 taxa across the order Boletales justify the recognition of the three new genera. PMID:26732137

  18. Identificación del gen MALT1 como oncogén dominante en el linfoma MALT.

    OpenAIRE

    Sánchez Izquierdo, Maria Dolores

    2005-01-01

    RESUMEN En los linfomas no Hodgkin (LNH), las traslocaciones primarias características dan lugar a la desregulación de un protooncogén (debido en un gran número de casos al reordenamiento con el gen de las inmunoglobulinas) y cuya consecuencia funcional es la alteración de la expresión génica con potencial neoplásico. Otro tipo de lesión genética frecuente es la delección cromosómica que inactiva genes supresores tumorales, por ejemplo p53 ó p16. Estas anomalías se identifican junto a las ...

  19. Reflexión bioética sobre el uso de organismos genéticamente modificados

    Science.gov (United States)

    Yunta, Eduardo Rodríguez

    2011-01-01

    El presente artículo reflexiona desde los 4 principios de la bioética el uso comercial de organismos genéticamente modificados. Se cuestiona fundamentalmente la falta de transferencia de tecnología entre el mundo desarrollado y en desarrollo y el que el presente sistema de patentamiento de organismos vivos modificados fomenta intereses comerciales y no da debida importancia al desarrollo sostenible de la agricultura y ganadería en los países en desarrollo, donde más se necesita. Se reflexiona sobre la importancia que tiene evaluar los riesgos antes de introducirse en el mercado organismos genéticamente modificados y la necesidad de regulación en los países. PMID:21927675

  20. Banque Cantonale de Genève

    CERN Document Server

    Banque Cantonale de Genève

    2011-01-01

    7e Salon Immobilier BCGE le samedi 3 septembre 2011, de 8 h 30 à 13 h 00, au Centre de formation de Conches À cette occasion, les meilleurs spécialistes professionnels genevois de l’immobilier seront réunis en un seul et même lieu. Si vous le souhaitez, un conseiller spécialisé dans les financements hypothécaires évaluera vos possibilités d’investissement immobilier adaptées à votre situation personnelle. En parallèle, les plus importantes régies immobilières de Genève seront à votre disposition pour vous présenter leurs offres actuelles, ainsi que les projets immobiliers futurs et discuter avec vous de la meilleure stratégie à adopter pour trouver l’objet de vos rêves. De plus, vous aurez la possibilité...

  1. The CFTR Met 470 allele is associated with lower birth rates in fertile men from a population isolate.

    Directory of Open Access Journals (Sweden)

    Gülüm Kosova

    2010-06-01

    Full Text Available Although little is known about the role of the cystic fibrosis transmembrane regulator (CFTR gene in reproductive physiology, numerous variants in this gene have been implicated in etiology of male infertility due to congenital bilateral absence of the vas deferens (CBAVD. Here, we studied the fertility effects of three CBAVD-associated CFTR polymorphisms, the (TGm and polyT repeat polymorphisms in intron 8 and Met470Val in exon 10, in healthy men of European descent. Homozygosity for the Met470 allele was associated with lower birth rates, defined as the number of births per year of marriage (P = 0.0029. The Met470Val locus explained 4.36% of the phenotypic variance in birth rate, and men homozygous for the Met470 allele had 0.56 fewer children on average compared to Val470 carrier men. The derived Val470 allele occurs at high frequencies in non-African populations (allele frequency = 0.51 in HapMap CEU, whereas it is very rare in African population (Fst = 0.43 between HapMap CEU and YRI. In addition, haplotypes bearing Val470 show a lack of genetic diversity and are thus longer than haplotypes bearing Met470 (measured by an integrated haplotype score [iHS] of -1.93 in HapMap CEU. The fraction of SNPs in the HapMap Phase2 data set with more extreme Fst and iHS measures is 0.003, consistent with a selective sweep outside of Africa. The fertility advantage conferred by Val470 relative to Met470 may provide a selective mechanism for these population genetic observations.

  2. Impact of the CFTR-potentiator ivacaftor on airway microbiota in cystic fibrosis patients carrying a G551D mutation.

    Directory of Open Access Journals (Sweden)

    Cédric Bernarde

    Full Text Available Airway microbiota composition has been clearly correlated with many pulmonary diseases, and notably with cystic fibrosis (CF, an autosomal genetic disorder caused by mutation in the CF transmembrane conductance regulator (CFTR. Recently, a new molecule, ivacaftor, has been shown to re-establish the functionality of the G551D-mutated CFTR, allowing significant improvement in lung function.The purpose of this study was to follow the evolution of the airway microbiota in CF patients treated with ivacaftor, using quantitative PCR and pyrosequencing of 16S rRNA amplicons, in order to identify quantitative and qualitative changes in bacterial communities. Three G551D children were followed up longitudinally over a mean period of more than one year covering several months before and after initiation of ivacaftor treatment.129 operational taxonomy units (OTUs, representing 64 genera, were identified. There was no significant difference in total bacterial load before and after treatment. Comparison of global community composition found no significant changes in microbiota. Two OTUs, however, showed contrasting dynamics: after initiation of ivacaftor, the relative abundance of the anaerobe Porphyromonas 1 increased (p<0.01 and that of Streptococcus 1 (S. mitis group decreased (p<0.05, possibly in relation to the anti-Gram-positive properties of ivacaftor. The anaerobe Prevotella 2 correlated positively with the pulmonary function test FEV-1 (r=0.73, p<0.05. The study confirmed the presumed positive role of anaerobes in lung function.Several airway microbiota components, notably anaerobes (obligate or facultative anaerobes, could be valuable biomarkers of lung function improvement under ivacaftor, and could shed light on the pathophysiology of lung disease in CF patients.

  3. Frequency of CFTR, SPINK1, and Cathepsin B Gene Mutation in North Indian Population: Connections between Genetics and Clinical Data

    Directory of Open Access Journals (Sweden)

    Shweta Singh

    2014-01-01

    Full Text Available Objectives. Genetic mutations and polymorphisms have been correlated with chronic pancreatitis (CP. This study aims to investigate the association of genetic variants of cystic fibrosis transmembrane conductance regulator (CFTR and serine protease inhibitor Kazal type 1 (SPINK-1 genes and Cathepsin B gene polymorphisms with CP and to associate genetic backgrounds with clinical phenotypes. Methods. 150 CP patients and 150 normal controls were enrolled consecutively. We analyzed SPINK-1 N34S and IVS3+2T>C gene mutations by PCR-restriction-fragment length polymorphism (RFLP. The identification of DF508, G551D, G542X, R117H, and W1282X mutations was carried out by ARMS-PCR. S549N mutation, IVS8 polyTn polymorphism, and Cathepsin B Lec26Val were analysed by PCR-RFLP, nested PCR, and PCR-RFLP plus sequencing, respectively. Results. We found a significant association of SPINK1 (N34S gene polymorphism. IVS1−37T>C polymorphism shows linkage with 101A>G. 300 chromosomes belonging to the CFTR subgroup exhibited minor allele frequency of 0.04, 0.03, 0.03, 0.013, 0.006, and 0.02 for DF508, G452X, G551D, S549N, R117H, and IVS8 T5, respectively. Except for R117H and IVS8 T5 polymorphisms, all other mutations showed significant variation. Conclusion. Analysis of potential susceptibility variants is needed to support nature of the genes and environment in pancreatitis. This data may help establish genetic screening and prenatal setup for Indian population.

  4. Mutant cycles at CFTR's non-canonical ATP-binding site support little interface separation during gating.

    Science.gov (United States)

    Szollosi, Andras; Muallem, Daniella R; Csanády, László; Vergani, Paola

    2011-06-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel belonging to the adenosine triphosphate (ATP)-binding cassette (ABC) superfamily. ABC proteins share a common molecular mechanism that couples ATP binding and hydrolysis at two nucleotide-binding domains (NBDs) to diverse functions. This involves formation of NBD dimers, with ATP bound at two composite interfacial sites. In CFTR, intramolecular NBD dimerization is coupled to channel opening. Channel closing is triggered by hydrolysis of the ATP molecule bound at composite site 2. Site 1, which is non-canonical, binds nucleotide tightly but is not hydrolytic. Recently, based on kinetic arguments, it was suggested that this site remains closed for several gating cycles. To investigate movements at site 1 by an independent technique, we studied changes in thermodynamic coupling between pairs of residues on opposite sides of this site. The chosen targets are likely to interact based on both phylogenetic analysis and closeness on structural models. First, we mutated T460 in NBD1 and L1353 in NBD2 (the corresponding site-2 residues become energetically coupled as channels open). Mutation T460S accelerated closure in hydrolytic conditions and in the nonhydrolytic K1250R background; mutation L1353M did not affect these rates. Analysis of the double mutant showed additive effects of mutations, suggesting that energetic coupling between the two residues remains unchanged during the gating cycle. We next investigated pairs 460-1348 and 460-1375. Although both mutations H1348A and H1375A produced dramatic changes in hydrolytic and nonhydrolytic channel closing rates, in the corresponding double mutants these changes proved mostly additive with those caused by mutation T460S, suggesting little change in energetic coupling between either positions 460-1348 or positions 460-1375 during gating. These results provide independent support for a gating model in which ATP-bound composite site 1 remains

  5. Variabilidad genética en cepas de Plasmodium falciparum circulantes en regiones colombianas con riesgo diferente para malaria

    Directory of Open Access Journals (Sweden)

    Mauricio Arcos Burgos

    2000-02-01

    grado de variabilidad genética de la población de P. falciparum circulante en estas regiones se utilizarán como marcadores genéticos los genes que codifican para las proteínas superficiales del merozoito 1 y 2 (MSP-1 y MSP-2 y para la proteína rica en glutamato (GLURP. Por medio de una PCR anidada se amplificarán segmentos altamente polimórficos de estos genes; variantes de las familias alélicas de MSP-1 (MAD20, K1 y RO33 y MSP-2 (FC27 e IC se amplificarán en la segunda reacción. Los productos de la PCR se analizarán por electroforesis en geles de agarosa para determinar el número de alelos para cada gen y la presencia de infección multiclonal.

  6. Productividad acumulada y su relación genética con características reproductivas en hembras Brahman

    Directory of Open Access Journals (Sweden)

    Orlando Duitama C.

    2013-10-01

    Full Text Available Objetivo. Estimar parámetros genéticos para la característica de productividad acumulada (PAC y correlaciones genéticas con edad al primer parto (EPP, intervalo entre el primero y segundo parto (IEP1 y longevidad (LONG. Materiales y métodos. Fueron usados 8584 registros de hembras Brahman, utilizando un modelo animal en análisis multi-característico con el método de máxima verosimilitud restricta, implementado en el software WOMBAT. Los modelos consideraron los efectos fijos de grupo contemporáneo, número de partos, y la covariable peso al destete del primer ternero; el único efecto aleatorio fue el genético aditivo directo. El peso al destete (P240 fue incluido para disminuir el efecto de la selección en la estimación de los componentes de varianza. Resultados. Las estimativas de heredabilidad fueron de 0.3±0.04, 0.11±0.03, 0.07±0.03 y 0.24±0.04 para EPP, IEP1, LONG y PAC respectivamente. Las correlaciones entre PAC y las otras características se presentaron de moderadas a altas y en sentido favorable. Conclusiones. PAC puede ser incluida en los programas de mejoramiento genético para Brahman, y utilizada como criterio de selección por su heredabilidad moderada y correlación genética favorable con las características reproductivas en estudio.

  7. Caracterización de nuevos reguladores de los mecanismos de tolerancia al daño en el DNA en Saccharomyces cerevisiae

    OpenAIRE

    Gallego Sánchez, Alfonso

    2014-01-01

    [ES]Tanto los mecanismos de tolerancia al daño en el DNA como los checkpoints de integridad del material genético son sistemas esenciales en el mantenimiento de la estabilidad genómica en los organismos eucariotas, concepto este a su vez ligado con campos de estudio tan diversos como la oncogénesis o la evolución de las especies. Empleando la levadura de gemación, Saccharomyces cerevisiae, este trabajo de tesis doctoral ha contribuido, en primer lugar y mediante una aproximación genética, al ...

  8. Diversidad genética y filogenia molecular de poblaciones de Mauritia flexuosa L.f. “aguaje” de la Amazonía Peruana

    Directory of Open Access Journals (Sweden)

    Jorge Angulo-Quintanilla

    2014-07-01

    Full Text Available Mauritia flexuosa es una especie vegetal amazónica que forma extensas poblaciones denominadas “aguajales”. Como los pobladores amazónicos emplean varios órganos de M. flexuosa para suplir sus necesidades y con fines comerciales, se está ejerciendo un gran impacto negativo sobre esta especie. A pesar de ello, a la fecha no se conoce la diversidad genética de esta especie en la Amazonía peruana. Consecuentemente, los planes de manejo para la especie serían limitados sin este tipo de información. Por tanto, el objetivo de esta investigación fue determinar la diversidad genética y filogenia molecular de poblaciones de M. flexuosa aledañas a la carretera Iquitos-Nauta. Las hojas se colectaron en seis zonas contiguas a la carretera Iquitos–Nauta. El ADN purificado con protocolos estándares fue amplificado mediante la técnica de ADN Polimórfico Amplificado al Azar (RAPD con dos cebadores aleatorios.  En total se generaron 28 amplicones RAPD (26 polimórficos y 2 monomórficos. Dentro de los aguajales la diversidad genética fue tres veces mayor (75±19 % que la diversidad genética entre las seis poblaciones de M. flexuosa (25±19 %. La diferenciación genética entre las poblaciones varió de 0,0 a 0,6. Los aguajales que se agruparon en clados en el dendrograma por su mayor similitud genética tuvieron proximidad geográfica. La similitud genética entre las poblaciones de M. flexuosa depende de la distancia geográfica, de tal manera que las poblaciones con más similitud genética están más próximas entre sí que las que tienen menos similitud genética.

  9. Diversidad genética de la población colombiana de ganado Cebú Brahman Americano Bos Indicus (Bovidae

    Directory of Open Access Journals (Sweden)

    Novoa Bravo Miguel Adriano

    2006-12-01

    Full Text Available La raza Cebú Brahman Americano se encuentra en Colombia alrededor de 100 años. Todo ese tiempo, esta raza ha estado bajo un proceso continuo de selección artificial dirigida, reproducción endogámica, efectos de deriva genética causados por eventos fundadores, migraciones de ejemplares entre las fincas del país y animales importados desde otros países. Estos hechos hacen a esta raza interesante y particular desde el punto de vista de la genética de poblaciones. El objetivo de este trabajo es estudiar la estructura y diversidad genética de la raza Cebú Brahman americano. Se utilizaron 162 animales registrados en la asociación colombiana de criadores de ganado cebú (ASOCEBU de 20 departamentos de Colombia. La genotipificación de los animales se llevó a cabo con el kit StockMarks® for cattle bovine genotyping de Applied Biosystems®, empleando 10 microsatélites dinucleótidos. Los resultados de los distintos análisis multivariados (Análisis de componentes principales y análisis de correspondencias múltiples, de inferencia bayesiana y distancias genéticas interindividuales, demuestran que no se presenta subestructura en la población, lo cual se explica por una alta tasa de migración de animales entre las diferentes fincas y regiones, que homogeniza las frecuencias en todo el país. Además, esta población posee un alto grado de heterocigocidad y diversidad alélica, comparado con otras razas, lo cual refleja su origen de mezcla multiracial. También se encontraron diferencias genéticas entre sexos, lo cual es causado por un proceso reproductivo diferencial, donde actúan diferentes criterios de selección entre sexos. Finalmente, al realizar un análisis de componentes principales para analizar las relaciones genéticas de Cebú Brahman americano colombiano con las razas cebuinas y taurinas, se determinó que esta raza se diferencia genéticamente de las demás razas cebuinas, debido a un aporte genético de razas taurinas

  10. Description of Cellulophaga baltica gen. nov., sp. nov. and Cellulophaga fucicola gen. nov., sp. nov. and reclassification of [Cytophaga] lytica to Cellulophaga lytica gen. nov., comb. nov.

    Science.gov (United States)

    Johansen, J E; Nielsen, P; Sjøholm, C

    1999-07-01

    Phenotypic data indicate that gliding, yellow/orange-pigmented, agar-digesting bacterial strains were members of the Cytophaga-Flavobacterium-Bacteroides (CFB) group. The strains were isolated from the surface of the marine benthic macroalga Fucus serratus L. and the surrounding seawater at three localities in Danish waters. The bacteria were Gram-negative, flexirubin-negative, aerobic, catalase-positive and oxidase-negative and were psychrophilic and halophilic. All strains utilized D-fructose, L-fucose and alpha-ketobutyric acid and degraded alginic acid, carrageenan, starch and autoclaved yeast cells. Amplification with primers specific for repetitive extragenic palindromic elements by PCR divided the strains of this study into two groups. Both groups showed unique PCR amplification patterns compared to reference strains of the CFB group. Phylogenetic analysis of 16S rDNA sequences showed association of these organisms and [Cytophaga] lytica at the genus level. Hybridization of total chromosomal DNA revealed that the new strains and [Cytophaga] lytica ATCC 23178T were clearly distinct from each other and other previously described species of the CFB group. A new genus is described, Cellulophaga gen. nov. comprising two new species, Cellulophaga baltica gen. nov., sp. nov. (NN015840T = LMG 18535T) and Cellulophaga fucicola gen. nov., sp. nov. (NN015860T = LMG 18536T), as well as the emendation of [Cytophaga] lytica to Cellulophaga lytica gen. nov., comb. nov. PMID:10425785

  11. Study on occurrence of the IVS8-5T allele of the CFTR gene in Ukrainian males with spermatogenesis failure

    Directory of Open Access Journals (Sweden)

    Zinchenko V. M.

    2010-07-01

    Full Text Available Aim. To study the IVS8-5T allele of the CFTR gene and it is involvement in spermatogenesis failure in men with azoospermia and oligozoospermia. Methods. The IVS8-nT polymorphism was analyzed by PCR followed by «A.L.F.-express» fragment analysis in the infertile men group, consisting of 113 azoospermic and 217 oligozoospermic patients, and the control group of 150 fertile men with proven paternity. Results. The frequency of the IVS8-5T allele among infertile males was higher than in controls. A statistically significant difference (P < 0.05 was observed in the frequencies of the IVS8-5T allele in azoospermia patients (5.3 % when compared with the control group (2.0 %. Conclusions. The IVS8-5T allele of the CFTR gene contributes to spermatogenesis failure and/or sperm maturation.

  12. Generation of KCL021 research grade human embryonic stem cell line carrying a ΔF508 mutation in the CFTR gene.

    Science.gov (United States)

    Miere, Cristian; Hewitson, Heema; Wood, Victoria; Kadeva, Neli; Cornwell, Glenda; Codognotto, Stefano; Stephenson, Emma; Ilic, Dusko

    2016-01-01

    The KCL021 human embryonic stem cell line was derived from an embryo donated for research that carried a ΔF508 mutation affecting the CFTR gene encoding the cystic fibrosis transmembrane conductance regulator. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays. PMID:27345808

  13. Expression und Funktion des Na+/K+/2Cl-Kotransporters NKCC1 im Gastrointestinaltrakt von CFTR-Knockout-Mäusen

    OpenAIRE

    Wüchner, Katrin

    2004-01-01

    Die Chlorid-Sekretion stellt eine der zentralen physiologischen Funktionen im Rahmen des gastrointestinalen Verdauungsprozesses dar. Bei Aktivierung der intestinalen Chloridsekretion durch cAMP muss auch die basolaterale Aufnahme gesteigert werden um das zelluläre Elektrolytgleichgewicht aufrecht zu halten. Dabei bildet die basolaterale Chlorid-Aufnahme via dem Bumetanid-sensitiven Kotransporter NKCC1 den limitierenden Faktor für die hauptsächlich durch den Chlorid-Kanal CFTR erfolgende apika...

  14. Bases genéticas del dolor Genetic foundations of pain

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    P. Armero

    2004-11-01

    Full Text Available La percepción de la sensación dolorosa es un proceso complejo en el que intervienen mútiples procesos bioquímicos bien conocidos junto con otros de integración cortical desconocidos hasta el momento. La existencia de diferencias individuales en la respuesta al estímulo doloroso es una observación bien conocida que sugiere qué factores genéticos pueden estar implicados en la modulación de la respuesta a estímulos dolorosos. Existen dos aproximaciones experimentales para estudiar la implicación del genotipo en la respuesta al estímulo doloroso, los estudios de ligamiento y los estudios de asociación. Hasta el momento los estudios de ligamiento han permitido asociar mutaciones en el gen TRKA con el síndrome de insensibilidad congénita al dolor con anhidrosis (CIPA y el gen CACNL1A4 y la migraña hemipléjica familiar (FHM. Los estudios de asociación son escasos y se han centrado principalmente en el estudio de pacientes con migraña. En este trabajo revisamos los estudios llevados a cabo hasta el momento en diferentes laboratorios y planteamos nuevas perspectivas de futuro.Perception of pain is a complex process which implies multiple biochemical pathways together with unknown processes of cortical integration. The existence of individual differences in the response to painful stimuli suggests that genetic factors can be involved in its modulation. Two different experimental approaches have been developed to study the implication of genotype in the response to pain: linkage studies and the association studies. Up to now linkage studies have allowed the association of TRKA gene mutations with the syndrome of congenital insensitivity to pain with anhidrosis (CIPA and CACNL1A4 gene mutations with the familial hemiplegic migraine (FHM. Few association studies have been performed until now, and have been focused on the study of patients with migraine. Here we review the studies carried out up to now in different laboratories and suggest

  15. Apparent Homozygosity of p.Phe508del in CFTR due to a Large Gene Deletion of Exons 4–11

    Directory of Open Access Journals (Sweden)

    Vassos Neocleous

    2014-01-01

    Full Text Available We report a classic cystic fibrosis (CF boy with a large deletion of exons 4–11 in the cystic fibrosis transmembrane conductance regulator (CFTR gene on one allele and p.Phe508del in exon 10 on the second allele. Both parents of Georgian and Ukrainian background had no personal or family history of the disease. The initial molecular diagnostic investigation identified the patient as homozygous for the p.Phe508del and not compatible with his parent’s genetic status. The possibility of nonpaternity or uniparental disomy (UPD7 was investigated and excluded using microsatellite analysis of highly polymorphic markers on chromosome 7. Array-CGH was also performed on the patient and revealed a male profile with a subtle deletion within the CFTR gene on the long arm (q-arm of chromosome 7 (7q31.2. The deletion was confirmed by MLPA extending from probe L02380 to probe L14978 (28.7 kb and that was inherited from his father, while p.PheF508del was inherited from his mother. These data highlight the need for additional testing for large deletions in patients with apparent homozygosity for a mutated CFTR allele that do not match the carrier status of the parents. Not testing can lead to misdiagnosis and misinterpretation of mutation carrier status and the expected penetrance of the disorder.

  16. Diversidad genética, entre y dentro de los mayores grupos humanos

    Directory of Open Access Journals (Sweden)

    Barbujani, G.

    2003-01-01

    Full Text Available Varios estudios están de acuerdo cuando reportan que cerca del 85% de la diversidad del ADN autosomal y de los loci de las proteínas se debe a diferencias entre individuos dentro de la misma población, mientras que las diferencias entre los grupos de diferentes continentes son responsables de solamente 10% de la variación genética total. Estos resultados están en conflicto con nociones populares de razas humanas claramente distintas y relativamente homogéneas, y nos hacen cuestionar la utilidad de clasificaciones étnicas en diagnósticos médicos, en el campo forense y en genética farmacológica. Nuevos datos obtenidos de inserciones polimórficas de Alu y del cromosoma Y confirman los resultados previos, aunque indican una diversidad mayor en algunos (pero no todos los loci del cromosoma Y. Estos datos nos permiten investigar dos preguntas: (1 si las diferencias continentales, aunque pequeñas, son suficientemente grandes como para asignar a individuos a sus continentes basados en sus genotipos; (2 si los genotipos observados se agrupan en grupos de población o continentales cuando el origen de la muestra se ignora. Usando varios métodos estadísticos, veremos que los errores de clasificación son por lo menos de un 30% para los polimorfismos autosomales bi-alélicos, y de un 27% para el cromosoma Y. Cuatro series de datos genéticos de todo el mundo sugieren la existencia de grupos de genotipos diferentes, pero que éstos cuatro grupos no coinciden el uno con el otro. Adicionalmente, estudios de bloques de ADN del genoma humano indican que la mayor parte de dichos bloques es compartida entre los continentes, con solamente un pequeño porcentaje siendo específico a ciertos continentes. Estos resultados no indican que haya una base clara para subdividir a los humanos en grupos biológicamente definidos. Este puede no ser un problema en áreas aplicadas de genéticas, dado que los métodos rápidos para obtener genotipos individuales

  17. Diversidad genética en poblaciones de agaves pulqueros (Agave spp. del nororiente del Estado de México

    Directory of Open Access Journals (Sweden)

    Guillermo Alfaro Rojas

    2007-01-01

    Full Text Available Se analizaron seis poblaciones de magueyes pulqueros: "Manso" (Agave salmiana var. "Salmiana", "Ayoteco" (Agave salmiana var. "Ayoteco", "Verde", "Carrizo" (Agave mapisaga Trel, "Negro" y "Xilometl", del Nororiente del Estado de México, para determinar la diversidad genética entre y dentro de las mismas, obtener las huellas genéticas correspondientes, y hacer comparaciones para diferenciar sus variantes genéticas. Se utilizaron marcadores moleculares tipo RAPD (Polimorfismos en el ADN Amplificados al Azar y algunas variables morfológicas. El análisis de varianza mostró diferencias significativas (P ≤ 0.01 entre poblaciones para: altura de planta, número de hojas, largo de hojas, ancho de hojas, número de espinas laterales y longitud de espina principal. Los discriminantes morfológicos que permitieron separar a las poblaciones en cuatro grupos, fueron: longitud de espina principal y número de espinas laterales, mismas que explicaron 94 % de la variación. El porcentaje total de loci polimórficos fue de 73.2 % pero dentro de poblaciones hubo reducida variabilidad genética, con porcentajes de loci polimórficos que variaron de 12.2 % para el maguey "Verde" hasta 32.5 % en los magueyes "Manso" y "Ayoteco". La diversidad genética de Nei (H promedio fue de 0.28 ± 0.20, lo que confirma la baja diversidad en las poblaciones. El grado de flujo genético fue bajo (Nm = 0.24, lo que indica que hay menos de un migrante por generación entre las seis poblaciones. El dendrograma obtenido con RAPD mostró dos grandes grupos, cada uno de los cuales incluyó tres variantes relacionadas. En el primer grupo Agave mapisaga Trel ("Carrizo" se relacionó con "Verde" y con "Xilometl", con valores de identidad de 0.91 y 0.82, respectivamente. En el segundo grupo hubo una asociación muy cercana entre el maguey "Manso" (Agave salmiana var. "Salmiana" y los magueyes "Negro" y "Ayoteco" (0.89 y 0.83.

  18. Anemia Falciforme: Um Problema Nosso. Uma abordagem bioética sobre a nova genética

    Directory of Open Access Journals (Sweden)

    Diniz Debora

    2003-01-01

    Full Text Available Este artigo analisa uma das ações educativas adotadas pelo Ministério da Saúde no campo das hemoglobinopatias: o folheto informativo Anemia Falciforme: Um Problema Nosso. O objetivo é discutir as premissas e os valores morais que se encontram associados a iniciativas no campo da educação genética, tendo as políticas públicas sobre anemia falciforme no Brasil como estudo de caso. A análise mostra que o conteúdo do folheto oscila entre políticas de prevenção para doenças e promoção de direitos fundamentais, uma característica da nova genética. Além disso, o excesso de informação biomédica especializada no folheto dificulta sua divulgação em massa. Os resultados encontrados foram discutidos à luz do debate bioético contemporâneo sobre a nova genética.

  19. Relaciones genéticas en localidades de Salta, Argentina: ¿qué reflejan las medidas de distancia?

    Directory of Open Access Journals (Sweden)

    Albeza, María V.

    2010-01-01

    Full Text Available A partir de datos de once STRs autosómicos se calcularon diferentes coeficientes de distancia genética en cinco poblaciones de la provincia de Salta y se elaboraron los dendrogramas correspondientes. Cuando se incorporaron datos de la bibliografía, se amplió el análisis a 16 poblaciones (siete de la provincia y nueve de diferentes regiones del país, reduciéndose a 9 el número de loci comunes utilizados en la estimación de las diferentes medidas de distancia. Las representaciones gráficas no reflejaron las vinculaciones esperadas en función de sus proximidades geográficas, pues hubo una amplia gama de asociaciones, reflejando inconsistencias mutuas. De acuerdo al test de Mantel, no existe correlación con la matriz de distancia geográfica en cada caso (0,23 < r < 0,40 con niveles de significación que variaron entre 0,22 y 0,98, por lo que las distancias genéticas no tuvieron relación con la proximidad geográfica entre poblaciones. Tampoco existió correlación entre STRs y otros marcadores genéticos para localidades de la provincia (r = 0,10163; t = 2,3918, p = 0,9916

  20. Türkiye’de ortaöğretim okullarındaki öğrencilerin gençlik liderlik özelliklerinin incelenmesi

    OpenAIRE

    Cansoy, Ramazan

    2015-01-01

    Amaç: Bu çalışmada ortaöğretim okullarındaki öğrencilerin gençlik liderlik özelliklerinin incelenmesi amaçlanmıştır. Yöntem: Bu çalışma tarama modelindedir. Veri toplama aracı olarak Gençlik Liderlik Özelikleri Ölçeği kullanılmıştır. Araştırmaya Üsküdar ve Beykoz ilçelerinde bulunan 6 tane ortaöğretim kurumuna devam eden 1123 tane öğrenci katılmıştır. Toplanan veriler, parametrik olmayan testlerden Mann Whitney-U ve Kruskal Wallis kullanılarak analiz edilmiştir. Bulgular: Öğrencilerin, güv...

  1. ANALISIS GEN HAEMAGGLUTININ PADA VIRUS CAMPAK LIAR

    Directory of Open Access Journals (Sweden)

    Subangkit Subangkit

    2015-05-01

    Full Text Available AbstrakPenyakit Campak disebabkan oleh virus campak yang termasuk genus Morbilivirus dan Family Paramyxoviridae. Penyakit campak masih menjadi masalah kesehatan karena masih ditemukan Kejadian Luar Biasa (KLB di Indonesia. Salah satu penyebab terjadinya KLB tersebut diduga sebagaiakibat perbedaan antigenesitas antara strain vaksin yang digunakan dengan strain virus campak liar yang beredar di Indonesia. Penelitian ini bertujuan mendapatkan gambaran tentang karakteristik genetik gen Haemagglutinin virus campak liar yang ada di Indonesia. Spesimen yang digunakan sebanyak 27 isolat virus penyebab KLB dari 17 propinsi selama periode tahun 2003-2010. Isolat virus dilakukan pemeriksaan secara RT-PCR dan sekuensing dengan metode Sanger. Hasil sekuensing dianalisis dengan menggunakan perangkat lunak Bioedit 7.0 dan MEGA 4.0. Hasil penelitian didapatkan perbedaan 10 asam amino antara virus campak strain vaksin CAM-70 dan virus campak liar pada posisi D416N; K424T; V451M; N455T; V466I; I473T; F476L; Y481S atau Y481N; H495N; G505D. Kesimpulan penelitian ini adalah terdapat perbedaan karakteristik genetik antara virus campak liar di Indonesia berbeda dengan strain virus vaksin CAM-70.Kata kunci : Campak, Analisis Molekuler, Hemagglutinin, CD46AbstractMeasles is caused by virus belonging to the genus Morbilivirus and Family Paramyxoviridae. Measles is still a public health problem because outbreak of measles still found in Indonesia. Outbreak is suspected as a result of differences in antigenicity between vaccine strains used with wild-type measles virus strains circulating in Indonesia. This study aims to get genetic characteristics of wild-type measles virus haemagglutinin gene in Indonesia. The specimens were used 27 viral isolates from 17 provinces period 2003-2010. Viral isolates examined by RT-PCR and sequencing with Sanger method. Sequencing analysis were conducted using Bioedit 7.0 and MEGA 4.0 software. The results showed 10 amino acid differences

  2. Identificación de los polimorfismos G1 y G8 del gen GDF9 en ovinos criollos Araucanos

    Directory of Open Access Journals (Sweden)

    E Paz

    2014-01-01

    Full Text Available En la especie ovina se han descrito una serie de polimorfismos en genes de efecto mayor relacionados con la actividad reproductiva. Las mutaciones ubicadas en el gen de efecto mayor GDF9 se han asociado con el incremento de la tasa ovulatoria y el tamaño de la camada. GDF9 es un factor celular secretado por el ovocito y es miembro de la familia de factores de crecimiento transformante (TGF-β localizado en el cromosoma 5 ovino. El objetivo de este trabajo fue identificar la presencia de los polimorfismos en los sitios G1 y G8 en el gen GDF9 en ovinos criollos Araucanos. Se extrajo el ADN de 100 muestras sanguíneas para posteriormente determinar la presencia de las mutaciones utilizando la técnica PCR-RFLP. Fue amplificada una región de 462 pb correspondiente al exón 1, la cuál fue digerida con la enzima de restricción HhaI, el siguiente fragmento amplificado fue de 139 pb correspondiente al exón 2, siendo digerido con la enzima Ddel. Se identificó la presencia del polimorfismo G1 con una frecuencia genotípica del 0,56 (genotipo GG, 0,44 (genotipo GA y una frecuencia alélica de 0,78 para el alelo (G y 0,22 para el alelo (A. No se detectó la presencia de polimorfismo G8. Este es el primer reporte de este polimorfismo en ovinos en Chile que podría servir como un marcador genético de prolificidad para la selección de ovinos criollos Araucanos.

  3. Genetic Analysis of Relative Cell Injury Percentage and Some Yield Contributing Traits in Wheat Under Nomral and Heat Stress Conditions Análisis Genético del Porcentaje Relativo de Daño celular y algún Rasgo que Contribuye al Rendimiento en Trigo bajo Condiciones Normales y de Estrés Térmico

    Directory of Open Access Journals (Sweden)

    Jehanzeb Farooq

    2011-12-01

    Full Text Available Several wheat genotypes were screened against heat stress. Seven wheat (Triticum aestivum L. cultivars obtained after screening against heat classified as tolerant, moderately tolerant, and susceptible to heat stress, were mated in a complete diallel mating system to study the inheritance pattern of relative cell injury percentage (cell injury % and some yield contributing parameters under normal and heat stress conditions. Significant genotypic differences were found (P Varios genotipos de trigo se discriminaron para estrés térmico. De ellos siete cultivares de trigo (Triticum aestivum L. incluyendo tolerantes, moderadamente tolerantes, y susceptibles a estrés térmico obtenidos después de discriminar con calor se aparearon en un sistema de apareamiento de dialelo completo para estudiar el patrón de heredabilidad del porcentaje de daño celular relativo (daño celular % y algunos parámetros que contribuyen al rendimiento bajo condiciones normales o de estrés térmico. Se encontraron diferencias genotípicas significativas (P < 0.01 para todos los rasgos estudiados bajo ambas condiciones. Pruebas de ajuste revelaron adecuación parcial para rasgos como días a espigadura y días a madurez en ambas condiciones pero mostraron suficiencia completa para área hoja bandera y daño celular % en ambas condiciones. El modelo para producción de grano por planta y biomasa por planta fue completamente suficiente bajo condiciones normales y completamente adecuado en estrés. El componente aditivo de variación genotípica (D fue significativo para todos los rasgos estudiados y más que los componentes de dominancia H1 y H2. Los valores de la proporción de genes con efectos positivos y negativos en los progenitores (H2/4H1 demostraron distribución desigual de genes dominantes en los progenitores para casi todos los rasgos excepto área de hoja bandera, producción de grano, y daño celular % en ambas condiciones. Se encontraron estimaciones

  4. Chancen der Beteiligung privater Haushalte am Produktivvermögen

    OpenAIRE

    Thiele, Silke

    2000-01-01

    Unter den jetzigen Rahmenbedingungen halten im Durchschnitt lediglich 12 % der Haushalte der alten bzw. 3 % der Haushalte der neuen Bundesländer Aktien, d.h. dieser Anteil an Haushalten beteiligt sich bereits freiwillig am Produktivvermögen. Um Beteiligungen in größerem Umfang zu realisieren, müßten folglich hohe Anreize geschaffen werden. Die geringsten Präferenzen Aktien zu bilden, haben Haushalte mit geringem Einkommen, geringer Vermögensstreuung und geringem Vermögen. Die Präferenzen dies...

  5. History of the Juliusruh ionospheric observatory on Rügen

    Science.gov (United States)

    Weiß, J.

    2016-02-01

    The history of the Juliusruh ionospheric observatory on Rügen is closely connected to the history of ground-based ionospheric sounding. After a short introduction to the ionospheric research and the sounding technique, the founding of the Juliusruh station in 1954 and its development until today are described. The different methods of ground-based sounding - as far as they apply to Juliusruh - are briefly discussed. The condition of life and work in a small team on the island of Rügen, remote from the respective parent institute, is also the subject of this article, whose author headed Juliusruh Station from 1965 to 2004.

  6. Craniostenose em gêmeos: estudo genético

    Directory of Open Access Journals (Sweden)

    Walter Carlos Pereira

    1968-09-01

    Full Text Available É relatada a ocorrência de formas clínicas diversas de craniostenose em gêmeos de sexo diferente. A menina apresentava obliteração completa da sutura coronaria e dos dois terços anteriores da sutura sagital; no menino a sutura sagital era a única afetada. O estudo genético mostrou que a craniostenose independe de aberrações cromossômicas, indicando ser transmitida por gens recessivos raros de natureza autossômica.

  7. Algoritmos genéticos para planejamento em inteligencia artificial

    OpenAIRE

    Lecheta, Edson Martins

    2011-01-01

    Este trabalho apresenta uma revisão bibliográfica atualizada sobre duas grandes áreas da Inteligência Artificial: Planejamento e Algoritmos Genéticos. A pesquisa se estende pela criação de modelos genéticos implementados em um sistema planejador dedicado à resolução de uma conhecida classe de problemas de planejamento, usando bibliotecas de código de domínio público em ambas as áreas. Uma análise dos resultados motivou a remodelagem e nova implementação, alterando a plataforma e o sistema ope...

  8. Rescue of NBD2 mutants N1303K and S1235R of CFTR by small-molecule correctors and transcomplementation.

    Directory of Open Access Journals (Sweden)

    Daniele Rapino

    Full Text Available Although, the most common Cystic Fibrosis mutation, ΔF508, in the cystic fibrosis transmembrane regulator. (CFTR, is located in nucleotide binding domain (NBD1, disease-causing mutations also occur in NBD2. To provide information on potential therapeutic strategies for mutations in NBD2, we studied, using a combination of biochemical approaches and newly created cell lines, two disease-causing NBD2 mutants, N1303K and S1235R. Surprisingly, neither was rescued by low temperature. Inhibition of proteasomes with MG132 or aggresomes with tubacin rescued the immature B and mature C bands of N1303K and S1235R, indicating that degradation occurs via proteasomes and aggresomes. We found no effect of the lysosome inhibitor E64. Thus, our results show that these NBD2 mutants are processing mutants with unique characteristics. Several known correctors developed to rescue ΔF508-CFTR, when applied either alone or in combination, significantly increased the maturation of bands B and C of both NBD 2 mutants. The best correction occurred with the combinations of C4 plus C18 or C3 plus C4. Co-transfection of truncated CFTR (∆27-264 into stably transfected cells was also able to rescue them. This demonstrates for the first time that transcomplementation with a truncated version of CFTR can rescue NBD2 mutants. Our results show that the N1303K mutation has a more profound effect on NBD2 processing than S1235R and that small-molecule correctors increase the maturation of bands B and C in NBD2 mutants. In addition, ∆27-264 was able to transcomplement both NDB2 mutants. We conclude that differences and similarities occur in the impact of mutations on NBD2 when compared to ΔF508-CFTR suggesting that individualized strategies may be needed to restore their function. Finally our results are important because they suggest that gene or corrector molecule therapies either alone or in combination individualized for NBD2 mutants may be beneficial for patients bearing

  9. Correlation of the level of full-length CFTR transcript with pulmonary phenotype in patients carrying R117H and 1342-1,-2delAG mutations

    Energy Technology Data Exchange (ETDEWEB)

    Hamosh, A.; Cutting, G.R. [Johns Hopkins Univ. School of Medicine, Balitmore, MD (United States); Oates, R.; Amos, J. [Boston Univ. School of Medicine, Boston, MA (United States)

    1994-09-01

    The R117H mutation occurs on two chromosome backgrounds, one associated with a 7 thymidine tract (7T-R11H) in the splice-acceptor site of intron 8, the other with a 5 thymidine tract (5T-R117H). We examined exon 9 splicing efficiency in 5 patients of genotype R117H/{delta}F508 and one carrying 1342-1,-2delAG{delta}F508, an obligate exon 9 slice site mutation. Four patients carried R117H on a 7T background -- three adult men with congenital bilateral absence of the vas deferens and one adolescent female with pancreatitis and borderline sweat chloride concentration. The patient with R117H on a 5T background had pancreatic sufficient CF (PS-CF). The 1342-1,-2delAG patient has classic pancreatic insufficient CF (PI-CF). cDNA was synthesized from total RNA extracted from nasal epithlial cells and analyzed for CFTR splicing by 35 cycle PCR using primers in exon 7 and 11. The quantity of full length transcript derived from the R117H or {delta}F508 alleles was assessed by allele-specific oligonucleotide hybridization. While 91.4% of transcript from the 5T-R117H allele was full-length, only 42.2% of CFTR transcript from the 5T-R117H allele was full length. Since CBAVD patients have no lung disease and PS-CF patients do, this indicates that the threshold of developing CF lung disease is crossed when the amount of CFTR transcript bearing R117H is reduced by half. Interestingly, 17.1% of transcript derived from the 1342-1,-2delAG allele (or 8.6% of total CFTR transcript) was normal and full length. This suggests that up to 9% of full length wild-type CFTR transcript may be inadequate to escape the lung disease of CF and that a 9 thymidine tract followed by AAC (the result of the AG deletion) can be used as a splice donor with 2-9% efficiency.

  10. A Virtual Reality Framework to Optimize Design, Operation and Refueling of GEN-IV Reactors

    International Nuclear Information System (INIS)

    Many GEN-IV candidate designs are currently under investigation. Technical issues related to material, safety and economics are being addressed at research laboratories, industry and in academia. After safety, economic feasibility is likely to be the most important criterion in the success of GEN-IV design(s). Lessons learned from the designers and operators of GEN-II (and GEN-III) reactors must play a vital role in achieving both safety and economic feasibility goals

  11. A Virtual Reality Framework to Optimize Design, Operation and Refueling of GEN-IV Reactors.

    Energy Technology Data Exchange (ETDEWEB)

    Rizwan-uddin; Nick Karancevic; Stefano Markidis; Joel Dixon; Cheng Luo; Jared Reynolds

    2008-04-23

    many GEN-IV candidate designs are currently under investigation. Technical issues related to material, safety and economics are being addressed at research laboratories, industry and in academia. After safety, economic feasibility is likely to be the most important crterion in the success of GEN-IV design(s). Lessons learned from the designers and operators of GEN-II (and GEN-III) reactors must play a vital role in achieving both safety and economic feasibility goals.

  12. Analisis Kata Penghubung He (和), Gen ( 跟) Dan Yu (与) Dalam Kalimat Bahasa Mandarin

    OpenAIRE

    Tarigan, Cicilia Aprilina Kartika

    2013-01-01

    The tittle of this paper is “Analisis Kata Penghubung He, Gen, dan Yu dalam Kalimat Bahasa Mandarin. Researcher analyzes the use of Chinese cunjuctions, he, gen, yu in Chinese sentences. Actually he, gen and yu have the same meaning but differently in their own characteristics. The concepts of the thesis are The methodology on the thesis is descriptive method. The author analyze about characteristic and similarities and differences of cunjuctions, hen, gen, and yu. By this, author is try to ...

  13. Estimación de parámetros genéticos para características de crecimiento en borregos Katahdin usando diferentes modelos

    Directory of Open Access Journals (Sweden)

    Coralia Inés V. Manzanilla Pech

    2012-01-01

    Full Text Available Se estimaron parámetros genéticos para características de crecimiento en corderos Katahdin, usando seis variantes del modelo animal. Se usó información de pesos al nacimiento (BW; n= 13,099, al destete ajustado a 75 d (WW; n= 11,509 y posdestete ajustado a 120 d (AW; n= 6,886 tomada durante 7 años (2004-2010 en 20 estados de la República Mexicana. Los análisis se hicieron ignorando o incluyendo efectos maternos. El modelo más sencillo incluyó el efecto genético aditivo directo como el único efecto aleatorio. El modelo más completo incluyó los efectos genéticos directo y materno, la covarianza entre ellos, y el efecto del ambiente permanente materno. Para seleccionar el mejor modelo se usó la prueba de razón de verosimilitud. Cuando los efectos maternos no fueron incluidos en el modelo, los estimadores de la heredabilidad directa y de la varianza genética directa resultaron sobreestimados. Las heredabilidades directas con el mejor modelo fueron 0.18 ± 0.03, 0.30 ± 0.04 y 0.20 ± 0.05 para BW, WW y AW, respectivamente. Las heredabilidades maternas también variaron dependiendo del modelo, de 0.05 a 0.23, 0.00 a 0.12, y 0.09 a 0.25 para BW, WW y AW. El ignorar los efectos maternos en el modelo resultaría en una evaluación genética equivocada para las características de crecimiento en borregos Katahdin.

  14. Ambiente genético del Gen blaCTX-M-12 en aislamientos hospitalarios de Klebsiella pneumoniae

    Directory of Open Access Journals (Sweden)

    Yamile Adriana Celis Bustos

    2009-06-01

    Full Text Available The blaCTX-M-12 gene’s genetic environmnt in Klebsiella pneumoniae hospital isolates Resumen: En Colombia se han detectado genes del grupo CTX-M-1 con alta frecuencia en aislamientos de Klebsiella pneumoniae causantes de infección intrahospitalaria. El conocimiento de los factores genéticos que pueden favorecer la diseminación de estos genes entre especies bacterianas es un aspecto importante para el control de la resistencia. En este estudio se identificaron los plásmidos portadores del gen blaCTX-M-12 en 21 aislamientos clínicos de K. pneumoniae. Se evaluó por conjugación la transferencia de resistencia a antibióticos. Integrones, secuencias de inserción y otros elementos genéticos fueron detectados por amplificación del ADN plasmídico con la reacción en cadena de la polimerasa (PCR. Mediante análisis por PCR se determinó la relación entre el gen blaCTX-M-12 y los elementos genéticos detectados. En todos los aislamientos, el gen blaCTX-M-12 se encontró en plásmidos conjugativos de tamaños entre 65 y 106 kpb. La transferencia por conjugación de estos elementos móviles puede explicar la amplia diseminación de este gen entre enterobacterias causantes de infección nosocomial en hospitales de Bogotá, Colombia. El gen blaCTX-M-12 se encontró corriente abajo de ISEcp1, secuencia de inserción que se ha asociado con la movilización de determinantes genéticos de resistencia. Los promotores de ISEcp1, detectados por análisis de secuencia, pueden facilitar la expresión de la cefotaximasa codificada por este gen.Palabras clave: resistencia a antibióticos; elementos genéticos móviles; gen blaCTX-M-12; plásmidos conjugativos;  Klebsiella pneumoniae. Abstract: Genes from CTX-M-1 group have been detected with great frequency in Colombia in intrahospital infection-causing Klebsiella pneumoniae isolates. Knowledge regarding the genetic factors favouring such genes’ dissemination amongst bacterial species is an

  15. Estructura genética del sistema de asimilación de nitrato y regulación global de la asimilación de nitrógeno en la cianobacteria synechococcus sp.

    OpenAIRE

    Luque Romero, Ignacio

    1994-01-01

    Los objetivos de este trabajo han sido profundizar en el estudio de la estructura genética de los genes implicados en la asimilación de nitrato en Synechococcus sp. PCC 7942, y contribuir al esclarecimiento del mecanismo de regulación de dichos genes mediante la proteína NtcA.

  16. Diversidad genética de aislamientos de Phytophthora infestans en plantaciones de papa en Costa Rica con el uso de RAPDs

    OpenAIRE

    Oswaldo P\\u00E1ez; Roberto Valverde; Luis G\\u00F3mez; Arturo Brenes

    2005-01-01

    Para determinar la diversidad genética de Phytophthora infestans en Costa Rica, 62 aislamientos de este patógeno fueron recolectados en plantaciones de papa en las zonas de Cartago, Zarcero, Fraijanes y Heredia durante 1999-2001 y analizados con el uso de RAPDs. Todos los aislamientos fueron previamente evaluados para el tipo de apareamiento y la resistencia al metalaxyl. Se seleccionó 11 imprimadores con los cuales fue posible formar 17 grupos RAPD cuyas frecuen...

  17. Reação de genótipos de soja ao alumínio em hidroponia e no solo

    Directory of Open Access Journals (Sweden)

    Souza Luiz Augusto Copati

    2001-01-01

    Full Text Available O objetivo deste trabalho foi avaliar os genótipos de soja BR86-5974, BR86-7396, Dourados, Doko RC, EMGOPA 305, IAC-9, BR-9 (Savana, UFV-1, UFV-9 e UFV Araguaia em relação à tolerância ao alumínio (Al em hidroponia e em solo. Na solução com Al foi medido o comprimento radicular. Em solo com 49% de saturação de Al avaliou-se área foliar, altura de planta, altura de inserção da primeira vagem, produção de matéria seca, produção de grãos e índice de colheita. Os genótipos BR86-7396 e IAC-9 são os de maior tolerância ao Al, e UFV-1 mostrou o pior desempenho. Houve correlação significativa entre alongamento radicular e produção de grãos (r = 0,705, área foliar (r = 0,645 e produção de matéria seca (r = 0,634. Isto indica que experimentos em hidroponia e solo são igualmente eficientes na seleção de soja tolerante ao alumínio. A variabilidade detectada sugere que o conjunto de genótipos de soja possui ampla variabilidade genética, o que é desejável em programas de melhoramento com o objetivo de elevar estabilidade de produção no Cerrado.

  18. Consecuencias genéticas de igualar las contribuciones familiares en programas de conservación

    OpenAIRE

    Sánchez Molano, Enrique

    2010-01-01

    En poblaciones en riesgo de extinción, puede ser necesario establecer un programa de conservación ex situ que, desde el punto de vista genético, permita controlar su estructura reproductiva para minimizar los efectos de la consanguinidad y la deriva. En tales programas, a menudo se recomienda igualar las contribuciones familiares al grupo reproductor (estrategia CI) para maximizar el censo efectivo. Este procedimiento causa una relajación parcial de la selección natural, impidiendo su acción ...

  19. Mejoramiento genético para tolerancia a altas temperaturas y resistencia a mosaico dorado en frijol común

    OpenAIRE

    Juan Carlos Rosas; Aracely Castro; James S. Beaver; Carlos A. P\\u00E9rez; Adri\\u00E1n Morales; Rogelio Lepiz

    2000-01-01

    Mejoramiento genético para tolerancia a altas temperaturas y resistencia a mosaico dorado en frijol común. Entre 1994-95 se identificaron fuentes de frijol tolerantes a altas temperaturas mediante la evaluación de germoplasma y líneas mejoradas en Choluteca y Nacaome (≤ 50 msnm), en la región Sur de Honduras. La tolerancia al calor de los mejores genotipos fue confirmada en Geneva, Nueva York, EE.UU., bajo condiciones de invernadero con temperatura controlada (35...

  20. Organización de los nucleosomas y regulación genómica en Schizosaccharomyces pombe

    OpenAIRE

    Serrano Pérez, Rebeca

    2014-01-01

    [ES]El genoma de los organismos eucariotas está organizado en cromatina, estructura cuya unidad fundamental es el nucleosoma. Éstos, facilitan el empaquetamiento del DNA dentro del núcleo y son esenciales en su mantenimiento, permitiendo la regulación de procesos genómicos básicos para la función celular. El trabajo presentado en esta tesis doctoral pretende contribuir al conocimiento de la organización de los nucleosomas y su papel en procesos relacionados con la regulación del genoma en ...

  1. Tipificación de marcadores genéticos sanguíneos en raza Hereford

    OpenAIRE

    Quinteros, Indalecio Rodolfo; Tejedor, Eugenio Daniel; Poli, Mario Andrés; Antonini de Ruiz, Alicia Graciela

    1981-01-01

    El paso inicial de esta investigación ha sido tipificar al Bovino Hereford de Argentina para definirlo mediante la metodología de la Inmunogenética. Se buscaron "expresiones" propias y coincidencias con los "marcadores genéticos sanguíneos" descubiertos en esta raza por otros países. Su gran adaptabilidad a "hábitats" diferentes induce a mantener intacto su germoplasma y enriquecerlo con el agregado de nuevos genes. No obstante su homogeneidad racial, el Hereford Argentino presenta destacado ...

  2. Correlaciones fenotípicas, genéticas y ambientales en cucurbita moschata duch. ex poir

    OpenAIRE

    Baena García D.; Espitia Camacho M.; Vallejo Cabrera F. A.

    2006-01-01

    El estudio estimó las correlaciones fenotípicas, genéticas y ambientales entre doce caracteres agronómicos en dos dialélicos de zapallo (uno entre cinco variedades y otro entre cinco líneas S1, originadas de las anteriores). Se usó un diseño en bloques completos al azar con 15 tratamientos (5 progenitores + 10 cruzamientos F1) para cada dialélico y cinco repeticiones. Los resultados señalaron mayor estimación de las correlaciones (en magnitud y significancia estadística) a favor del dialélico...

  3. Genética molecular y biogerontología en la era postgenómica

    OpenAIRE

    Michán Aguirre Shaday

    2011-01-01

    RESUMEN La forma de estudiar la genética ha progresado notablemente en las últimas decadas. Sus orígenes se remontan al estudio de los caracteres hereditarios, seguido por el descubrimiento de los genes y los cromosomas hasta conocer la estructura del ADN. Este último evento impulsó el desarrollo de la tecnología del ADN recombinante y de la secuenciación masiva y automatizada, los cuales permitieron posteriormente determinar la anatomía de los genomas. Todos estos descubrimientos ...

  4. Genética y Conducta en el Síndrome de Déficit Atencional e Hiperactividad:

    OpenAIRE

    Francisco Aboitiz; Carolina Schroter

    2006-01-01

    El síndrome de déficit atencional e hiperactividad es una condición de alta prevalencia entre niños y adultos, que puede afectar seriamente el desempeño escolar y laboral. En este artículo revisamos los aspectos genéticos de este síndrome y los interpretamos en función de una falla en el sistema de señalización dopaminérgica, que se caracteriza por un bajo coeficiente señal/ruido y una excesiva recaptación presináptica del neurotransmisor liberado.

  5. GA-Ensemble : optimización de conjuntos de clasificadores mediante algoritmos genéticos

    OpenAIRE

    Ordóñez Morales, Francisco Javier

    2007-01-01

    El objetivo general de este Proyecto Fin de Carrera es desarrollar un algoritmo optimizado de generación de conjuntos de clasificadores heterogéneos basándose en el algoritmo GA-Stacking. Los objetivos específicos son: - Determinar los métodos de generación de clasificadores más adecuados al proceso de creación del conjunto de clasificadores. - Definir los parámetros adecuados de los algoritmos genéticos a utilizar para la generación del conjunto de clasificadores, haciendo ...

  6. Diversidade genética entre progênies e matrizes de rambutan

    OpenAIRE

    Renata Aparecida de Andrade; Ester Wickert; Antonio Baldo Geraldo Martins; Eliana Gertrudes de Macedo Lemos

    2012-01-01

    O rambutan é uma frutífera exótica que apresenta alto potencial de mercado, e suas mudas podem ser obtidas por sementes ou vegetativamente. A produção de mudas via sementes é rotineiramente feita no Estado de São Paulo, tendo-se alta variabilidade no pomar, além de demorar mais tempo para entrar em produção. Embora caracteres morfológicos sejam amplamente usados na diferenciação de variedades, as técnicas moleculares permitem a comparação e a identificação genética dos materiais. Diante disso...

  7. Bases Genéticas de la Ataxia Espinocerebelosa tipo 2

    Directory of Open Access Journals (Sweden)

    Luis E. Almaguer Mederos

    2008-01-01

    Full Text Available Aborda la genética clásica y molecular de la ataxia espinocerebelosa tipo 2, enfermedad neuro degenerativa que alcanza las mayores tasas de prevalencia e incidencia del mundo en Holguín, y que constituye un serio problema de salud. Desde el descubrimiento de la mutación causal de la enfermedad en 1996, se han obtenido avances significativos en la comprensión de los mecanismos moleculares involucrados en el proceso patológico, y en sus implicaciones clínicas. Tales hallazgos han permitido el desarrollo de modelos celulares y animales de la enfermedad, que constituyen herramientas indispensables para la búsqueda de alternativas terapéuticas orientadas al tratamiento de las personas afectadas.

  8. Disseny i implementació d'un robot 3R genèric

    OpenAIRE

    Ballbè Chaler, Marc

    2015-01-01

    Aquest projecte s'ha realitzat al laboratori del Institut de Robòtica i Informàtica Industrial ubicat en la Facultat de Matemàtiques i Estadística de la Universitat Politècnica de Catalunya. Els objectius d'aquest projecte han sigut diversos, encara que tots s'engloben en l'anàlisi genèrica d'un robot amb tres graus de llibertat de rotació i la implementació d'un prototip. Un dels objectius principals ha sigut la resolució de la cinemàtica directa i inversa d'un robot amb...

  9. Divergência genética em genótipos de girassol Genetic divergence in sunflower genotypes

    Directory of Open Access Journals (Sweden)

    Edson Perito Amorim

    2007-12-01

    Full Text Available Uma investigação sobre a diversidade genética entre 15 genótipos de girassol, por meio de 12 características agronômicas, foi implementada no Instituto Agronômico, Campinas, Brasil. Análises de variância univariada e multivariada revelaram diferenças entre os genótipos. A distância generalizada de Mahalanobis indicou um alto grau de divergência genética. Os genótipos foram agrupados em três grupos. As características início do florescimento, 50% do florescimento, número de folhas e altura da inserção do capítulo contribuíram com grande parte da divergência genética observada. Por meio desses resultados, é possível identificar materiais divergentes e com características agronômicas complementares para o desenvolvimento de novos cultivares superiores.An investigation about the genetical diversity among fifteen sunflower genotypes using twelve agronomical characteristics was implanted at the Agronomic Institute, Campinas Brazil. Univariate and multivariate analyses of variance revealed the presence of differences among the genotypes. The generalized distance of Mahalanobis indicated the presence of genetic diversity. The genotypes were grouped into tree clusters. Among the investigated characteristics, the beginning of flowering, 50% flowering, leaf number and head height of chapter insertion exhibited high contribution towards genetic divergence. Through these studies it is possible to identify divergent material with further agronomical features for the development of new superior sunflower cultivars.

  10. Linfohistiocitosis hemofagocítica, el espectro desde la enfermedad genética al síndrome de activación macrofágica Hemophagocityc Lymphohistiocytosis: A Spectrum from the Genetic Disorder to the Macrophage Activation Syndrome

    Directory of Open Access Journals (Sweden)

    Oscar Porras

    2011-06-01

    Full Text Available El compromiso de la regulación de la inflamación produce activación excesiva y expansión de macrófagos y linfocitos T que desencadenan una reacción inflamatoria severa, sin vías naturales de control. Los trastornos hemofagocíticos son la traducción clínica de este proceso inflamatorio. La linfohistiocitosis hemofagocítica se refiere a todas las variantes de esta patología, y el síndrome de activación macrofágica, a la variante asociada con enfermedad autoinmune. Los casos primarios se asocian con la forma familiar autosómica recesiva y los secundarios con inmunodeficiencias primarias, infección, malignidad y enfermedades autoinmunes. El principal distintivo de este grupo de patologías es la proliferación agresiva de macrófagos e histiocitos que fagocitan otras células sanguíneas. La reducción en la actividad de las células NK produce un aumento en la activación y expansión de linfocitos T, los cuales producen grandes cantidades de citoquinas. Las citoquinas inducen activación de macrófagos y células dendríticas, infiltración tisular y producción de interleuquinas, lo que genera una reacción inflamatoria severa, responsable del daño tisular y de las manifestaciones clínicas. El curso clínico se caracteriza principalmente por fiebre prolongada, hepatoesplenomegalia y citopenias. Los estudios de laboratorio muestran aumento de ferritina, triglicéridos e hipofibrinogenemia. La hemofagocitosis en médula ósea está presente en más del 80% de los casos al diagnóstico. El tratamiento está dirigido contra el linfocito T y los histiocitos hiperactivados, combinando quimioterapia con inmunosupresores y, en algunos casos, trasplante de células madre hematopoyéticas. Este tratamiento ha producido un cambio en la sobrevida de los pacientes. El protocolo de tratamiento HLH-2004 es una guía que estandariza el tratamiento, combinando etopósido, dexametazona y ciclosporina A. En Costa Rica se han reportado 60 casos en

  11. Los usuarios ante los alimentos genéticamente modificados y su información en el etiquetado

    Directory of Open Access Journals (Sweden)

    Miren Itxaso Sebastian-Ponce

    2014-02-01

    Full Text Available OBJETIVO : Analizar la opinión que los usuarios tienen sobre alimentos genéticamente modificados y su información en el etiquetado. MÉTODOS : Realizada revisión sistemática de la literatura científica sobre los alimentos transgénicos y el etiquetado a partir de la consulta de las bases de datos bibliográficas: Medline (vía PubMed, EMBASE, ISI-Web of Knowledge, Cochrane Library Plus, FSTA, LILACS, CINAHL y AGRICOLA. Los descriptores seleccionados fueron: «organisms, genetically modified » y «food labeling». La búsqueda se realizó desde la primera fecha disponible hasta junio de 2012, seleccionando los artículos pertinentes escritos en inglés, portugués y castellano. RESULTADOS : Se seleccionaron 40 artículos. En todos ellos, se debía haber realizado una intervención poblacional enfocada al conocimiento de los consumidores sobre los alimentos genéticamente modificados y su necesidad, o no, de incluir información en el etiquetado. El consumidor expresa su preferencia por el producto no-genéticamente modificado, y apunta que está dispuesto a pagar algo más por él, pero, en definitiva compra el artículo que está a mejor precio en un mercado que acoge las nuevas tecnologías. En 18 artículos la población se mostraba favorable a su etiquetado obligatorio y seis al etiquetado voluntario; siete trabajos demostraban el poco conocimiento de la población sobre los transgénicos y, en tres, la población subestimó la cantidad que consumía. En todo caso, se observó la influencia del precio del producto genéticamente modificado. CONCLUSIONES : La etiqueta debe ser homogénea y aclarar el grado de tolerancia en humanos de alimentos genéticamente modificados en comparación con los no modificados. Asimismo, debe dejar claro su composición, o no, de alimento genéticamente modificado y la forma de producción de estos artículos de consumo. La etiqueta también debe ir acompañada de un sello de certificación de una agencia

  12. El uso de alteraciones genéticas en la estratificación por riesgo del mieloma múltiple

    Directory of Open Access Journals (Sweden)

    Esteban Braggio

    2013-08-01

    Full Text Available Los estudios genéticos han alcanzado un papel central en el estudio del mieloma múltiple (MM, al convertirse en un componente crítico en la estratificación basada en el riesgo de la enfermedad. Se han hecho grandes esfuerzos para identificar cambios genéticos que puedan predecir el resultado clínico e incluirlos en la práctica clínica diaria. La hibridización in situ fluorescente (FISH es todavía la técnica genética más utilizada en la práctica clínica, mayormente debido a su sencilla implementación y su simplicidad para el análisis de datos. El advenimiento de la genómica (hibridización genómica comparativa, secuenciación exónica o genómica completa y del transcriptoma de alta resolución (perfiles de expresión de genes - GEP y secuenciación de ARNm proveen un análisis exhaustivo de los ya definidos factores pronósticos genéticos y son herramientas útiles para la identificación de potenciales nuevos marcadores pronósticos de enfermedad en el clon tumoral de MM. Más aún, GEP ha sido exitosamente implementado en MM como una herramienta de estratificación de riesgo, siendo la de mayor poder de discriminación de resultados. De todas maneras, algunos aspectos técnicos y logísticos complejos (necesidad de una elevada purificación del clon tumoral, costo de los ensayos y complejidad en los análisis de los datos deben ser considerados antes de la incorporación definitiva de estas tecnologías de alto rendimiento dentro de los ensayos clínicos de rutina. Hasta entonces, FISH continúa siendo la herramienta estándar para la detección de anormalidades genéticas y de valoración pronóstico de enfermedad.

  13. Caracterização físico-química de frutos de genótipos de aceroleira (Malpighia emarginata D.C.

    Directory of Open Access Journals (Sweden)

    Maria Inês Sucupira Maciel

    2010-12-01

    Full Text Available No Brasil, a aceroleira, decorrente principalmente da propagação por sementes, tem dado origem a plantios comerciais cujos frutos apresentam parâmetros de qualidade diferenciados. Características físico-químicas de frutos de 18 genótipos de aceroleira (Malpighia emarginata DC. do Banco Ativo de Germoplasma da Universidade Federal Rural de Pernambuco - UFRPE foram avaliados. O rendimento em polpa variou de 41,06% (PL 40 a 72,54% (PL 43 e, com exceção do genótipo PL 37, os demais apresentaram frutos com teores de ácido ascórbico superiores a 1000 mg.100 g-1. O genótipo PL 39 destacou-se por apresentar o maior teor de ácido ascórbico (1667 mg.100 g-1, SST e flavonóis (15,04 mg.100 g-1, além de elevado teor de antocianinas, sendo, portanto, o mais promissor. Os frutos do genótipo PL 34 revelaram o maior valor de SST/ATT, indicando ser o mais doce

  14. Meet Mr. and Mrs. Gen X: A New Parent Generation

    Science.gov (United States)

    Howe, Neil

    2010-01-01

    Slowly but surely, Generation Xers have been taking over from Baby Boomers as the majority of parents in elementary and secondary education. In the early 1990s, Gen Xers began joining parent-teacher associations in the nation's elementary schools. Around 2005, they became the majority of middle school parents. By the fall of 2008, they took over…

  15. PowerGen plc report and accounts 1994

    International Nuclear Information System (INIS)

    The annual report and accounts of PowerGen plc for the year 1994 are presented. Financial highlights are quoted, followed by the Chairman's statement, reviews by the Chief Executive and Financial Directors, reports by the Auditors and Directors, balance sheets and details of the consolidated profit and loss account and principal accounting policies. A four year summary and shareholder information are included. (UK)

  16. GenBank blastx search result: AK058467 [KOME

    Lifescience Database Archive (English)

    Full Text Available ome 1p34.2-36.11 Contains the 3' end of the KPNA6 gene for karyopherin alpha 6 (importin alpha 7), a novel g...ene (DKFZp451J0118), a novel gene (MGC1203) a novel gene, a novel gene (FLJ10547), a novel gene, a novel gen

  17. GenBank blastn search result: AK288295 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288295 J090019J01 AM157075.1 AM157075 Echinolittorina reticulata mitochondrial partial COI gen ... for cytochrome oxidase subunit I, specimen voucher Philippines ... C.2, NRET.GUI.2. INV 3e-18 1 -1 ...

  18. El desarrollo de la investigación en genética en Colombia

    Directory of Open Access Journals (Sweden)

    Carolina Isaza

    2008-06-01

    Full Text Available Durante la última década la genética y la biología molecular en Colombia han dado un gran salto en su desarrollo. Al revisar las convocatorias nacionales de salud que realiza anualmente el Ministerio de Protección Social a través de Colciencias es posible visualizarlo. Al dar una breve mirada a las cifras estadísticas desde el año 2000, encontramos que el país muestra un claro aumento de la capacidad científica y el recurso humano para investigación en salud, reflejado en el incremento del número de proyectos presentados en cada convocatoria y el aumento considerable en la inversión del país para financiar investigación en salud, llegando en el año 2008 a la cifra de 28,000 millones de pesos.En el año 2000 se presentaron para evaluación a Colciencias 133 proyectos y se aprobaron para ejecución solamente 43 por un monto de 1,585 millones de pesos, mientras que en el 2007 se presentaron 690 y aprobados 103 por un monto de 21,223 millones de pesos. Este año se han presentado 711 proyectos que se encuentran actualmente en evaluación y se dispone de 28,000 millones para financiación. Estas cifras muestran un incremento de siete veces en el número de propuestas y un crecimiento de cerca de 27 veces en el presupuesto durante los últimos 7 años. Es muy claro que los proyectos seleccionados son cada vez de mayor impacto para la salud de los colombianos y ahora entre 20% y 30% de las propuestas presentadas en la modalidad nacional, incluyen temas relacionados con genética o utilizan herramientas de biología molecular. Sin embargo, este porcentaje es mucho menor en la convocatoria regional, lo que muestra una concentración de científicos con preparación en genética y biología molecular en las principales ciuda-des del Colombia, que es lo que se ha denominado el triángulo de oro de la ciencia en Colombia (Medellín, Bogotá y Cali. Es interesante ver los desarrollos de la genética molecular para el estudio de las

  19. Analysis of gene repair tracts from Cas9/gRNA double-stranded breaks in the human CFTR gene.

    Science.gov (United States)

    Hollywood, Jennifer A; Lee, Ciaran M; Scallan, Martina F; Harrison, Patrick T

    2016-01-01

    To maximise the efficiency of template-dependent gene editing, most studies describe programmable and/or RNA-guided endonucleases that make a double-stranded break at, or close to, the target sequence to be modified. The rationale for this design strategy is that most gene repair tracts will be very short. Here, we describe a CRISPR Cas9/gRNA selection-free strategy which uses deep sequencing to characterise repair tracts from a donor plasmid containing seven nucleotide differences across a 216 bp target region in the human CFTR gene. We found that 90% of the template-dependent repair tracts were >100 bp in length with equal numbers of uni-directional and bi-directional repair tracts. The occurrence of long repair tracts suggests that a single gRNA could be used with variants of the same template to create or correct specific mutations within a 200 bp range, the size of ~80% of human exons. The selection-free strategy used here also allowed detection of non-homologous end joining events in many of the homology-directed repair tracts. This indicates a need to modify the donor, possibly by silent changes in the PAM sequence, to prevent creation of a second double-stranded break in an allele that has already been correctly edited by homology-directed repair. PMID:27557525

  20. The progress in CFTR chloride ion channel%CFTR型氯离子通道研究进展

    Institute of Scientific and Technical Information of China (English)

    郭晓强

    2007-01-01

    囊性纤维化跨膜传导调节因子(CFTR)是一种重要的氯离子通道,突变易引起囊性纤维化病变,故得名.一系列研究表明,CFTR由5个结构域组成:两个跨膜结构域形成氯离子通道;两个核苷酸结合结构域调节通道的开闭;一个调节结构域主要影响氯通道的活动.这些结构域通过协同作用共同控制了氯离子的跨膜流动,而一些突变可以影响细胞功能而导致囊性纤维化的发生.本文通过介绍CFTR基本结构、调节机制、与囊性纤维化病变的关系及针对CFTR的治疗而对CFTR型氯离子通道有一个的全面的理解.

  1. Caracterización genética y funcional del gen causante de la Epilepsia Lateral Temporal Autosómica Dominante

    OpenAIRE

    Morante Redolat, José Manuel

    2006-01-01

    En el primer capítulo se presenta el trabajo de identificación del gen causante de la Epilepsia Lateral Temporal Autosómica Dominante (ADLTE) mediante un rastreo de genes candidatos en familias afectadas. Una vez identificado dicho gen se procedió a diseñar las estrategias necesarias para su caracterización según la información disponible sobre el gen en aquel momento. El segundo capítulo se centra en los resultados obtenidos sobre la naturaleza de la proteína codificada por el gen iden...

  2. Propuesta de consulta para desregulación de alimentos derivados de cultivos genéticamente mejorados, desarrollados en Costa Rica Request for consultation for deregulation of foods derived from genetically enhanced crops developed in Costa Rica

    OpenAIRE

    M-Eugenia Villalobos Hernández; Ana M Espinoza Esquivel

    2009-01-01

    Objetivo: Proponer un proceso de desregulación de alimentos derivados de cultivos genéticamente mejorados desarrollados en Costa Rica, plantea un trámite a la Consulta de desregulación de un nuevo alimento desarrollado por ingeniería genética, labor que le competerá al Ministerio de Salud principalmente. Material y Métodos: La propuesta incluye procedimientos de iniciar la Consulta, conformación del equipo de evaluadores, información que se solicita en el dossier sobre alimentos derivados de ...

  3. Caracterización molecular, genética poblacional y relación biológica del nemertino Malacobdella arrokeana endocomensal de la almeja gigante Panopea abbreviata

    OpenAIRE

    Alfaya, José E. F.

    2014-01-01

    [ES] Malacobdella arrokeana Ivanov et al., 2002 es un nemertino que vive en la cavidad del manto de la almeja gigante Panopea abbreviata, ambas especies endemicas del Mar Argentino. Mediante la secuenciación y análisis del gen mitocondrial citocromo oxidasa I (COI), se corroboró que M arrokeana diverge genéticamente de M. grossa en un 11,6%, y 10,4% con respecto a M. japonica. Tanto M. arrokeana como P. abbreviata comparten el mismo nivel trófico y los mismos ítems alimentarios, compuestos en...

  4. La Enseñanza de la genética: Una propuesta didáctica para la educación secundaria obligatoria desde una perspectiva constructivista

    OpenAIRE

    Iñiguez Porras, Francisco Javier

    2006-01-01

    [spa] Frente al modelo tradicional de la enseñanza-aprendizaje de la genética, basado en la transmisión por parte del profesorado de conocimientos ya elaborados, y que no tiene en cuenta las concepciones del alumnado, nosotros hemos propuesta una secuencia didáctica basada en los principios del constructivismo. Dicha secuencia tiene en consideración las ideas de los estudiantes, tiene una cuenta las propuestas de los investigadores en didáctica de la genética y consta de diferentes actividade...

  5. Desarrollo de un sistema de expresión condicional del gen RHBDD2 para su sobreexpresión en un ratón transgénico

    OpenAIRE

    Lacunza, Ezequiel; Abba, Martin Carlos; Aldaz, C. Marcelo

    2011-01-01

    En estudios previos hemos identificado al gen RHBDD2 sobreexpresado en carcinomas invasores de mama. Hasta el momento se desconoce el rol funcional de dicha alteración aunque existen evidencias de que podría determinar un fenotipo favorable para las células tumorales. A fin de analizar in vivo las consecuencias biológicas de la sobreexpresión del gen RHBDD2, se llevó a cabo la construcción de un sistema de expresión condicional, para su inyección en células madres emb...

  6. Evaluación genética del recurso animal de los sistemas de producción de bovinos en doble propósito en Colombia / Genetic evaluation of dual purpose production systems in Colombia

    OpenAIRE

    Galeano Rivera, Adriana Patricia

    2010-01-01

    Con el objetivo de determinar el verdadero potencial genético del recurso animal disponible en los sistemas de producción bovina de doble propósito del trópico bajo colombiano, se evaluaron los registros de producción de leche por lactancia (Kg), peso al destete (Kg), intervalo entre partos (d) e Índice de Vaca, de 1.687 hembras reproductoras, durante los años 1998 y 2007. Se empleó un modelo animal mixto que incluyó los efectos genéticos aleatorios del animal, el medio ambiente permanente y ...

  7. Flexibilität organisieren: Callcenter als kundenorientierte Organisationen

    OpenAIRE

    Holtgrewe, Ursula; Kerst, Christian

    2003-01-01

    "Das Aufkommen und Wachstum der Callcenter wird häufig entlang der Frage diskutiert, ob sich damit der Taylorismus in neua rtiger und durchschlagender Weise der Dienstleistungsarbeit bemächtigt. Vor allem der Einsatz tayloristischer Kontrolltechnologien (Fernie/Metcalf 1998) und der Einsatz niedrig qualifizierter Arbeitskräfte, vorwiegend mit befristeten Verträgen oder geringfügig beschäftigt, lässt Callcenter als die Vorreiter atypischer Beschäftigung erscheinen (vgl. Flecker ...

  8. Logos versus Aisthesis. Die kunsthistorische Diaprojektion als codierendes Instrument

    OpenAIRE

    Hiller-Norouzi, Julica

    2009-01-01

    Seit die akademische Kunstgeschichte am Ende des 19. Jahrhunderts die Diaprojektion als bevorzugtes Lehrmedium für sich bestimmte, änderte sich die kunsthistorische Lehre maßgeblich. Fotografische Abbildungen von Kunstwerken prägen seitdem die visuelle Wahrnehmung von Kunst und geben eine bestimmte Sehrichtung der Wissenschaft vor. Einhergehend damit veränderte sich auch die Wahrnehmung der Medialität von Kunstwerken. Ich lote aus, welchen Einfluss die Diaprojektion mit ihren zentralperspekti...

  9. Estimación mediante RAPD's de la diversidad genética en Guadua en el departamento del Cauca, Colombia

    Directory of Open Access Journals (Sweden)

    Palacio M. Juan Diego

    2006-06-01

    Full Text Available

    Mediante RAPD's se analizaron 120 muestras foliares de 12 biotipos de Guadua angustifolia Kunth clasificados morfológicamente, procedentes de la cuenca del río Cauca, en el departamento del Cauca, Colombia, para determinar diversidad genética. El ADN se extrajo mediante el protocolo modificado de Dellaporta (1983. Se emplearon los cebadores; OPF-12, OPG-19, OPN-19 y OPP-16 con mayor número de bandas polimórficas. El índice de Shannon (HT = 0.4556 ± 0.1849 señaló diversidad genética total alta y diversidad entre los biotipos y al interior de ellos. El Índice de estructura genética (Gst = 0.5200 e Indice de migración efectiva (Nm = 0.4615 definieron biotipos bien diferenciados. El análisis de similaridad conformó tres grupos a un coeficiente de 0.64. El grupo G1 incluyó los biotipos Curvado, Rayada frecuente, Amarilla Playón, Rayada ancha, Rayada escasa, Convexa, Amarilla, Hembra, Verde irregular y algunos individuos de verde alta. El grupo G2, Verde alta y Macho. El grupo G3, Rayada negra. El estudio molecular agrupó los individuos de forma similar al estudio morfológico, con excepción de los individuos del biotipo Hembra.

    Palabras claves: Guadua angustifolia, caracterización molecular, variación genética.

  10. Comparación de metodologías moleculares para identificar el gen de la kappa caseína en ganado Holstein

    OpenAIRE

    Carlos Solarte P.; Carol Rosero G.; Yohana Eraso C.

    2012-01-01

    RESUMENObjetivo. Comparar las metodologías moleculares, PCR-RFLPs y PCR-SSCP, para identificar las variantes alélicas del gen de la kappa caseína (CSN3) en bovinos Holstein del trópico alto de Nariño-Colombia. Materiales y métodos. Se escogieron al azar 50 vacas Holstein y mediante punción en la vena coxígea media se tomaron muestras de 5cc de sangre, que se almacenaron y preservaron en tarjetas FTA® para su posterior análisis en el laboratorio. El ADN se amplificó por PCR utilizando cebadore...

  11. Variabilidade genética de Eugenia uniflora L. em remanescentes florestais em diferentes estádios sucessionais

    Directory of Open Access Journals (Sweden)

    Roberto Valmorbida Aguiar

    2013-04-01

    Full Text Available A compreensão da diversidade genética fornece elementos básicos sobre a dinâmica e funcionamento de populações, auxiliando na conservação e uso sustentável das espécies. Supõe-se que populações sucessionais precoces poderiam ser geneticamente mais diferenciadas do que populações sucessionais mais tardias. Visando testar esta hipótese, o presente trabalho teve como objetivo analisar a variabilidade genética de populações de Eugenia uniflora L. em manchas florestais em diferentes estádios sucessionais. Foram selecionadas duas áreas em diferentes estádios de sucessão, sendo a primeira em estádio inicial e a segunda em estádio avançado. A área de estudo apresenta um remanescente florestal em transição de Floresta Ombrófila Mista e Floresta Estacional Semidecídua. Por meio da técnica de RAPD (Random Amplified Polymorphic DNA e análise multivariada, a diversidade gênica esperada e a porcentagem de loci polimórficos foram estimadas, além da similaridade genética entre as populações de cada mancha florestal e a diversidade de cada área por meio do índice de diversidade de Simpson. Os resultados indicaram 79% de loci polimórficos para a área em estádio avançado e 70% para a área em estádio inicial de sucessão. A similaridade genética entre pares de indivíduos variou entre 0,55 e 0,86 na área em estádio inicial de sucessão e entre 0,45 e 0,78 para a área em estádio avançado. Não houve diferenças significativas entre a diversidade das duas áreas (P = 89. Um escalonamento multidimensional não-métrico indicou menor distância genética entre os indivíduos da área em estádio inicial. Da mesma forma, uma análise de similaridade - ANOSIM indicou separação entre os indivíduos das duas áreas.

  12. Trifurcatia flabellata n. gen. n. sp., a putative monocotyledon angiosperm from the Lower Cretaceous Crato Formation (Brazil

    Directory of Open Access Journals (Sweden)

    B. Mohr

    2002-01-01

    Full Text Available The Lower Cretaceous Crato Formation (northeast Brazil contains plant remains, here described as Trifurcatia flabellata n. gen. and n. sp., consisting of shoot fragments with jointed trifurcate axes, each axis bearing a single amplexicaul serrate leaf at the apex. The leaves show a flabellate acrodromous to parallelodromous venation pattern, with several primary, secondary and higher order cross-veins. This very unique fossil taxon shares many characters with monocots. However, this fossil taxon exhibits additional features which point to a partly reduced, and specialized plant, which probably enabled this plant to grow in (seasonally dry, even salty environments. In der unterkretazischen Cratoformation (Nordostbrasilien sind Pflanzenfossilien erhalten, die hier als Trifurcatia flabellata n. gen. n. sp. beschrieben werden. Sie bestehen aus trifurcaten Achsen, mit einem apikalen amplexicaulen fächerförmigen serraten Blatt. Diese Blätter zeigen eine flabellate bis acrodrome-paralellodrome Aderung mit Haupt- und Nebenadern und transversale Adern 3. Ordnung. Diese Merkmale sind typisch für Monocotyledone. Allerdings weist dieses Taxon einige Merkmale auf, die weder bei rezenten noch fossilen Monocotyledonen beobachtet werden. Sie müssen als besondere Anpassungen an einen (saisonal trockenen und vielleicht übersalzenen Lebensraum dieser Pflanze interpretiert werden. doi:10.1002/mmng.20020050121

  13. Molecular mechanisms of reduced glutathione transport: role of the MRP/CFTR/ABCC and OATP/SLC21A families of membrane proteins

    International Nuclear Information System (INIS)

    The initial step in reduced glutathione (GSH) turnover in all mammalian cells is its transport across the plasma membrane into the extracellular space; however, the mechanisms of GSH transport are not clearly defined. GSH export is required for the delivery of its constituent amino acids to other tissues, detoxification of drugs, metals, and other reactive compounds of both endogenous and exogenous origin, protection against oxidant stress, and secretion of hepatic bile. Recent studies indicate that some members of the multidrug resistance-associated protein (MRP/CFTR or ABCC) family of ATP-binding cassette (ABC) proteins, as well as some members of the organic anion transporting polypeptide (OATP or SLC21A) family of transporters contribute to this process. In particular, five of the 12 members of the MRP/CFTR family appear to mediate GSH export from cells namely, MRP1, MRP2, MRP4, MRP5, and CFTR. Additionally, two members of the OATP family, rat Oatp1 and Oatp2, have been identified as GSH transporters. For the Oatp1 transporter, efflux of GSH may provide the driving force for the uptake of extracellular substrates. In humans, OATP-B and OATP8 do not appear to transport GSH; however, other members of this family have yet to be characterized in regards to GSH transport. In yeast, the ABC proteins Ycf1p and Bpt1p transport GSH from the cytosol into the vacuole, whereas Hgt1p mediates GSH uptake across the plasma membrane. Because transport is a key step in GSH homeostasis and is intimately linked to its biological functions, GSH export proteins are likely to modulate essential cellular functions

  14. New PRSS1 and common CFTR mutations in a child with acute recurrent pancreatitis, could be considered an "Hereditary" form of pancreatitis ?

    Directory of Open Access Journals (Sweden)

    Galli Elena

    2010-10-01

    Full Text Available Abstract Background acute recurrent pancreatitis is a complex multigenic disease, the diagnosis is even more difficult when this disease develops in a child. Case Presentation a 6-years old boy, hospitalized with epigastric pain radiating to the back showed high serum levels of serum amylase, lipase, CRP and erythrosedimentation rate. Several similar milder episodes of pain, followed by quick recovery and complete disappearance of symptoms were reported during the previous 13 months. The child was medically treated and after 7 days with normal clinic and laboratory tests was discharged with a hypolipidic diet. All the known aetiologic hypotheses were excluded by anamnestic investigation, clinical observation and biochemical evaluation, whereas, anatomic abnormality were excluded by a secretin stimulated magnetic resonance (MRI. At the last follow-up visit, (11 months later, the child showed a normal body weight and anthropometric profile, without further abdominal pain. Mutation screening for coding regions of PRSS1, SPINK1, CFTR and the new hereditary pancreatitis-associated chymotrypsin C (CTRC genes showed a novel variation, c.541A > G (p.S181G, in the exon 4 of PRSS1 gene and the classical CF p.F508del mutation in the CFTR. Both mutations were present in his clinically normal mother and absent in the patient's father. Conclusions this report extend the spectrum of PRSS1 mutations, however, the absence of family history of pancreatitis leaves the present case without the hallmark of the hereditary origin of pancreatitis. At the present knowledge it can be only stated that the combined genotype CFTR (F508del/PRSS1 (S181G is associated to a mild phenotype of acute recurrent pancreatitis in this child without any further conclusion on its pathogenetic role or prediction on the course of the disease.

  15. Applicability and Efficiency of NGS in Routine Diagnosis: In-Depth Performance Analysis of a Complete Workflow for CFTR Mutation Analysis.

    Directory of Open Access Journals (Sweden)

    Adrien Pagin

    Full Text Available Actually, about 2000 sequence variations have been documented in the CFTR gene requiring extensive and multi-step genetic testing in the diagnosis of cystic fibrosis and CFTR-related disorders. We present a two phases study, with validation and performance monitoring, of a single experiment methodology based on multiplex PCR and high throughput sequencing that allows detection of all variants, including large rearrangements, affecting the coding regions plus three deep intronic loci.A total of 340 samples, including 257 patients and 83 previously characterized control samples, were sequenced in 17 MiSeq runs and analyzed with two bioinformatic pipelines in routine diagnostic conditions. We obtained 100% coverage for all the target regions in every tested sample.We correctly identified all the 87 known variants in the control samples and successfully confirmed the 62 variants identified among the patients without observing false positive results. Large rearrangements were identified in 18/18 control samples. Only 17 patient samples showed false positive signals (6.6%, 12 of which showed a borderline result for a single amplicon. We also demonstrated the ability of the assay to detect allele specific dropout of amplicons when a sequence variation occurs at a primer binding site thus limiting the risk for false negative results.We described here the first NGS workflow for CFTR routine analysis that demonstrated equivalent diagnostic performances compared to Sanger sequencing and multiplex ligation-dependent probe amplification. This study illustrates the advantages of NGS in term of scalability, workload reduction and cost-effectiveness in combination with an improvement of the overall data quality due to the simultaneous detection of SNVs and large rearrangements.

  16. Na+/K+/2Cl- cotransporter and CFTR gill expression after seawater transfer in smolts (0+) of different Atlantic salmon (Salmo salar) families

    Science.gov (United States)

    Mackie, P.M.; Gharbi, K.; Ballantyne, J.S.; McCormick, S.D.; Wright, P.A.

    2007-01-01

    Smoltification involves morphological and physiological changes in the gills that prepare anadromous salmonids to osmoregulate efficiently in seawater. In a previous study, we found that different families of Atlantic salmon (Salmo salar) smolts vary in their ability to osmoregulate when abruptly transferred to cold seawater and that these differences are correlated with gill Na+/K+ ATPase activity. Here we extend these findings to test whether other key transport proteins, namely Na+/K+/2Cl- contransporter (NKCC) and the Cl- channel or cystic fibrosis transmembrane conductance regulator (CFTR), play a significant role in osmoregulatory differences between families. To facilitate molecular analysis of NKCC, we first isolated a gill cDNA containing the complete coding region (1147 aa) of an isoform previously reported as a partial sequence. Phylogenetic analysis showed that this isoform is most closely related to isoforms of the NKCC1a subfamily found in European eel and Mozambique tilapia. In a second step, we quantified NKCC protein abundance as well as mRNA expression levels for NKCC1a and two CFTR isoforms (CFTRI and CFTRII) in 0+ smolts from three families prior to and following seawater transfer. The family with the lowest salinity tolerance also showed significant increases in gill NKCC1a mRNA after seawater transfer. Taken together with our previous study, these data indicate that family differences in expression of transport proteins are in part related to salinity tolerance, although the best indicator of osmoregulatory performance between families may be gill Na+/K+ ATPase activity and CFTR I mRNA levels, rather than Na+/K+ ATPase and NKCC1a mRNA levels or NKCC protein abundance. ?? 2007 Elsevier B.V. All rights reserved.

  17. Comprehensive screening for PRSS1, SPINK1, CFTR, CTRC and CLDN2 gene mutations in Chinese paediatric patients with idiopathic chronic pancreatitis: a cohort study

    Science.gov (United States)

    Wang, Wei; Sun, Xiao-Tian; Weng, Xiao-Ling; Zhou, Dai-Zhan; Sun, Chang; Xia, Tian; Hu, Liang-Hao; Lai, Xiao-Wei; Ye, Bo; Liu, Mu-Yun; Jiang, Fei; Gao, Jun; Bo, Lu-Min; Liu, Yun; Liao, Zhuan; Li, Zhao-Shen

    2013-01-01

    Objective Genetic alterations may contribute to chronic pancreatitis (CP) in Chinese young patients. This study was designed to investigate mutations of cationic trypsinogen (PRSS1), pancreatic secretory trypsin inhibitor or serine protease inhibitor Kazal type 1 (SPINK1), cystic fibrosis transmembrane conductance regulator (CFTR), chymotrypsin C (CTRC) and CLDN2 genes and the copy number variations (CNVs) of PRSS1 and asses associations with the development of idiopathic CP (ICP) in Chinese children. Design Retrospective. Setting A single center. Participants 75 ICP Chinese children (40 boys and 35 girls). Primary and secondary outcome measures Mutations of PRSS1, SPINK1, CFTR, CTRC and CLDN2 genes and CNVs. Results 7 patients had heterozygous mutations in PRSS1, that is, N29I (n=1), R122H or R122C (n=6). The CNVs of PRSS1 in five patients had abnormal copies (1 copy (n=4), five copies (n=1)). 43 patients had IVS3+2T>C (rs148954387) (10 homozygous and 33 heterozygous) in SPINK1. None of the PRSS1 mutation patients carried a SPINK1 mutation. Frequency of PRSS1 and SPINK1 mutations was 9.3% and 57.3%, respectively, with an overall frequency of 66.6% (50/75). In addition, one patient had a novel deletion of CFTR (GCTTCCTA from c.500 to c.508 leading to the shortened polypeptide molecule via a stop codon). Another patient had a novel missense in CLDN2 exon 2 (c.592A>C mutation). Clinically, patients with SPINK1 mutations had a higher rate of pancreatic duct stones, pancreatic pseudocyst and pancreatic calcification than those without SPINK1 mutations (pC mutation may play an important role in the pathogenesis of Chinese paediatric ICP. However, further study is needed to confirm and to investigate the role of these genes in the development of Chinese ICP. PMID:24002981

  18. Immunologisches Reaktionsvermögen als Indikator für Belastungen bei der Milchkuh

    OpenAIRE

    Träbing, Claudia

    2006-01-01

    Mit ansteigenden Jahresmilchleistungen werden die Kühe einer hohen metabolischen Belastung ausgesetzt, die bei einer suboptimalen Gestaltung der Lebensbedingungen mit einer Erhöhung der Inzidenzrate von Faktorenkrankheiten einhergehen kann. Im Vordergrund der Untersuchungen stand die Frage, inwieweit metabolische Belastungszustände von Milchkühen in der Phase des Puerperiums und in Abhängigkeit von der Milchleistung sowie Belastungen durch eine klinische Erkrankung zu Beginn der Laktation mit...

  19. Análisis de la diversidad genética de 21 aislamientos del hongo Moniliophthora roreri basado en marcadores RAPD

    Directory of Open Access Journals (Sweden)

    Boris Gutarra Castillo

    2013-12-01

    Full Text Available Objetivos: Estudiar la diversidad genética de 21 aislamientos del hongo que afecta al cultivo del cacao, Moniliophthora roreri, en tres zonas cacaoteras del Perú (Tocache, Mariscal Cáceres y Leoncio Prado. Métodos: Se utilizó 14 iniciadores RAPD (random amplified polymorphic DNA polimórficos y una pareja de oligonucleótidos, los que fueron empleados bajo condiciones de amplificación estandarizadas. Con los datos obtenidos se construyó un dendograma utilizando el coeficiente de Jaccard y el algoritmo UPGMA (Unweighted Pair-Group Method using Arithmetic Average. La estructura genética fue estimada en función del análisis molecular de variancia (AMOVA y la diversidad mediante los índices de Shannon y Nei. Resultados: Fueron conseguidas 59 bandas RAPD con un 73% de polimorfismo. El dendograma obtenido a un índice de similitud de 0,70, claramente dividió los individuos en tres grupos. El análisis de la diversidad genética mostró altos valores en las zonas estudiadas de acuerdo con el índice de Shannon (0,3936 y de Nei (0,2622, con mayor riqueza en Leoncio Prado. Estas zonas presentan alta variabilidad, y según el AMOVA realizado: 88% entre accesiones por zona y solo 12% entre zonas. Conclusiones: Existe más de un grupo genético de Moniliophthora roreri en la Amazonía del Perú. Estos grupos, provenientes del Ecuador, pudieron haber ingresado por el intercambio de semillas y/o de forma natural por medio de los ríos en común y estarían originando nuevos grupos genéticos locales.

  20. Variabilidad genética de Moniliophthora perniciosa (Stahel Aime y Phillips-Mora, comb. nov. (Agaricales - Marasmiaceae en variedades de cacao (Theobroma cacao L.

    Directory of Open Access Journals (Sweden)

    Carolina Osorio-Solano

    2012-04-01

    Full Text Available Moniliophthora perniciosa, agente causante de la ‘escoba de bruja’ en cacao (Theobroma cacao, presenta una elevada variabilidad genética y discrepancias en su taxonomía y es una de las enfermedades más importantes en plantaciones cacaoteras que ocasiona pérdidas económicas a nivel mundial cercanas a 70%, y de 40% a nivel nacional. La caracterización de la diversidad genética de los biotipos es importante para la ejecución de proyectos encaminados al manejo de este patógeno y el desarrollo de materiales resistentes de cacao. En este estudio se analizaron 12 aislamientos del hongo obtenidos de diferentes materiales de cacao. Cada una de las muestras se evaluó con marcadores moleculares que tienen como blanco una región del ADN ribosomal (ADNr nuclear conocida como ITS (Internal Transcribed Spacer, una región intergénica (IGS-1 y cinco secuencias simples repetidas (SSR. El marcador IGS-1 permitió la determinación del biotipo C, no obstante se encontró una variabilidad genética evidente dentro de este biotipo, aún no registrada. El análisis de la diversidad genética de M. perniciosa por medio de marcadores microsatélite arrojó un valor total de 0.4260, una heterocigosidad total de 0.6143 y un índice de información polimórfica (PIC de 0.3407, valores considerados de rango medio a alto para los aislamientos estudiados y que estiman la variabilidad genética presente en M. perniciosa.