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Sample records for linee guida esc

  1. [Linee guida italiane per il trattamento dell'alcolismo].

    Science.gov (United States)

    Ceccanti, Mauro; Iannitelli, Angela; Fiore, Marco

    2018-01-01

    RIASSUNTO. L'Europa è la regione con il più forte consumo di alcol al mondo, con la più elevata percentuale di malattie totali e morti premature alcol-correlate. In Italia, questo fenomeno coinvolge circa il 13% della popolazione oltre i 18 anni e oltre il 25% dei pazienti ospedalizzati. Sfortunatamente, solo il 5% di questi pazienti vengono riconosciuti come persone affette da disturbo da uso di alcol (DUA). Una scarsa conoscenza della malattia, la mancanza di accesso alle cure e le poche risorse messe a disposizione nella gestione del problema sono tra le principali cause di ritardo nella diagnosi, alla fine con conseguenze cliniche molto più importanti e costose da gestire. Alla luce di ciò, abbiamo deciso di dedicare questo numero speciale della Rivista di psichiatria a una serie di articoli riguardanti le linee guida italiane per il trattamento della dipendenza da alcol. Realizzate in collaborazione con numerosi esperti e importanti società scientifiche italiane come il Centro di Riferimento Alcologico della Regione Lazio (CRARL), la Società Italiana Tossicodipendenze (SITD), la Società italiana per il Trattamento dell'Alcolismo e le sue Complicanze (SITAC), la Società Italiana Psichiatria delle Dipendenze (SIPDip), la Società Italiana Patologie da Dipendenza (SIPaD) e l'Istituto di Biologia Cellulare e Neurobiologia (IBCN-CNR), queste linee guida forniscono agli operatori una serie di raccomandazioni basate su prove di efficacia volte ad aumentare la conoscenza e un uso appropriato dei farmaci per le persone affette da DUA. Con la possibilità di un continuo aggiornamento, l'obiettivo principale di queste linee guida sarà quello di garantire l'omogeneità dei trattamenti e un incremento qualitativo nell'assistenza dei pazienti affetti da DUA, così da ridurre le conseguenze psicosociali e sulla salute pubblica di questa importante malattia psichiatrica.

  2. Applicazione delle linee guida nazionali nell'attivitá di notifica di casi di legionellosi

    Directory of Open Access Journals (Sweden)

    E. Corsano

    2003-05-01

    Full Text Available

    Obiettivi: é noto che effettuare diagnosi di Legionellosi risulta difficile a causa della sovrapposizione della sintomatologia con altre forme di polmonite. Questo spiega, in parte, perché i casi di Legionellosi siano sottostimati. I metodi diagnostici disponibili, sebbene richiedano tempi relativamente lunghi, sono necessari per l’esatta stima di prevalenza della Legionellosi e per gli eventuali interventi preventivi.

    Metodi: nell’Azienda Ospedaliera “G. Panico” di Tricase, seguendo i criteri delle Linee Guida della Legionellosi (G.U. 5 maggio 2000 n. 103 durante la degenza dei pazienti che presentavano segni clinici ed obiettivi di polmonite sono state eseguite indagini radiologiche (Rx Torace, TAC, esami di laboratorio (espettorato, sangue, urina ed avviata terapia antibiotica (macrolidi e cefalosporine di III generazione. L’antigene urinario e le IgG anti-Legionella (metodo IFA, sono stati determinati in collaborazione con il Laboratorio di riferimento Regionale.

    Risultati: nel periodo ottobre 2001 gennaio 2003 sono stati notificati nel nostro ospedale 11 casi di Legionellosi accertati da indagini clinico-strumentali e di laboratorio: tutti i casi avevano titoli anticorpali abbastanza elevati, confermati poi da un secondo controllo a distanza di tempo. Solo due hanno presentato una positività per l’antigene urinario. All’atto della dimissione, per il 63.6% dei casi è stato documentato un miglioramento delle condizioni clinico-laboratoristico-strumentali, nel 18.2% si è avuta una effettiva guarigione. La letalità è stata pari al 18.2%. In tutti casi si è provveduto alla sorveglianza ambientale.

    Considerazioni: i nostri dati evidenziano che i casi di Legionellosi registrati sono stati tutti di tipo comunitario. Una corretta applicazione delle Linee Guida nel sistema di notifica permette l’avvio dell’attività di prevenzione oltre

  3. Wordstar professional : breve guida di consultazione

    OpenAIRE

    Biagioni, Stefania

    1990-01-01

    La presente nota ? una breve guida all'uso di WordStar Professional 4.0, utile per una rapida consultazione. Non intende esaurire l'argomento e neppure riportare tutto quello che pu? essere trovato nei manuali citati nella bibliografia presente nel testo. I comandi sono trattati secondo il raggruppamento dei MENU guida che WordStar offre agli utenti, mentre alcune delle funzioni pi? complesse sono spiegate pi? in dettaglio unitamente ad alcuni esempi.

  4. Quality assurance in brachytherapy. Proposal of guidelines concerning clinical, technological and physical dosimetric aspects; Assicurazione di qualita' in brachiterapia: proposta di linee guida in relazione agli aspetti clinici, tecnologici e fisico-sanitari

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-07-01

    The report is an up-dating of the guidelines presented by the Italian National Institute of Health study group on quality assurance in radiotherapy (ISTISAN report 96/39). The study points out the necessity of a strong cooperation among the different professionals involved in brachytherapy procedures. Minimal requirements for quality control institutional protocols are also presented and discussed. [Italian] Il documento costituisce un'integrazione delle linee guida italiane elaborate dal gruppo di studio Assicurazione di qualita' in radioterapia dell'Istituto Superiore di Sanita' (ISTISAN 96/39). Un rilievo ancora maggiore e' stato dato alla necessita' di una stretta collaborazione tra le figure professionali operanti nel campo della brachiterapia. Queste linee guida riportano anche i requisiti essenziali di un programma locale di controlli di qualita' in brachiterapia.

  5. Quality assurance in brachytherapy. Proposal of guidelines concerning clinical, technological and physical dosimetric aspects; Assicurazione di qualita' in brachiterapia: proposta di linee guida in relazione agli aspetti clinici, tecnologici e fisico-sanitari

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-07-01

    The report is an up-dating of the guidelines presented by the Italian National Institute of Health study group on quality assurance in radiotherapy (ISTISAN report 96/39). The study points out the necessity of a strong cooperation among the different professionals involved in brachytherapy procedures. Minimal requirements for quality control institutional protocols are also presented and discussed. [Italian] Il documento costituisce un'integrazione delle linee guida italiane elaborate dal gruppo di studio Assicurazione di qualita' in radioterapia dell'Istituto Superiore di Sanita' (ISTISAN 96/39). Un rilievo ancora maggiore e' stato dato alla necessita' di una stretta collaborazione tra le figure professionali operanti nel campo della brachiterapia. Queste linee guida riportano anche i requisiti essenziali di un programma locale di controlli di qualita' in brachiterapia.

  6. Stress indotto dalla guida di autoveicoli: studio di parametri psicofisiologici

    Directory of Open Access Journals (Sweden)

    R. Vivoli

    2003-05-01

    Full Text Available

    La guida di un autoveicolo, specie in condizioni di intenso traffico o di cattive condizioni meteoclimatiche, è considerata uno dei principali fattori stressanti della vita odierna. Durante la guida si possono infatti osservare significative risposte simpatoadrenergiche e cardiovascolari con la comparsa anche di episodi coronarici. Tali risposte presentano un’ampia variabilità interindividuale in quanto sono influenzate da vari fattori (stile di vita, personalità etc..

    In questa rassegna, vengono presentati i principali risultati degli studi da noi condotti sulle risposte psicofisiologiche indotte dalla guida di diversi tipi di autoveicoli. In conducenti di autobus urbani è stato registrato un sensibile aumento del tasso urinario di adrenalina durante il turno lavorativo rispetto alla stessa fascia oraria di un giorno di riposo. Tale risposta adrenergica era esaltata dall’abitudine tabagica e dall’assunzione di caffè e bevande alcoliche. In soggetti che alla guida di camion coprono lunghe distanze, i più elevati tassi urinari di catecolamine e i più alti livelli di frequenza cardiaca sono stati registrati quando la guida era particolarmente stressante per la presenza di nebbia o di intenso traffico. In soggetti che partecipano, a livello amatoriale, a manifestazioni agonistiche di velocità su pista, abbiamo osservato che durante la gara, rispetto al periodo pre-gara, l’escrezione urinaria di catecolamine aumenta in misura molto rilevante. Analogamente a quanto osservato in autisti di camion, l’attivazione del sistema adrenergico è risultata direttamente associata ai livelli di ansietà. Dall’elettrocardiogramma dinamico è emerso che durante la gara la frequenza cardiaca raggiunge un valore medio di 163.5±7.4 battiti/min. (range: 146,180 battiti/min..

    Un significativo peggioramento della percezione visiva stereoscopica, con potenziale ricaduta negativa sulla performance dei guidatori, è stato osservato in

  7. Effect of Chromatin Structure on the Extent and Distribution of DNA Double Strand Breaks Produced by Ionizing Radiation; Comparative Study of hESC and Differentiated Cells Lines.

    Science.gov (United States)

    Venkatesh, Priyanka; Panyutin, Irina V; Remeeva, Evgenia; Neumann, Ronald D; Panyutin, Igor G

    2016-01-02

    Chromatin structure affects the extent of DNA damage and repair. Thus, it has been shown that heterochromatin is more protective against DNA double strand breaks (DSB) formation by ionizing radiation (IR); and that DNA DSB repair may proceed differently in hetero- and euchromatin regions. Human embryonic stem cells (hESC) have a more open chromatin structure than differentiated cells. Here, we study the effect of chromatin structure in hESC on initial DSB formation and subsequent DSB repair. DSB were scored by comet assay; and DSB repair was assessed by repair foci formation via 53BP1 antibody staining. We found that in hESC, heterochromatin is confined to distinct regions, while in differentiated cells it is distributed more evenly within the nuclei. The same dose of ionizing radiation produced considerably more DSB in hESC than in differentiated derivatives, normal human fibroblasts; and one cancer cell line. At the same time, the number of DNA repair foci were not statistically different among these cells. We showed that in hESC, DNA repair foci localized almost exclusively outside the heterochromatin regions. We also noticed that exposure to ionizing radiation resulted in an increase in heterochromatin marker H3K9me3 in cancer HT1080 cells, and to a lesser extent in IMR90 normal fibroblasts, but not in hESCs. These results demonstrate the importance of chromatin conformation for DNA protection and DNA damage repair; and indicate the difference of these processes in hESC.

  8. Recommendation for environmental management and safety systems in high risk industrial accidents; Linee guida per l'integrazione dei sistemi di gestione ambiente e sicurezza nelle aziende a rischio di incidente rilevante

    Energy Technology Data Exchange (ETDEWEB)

    Luciani, R; Andriola, L; Caropresso, G; Paci, S; Vignati, S; Vignoni, P C [ENEA Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    2000-07-01

    This document presents the IMPREMAS Project from Italian ENEA (National Agency for New Technology, Energy and the Environment) for the eco-management of small and medium enterprises at risk for se and manipulation of hazardous substances. [Italian] Il presente documento e' stato elaborato nell'ambito del progetto IMPREMAS condotto dall'ENEA, in collaborazione con il Ministero dell'Ambiente, Regione Lombardia, Federchimica e Finlombarda, per la realizzazione di un progetto pilota per l'eco-gestione delle piccole e medie imprese classificate a rischio per l'uso e lo stoccaggio di sostanze pericolose previste dal DPR. n. 175 del 17-5-1988. L'azione ha riguardato l'applicazione del regolamento EMAS 1836/93 congiuntamente alla implementazione di un sistema di sicurezza per piccole e medie imprese a rischio di incidente rilevante del settore chimico-farmaceutico con l'obiettivo di mettere a punto le linee guida di riferimento per l'integrazione dei sistemi ambiente e sicurezza.

  9. Recommendation for environmental management and safety systems in high risk industrial accidents; Linee guida per l'integrazione dei sistemi di gestione ambiente e sicurezza nelle aziende a rischio di incidente rilevante

    Energy Technology Data Exchange (ETDEWEB)

    Luciani, R.; Andriola, L.; Caropresso, G.; Paci, S.; Vignati, S.; Vignoni, P.C. [ENEA Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    2000-07-01

    This document presents the IMPREMAS Project from Italian ENEA (National Agency for New Technology, Energy and the Environment) for the eco-management of small and medium enterprises at risk for se and manipulation of hazardous substances. [Italian] Il presente documento e' stato elaborato nell'ambito del progetto IMPREMAS condotto dall'ENEA, in collaborazione con il Ministero dell'Ambiente, Regione Lombardia, Federchimica e Finlombarda, per la realizzazione di un progetto pilota per l'eco-gestione delle piccole e medie imprese classificate a rischio per l'uso e lo stoccaggio di sostanze pericolose previste dal DPR. n. 175 del 17-5-1988. L'azione ha riguardato l'applicazione del regolamento EMAS 1836/93 congiuntamente alla implementazione di un sistema di sicurezza per piccole e medie imprese a rischio di incidente rilevante del settore chimico-farmaceutico con l'obiettivo di mettere a punto le linee guida di riferimento per l'integrazione dei sistemi ambiente e sicurezza.

  10. The growth hormone-releasing hormone (GHRH) antagonist JV-1-36 inhibits proliferation and survival of human ectopic endometriotic stromal cells (ESCs) and the T HESC cell line.

    Science.gov (United States)

    Annunziata, Marta; Grande, Cristina; Scarlatti, Francesca; Deltetto, Francesco; Delpiano, Elena; Camanni, Marco; Ghigo, Ezio; Granata, Riccarda

    2010-08-01

    To determine the effect of the GHRH antagonist JV-1-36 on proliferation and survival of primary ectopic human endometriotic stromal cells (ESCs) and the T HESC cell line. Prospective laboratory study. University hospital. 22 women with endometriosis (aged 34.8+/-5.7 years) undergoing therapeutic laparoscopy. Eutopic (n=10) and ectopic (n=22) endometrial tissues were collected from women who underwent therapeutic laparoscopic surgery for endometriosis (stage III/IV). Expression of GHRH, GHRH receptor (GHRH-R) and GHRH-R splice variant (SV) 1 mRNA was determined by reverse-transcription polymerase chain reaction (RT-PCR). The ESC proliferation was assessed by 5-bromo-2-deoxyuridine incorporation, cell survival by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and Trypan blue assay. The T HESC survival was evaluated by MTT, cyclic adenosine monophosphate (cAMP) levels by ELISA, extracellular signal-regulated kinases 1 and 2 (ERK1/2) phosphorylation by Western blot, and insulin-like growth factor (IGF)-2 mRNA by real-time PCR. The ESCs and T HESCs, but not normal endometrial tissues, expressed GHRH-R mRNA; SV1 mRNA was determined in normal endometrial tissues, ESCs, and T HESCs; GHRH mRNAwas found in T HESCs; JV-1-36 inhibited ESC proliferation and ESC and T HESC survival. In T HESCs, JV-1-36 reduced cAMP production and ERK1/2 phosphorylation but had no effect on IGF-2 mRNA expression. The GHRH antagonist JV-1-36 inhibits endometriotic cell proliferation and survival, suggesting that GHRH antagonist may represent promising tools for treatment of endometriosis. Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  11. The transcriptomes of novel marmoset monkey embryonic stem cell lines reflect distinct genomic features.

    Science.gov (United States)

    Debowski, Katharina; Drummer, Charis; Lentes, Jana; Cors, Maren; Dressel, Ralf; Lingner, Thomas; Salinas-Riester, Gabriela; Fuchs, Sigrid; Sasaki, Erika; Behr, Rüdiger

    2016-07-07

    Embryonic stem cells (ESCs) are useful for the study of embryonic development. However, since research on naturally conceived human embryos is limited, non-human primate (NHP) embryos and NHP ESCs represent an excellent alternative to the corresponding human entities. Though, ESC lines derived from naturally conceived NHP embryos are still very rare. Here, we report the generation and characterization of four novel ESC lines derived from natural preimplantation embryos of the common marmoset monkey (Callithrix jacchus). For the first time we document derivation of NHP ESCs derived from morula stages. We show that quantitative chromosome-wise transcriptome analyses precisely reflect trisomies present in both morula-derived ESC lines. We also demonstrate that the female ESC lines exhibit different states of X-inactivation which is impressively reflected by the abundance of the lncRNA X inactive-specific transcript (XIST). The novel marmoset ESC lines will promote basic primate embryo and ESC studies as well as preclinical testing of ESC-based regenerative approaches in NHP.

  12. Electrophysiological Characteristics of Embryonic Stem Cell-Derived Cardiomyocytes are Cell Line-Dependent

    Directory of Open Access Journals (Sweden)

    Tobias Hannes

    2015-01-01

    Full Text Available Background: Modelling of cardiac development, physiology and pharmacology by differentiation of embryonic stem cells (ESCs requires comparability of cardiac differentiation between different ESC lines. To investigate whether the outcome of cardiac differentiation is consistent between different ESC lines, we compared electrophysiological properties of ESC-derived cardiomyocytes (ESC-CMs of different murine ESC lines. Methods: Two wild-type (D3 and R1 and two transgenic ESC lines (D3/aPIG44 and CGR8/AMPIGX-7 were differentiated under identical culture conditions. The transgenic cell lines expressed enhanced green fluorescent protein (eGFP and puromycin-N-acetyltransferase under control of the cardiac specific α-myosin heavy chain (αMHC promoter. Action potentials (APs were recorded using sharp electrodes and multielectrode arrays in beating clusters of ESC-CMs. Results: Spontaneous AP frequency and AP duration (APD as well as maximal upstroke velocity differed markedly between unpurified CMs of the four ESC lines. APD heterogeneity was negligible in D3/aPIG44, moderate in D3 and R1 and extensive in CGR8/AMPIGX-7. Interspike intervals calculated from long-term recordings showed a high degree of variability within and between recordings in CGR8/AMPIGX-7, but not in D3/aPIG44. Purification of the αMHC+ population by puromycin treatment posed only minor changes to APD in D3/aPIG44, but significantly shortened APD in CGR8/AMPIGX-7. Conclusion: Electrophysiological properties of ESC-CMs are strongly cell line-dependent and can be influenced by purification of cardiomyocytes by antibiotic selection. Thus, conclusions on cardiac development, physiology and pharmacology derived from single stem cell lines have to be interpreted carefully.

  13. Transcriptome variations among human embryonic stem cell lines are associated with their differentiation propensity.

    Directory of Open Access Journals (Sweden)

    Changbin Sun

    Full Text Available Human embryonic stem cells (hESCs have the potential to form any cell type in the body, making them attractive cell sources in drug screening, regenerative medicine, disease and developmental processes modeling. However, not all hESC lines have the equal potency to generate desired cell types in vitro. Significant variations have been observed for the differentiation efficiency of various human ESC lines. The precise underpinning molecular mechanisms are still unclear. In this work, we compared transcriptome variations of four hESC lines H7, HUES1, HUES8 and HUES9. We found that hESC lines have different gene expression profiles, and these differentially expressed genes (DEGs are significantly enriched in developmental processes, such as ectodermal, mesodermal and endodermal development. The enrichment difference between hESC lines was consistent with its lineage bias. Among these DEGs, some pluripotency factors and genes involved in signaling transduction showed great variations as well. The pleiotropic functions of these genes in controlling hESC identity and early lineage specification, implicated that different hESC lines may utilize distinct balance mechanisms to maintain pluripotent state. When the balance is broken in a certain environment, gene expression variation between them could impact on their different lineage specification behavior.

  14. Derivation and characterization of human embryonic stem cell lines from the Chinese population

    Institute of Scientific and Technical Information of China (English)

    Zhao Wu; Huimin Dai; Lei Qian; Qing Tian; Lei Xiao; Xiaojun Tan; Hui Li; Lingjun Rao; Lixiazi He; Lei Bao; Jing Liao; Chun Cui; Zhenyu Zuo; Qiao Li

    2011-01-01

    Human embryonic stem cells (hESCs) can self-renew indefinitely and differentiate into all cell types in the human body. Therefore, they are valuable in regenerative medicine, human developmental biology and drug discovery. A number of hESC lines have been derived from the Chinese population,but limited of them are available for research purposes. Here we report the derivation and characterization of two hESC lines derived from human blastocysts of Chinese origin. These hESCs express alkaline phosphatase and hESC-specific markers, including Oct4, Nanog, SSEA-3, SSEA-4,TRA-1-60 and TRA-1-81. They also have high levels of telomerase activity and normal karyotypes. These cells can form embryoid body in vitro and can be differentiated into all three germ layers in vivo by teratoma formation. The newly established hESCs will be distributed for research purposes.The availability of hESC lines from the Chinese population will facilitate studies on the differences in hESCs from different ethnic groups.

  15. No Relationship between Embryo Morphology and Successful Derivation of Human Embryonic Stem Cell Lines

    Science.gov (United States)

    Ström, Susanne; Rodriguez-Wallberg, Kenny; Holm, Frida; Bergström, Rosita; Eklund, Linda; Strömberg, Anne-Marie; Hovatta, Outi

    2010-01-01

    Background The large number (30) of permanent human embryonic stem cell (hESC) lines and additional 29 which did not continue growing, in our laboratory at Karolinska Institutet have given us a possibility to analyse the relationship between embryo morphology and the success of derivation of hESC lines. The derivation method has been improved during the period 2002–2009, towards fewer xeno-components. Embryo quality is important as regards the likelihood of pregnancy, but there is little information regarding likelihood of stem cell derivation. Methods We evaluated the relationship of pronuclear zygote stage, the score based on embryo morphology and developmental rate at cleavage state, and the morphology of the blastocyst at the time of donation to stem cell research, to see how they correlated to successful establishment of new hESC lines. Results Derivation of hESC lines succeeded from poor quality and good quality embryos in the same extent. In several blastocysts, no real inner cell mass (ICM) was seen, but permanent well growing hESC lines could be established. One tripronuclear (3PN) zygote, which developed to blastocyst stage, gave origin to a karyotypically normal hESC line. Conclusion Even very poor quality embryos with few cells in the ICM can give origin to hESC lines. PMID:21217828

  16. Derivation of novel genetically diverse human embryonic stem cell lines.

    Science.gov (United States)

    Stefanova, Valentina T; Grifo, James A; Hansis, Christoph

    2012-06-10

    Human embryonic stem cells (hESCs) have the potential to revolutionize many biomedical fields ranging from basic research to disease modeling, regenerative medicine, drug discovery, and toxicity testing. A multitude of hESC lines have been derived worldwide since the first 5 lines by Thomson et al. 13 years ago, but many of these are poorly characterized, unavailable, or do not represent desired traits, thus making them unsuitable for application purposes. In order to provide the scientific community with better options, we have derived 12 new hESC lines at New York University from discarded genetically normal and abnormal embryos using the latest techniques. We examined the genetic status of the NYUES lines in detail as well as their molecular and cellular features and DNA fingerprinting profile. Furthermore, we differentiated our hESCs into the tissues most affected by a specific condition or into clinically desired cell types. To our knowledge, a number of characteristics of our hESCs have not been previously reported, for example, mutation for alpha thalassemia X-linked mental retardation syndrome, linkage to conditions with a genetic component such as asthma or poor sperm morphology, and novel combinations of ethnic backgrounds. Importantly, all of our undifferentiated euploid female lines tested to date did not show X chromosome inactivation, believed to result in superior potency. We continue to derive new hESC lines and add them to the NIH registry and other registries. This should facilitate the use of our hESCs and lead to advancements for patient-benefitting applications.

  17. A Panel of Embryonic Stem Cell Lines Reveals the Variety and Dynamic of Pluripotent States in Rabbits

    Directory of Open Access Journals (Sweden)

    Pierre Osteil

    2016-09-01

    Full Text Available Conventional rabbit embryonic stem cell (ESC lines are derived from the inner cell mass (ICM of pre-implantation embryos using methods and culture conditions that are established for primate ESCs. In this study, we explored the capacity of the rabbit ICM to give rise to ESC lines using conditions similar to those utilized to generate naive ESCs in mice. On single-cell dissociation and culture in fibroblast growth factor 2 (FGF2-free, serum-supplemented medium, rabbit ICMs gave rise to ESC lines lacking the DNA-damage checkpoint in the G1 phase like mouse ESCs, and with a pluripotency gene expression profile closer to the rabbit ICM/epiblast profiles. These cell lines can be converted to FGF2-dependent ESCs after culture in conventional conditions. They can also colonize the rabbit pre-implantation embryo. These results indicate that rabbit epiblast cells can be coaxed toward different types of pluripotent stem cells and reveal the dynamics of pluripotent states in rabbit ESCs.

  18. Combats escènics

    Directory of Open Access Journals (Sweden)

    Pawel Rouba Billowicz

    2016-04-01

    Full Text Available “Combats Escènics” és un treball que tracta sobre la interpretació artística de la violència d’artistes de l’espectacle per tal de divertir el públic i emetre un missatge humanista mitjançant una coreografia ritual. En aquest estudi es presenta una classificació del combat escènic des del doble vessant agonista/antagonista, es realitza un passeig històric de la representació artística del combat a través de les diferents etapes i de les diverses cultures, s’aborda la preparació escènica de l’actor i del coreògraf, i s’entreveuen les perspectives de futur d’aquesta modalitat artística. Estudi realitzat per Pawel Rouba Billewicz (Inowroclaw, Polònia, 1939 - Barcelona, 2007, director, coreògraf, actor, mestre d’armes, mestre del gest i de la pantomima i professor de l’INEF de Catalunya. Aquest article, editorialment inèdit, es publica postmortem per Apunts. Educació Física i Esports com a homenatge i reconeixement de l’autor per la seva extraordinària i polivalent aportació al camp de l’art i l’Activitat Física i l’Esport.

  19. Car drivers' perceptions of electronic stability control (ESC) systems.

    Science.gov (United States)

    Vadeby, Anna; Wiklund, Mats; Forward, Sonja

    2011-05-01

    As a way to reduce the number of car crashes different in-car safety devices are being introduced. In this paper one such application is being investigated, namely the electronic stability control system (ESC). The study used a survey method, including 2000 private car drivers (1000 driving a car with ESC and 1000 driving a car without ESC). The main objective was to investigate the effect of ESC on driver behaviour. Results show that drivers report that they drive even more carelessly when they believe that they have ESC, than when they do not. Men are more risk prone than women and young drivers more than older drivers. Using the theory of planned behaviour the results show that attitude, subjective norm and perceived control explain between 62% and 67% of driver's variation of intentions to take risks. When descriptive norm was added to the model a small but statistically significant increase was found. The study also shows that more than 35% erroneously believe that their car is equipped with an ESC system. These findings may suggest that driver behaviour could reduce the positive effect ESC has on accidents. It also shows that drivers who purchase a new car are not well informed about what kind of safety devices the car is equipped with. These findings highlight the need for more targeted information to drivers. Copyright © 2010 Elsevier Ltd. All rights reserved.

  20. A model-based analysis of culture-dependent phenotypes of mESCs.

    Directory of Open Access Journals (Sweden)

    Maria Herberg

    Full Text Available Mouse embryonic stem cells (mESCs can be maintained in a proliferative and undifferentiated state over many passages (self-renewal while retaining the potential to give rise to every cell type of the organism (pluripotency. Autocrine FGF4/Erk signalling has been identified as a major stimulus for fate decisions and lineage commitment in these cells. Recent findings on serum-free culture conditions with specific inhibitors (known as 2i demonstrate that the inhibition of this pathway reduces transcription factor heterogeneity and is vital to maintain ground state pluripotency of mESCs. We suggest a novel mathematical model to explicitly integrate FGF4/Erk signalling into an interaction network of key pluripotency factors (namely Oct4, Sox2, Nanog and Rex1. The envisaged model allows to explore whether and how proposed mechanisms and feedback regulations can account for different expression patterns in mESC cultures. We demonstrate that an FGF4/Erk-mediated negative feedback is sufficient to induce molecular heterogeneity with respect to Nanog and Rex1 expression and thus critically regulates the propensity for differentiation and the loss of pluripotency. Furthermore, we compare simulation results on the transcription factor dynamics in different self-renewing states and during differentiation with experimental data on a Rex1GFPd2 reporter cell line using flow cytometry and qRT-PCR measurements. Concluding from our results we argue that interaction between FGF4/Erk signalling and Nanog expression qualifies as a key mechanism to manipulate mESC pluripotency. In particular, we infer that ground state pluripotency under 2i is achieved by shifting stable expression pattern of Nanog from a bistable into a monostable regulation impeding stochastic state transitions. Furthermore, we derive testable predictions on altering the degree of Nanog heterogeneity and on the frequency of state transitions in LIF/serum conditions to challenge our model assumptions.

  1. Derivation and characterization of novel nonhuman primate embryonic stem cell lines from in vitro-fertilized baboon preimplantation embryos.

    Science.gov (United States)

    Chang, Tien-Cheng; Liu, Ya-Guang; Eddy, Carlton A; Jacoby, Ethan S; Binkley, Peter A; Brzyski, Robert G; Schenken, Robert S

    2011-06-01

    The development of nonhuman primate (NHP) embryonic stem cell (ESC) models holds great promise for cell-mediated treatment of debilitating diseases and to address numerous unanswered questions regarding the therapeutic efficacy of ESCs while supplanting ethical considerations involved with human studies. Here we report successful establishment and characterization of 3 novel baboon (Papio cynocephalus) ESC lines from the inner cell mass of intracytoplasmic sperm injection-derived blastocysts. Embryos were cultured in an improved baboon embryo in vitro culture protocol. The inner cell mass of blastocyst was laser-dissected and plated on mouse embryonic fibroblast feeder cell monolayer in the NHP ESC culture medium. Three cell lines with characteristic ESC morphology have been cultured through an extended period (>14 months), with 2 male cell lines (UT-1 and -2) and 1 female cell line (UT-3) displaying normal baboon karyotypes. Reverse transcription-polymerase chain reaction analysis confirmed that all 3 lines express primate ESC pluripotency markers, including OCT-4, NANOG, SOX-2, TERT, TDGF, LEFTYA, and REX-1. All 3 lines demonstrated positive immunocytochemical staining for OCT-4, stage-specific embryonic antigen-3, stage-specific embryonic antigen-4, TRA-1-60, and TRA-1-81. Baboon ESCs injected into NOD/SCID mice formed teratomas with all 3 germ layers. In addition, embryoid body-like spherical structures were derived and initial outgrowth was observed when embedded into extracellular matrix Matrigel. The ESC lines established in this NHP model have the potential to extend our knowledge in the fields of developmental biology, regenerative medicine, and future applications, including preclinical safety assessment of in vivo stem cell therapy.

  2. File list: ALL.PSC.50.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  3. File list: ALL.PSC.10.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  4. File list: ALL.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.20.AllAg.mESCs,_differentiated mm9 All antigens Pluripotent stem cell mESCs, differentia...barchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.20.AllAg.mESCs,_differentiated.bed ...

  5. File list: ALL.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.05.AllAg.mESCs,_differentiated mm9 All antigens Pluripotent stem cell mESCs, differentia...barchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.05.AllAg.mESCs,_differentiated.bed ...

  6. File list: Unc.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.05.AllAg.mESCs,_differentiated mm9 Unclassified Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.05.AllAg.mESCs,_differentiated.bed ...

  7. File list: Unc.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.20.AllAg.mESCs,_differentiated mm9 Unclassified Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.20.AllAg.mESCs,_differentiated.bed ...

  8. File list: DNS.PSC.50.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.50.AllAg.mESCs,_differentiated mm9 DNase-seq Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.50.AllAg.mESCs,_differentiated.bed ...

  9. File list: DNS.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.05.AllAg.mESCs,_differentiated mm9 DNase-seq Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.05.AllAg.mESCs,_differentiated.bed ...

  10. File list: DNS.PSC.10.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.10.AllAg.mESCs,_differentiated mm9 DNase-seq Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.10.AllAg.mESCs,_differentiated.bed ...

  11. File list: DNS.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.20.AllAg.mESCs,_differentiated mm9 DNase-seq Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.20.AllAg.mESCs,_differentiated.bed ...

  12. File list: Pol.PSC.10.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.10.AllAg.mESCs,_differentiated mm9 RNA polymerase Pluripotent stem cell mESCs, differentia...ted SRX590276 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.10.AllAg.mESCs,_differentiated.bed ...

  13. File list: Pol.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.05.AllAg.mESCs,_differentiated mm9 RNA polymerase Pluripotent stem cell mESCs, differentia...ted SRX590276 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.05.AllAg.mESCs,_differentiated.bed ...

  14. File list: Pol.PSC.50.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.50.AllAg.mESCs,_differentiated mm9 RNA polymerase Pluripotent stem cell mESCs, differentia...ted SRX590276 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.50.AllAg.mESCs,_differentiated.bed ...

  15. File list: Pol.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.20.AllAg.mESCs,_differentiated mm9 RNA polymerase Pluripotent stem cell mESCs, differentia...ted SRX590276 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.20.AllAg.mESCs,_differentiated.bed ...

  16. Tet Enzymes Regulate Telomere Maintenance and Chromosomal Stability of Mouse ESCs

    Directory of Open Access Journals (Sweden)

    Jiao Yang

    2016-05-01

    Full Text Available Ten-eleven translocation (Tet family proteins convert 5-methylcytosine to 5-hydroxymethylcytosine. We show that mouse embryonic stem cells (ESCs depleted of Tet1 and/or Tet2 by RNAi exhibit short telomeres and chromosomal instability, concomitant with reduced telomere recombination. Tet1 and Tet2 double-knockout ESCs also display short telomeres but to a lesser extent. Notably, Tet1/2/3 triple-knockout ESCs show heterogeneous telomere lengths and increased frequency of telomere loss and chromosomal fusion. Mechanistically, Tets depletion or deficiency increases Dnmt3b and decreases 5hmC levels, resulting in elevated methylation levels at sub-telomeres. Consistently, knockdown of Dnmt3b or addition of 2i (MAPK and GSK3β inhibitors, which also inhibits Dnmt3b, reduces telomere shortening, partially rescuing Tet1/2 deficiency. Interestingly, Tet1/2 double or Tet1/2/3 triple knockout in ESCs consistently upregulates Zscan4, which may counteract telomere shortening. Together, Tet enzymes play important roles in telomere maintenance and chromosomal stability of ESCs by modulating sub-telomeric methylation levels.

  17. Aberrant patterns of X chromosome inactivation in a new line of human embryonic stem cells established in physiological oxygen concentrations.

    Science.gov (United States)

    de Oliveira Georges, Juliana Andrea; Vergani, Naja; Fonseca, Simone Aparecida Siqueira; Fraga, Ana Maria; de Mello, Joana Carvalho Moreira; Albuquerque, Maria Cecília R Maciel; Fujihara, Litsuko Shimabukuro; Pereira, Lygia Veiga

    2014-08-01

    One of the differences between murine and human embryonic stem cells (ESCs) is the epigenetic state of the X chromosomes in female lines. Murine ESCs (mESCs) present two transcriptionally active Xs that will undergo the dosage compensation process of XCI upon differentiation, whereas most human ESCs (hESCs) spontaneously inactivate one X while keeping their pluripotency. Whether this reflects differences in embryonic development of mice and humans, or distinct culture requirements for the two kinds of pluripotent cells is not known. Recently it has been shown that hESCs established in physiological oxygen levels are in a stable pre-XCI state equivalent to that of mESCs, suggesting that culture in low oxygen concentration is enough to preserve that epigenetic state of the X chromosomes. Here we describe the establishment of two new lines of hESCs under physiological oxygen level and the characterization of the XCI state in the 46,XX line BR-5. We show that a fraction of undifferentiated cells present XIST RNA accumulation and single H3K27me foci, characteristic of the inactive X. Moreover, analysis of allele specific gene expression suggests that pluripotent BR-5 cells present completely skewed XCI. Our data indicate that physiological levels of oxygen are not sufficient for the stabilization of the pre-XCI state in hESCs.

  18. European Society of Cardiology (ESC) Annual Congress Report From Barcelona 2017.

    Science.gov (United States)

    Satoh, Kimio; Takahashi, Jun; Matsumoto, Yasuharu; Tatebe, Shunsuke; Aoki, Tatsuo; Kikuchi, Yoku; Hao, Kiyotaka; Ohyama, Kazuma; Nogi, Masamichi; Suda, Akira; Kasahara, Shintaro; Sato, Koichi; Ichijo, Sadamitsu; Shimokawa, Hiroaki

    2017-11-24

    From August 26th to 30th, the 2017 Annual Congress of the European Society of Cardiology (ESC 2017) was held in Barcelona, Spain. Despite the terrorism tradegy just before the ESC congress, the congress attracted many medical professionals from all over the world to discuss the recent topics in cardiovascular medicine in more than 500 sessions, including COMPASS (Cardiovascular OutcoMes for People using Anticoagulation StrategieS Trial), CANTOS (Canakinumab Anti-Inflammatory Thrombosis Outcomes Study), and ORION (which assessed the effect of a novel siRNA inhibitor to PCSK9 on reductions in low-density lipoprotein cholesterol). Japanese cardiologists and the Japanese Circulation Society greatly contributed to the congress. This report briefly introduces some late-breaking registry results, late-breaking clinical trials, and ESC Guidelines from the ESC 2017 Congress.

  19. File list: Oth.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.20.AllAg.mESCs,_differentiated mm9 TFs and others Pluripotent stem cell mESCs, differentia...4,SRX754570,SRX065538,SRX523453,SRX523451,SRX065537 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.20.AllAg.mESCs,_differentiated.bed ...

  20. File list: Oth.PSC.50.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.50.AllAg.mESCs,_differentiated mm9 TFs and others Pluripotent stem cell mESCs, differentia...8,SRX523453,SRX523451,SRX754570,SRX882858,SRX065537 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.50.AllAg.mESCs,_differentiated.bed ...

  1. File list: Oth.PSC.10.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.10.AllAg.mESCs,_differentiated mm9 TFs and others Pluripotent stem cell mESCs, differentia...2,SRX065538,SRX523453,SRX754570,SRX065537,SRX523451 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.10.AllAg.mESCs,_differentiated.bed ...

  2. File list: Oth.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.AllAg.mESCs,_differentiated mm9 TFs and others Pluripotent stem cell mESCs, differentia...8,SRX209322,SRX065538,SRX065537,SRX523453,SRX523451 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.05.AllAg.mESCs,_differentiated.bed ...

  3. The perfect host: a mouse host embryo facilitating more efficient germ line transmission of genetically modified embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Robert A Taft

    Full Text Available There is a continual need to improve efficiency in creating precise genetic modifications in mice using embryonic stem cells (ESCs. We describe a novel approach resulting in 100% germline transmission from competent injected ESCs. We developed an F1 mouse host embryo (Perfect Host, PH that selectively ablates its own germ cells via tissue-specific induction of diphtheria toxin. This approach allows competent microinjected ESCs to fully dominate the germline, eliminating competition for this critical niche in the developing and adult animal. This is in contrast to conventional methods, where competition from host germ cells results in offspring derived from host cells and ESCs, necessitating extensive breeding of chimeras and genotyping to identify germline. The germline transmission process is also complicated by variability in the actual number of ESCs that colonize the germline niche and the proportion that are germline competent. To validate the PH approach we used ESC lines derived from 129 F1, BALB/cByJ, and BTBR backgrounds as well as an iPS line. Resulting chimeric males produced 194 offspring, all paternally derived from the introduced stem cells, with no offspring being derived from the host genome. We further tested this approach using eleven genetically modified C57BL/6N ESC lines (International Knockout Mouse Consortium. ESC germline transmission was observed in 9/11 (82% lines using PH blastocysts, compared to 6/11 (55% when conventional host blastocysts were used. Furthermore, less than 35% (83/240 of mice born in the first litters from conventional chimeras were confirmed to be of ESC-origin. By comparison, 100% (137/137 of the first litter offspring of PH chimeras were confirmed as ESC-derived. Together, these data demonstrate that the PH approach increases the probability of germline transmission and speeds the generation of ESC derived animals from chimeras. Collectively, this approach reduces the time and costs inherent in the

  4. File list: NoD.PSC.05.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.05.AllAg.mESCs,_differentiated mm9 No description Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.PSC.05.AllAg.mESCs,_differentiated.bed ...

  5. File list: InP.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.20.AllAg.mESCs,_differentiated mm9 Input control Pluripotent stem cell mESCs, differentia...,SRX388447 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.20.AllAg.mESCs,_differentiated.bed ...

  6. File list: NoD.PSC.20.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.20.AllAg.mESCs,_differentiated mm9 No description Pluripotent stem cell mESCs, differentia...ted http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.PSC.20.AllAg.mESCs,_differentiated.bed ...

  7. File list: InP.PSC.50.AllAg.mESCs,_differentiated [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.50.AllAg.mESCs,_differentiated mm9 Input control Pluripotent stem cell mESCs, differentia...,SRX388447 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.50.AllAg.mESCs,_differentiated.bed ...

  8. KAT-Independent Gene Regulation by Tip60 Promotes ESC Self-Renewal but Not Pluripotency

    Directory of Open Access Journals (Sweden)

    Diwash Acharya

    2017-04-01

    Full Text Available Although histone-modifying enzymes are generally assumed to function in a manner dependent on their enzymatic activities, this assumption remains untested for many factors. Here, we show that the Tip60 (Kat5 lysine acetyltransferase (KAT, which is essential for embryonic stem cell (ESC self-renewal and pre-implantation development, performs these functions independently of its KAT activity. Unlike ESCs depleted of Tip60, KAT-deficient ESCs exhibited minimal alterations in gene expression, chromatin accessibility at Tip60 binding sites, and self-renewal, thus demonstrating a critical KAT-independent role of Tip60 in ESC maintenance. In contrast, KAT-deficient ESCs exhibited impaired differentiation into mesoderm and endoderm, demonstrating a KAT-dependent function in differentiation. Consistent with this phenotype, KAT-deficient mouse embryos exhibited post-implantation developmental defects. These findings establish separable KAT-dependent and KAT-independent functions of Tip60 in ESCs and during differentiation, revealing a complex repertoire of regulatory functions for this essential chromatin remodeling complex.

  9. O Escândalo Político como experiência narrativa

    Directory of Open Access Journals (Sweden)

    Helder Prior

    2015-12-01

    Full Text Available O trabalho sobre a experiência narrativa do escândalo político procura abordar as relações entre o campo do jornalismo e os pressupostos da narratologia. Procuraremos estabelecer um quadro teórico e conceptual de análise dos escândalos político-mediáticos tendo em conta o papel dos meios de comunicação na reconfiguração do acontecimento em intrigas inteligíveis para o leitor. Na nossa perspectiva, os escândalos políticos são narrativas complexas que se desenvolvem na imprensa, podendo ser interpretados como “estórias” que têm um enredo, episódios principais e secundários, personagens, e efeitos de sentido inerentes ao trabalho plásticos do medium no momento de converter o fenómeno numa experiência mediática. Na parte final, oferecemos ao leitor um esquema teórico e conceptual de desconstrução hermenêutica e pragmática do escândalo mediático, colocando em evidência o valor expressivo do escândalo e a componente esteticizante que ele revela.

  10. File list: ALL.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.50.AllAg.mESC_derived_pancreatic_cells mm9 All antigens Pluripotent stem cell mESC derived panc...cedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.50.AllAg.mESC_derived_pancreatic_cells.bed ...

  11. File list: ALL.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: ALL.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.20.AllAg.mESC_derived_pancreatic_cells mm9 All antigens Pluripotent stem cell mESC derived panc...cedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.20.AllAg.mESC_derived_pancreatic_cells.bed ...

  13. File list: His.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.05.AllAg.mESC_derived_neural_cells mm9 Histone Pluripotent stem cell mESC derived neural...tp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  14. File list: His.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.20.AllAg.mESC_derived_neural_cells mm9 Histone Pluripotent stem cell mESC derived neural...tp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  15. Rtt107/Esc4 binds silent chromatin and DNA repair proteins using different BRCT motifs

    Directory of Open Access Journals (Sweden)

    Jockusch Rebecca A

    2006-11-01

    Full Text Available Abstract Background By screening a plasmid library for proteins that could cause silencing when targeted to the HMR locus in Saccharomyces cerevisiae, we previously reported the identification of Rtt107/Esc4 based on its ability to establish silent chromatin. In this study we aimed to determine the mechanism of Rtt107/Esc4 targeted silencing and also learn more about its biological functions. Results Targeted silencing by Rtt107/Esc4 was dependent on the SIR genes, which encode obligatory structural and enzymatic components of yeast silent chromatin. Based on its sequence, Rtt107/Esc4 was predicted to contain six BRCT motifs. This motif, originally identified in the human breast tumor suppressor gene BRCA1, is a protein interaction domain. The targeted silencing activity of Rtt107/Esc4 resided within the C-terminal two BRCT motifs, and this region of the protein bound to Sir3 in two-hybrid tests. Deletion of RTT107/ESC4 caused sensitivity to the DNA damaging agent MMS as well as to hydroxyurea. A two-hybrid screen showed that the N-terminal BRCT motifs of Rtt107/Esc4 bound to Slx4, a protein previously shown to be involved in DNA repair and required for viability in a strain lacking the DNA helicase Sgs1. Like SLX genes, RTT107ESC4 interacted genetically with SGS1; esc4Δ sgs1Δ mutants were viable, but exhibited a slow-growth phenotype and also a synergistic DNA repair defect. Conclusion Rtt107/Esc4 binds to the silencing protein Sir3 and the DNA repair protein Slx4 via different BRCT motifs, thus providing a bridge linking silent chromatin to DNA repair enzymes.

  16. File list: Unc.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.05.AllAg.mESC_derived_pancreatic_cells mm9 Unclassified Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.05.AllAg.mESC_derived_pancreatic_cells.bed ...

  17. File list: Unc.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.50.AllAg.mESC_derived_pancreatic_cells mm9 Unclassified Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.50.AllAg.mESC_derived_pancreatic_cells.bed ...

  18. File list: Unc.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 Unclassified Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  19. File list: Unc.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.20.AllAg.mESC_derived_pancreatic_cells mm9 Unclassified Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.20.AllAg.mESC_derived_pancreatic_cells.bed ...

  20. File list: Oth.PSC.50.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.50.AllAg.hESC_derived_neural_cells hg19 TFs and others Pluripotent stem cell hESC derived neural...http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.PSC.50.AllAg.hESC_derived_neural_cells.bed ...

  1. File list: Oth.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.AllAg.hESC_derived_neural_cells hg19 TFs and others Pluripotent stem cell hESC derived neural...http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  2. File list: Pol.PSC.20.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.20.AllAg.hESC_derived_neural_crests hg19 RNA polymerase Pluripotent stem cell hESC derived neural... crests http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.20.AllAg.hESC_derived_neural_crests.bed ...

  3. File list: Pol.PSC.05.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.05.AllAg.hESC_derived_neural_crests hg19 RNA polymerase Pluripotent stem cell hESC derived neural... crests http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.05.AllAg.hESC_derived_neural_crests.bed ...

  4. File list: DNS.PSC.50.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.50.AllAg.mESC_derived_neural_cells mm9 DNase-seq Pluripotent stem cell mESC derived neural... cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.50.AllAg.mESC_derived_neural_cells.bed ...

  5. File list: DNS.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.20.AllAg.mESC_derived_neural_cells mm9 DNase-seq Pluripotent stem cell mESC derived neural... cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  6. File list: Pol.PSC.10.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.10.AllAg.hESC_derived_neural_crests hg19 RNA polymerase Pluripotent stem cell hESC derived neural... crests http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.10.AllAg.hESC_derived_neural_crests.bed ...

  7. Traditional Chinese Medicine Baicalin Suppresses mESCs Proliferation through Inhibition of miR-294 Expression

    Directory of Open Access Journals (Sweden)

    Jian Wang

    2015-03-01

    Full Text Available Background: Traditional Chinese herbal medicines (TCMs have been widely used against a broad spectrum of biological activities, including influencing the cardiac differentiation from mouse embryonic stem cells (mESCs. However, their effects and mechanisms of action on ESCs proliferation remain to be determined. The present study aimed to determine the effect of three TCMs, baicalin, ginsenoside Rg1, and puerarin, on mESCs proliferation and to elucidate the possible mechanism of their action. Methods: Cell proliferation was examined with a cell proliferation assay Cell Counting Kit-8 (CCK-8, propidium iodide (PI staining was used to visualize cell cycle. The mRNA expression level of c-myc, c-fos, c-jun, GAPDH and microRNAs were measured by quantitative real time RT-PCR. Results: We found that baicalin 50 μM suppressed the proliferation of mESCs as observations in more cells in G1 phase and less cells in either S phase or G2/M phase. Moreover, baicalin suppressed the expressions of c-jun and c-fos in mESCs and down-regulated the expression of miR-294. Overexpression of miR-294 in mESCs significantly reversed the effects of baicalin both on mESC proliferation and c-fos/c-jun expression. Conclusions: Baicalin down-regulation of miR-294 may be its key mechanism of action in decreasing mESCs proliferation.

  8. File list: Pol.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 RNA polymerase Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  9. File list: Pol.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.05.AllAg.mESC_derived_pancreatic_cells mm9 RNA polymerase Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.05.AllAg.mESC_derived_pancreatic_cells.bed ...

  10. File list: Oth.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.20.AllAg.mESC_derived_pancreatic_cells mm9 TFs and others Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.20.AllAg.mESC_derived_pancreatic_cells.bed ...

  11. File list: Oth.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.AllAg.mESC_derived_pancreatic_cells mm9 TFs and others Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.05.AllAg.mESC_derived_pancreatic_cells.bed ...

  12. File list: Oth.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 TFs and others Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  13. File list: Pol.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.50.AllAg.mESC_derived_pancreatic_cells mm9 RNA polymerase Pluripotent stem cell mESC derived panc...reatic cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.50.AllAg.mESC_derived_pancreatic_cells.bed ...

  14. File list: Unc.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.05.AllAg.hESC_derived_neural_cells hg19 Unclassified Pluripotent stem cell hESC derived neural... cells SRX378284 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  15. File list: Unc.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.20.AllAg.mESC_derived_neural_cells mm9 Unclassified Pluripotent stem cell mESC derived neural...,SRX213761,SRX213757 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  16. File list: Unc.PSC.20.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.20.AllAg.hESC_derived_neural_crests hg19 Unclassified Pluripotent stem cell hESC derived neural... crests SRX059366 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.PSC.20.AllAg.hESC_derived_neural_crests.bed ...

  17. File list: Unc.PSC.05.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.PSC.05.AllAg.hESC_derived_neural_crests hg19 Unclassified Pluripotent stem cell hESC derived neural... crests SRX059366 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.PSC.05.AllAg.hESC_derived_neural_crests.bed ...

  18. File list: Pol.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.05.AllAg.hESC_derived_neural_cells hg19 RNA polymerase Pluripotent stem cell hESC derived neural... cells SRX190259 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  19. File list: Pol.PSC.10.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.10.AllAg.hESC_derived_neural_cells hg19 RNA polymerase Pluripotent stem cell hESC derived neural... cells SRX190259 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.10.AllAg.hESC_derived_neural_cells.bed ...

  20. File list: Pol.PSC.50.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.50.AllAg.hESC_derived_neural_cells hg19 RNA polymerase Pluripotent stem cell hESC derived neural... cells SRX190259 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.PSC.50.AllAg.hESC_derived_neural_cells.bed ...

  1. File list: DNS.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.50.AllAg.mESC_derived_pancreatic_cells mm9 DNase-seq Pluripotent stem cell mESC derived panc...reatic cells SRX404487 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.50.AllAg.mESC_derived_pancreatic_cells.bed ...

  2. File list: His.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.AllAg.mESC_derived_pancreatic_cells mm9 Histone Pluripotent stem cell mESC derived panc...reatic cells SRX146012,SRX146011 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.50.AllAg.mESC_derived_pancreatic_cells.bed ...

  3. File list: His.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.05.AllAg.mESC_derived_pancreatic_cells mm9 Histone Pluripotent stem cell mESC derived panc...reatic cells SRX146012,SRX146011 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.05.AllAg.mESC_derived_pancreatic_cells.bed ...

  4. File list: His.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 Histone Pluripotent stem cell mESC derived panc...reatic cells SRX146012,SRX146011 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  5. File list: DNS.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 DNase-seq Pluripotent stem cell mESC derived panc...reatic cells SRX404487 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  6. File list: DNS.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.20.AllAg.mESC_derived_pancreatic_cells mm9 DNase-seq Pluripotent stem cell mESC derived panc...reatic cells SRX404487 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.20.AllAg.mESC_derived_pancreatic_cells.bed ...

  7. File list: DNS.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.05.AllAg.mESC_derived_pancreatic_cells mm9 DNase-seq Pluripotent stem cell mESC derived panc...reatic cells SRX404487 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.PSC.05.AllAg.mESC_derived_pancreatic_cells.bed ...

  8. File list: Pol.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.20.AllAg.mESC_derived_neural_cells mm9 RNA polymerase Pluripotent stem cell mESC derived neural...RX213760,SRX213764 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  9. File list: ALL.PSC.05.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.05.AllAg.hESC_derived_neural_crests hg19 All antigens Pluripotent stem cell hESC derived neural...RX059366,SRX059364 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.PSC.05.AllAg.hESC_derived_neural_crests.bed ...

  10. File list: DNS.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.05.AllAg.hESC_derived_neural_cells hg19 DNase-seq Pluripotent stem cell hESC derived neural... cells SRX121241,SRX134721 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  11. File list: Pol.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.05.AllAg.mESC_derived_neural_cells mm9 RNA polymerase Pluripotent stem cell mESC derived neural...RX213764,SRX213760 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  12. File list: DNS.PSC.50.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.50.AllAg.hESC_derived_neural_cells hg19 DNase-seq Pluripotent stem cell hESC derived neural... cells SRX121241,SRX134721 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.PSC.50.AllAg.hESC_derived_neural_cells.bed ...

  13. File list: Pol.PSC.50.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.50.AllAg.mESC_derived_neural_cells mm9 RNA polymerase Pluripotent stem cell mESC derived neural...RX213760,SRX213764 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.50.AllAg.mESC_derived_neural_cells.bed ...

  14. File list: Pol.PSC.10.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.PSC.10.AllAg.mESC_derived_neural_cells mm9 RNA polymerase Pluripotent stem cell mESC derived neural...RX213760,SRX213764 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.PSC.10.AllAg.mESC_derived_neural_cells.bed ...

  15. File list: DNS.PSC.10.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.PSC.10.AllAg.hESC_derived_neural_cells hg19 DNase-seq Pluripotent stem cell hESC derived neural... cells SRX121241,SRX134721 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.PSC.10.AllAg.hESC_derived_neural_cells.bed ...

  16. File list: ALL.PSC.50.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.50.AllAg.hESC_derived_neural_crests hg19 All antigens Pluripotent stem cell hESC derived neural...X1091539,SRX059364 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.PSC.50.AllAg.hESC_derived_neural_crests.bed ...

  17. Image-based quantification and mathematical modeling of spatial heterogeneity in ESC colonies.

    Science.gov (United States)

    Herberg, Maria; Zerjatke, Thomas; de Back, Walter; Glauche, Ingmar; Roeder, Ingo

    2015-06-01

    Pluripotent embryonic stem cells (ESCs) have the potential to differentiate into cells of all three germ layers. This unique property has been extensively studied on the intracellular, transcriptional level. However, ESCs typically form clusters of cells with distinct size and shape, and establish spatial structures that are vital for the maintenance of pluripotency. Even though it is recognized that the cells' arrangement and local interactions play a role in fate decision processes, the relations between transcriptional and spatial patterns have not yet been studied. We present a systems biology approach which combines live-cell imaging, quantitative image analysis, and multiscale, mathematical modeling of ESC growth. In particular, we develop quantitative measures of the morphology and of the spatial clustering of ESCs with different expression levels and apply them to images of both in vitro and in silico cultures. Using the same measures, we are able to compare model scenarios with different assumptions on cell-cell adhesions and intercellular feedback mechanisms directly with experimental data. Applying our methodology to microscopy images of cultured ESCs, we demonstrate that the emerging colonies are highly variable regarding both morphological and spatial fluorescence patterns. Moreover, we can show that most ESC colonies contain only one cluster of cells with high self-renewing capacity. These cells are preferentially located in the interior of a colony structure. The integrated approach combining image analysis with mathematical modeling allows us to reveal potential transcription factor related cellular and intercellular mechanisms behind the emergence of observed patterns that cannot be derived from images directly. © 2015 International Society for Advancement of Cytometry.

  18. Epigenetic status of H19/IGF2 and SNRPN imprinted genes in aborted and successfully derived embryonic stem cell lines in non-human primates

    Directory of Open Access Journals (Sweden)

    Florence Wianny

    2016-05-01

    Full Text Available The imprinted genes of primate embryonic stem cells (ESCs often show altered DNA methylation. It is unknown whether these alterations emerge while deriving the ESCs. Here we studied the methylation patterns of two differentially methylated regions (DMRs, SNRPN and H19/IGF2 DMRs, during the derivation of monkey ESCs. We show that the SNRPN DMR is characteristically methylated at maternal alleles, whereas the H19/IGF2 DMR is globally highly methylated, with unusual methylation on the maternal alleles. These methylation patterns remain stable from the early stages of ESC derivation to late passages of monkey ESCs and following differentiation. Importantly, the methylation status of H19/IGF2 DMR and the expression levels of IGF2, H19, and DNMT3B mRNAs in early embryo-derived cells were correlated with their capacity to generate genuine ESC lines. Thus, we propose that these markers could be useful to predict the outcomes of establishing an ESC line in primates.

  19. File list: ALL.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.05.AllAg.hESC_derived_neural_cells hg19 All antigens Pluripotent stem cell hESC derived neural...RX729682,SRX729698,SRX729709 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  20. File list: His.PSC.10.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.10.AllAg.hESC_derived_neural_crests hg19 Histone Pluripotent stem cell hESC derived neural...3,SRX1091531,SRX059364,SRX1091530 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.10.AllAg.hESC_derived_neural_crests.bed ...

  1. File list: Oth.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.AllAg.mESC_derived_neural_cells mm9 TFs and others Pluripotent stem cell mESC derived neural...10563,SRX213763,SRX352996 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  2. File list: His.PSC.20.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.20.AllAg.hESC_derived_neural_crests hg19 Histone Pluripotent stem cell hESC derived neural...30,SRX059362,SRX1091539,SRX059364 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.20.AllAg.hESC_derived_neural_crests.bed ...

  3. File list: His.PSC.50.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.AllAg.hESC_derived_neural_crests hg19 Histone Pluripotent stem cell hESC derived neural...30,SRX059362,SRX1091539,SRX059364 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.50.AllAg.hESC_derived_neural_crests.bed ...

  4. File list: Oth.PSC.50.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.50.AllAg.mESC_derived_neural_cells mm9 TFs and others Pluripotent stem cell mESC derived neural...13762,SRX213759,SRX352995 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.50.AllAg.mESC_derived_neural_cells.bed ...

  5. Icaritin enhances mESC self-renewal through upregulating core pluripotency transcription factors mediated by ERα.

    Science.gov (United States)

    Tsang, Wing Pui; Zhang, Fengjie; He, Qiling; Cai, Waijiao; Huang, Jianhua; Chan, Wai Yee; Shen, Ziyin; Wan, Chao

    2017-01-16

    Utilization of small molecules in modulation of stem cell self-renewal is a promising approach to expand stem cells for regenerative therapy. Here, we identify Icaritin, a phytoestrogen molecule enhances self-renewal of mouse embryonic stem cells (mESCs). Icaritin increases mESCs proliferation while maintains their self-renewal capacity in vitro and pluripotency in vivo. This coincides with upregulation of key pluripotency transcription factors OCT4, NANOG, KLF4 and SOX2. The enhancement of mESCs self-renewal is characterized by increased population in S-phase of cell cycle, elevation of Cylin E and Cyclin-dependent kinase 2 (CDK2) and downregulation of p21, p27 and p57. PCR array screening reveals that caudal-related homeobox 2 (Cdx2) and Rbl2/p130 are remarkably suppressed in mESCs treated with Icaritin. siRNA knockdown of Cdx2 or Rbl2/p130 upregulates the expression of Cyclin E, OCT4 and SOX2, and subsequently increases cell proliferation and colony forming efficiency of mESCs. We then demonstrate that Icaritin co-localizes with estrogen receptor alpha (ERα) and activates its nuclear translocation in mESCs. The promotive effect of Icaritin on cell cycle and pluripotency regulators are eliminated by siRNA knockdown of ERα in mESCs. The results suggest that Icaritin enhances mESCs self-renewal by regulating cell cycle machinery and core pluripotency transcription factors mediated by ERα.

  6. File list: Oth.PSC.05.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.AllAg.hESC_derived_neural_crests hg19 TFs and others Pluripotent stem cell hESC derived neural...X1091546,SRX1091550,SRX059360,SRX059368,SRX059367 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.PSC.05.AllAg.hESC_derived_neural_crests.bed ...

  7. File list: Oth.PSC.10.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  8. File list: Oth.PSC.50.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.50.AllAg.hESC_derived_neural_crests hg19 TFs and others Pluripotent stem cell hESC derived neural...X1091550,SRX059360,SRX1091547,SRX059367,SRX059368 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.PSC.50.AllAg.hESC_derived_neural_crests.bed ...

  9. File list: His.PSC.20.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. File list: ALL.PSC.50.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.50.AllAg.mESC_derived_neural_cells mm9 All antigens Pluripotent stem cell mESC derived neural...3762,SRX213759,SRX276676,SRX018672,SRX352995,SRX276677,SRX298194,SRX213757 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.50.AllAg.mESC_derived_neural_cells.bed ...

  11. File list: His.PSC.50.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.AllAg.hESC_derived_neural_cells hg19 Histone Pluripotent stem cell hESC derived neural...692,SRX729710,SRX729684,SRX729689,SRX729699,SRX729704,SRX729694 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.50.AllAg.hESC_derived_neural_cells.bed ...

  12. File list: ALL.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.05.AllAg.mESC_derived_neural_cells mm9 All antigens Pluripotent stem cell mESC derived neural...8051,SRX810565,SRX810562,SRX810563,SRX810564,SRX213763,SRX352996,SRX604259 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  13. File list: His.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.05.AllAg.hESC_derived_neural_cells hg19 Histone Pluripotent stem cell hESC derived neural...707,SRX027494,SRX698181,SRX729681,SRX729682,SRX729698,SRX729709 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  14. File list: ALL.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.PSC.20.AllAg.mESC_derived_neural_cells mm9 All antigens Pluripotent stem cell mESC derived neural...3762,SRX352996,SRX604259,SRX213763,SRX213761,SRX213764,SRX213758,SRX213757 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  15. File list: ALL.PSC.10.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: NoD.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.05.AllAg.hESC_derived_neural_cells hg19 No description Pluripotent stem cell hESC derived neural...edbc.jp/kyushu-u/hg19/assembled/NoD.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  17. File list: NoD.PSC.50.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.50.AllAg.hESC_derived_neural_cells hg19 No description Pluripotent stem cell hESC derived neural...edbc.jp/kyushu-u/hg19/assembled/NoD.PSC.50.AllAg.hESC_derived_neural_cells.bed ...

  18. File list: NoD.PSC.20.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.20.AllAg.hESC_derived_neural_cells hg19 No description Pluripotent stem cell hESC derived neural...edbc.jp/kyushu-u/hg19/assembled/NoD.PSC.20.AllAg.hESC_derived_neural_cells.bed ...

  19. File list: NoD.PSC.10.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  20. File list: NoD.PSC.10.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.10.AllAg.hESC_derived_neural_crests hg19 No description Pluripotent stem cell hESC derived neural... crests http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.PSC.10.AllAg.hESC_derived_neural_crests.bed ...

  1. File list: NoD.PSC.20.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.20.AllAg.hESC_derived_neural_crests hg19 No description Pluripotent stem cell hESC derived neural... crests http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.PSC.20.AllAg.hESC_derived_neural_crests.bed ...

  2. Establishing clonal cell lines with endothelial-like potential from CD9(hi, SSEA-1(- cells in embryonic stem cell-derived embryoid bodies.

    Directory of Open Access Journals (Sweden)

    Qizhou Lian

    Full Text Available BACKGROUND: Differentiation of embryonic stem cells (ESCs into specific cell types with minimal risk of teratoma formation could be efficiently directed by first reducing the differentiation potential of ESCs through the generation of clonal, self-renewing lineage-restricted stem cell lines. Efforts to isolate these stem cells are, however, mired in an impasse where the lack of purified lineage-restricted stem cells has hindered the identification of defining markers for these rare stem cells and, in turn, their isolation. METHODOLOGY/PRINCIPAL FINDINGS: We describe here a method for the isolation of clonal lineage-restricted cell lines with endothelial potential from ESCs through a combination of empirical and rational evidence-based methods. Using an empirical protocol that we have previously developed to generate embryo-derived RoSH lines with endothelial potential, we first generated E-RoSH lines from mouse ESC-derived embryoid bodies (EBs. Despite originating from different mouse strains, RoSH and E- RoSH lines have similar gene expression profiles (r(2 = 0.93 while that between E-RoSH and ESCs was 0.83. In silico gene expression analysis predicted that like RoSH cells, E-RoSH cells have an increased propensity to differentiate into vasculature. Unlike their parental ESCs, E-RoSH cells did not form teratomas and differentiate efficiently into endothelial-like cells in vivo and in vitro. Gene expression and FACS analysis revealed that RoSH and E-RoSH cells are CD9(hi, SSEA-1(- while ESCs are CD9(lo, SSEA-1(+. Isolation of CD9(hi, SSEA-1(- cells that constituted 1%-10% of EB-derived cultures generated an E-RoSH-like culture with an identical E-RoSH-like gene expression profile (r(2 = 0.95 and a propensity to differentiate into endothelial-like cells. CONCLUSIONS: By combining empirical and rational evidence-based methods, we identified definitive selectable surface antigens for the isolation and propagation of lineage-restricted stem cells

  3. The effect of cephalosporin usage on the occurrence of ESCs producing E. coli in pig herds

    DEFF Research Database (Denmark)

    Dalhoff Andersen, Vibe; Jensen, Vibeke Frøkjær; Agersø, Yvonne

    in 19 pig herds which have had five to fourteen prescriptions of ceph. and 20 pig herds without prescribed ceph. in a previous 12 month period. The 39 herds were all integrated and represent typical Danish pig farms. The occurrence of ESCs producing E. coli in the herds were tested in a total of 9...... pooled samples per herd. A pig herd was considered positive if one or more of the nine samples contained ESCs producing E. coli. Initially, the association between usages of ceph. and occurrence of ESCs producing E. coli in the pig herds was analyzed using logistic regression, and the effect was adjusted...... of ESCs producing E. coli was estimated as risk ratio(RR). The results showed that consumption of ceph. increased the risk of occurrence of ESCs producing E. coli significantly with a RR of 5 (95% CI: 2-11). This demonstrates that ceph. usage significant affect the occurrence of ESCs resistance...

  4. File list: NoD.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  5. File list: NoD.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  6. File list: NoD.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  7. File list: NoD.PSC.10.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.10.AllAg.mESC_derived_neural_cells mm9 No description Pluripotent stem cell mESC derived neural... cells SRX440736,SRX440731 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.PSC.10.AllAg.mESC_derived_neural_cells.bed ...

  8. File list: NoD.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.05.AllAg.mESC_derived_neural_cells mm9 No description Pluripotent stem cell mESC derived neural... cells SRX440731,SRX440736 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  9. File list: NoD.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.PSC.20.AllAg.mESC_derived_neural_cells mm9 No description Pluripotent stem cell mESC derived neural... cells SRX440731,SRX440736 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.PSC.20.AllAg.mESC_derived_neural_cells.bed ...

  10. File list: InP.PSC.50.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  11. File list: InP.PSC.20.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: InP.PSC.10.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.10.AllAg.mESC_derived_pancreatic_cells mm9 Input control Pluripotent stem cell mESC derived panc...reatic cells SRX146010,SRX146009 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.10.AllAg.mESC_derived_pancreatic_cells.bed ...

  13. File list: InP.PSC.05.AllAg.mESC_derived_pancreatic_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  14. File list: InP.PSC.05.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.05.AllAg.hESC_derived_neural_crests hg19 Input control Pluripotent stem cell hESC derived neural... crests SRX1091573,SRX059369,SRX059361 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.PSC.05.AllAg.hESC_derived_neural_crests.bed ...

  15. File list: InP.PSC.50.AllAg.hESC_derived_neural_crests [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.50.AllAg.hESC_derived_neural_crests hg19 Input control Pluripotent stem cell hESC derived neural... crests SRX1091573,SRX059369,SRX059361 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.PSC.50.AllAg.hESC_derived_neural_crests.bed ...

  16. File list: InP.PSC.05.AllAg.hESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.05.AllAg.hESC_derived_neural_cells hg19 Input control Pluripotent stem cell hESC derived neural...RX698183,SRX729711,SRX729701 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.PSC.05.AllAg.hESC_derived_neural_cells.bed ...

  17. Heterogeneity of osteosarcoma cell lines led to variable responses in reprogramming.

    Science.gov (United States)

    Choong, Pei Feng; Teh, Hui Xin; Teoh, Hoon Koon; Ong, Han Kiat; Choo, Kong Bung; Sugii, Shigeki; Cheong, Soon Keng; Kamarul, Tunku

    2014-01-01

    Four osteosarcoma cell lines, Saos-2, MG-63, G-292 and U-2 OS, were reprogrammed to pluripotent state using Yamanaka factors retroviral transduction method. Embryonic stem cell (ESC)-like clusters started to appear between 15 to 20 days post transduction. Morphology of the colonies resembled that of ESC colonies with defined border and tightly-packed cells. The reprogrammed sarcomas expressed alkaline phosphatase and pluripotency markers, OCT4, SSEA4, TRA-1-60 and TRA-1-81, as in ESC up to Passage 15. All reprogrammed sarcomas could form embryoid body-like spheres when cultured in suspension in a low attachment dish for up to 10 days. Further testing on the directed differentiation capacity of the reprogrammed sarcomas showed all four reprogrammed sarcoma lines could differentiate into adipocytes while reprogrammed Saos-2-REP, MG-63-REP and G-292-REP could differentiate into osteocytes. Among the 4 osteosarcoma cell lines, U-2 OS reported the highest transduction efficiency but recorded the lowest reprogramming stability under long term culture. Thus, there may be intrinsic differences governing the variable responses of osteosarcoma cell lines towards reprogramming and long term culture effect of the reprogrammed cells. This is a first report to associate intrinsic factors in different osteosarcoma cell lines with variable reprogramming responses and effects on the reprogrammed cells after prolonged culture.

  18. Heterogeneity of Osteosarcoma Cell Lines Led to Variable Responses in Reprogramming

    Science.gov (United States)

    Choong, Pei Feng; Teh, Hui Xin; Teoh, Hoon Koon; Ong, Han Kiat; Choo, Kong Bung; Sugii, Shigeki; Cheong, Soon Keng; Kamarul, Tunku

    2014-01-01

    Four osteosarcoma cell lines, Saos-2, MG-63, G-292 and U-2 OS, were reprogrammed to pluripotent state using Yamanaka factors retroviral transduction method. Embryonic stem cell (ESC)-like clusters started to appear between 15 to 20 days post transduction. Morphology of the colonies resembled that of ESC colonies with defined border and tightly-packed cells. The reprogrammed sarcomas expressed alkaline phosphatase and pluripotency markers, OCT4, SSEA4, TRA-1-60 and TRA-1-81, as in ESC up to Passage 15. All reprogrammed sarcomas could form embryoid body-like spheres when cultured in suspension in a low attachment dish for up to 10 days. Further testing on the directed differentiation capacity of the reprogrammed sarcomas showed all four reprogrammed sarcoma lines could differentiate into adipocytes while reprogrammed Saos-2-REP, MG-63-REP and G-292-REP could differentiate into osteocytes. Among the 4 osteosarcoma cell lines, U-2 OS reported the highest transduction efficiency but recorded the lowest reprogramming stability under long term culture. Thus, there may be intrinsic differences governing the variable responses of osteosarcoma cell lines towards reprogramming and long term culture effect of the reprogrammed cells. This is a first report to associate intrinsic factors in different osteosarcoma cell lines with variable reprogramming responses and effects on the reprogrammed cells after prolonged culture. PMID:25170299

  19. International Earth Science Constellation (ESC) Introduction

    Science.gov (United States)

    Guit, William J.; Machado, Michael J.

    2016-01-01

    This is the Welcome and Introduction presentation for the International Earth Science Constellation (ESC) Mission Operations Working Group (MOWG) meeting held in Albuquerque NM from September 27-29. It contains an org chart, charter, history, significant topics to be discussed, AquaAura 2017 inclination adjust maneuver calendar, a-train long range plans, upcoming events, and action items.

  20. File list: InP.PSC.20.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  1. File list: InP.PSC.50.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.50.AllAg.mESC_derived_neural_cells mm9 Input control Pluripotent stem cell mESC derived neural...65,SRX1214070,SRX810564,SRX101693,SRX604259,SRX018672,SRX276677 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.50.AllAg.mESC_derived_neural_cells.bed ...

  2. File list: InP.PSC.05.AllAg.mESC_derived_neural_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.PSC.05.AllAg.mESC_derived_neural_cells mm9 Input control Pluripotent stem cell mESC derived neural...4,SRX1214071,SRX1214070,SRX518051,SRX810565,SRX810564,SRX604259 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.05.AllAg.mESC_derived_neural_cells.bed ...

  3. Extracellular matrix aggregates from differentiating embryoid bodies as a scaffold to support ESC proliferation and differentiation.

    Directory of Open Access Journals (Sweden)

    Saik-Kia Goh

    Full Text Available Embryonic stem cells (ESCs have emerged as potential cell sources for tissue engineering and regeneration owing to its virtually unlimited replicative capacity and the potential to differentiate into a variety of cell types. Current differentiation strategies primarily involve various growth factor/inducer/repressor concoctions with less emphasis on the substrate. Developing biomaterials to promote stem cell proliferation and differentiation could aid in the realization of this goal. Extracellular matrix (ECM components are important physiological regulators, and can provide cues to direct ESC expansion and differentiation. ECM undergoes constant remodeling with surrounding cells to accommodate specific developmental event. In this study, using ESC derived aggregates called embryoid bodies (EB as a model, we characterized the biological nature of ECM in EB after exposure to different treatments: spontaneously differentiated and retinoic acid treated (denoted as SPT and RA, respectively. Next, we extracted this treatment-specific ECM by detergent decellularization methods (Triton X-100, DOC and SDS are compared. The resulting EB ECM scaffolds were seeded with undifferentiated ESCs using a novel cell seeding strategy, and the behavior of ESCs was studied. Our results showed that the optimized protocol efficiently removes cells while retaining crucial ECM and biochemical components. Decellularized ECM from SPT EB gave rise to a more favorable microenvironment for promoting ESC attachment, proliferation, and early differentiation, compared to native EB and decellularized ECM from RA EB. These findings suggest that various treatment conditions allow the formulation of unique ESC-ECM derived scaffolds to enhance ESC bioactivities, including proliferation and differentiation for tissue regeneration applications.

  4. SUV rollover in single vehicle crashes and the influence of ESC and SSF.

    Science.gov (United States)

    Kallan, Michael J; Jermakian, Jessica Steps

    2008-10-01

    The modern Sport Utility Vehicle (SUV) fleet continues to go through a transformation in response to the concern that they are at an increased risk of rollover. Our research objective was to look at changes in rollover rates for single vehicle crashes in the modern SUV fleet (corresponding to NCAP rollover testing model years) and the impact of electronic stability control (ESC) and lowered center of gravity. We looked at 2001-2006 NASS-GES data on a probability sample of 3,331 SUVs involved in single vehicle crashes, weighted to represent 324,149 crashes in the study population. Static Stability Factor (SSF) information from NCAP testing and ESC presence (from IIHS) were also incorporated. 20.2% of these SUVs were involved a rollover, which decreased by more than half from model year 2001 (25.3%) through 2006 (11.5%). Nearly 9% had ESC as a standard feature, including 47% in model year 2006. The majority of the late model year decline in rollover rates can be attributed to ESC presence and higher SSF. Rollover was two-thirds less likely (adjusted OR=0.33, 95% CI=0.20-0.55) in SUVs with ESC as a standard feature versus those known not to have ESC. Those SUVs with SSF > or = 1.20 were significantly less likely to rollover (adjusted OR=0.31, 95% CI=0.20-0.48). Additional significant predictors of rollover included SUV size, driver age and alcohol use. Our study builds on the previous work of NHTSA, IIHS, and others with regard to rollover risk by looking at an even wider array of late model year SUVs.

  5. ELABELA Is an Endogenous Growth Factor that Sustains hESC Self-Renewal via the PI3K/AKT Pathway.

    Science.gov (United States)

    Ho, Lena; Tan, Shawn Y X; Wee, Sheena; Wu, Yixuan; Tan, Sam J C; Ramakrishna, Navin B; Chng, Serene C; Nama, Srikanth; Szczerbinska, Iwona; Sczerbinska, Iwona; Chan, Yun-Shen; Avery, Stuart; Tsuneyoshi, Norihiro; Ng, Huck Hui; Gunaratne, Jayantha; Dunn, N Ray; Reversade, Bruno

    2015-10-01

    ELABELA (ELA) is a peptide hormone required for heart development that signals via the Apelin Receptor (APLNR, APJ). ELA is also abundantly secreted by human embryonic stem cells (hESCs), which do not express APLNR. Here we show that ELA signals in a paracrine fashion in hESCs to maintain self-renewal. ELA inhibition by CRISPR/Cas9-mediated deletion, shRNA, or neutralizing antibodies causes reduced hESC growth, cell death, and loss of pluripotency. Global phosphoproteomic and transcriptomic analyses of ELA-pulsed hESCs show that it activates PI3K/AKT/mTORC1 signaling required for cell survival. ELA promotes hESC cell-cycle progression and protein translation and blocks stress-induced apoptosis. INSULIN and ELA have partially overlapping functions in hESC medium, but only ELA can potentiate the TGFβ pathway to prime hESCs toward the endoderm lineage. We propose that ELA, acting through an alternate cell-surface receptor, is an endogenous secreted growth factor in human embryos and hESCs that promotes growth and pluripotency. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Derivation of mouse embryonic stem cell lines from tyrosine hydroxylase reporter mice crossed with a human SNCA transgenic mouse model of Parkinson's disease

    Directory of Open Access Journals (Sweden)

    Margarita Chumarina

    2017-03-01

    Full Text Available Mouse embryonic stem cell (mESC lines were derived by crossing heterozygous transgenic (tg mice expressing green fluorescent protein (GFP under the control of the rat tyrosine hydroxylase (TH promoter, with homozygous alpha-synuclein (aSYN mice expressing human mutant SNCAA53T under the control of the mouse Prion promoter (MoPrP, or wildtype (WT mice. The expression of GFP and human aSYN was validated by immunocytochemistry in midbrain neuron cultures upon differentiation of mESC lines using stromal cell-derived inducing activity. These mESC lines can help to study the impact of human aSYN expression in neurons and oligodendrocytes, and also trace GFP-expressing midbrain neurons.

  7. Characterization of a novel telomerase-immortalized human endometrial stromal cell line, St-T1b

    Directory of Open Access Journals (Sweden)

    Brosens Jan J

    2009-07-01

    Full Text Available Abstract Background Coordinated differentiation of the endometrial compartments in the second half of the menstrual cycle is a prerequisite for the establishment of pregnancy. Endometrial stromal cells (ESC decidualize under the influence of ovarian progesterone to accommodate implantation of the blastocyst and support establishment of the placenta. Studies into the mechanisms of decidualization are often hampered by the lack of primary ESC. Here we describe a novel immortalized human ESC line. Methods Primary ESC were immortalized by the transduction of telomerase. The resultant cell line, termed St-T1b, was characterized for its morphological and biochemical properties by immunocytochemistry, RT-PCR and immunoblotting. Its progestational response was tested using progesterone and medroxyprogesterone acetate with and without 8-Br-cAMP, an established inducer of decidualization in vitro. Results St-T1b were positive for the fibroblast markers vimentin and CD90 and negative for the epithelial marker cytokeratin-7. They acquired a decidual phenotype indistinguishable from primary ESC in response to cAMP stimulation. The decidual response was characterized by transcriptional activation of marker genes, such as PRL, IGFBP1, and FOXO1, and enhanced protein levels of the tumor suppressor p53 and the metastasis suppressor KAI1 (CD82. Progestins alone had no effect on St-T1b cells, but medroxyprogesterone acetate greatly enhanced the cAMP-stimulated expression of IGFBP-1 after 3 and 7 days. Progesterone, albeit more weakly, also augmented the cAMP-induced IGFBP-1 production but only after 7 days of treatment. The cell line remained stable in continuous culture for more than 150 passages. Conclusion St-T1b express the appropriate phenotypic ESC markers and their decidual response closely mimics that of primary cultures. Decidualization is efficiently induced by cAMP analog and enhanced by medroxyprogesterone acetate, and, to a lesser extent, by natural

  8. Label-free separation of human embryonic stem cells (hESCs) and their cardiac derivatives using Raman spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chan, J W; Lieu, D K; Huser, T R; Li, R A

    2008-09-08

    Self-renewable, pluripotent human embryonic stem cells (hESCs) can be differentiated into cardiomyocytes (CMs), providing an unlimited source of cells for transplantation therapies. However, unlike certain cell lineages such as hematopoietic cells, CMs lack specific surface markers for convenient identification, physical separation, and enrichment. Identification by immunostaining of cardiac-specific proteins such as troponin requires permeabilization, which renders the cells unviable and non-recoverable. Ectopic expression of a reporter protein under the transcriptional control of a heart-specific promoter for identifying hESC-derived CMs (hESC-CMs) is useful for research but complicates potential clinical applications. The practical detection and removal of undifferentiated hESCs in a graft, which may lead to tumors, is also critical. Here, we demonstrate a non-destructive, label-free optical method based on Raman scattering to interrogate the intrinsic biochemical signatures of individual hESCs and their cardiac derivatives, allowing cells to be identified and classified. By combining the Raman spectroscopic data with multivariate statistical analysis, our results indicate that hESCs, human fetal left ventricular CMs, and hESC-CMs can be identified by their intrinsic biochemical characteristics with an accuracy of 96%, 98% and 66%, respectively. The present study lays the groundwork for developing a systematic and automated method for the non-invasive and label-free sorting of (i) high-quality hESCs for expansion, and (ii) ex vivo CMs (derived from embryonic or adult stem cells) for cell-based heart therapies.

  9. PHB Associates with the HIRA Complex to Control an Epigenetic-Metabolic Circuit in Human ESCs.

    Science.gov (United States)

    Zhu, Zhexin; Li, Chunliang; Zeng, Yanwu; Ding, Jianyi; Qu, Zepeng; Gu, Junjie; Ge, Laixiang; Tang, Fan; Huang, Xin; Zhou, Chenlin; Wang, Ping; Zheng, Deyou; Jin, Ying

    2017-02-02

    The chromatin landscape and cellular metabolism both contribute to cell fate determination, but their interplay remains poorly understood. Using genome-wide siRNA screening, we have identified prohibitin (PHB) as an essential factor in self-renewal of human embryonic stem cells (hESCs). Mechanistically, PHB forms protein complexes with HIRA, a histone H3.3 chaperone, and stabilizes the protein levels of HIRA complex components. Like PHB, HIRA is required for hESC self-renewal. PHB and HIRA act together to control global deposition of histone H3.3 and gene expression in hESCs. Of particular note, PHB and HIRA regulate the chromatin architecture at the promoters of isocitrate dehydrogenase genes to promote transcription and, thus, production of α-ketoglutarate, a key metabolite in the regulation of ESC fate. Our study shows that PHB has an unexpected nuclear role in hESCs that is required for self-renewal and that it acts with HIRA in chromatin organization to link epigenetic organization to a metabolic circuit. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. MicroRNA-302/367 Cluster Governs hESC Self-Renewal by Dually Regulating Cell Cycle and Apoptosis Pathways

    Directory of Open Access Journals (Sweden)

    Zhonghui Zhang

    2015-04-01

    Full Text Available miR-302/367 is the most abundant miRNA cluster in human embryonic stem cells (hESCs and can promote somatic cell reprogramming. However, its role in hESCs remains poorly understood. Here, we studied functional roles of the endogenous miR-302/367 cluster in hESCs by employing specific TALE-based transcriptional repressors. We revealed that miR-302/367 cluster dually regulates hESC cell cycle and apoptosis in dose-dependent manner. Gene profiling and functional studies identified key targets of the miR-302/367 cluster in regulating hESC self-renewal and apoptosis. We demonstrate that in addition to its role in cell cycle regulation, miR-302/367 cluster conquers apoptosis by downregulating BNIP3L/Nix (a BH3-only proapoptotic factor and upregulating BCL-xL expression. Furthermore, we show that butyrate, a natural compound, upregulates miR-302/367 cluster expression and alleviates hESCs from apoptosis induced by knockdown of miR-302/367 cluster. In summary, our findings provide new insights in molecular mechanisms of how miR-302/367 cluster regulates hESCs.

  11. ESC-Derived Basal Forebrain Cholinergic Neurons Ameliorate the Cognitive Symptoms Associated with Alzheimer’s Disease in Mouse Models

    Directory of Open Access Journals (Sweden)

    Wei Yue

    2015-11-01

    Full Text Available Degeneration of basal forebrain cholinergic neurons (BFCNs is associated with cognitive impairments of Alzheimer’s disease (AD, implying that BFCNs hold potentials in exploring stem cell-based replacement therapy for AD. However, studies on derivation of BFCNs from embryonic stem cells (ESCs are limited, and the application of ESC-derived BFCNs remains to be determined. Here, we report on differentiation approaches for directing both mouse and human ESCs into mature BFCNs. These ESC-derived BFCNs exhibit features similar to those of their in vivo counterparts and acquire appropriate functional properties. After transplantation into the basal forebrain of AD model mice, ESC-derived BFCN progenitors predominantly differentiate into mature cholinergic neurons that functionally integrate into the endogenous basal forebrain cholinergic projection system. The AD mice grafted with mouse or human BFCNs exhibit improvements in learning and memory performances. Our findings suggest a promising perspective of ESC-derived BFCNs in the development of stem cell-based therapies for treatment of AD.

  12. Generation of a human embryonic stem cell line, NERCe003-A-1, with lentivirus vector-mediated inducible CTNNB1 overexpression

    Directory of Open Access Journals (Sweden)

    Yang Wang

    2018-04-01

    Full Text Available The human embryonic stem cell (hESC line NERCe003-A-1 was generated by introducing lentiviral-vector–mediated tetracycline-inducible β-catenin expression into a normal hESC line, NERCe003-A. The resulting cell line can overexpress the β-catenin protein, encoded by the CTNNB1 gene, after exposure to doxycycline (Dox. CTNNB1 gene expression was confirmed by quantitative PCR (qPCR and immunofluorescence assays. Further characterization confirmed that the NERCe003-A-1 cell line expresses typical pluripotency markers and has the ability to form the three germ layers both in vitro and in vivo.

  13. Interspecies chimera between primate embryonic stem cells and mouse embryos: monkey ESCs engraft into mouse embryos, but not post-implantation fetuses.

    Science.gov (United States)

    Simerly, Calvin; McFarland, Dave; Castro, Carlos; Lin, Chih-Cheng; Redinger, Carrie; Jacoby, Ethan; Mich-Basso, Jocelyn; Orwig, Kyle; Mills, Parker; Ahrens, Eric; Navara, Chris; Schatten, Gerald

    2011-07-01

    Unequivocal evidence for pluripotency in which embryonic stem cells contribute to chimeric offspring has yet to be demonstrated in human or nonhuman primates (NHPs). Here, rhesus and baboons ESCs were investigated in interspecific mouse chimera generated by aggregation or blastocyst injection. Aggregation chimera produced mouse blastocysts with GFP-nhpESCs at the inner cell mass (ICM), and embryo transfers (ETs) generated dimly-fluorescencing abnormal fetuses. Direct injection of GFP-nhpESCs into blastocysts produced normal non-GFP-fluorescencing fetuses. Injected chimera showed >70% loss of GFP-nhpESCs after 21 h culture. Outgrowths of all chimeric blastocysts established distinct but separate mouse- and NHP-ESC colonies. Extensive endogenous autofluorescence compromised anti-GFP detection and PCR analysis did not detect nhpESCs in fetuses. NhpESCs localize to the ICM in chimera and generate pregnancies. Because primate ESCs do not engraft post-implantation, and also because endogenous autofluorescence results in misleading positive signals, interspecific chimera assays for pluripotency with primate stem cells is unreliable with the currently available ESCs. Testing primate ESCs reprogrammed into even more naïve states in these inter-specific chimera assays will be an important future endeavor. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Relación entre ansiedad escénica, perfeccionismo y calificaciones en estudiantes del Título Superior de Música

    Directory of Open Access Journals (Sweden)

    Francisco Javier Zarza-Alzugaray

    2016-02-01

    Full Text Available La ansiedad escénica es uno de los principales problemas a los que los músicos, estudiantes y profesionales, deben enfrentarse. El perfeccionismo es un constructo asociado a problemas de fobia social como pueda ser la ansiedad escénica. La asociación entre perfeccionismo y ansiedad escénica no ha recibido especial atención dentro del ámbito investigador y formativo español. Objetivos: Estudiar la presencia de niveles de ansiedad escénica en una muestra de estudiantes del Conservatorio Superior de Música de Alicante y cómo se relaciona ésta con el perfeccionismo y las calificaciones obtenidas. Método: El estudio de los datos es cuantitativo mediante un diseño selectivo con un muestreo por disponibilidad. Las respuestas corresponden a dos instrumentos de medida específicos de la ansiedad escénica y del perfeccionismo por parte de 87 estudiantes del Conservatorio Superior de Música de Alicante. Resultados: Encontramos niveles de ansiedad escénica algo mayores a la media. La ansiedad escénica está relacionada significativamente con el perfeccionismo, y éste con las calificaciones del alumnado. Conclusiones: El perfeccionismo explica una parte tanto de la ansiedad escénica como de las calificaciones obtenidas por parte de los estudiantes. Es necesaria más investigación sobre la ansiedad escénica que suponga en un futuro implementar mejoras formativas.

  15. Fostering diffusion of scientific contents of National Society Cardiovascular Journals: The new ESC search engine

    Directory of Open Access Journals (Sweden)

    Fernando Alfonso

    2013-12-01

    Full Text Available European Society of Cardiology (ESC National Society Cardiovascular Journals (NSCJs are high-quality biomedical journals focused on cardiovascular diseases. The Editors’ Network of the ESC devises editorial initiatives aimed at improving the scientific quality and diffusion of NSCJ. In this article we will discuss on the importance of the Internet, electronic editions and open access strategies on scientific publishing. Finally, we will propose a new editorial initiative based on a novel electronic tool on the ESC web-page that may further help to increase the dissemination of contents and visibility of NSCJs.

  16. Lentiviral vectors and protocols for creation of stable hESC lines for fluorescent tracking and drug resistance selection of cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Hiroko Kita-Matsuo

    Full Text Available Developmental, physiological and tissue engineering studies critical to the development of successful myocardial regeneration therapies require new ways to effectively visualize and isolate large numbers of fluorescently labeled, functional cardiomyocytes.Here we describe methods for the clonal expansion of engineered hESCs and make available a suite of lentiviral vectors for that combine Blasticidin, Neomycin and Puromycin resistance based drug selection of pure populations of stem cells and cardiomyocytes with ubiquitous or lineage-specific promoters that direct expression of fluorescent proteins to visualize and track cardiomyocytes and their progenitors. The phospho-glycerate kinase (PGK promoter was used to ubiquitously direct expression of histone-2B fused eGFP and mCherry proteins to the nucleus to monitor DNA content and enable tracking of cell migration and lineage. Vectors with T/Brachyury and alpha-myosin heavy chain (alphaMHC promoters targeted fluorescent or drug-resistance proteins to early mesoderm and cardiomyocytes. The drug selection protocol yielded 96% pure cardiomyocytes that could be cultured for over 4 months. Puromycin-selected cardiomyocytes exhibited a gene expression profile similar to that of adult human cardiomyocytes and generated force and action potentials consistent with normal fetal cardiomyocytes, documenting these parameters in hESC-derived cardiomyocytes and validating that the selected cells retained normal differentiation and function.The protocols, vectors and gene expression data comprise tools to enhance cardiomyocyte production for large-scale applications.

  17. Extract of Stellerachamaejasme L(ESC) inhibits growth and metastasis of human hepatocellular carcinoma via regulating microRNA expression.

    Science.gov (United States)

    Liu, Xiaoni; Wang, Shuang; Xu, Jianji; Kou, Buxin; Chen, Dexi; Wang, Yajie; Zhu, Xiaoxin

    2018-03-20

    MicroRNAs(miRNAs)are involved in the initiation and progression of hepatocellular carcinoma. ESC, an extract of Stellerachamaejasme L, had been confirmed as a potential anti-tumor extract of Traditional Chinese Medicine. In light of the important role of miRNAs in hepatocellular carcinoma, we questioned whether the inhibitory effects of ESC on hepatocellular carcinoma (HCC) were associated with miRNAs. The proliferation inhibition of ESC on HCC cells was measured with MTT assay. The migration inhibition of ESC on HCC cells was measured with transwell assay. The influences of ESC on growth and metastasis inhibition were evaluated with xenograft tumor model of HCC. Protein expressions were measured with western blot and immunofluorescence methods and miRNA profiles were detected with miRNA array. Differential miRNA and target mRNAs were verified with real-time PCR. The results showed that ESC could inhibit proliferation and epithelial mesenchymal transition (EMT) in HCC cells in vitro and tumor growth and metastasis in xenograft models in vivo. miRNA array results showed that 69 differential miRNAs in total of 429 ones were obtained in MHCC97H cells treated by ESC. hsa-miR-107, hsa-miR-638, hsa-miR-106b-5p were selected to be validated with real-time PCR method in HepG2 and MHCC97H cells. Expressions of hsa-miR-107 and hsa-miR-638 increased obviously in HCC cells treated by ESC. Target genes of three miRNAs were also validated with real-time PCR. Interestingly, only target genes of hsa-miR-107 changed greatly. ESC downregulated the MCL1, SALL4 and BCL2 gene expressions significantly but did not influence the expression of CACNA2D1. The findings suggested ESC regressed growth and metastasis of human hepatocellular carcinoma via regulating microRNAs expression and their corresponding target genes.

  18. Comparing ESC and iPSC—Based Models for Human Genetic Disorders

    Directory of Open Access Journals (Sweden)

    Tomer Halevy

    2014-10-01

    Full Text Available Traditionally, human disorders were studied using animal models or somatic cells taken from patients. Such studies enabled the analysis of the molecular mechanisms of numerous disorders, and led to the discovery of new treatments. Yet, these systems are limited or even irrelevant in modeling multiple genetic diseases. The isolation of human embryonic stem cells (ESCs from diseased blastocysts, the derivation of induced pluripotent stem cells (iPSCs from patients’ somatic cells, and the new technologies for genome editing of pluripotent stem cells have opened a new window of opportunities in the field of disease modeling, and enabled studying diseases that couldn’t be modeled in the past. Importantly, despite the high similarity between ESCs and iPSCs, there are several fundamental differences between these cells, which have important implications regarding disease modeling. In this review we compare ESC-based models to iPSC-based models, and highlight the advantages and disadvantages of each system. We further suggest a roadmap for how to choose the optimal strategy to model each specific disorder.

  19. Comparing ESC and iPSC-Based Models for Human Genetic Disorders.

    Science.gov (United States)

    Halevy, Tomer; Urbach, Achia

    2014-10-24

    Traditionally, human disorders were studied using animal models or somatic cells taken from patients. Such studies enabled the analysis of the molecular mechanisms of numerous disorders, and led to the discovery of new treatments. Yet, these systems are limited or even irrelevant in modeling multiple genetic diseases. The isolation of human embryonic stem cells (ESCs) from diseased blastocysts, the derivation of induced pluripotent stem cells (iPSCs) from patients' somatic cells, and the new technologies for genome editing of pluripotent stem cells have opened a new window of opportunities in the field of disease modeling, and enabled studying diseases that couldn't be modeled in the past. Importantly, despite the high similarity between ESCs and iPSCs, there are several fundamental differences between these cells, which have important implications regarding disease modeling. In this review we compare ESC-based models to iPSC-based models, and highlight the advantages and disadvantages of each system. We further suggest a roadmap for how to choose the optimal strategy to model each specific disorder.

  20. Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs

    Directory of Open Access Journals (Sweden)

    Choo Andre

    2011-04-01

    Full Text Available Abstract Background Exosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes. Methods The hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining. Results MYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved. Conclusion Our results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time

  1. Publishing SNP genotypes of human embryonic stem cell lines: policy statement of the International Stem Cell Forum Ethics Working Party.

    Science.gov (United States)

    Knoppers, Bartha M; Isasi, Rosario; Benvenisty, Nissim; Kim, Ock-Joo; Lomax, Geoffrey; Morris, Clive; Murray, Thomas H; Lee, Eng Hin; Perry, Margery; Richardson, Genevra; Sipp, Douglas; Tanner, Klaus; Wahlström, Jan; de Wert, Guido; Zeng, Fanyi

    2011-09-01

    Novel methods and associated tools permitting individual identification in publicly accessible SNP databases have become a debatable issue. There is growing concern that current technical and ethical safeguards to protect the identities of donors could be insufficient. In the context of human embryonic stem cell research, there are no studies focusing on the probability that an hESC line donor could be identified by analyzing published SNP profiles and associated genotypic and phenotypic information. We present the International Stem Cell Forum (ISCF) Ethics Working Party's Policy Statement on "Publishing SNP Genotypes of Human Embryonic Stem Cell Lines (hESC)". The Statement prospectively addresses issues surrounding the publication of genotypic data and associated annotations of hESC lines in open access databases. It proposes a balanced approach between the goals of open science and data sharing with the respect for fundamental bioethical principles (autonomy, privacy, beneficence, justice and research merit and integrity).

  2. Relación entre ansiedad escénica, perfeccionismo y calificaciones en estudiantes del Título Superior de Música

    OpenAIRE

    Francisco Javier Zarza-Alzugaray; Óscar Casanova-López; José Elías Robles-Rubio

    2016-01-01

    La ansiedad escénica es uno de los principales problemas a los que los músicos, estudiantes y profesionales, deben enfrentarse. El perfeccionismo es un constructo asociado a problemas de fobia social como pueda ser la ansiedad escénica. La asociación entre perfeccionismo y ansiedad escénica no ha recibido especial atención dentro del ámbito investigador y formativo español. Objetivos: Estudiar la presencia de niveles de ansiedad escénica en una muestra de estudiantes del Conservatorio Superio...

  3. Summary of 2017 ESC guidelines on valvular heart disease, peripheral artery disease, STEMI and on dual antiplatelet therapy.

    Science.gov (United States)

    Van Camp, Guy; De Backer, Tine; Beauloye, Christophe; Desmet, Walter; Claeys, Marc J

    2017-12-11

    During the ESC congress in September 2017 in Barcelona, the new ESC guidelines were presented and are now available on the ESC website. The new guidelines cover management recommendations on following cardiovascular items: valvular heart disease, peripheral artery disease, ST elevation myocardial infarction (STEMI) and on dual antiplatelet therapy. The present document gives a summary of these guidelines and highlights the most important recommendations and changes in the management of these diseases. It will help to increase awareness about the new guidelines and may stimulate to consult the full document for specific items. Ultimately, the authors hope that this document will enhance implementation of new ESC guidelines in daily clinical practice.

  4. Fostering diffusion of scientific contents of National Society Cardiovascular Journals: The new ESC search engine.

    Science.gov (United States)

    Alfonso, Fernando; Gonçalves, Lino; Pinto, Fausto; Timmis, Adam; Ector, Hugo; Ambrosio, Giuseppe; Vardas, Panos

    2015-05-01

    European Society of Cardiology (ESC) National Society Cardiovascular Journals (NSCJs) are high-quality biomedical journals focused on cardiovascular diseases. The Editors' Network of the ESC devises editorial initiatives aimed at improving the scientific quality and diffusion of NSCJ. In this article we will discuss on the importance of the Internet, electronic editions and open access strategies on scientific publishing. Finally, we will propose a new editorial initiative based on a novel electronic tool on the ESC web-page that may further help to increase the dissemination of contents and visibility of NSCJs. Copyright © 2013 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  5. Formalization of 2007 ESH/ESC Hypertension Guidelines (HGL)

    Czech Academy of Sciences Publication Activity Database

    Peleška, Jan; Anger, Z.; Buchtela, David; Tomečková, Marie; Veselý, Arnošt; Zvárová, Jana; Zvolský, Miroslav

    2008-01-01

    Roč. 26, Suppl. 1 (2008), s. 302-302 ISSN 0952-1178. [ Hypertension 2008. 14.06.2008-19.06.2008, Berlin] R&D Projects: GA AV ČR 1ET200300413 Institutional research plan: CEZ:AV0Z10300504 Keywords : formalization of medical guidelines * 2007 ESH/ESC hypertension guidelines Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery

  6. ACC/AHA guidelines superior to ESC/EAS guidelines for primary prevention with statins in non-diabetic Europeans

    DEFF Research Database (Denmark)

    Mortensen, Martin Bødtker; Nordestgaard, Børge G; Afzal, Shoaib

    2017-01-01

    Aim We compared the 2013 American College of Cardiology/American Heart Association (ACC/AHA) and the 2016 European Society of Cardiology/European Atherosclerosis Society (ESC/EAS) guidelines on prevention of atherosclerotic cardiovascular disease (ASCVD) using different risk prediction models [US......-calibrated around decision thresholds for statin therapy. For a Class I recommendation, 42% of individuals qualified for statins using the ACC/AHA guidelines vs. 6% with the ESC/EAS guidelines. Using ACC/AHA- vs. ESC/EAS-defined statin eligibility led to a substantial gain in sensitivity (+62% for any ASCVD and+76......% for fatal ASCVD) with a smaller loss in specificity (-35% for any ASCVD and -36% for fatal ASCVD). Similar differences between the ACC/AHA and ESC/EAS guidelines were found for men and women separately, and for Class IIa recommendations. The sensitivity and specificity of a US-PCE risk of 5% were similar...

  7. Chromosome togetherness at the onset of ESC differentiation.

    Science.gov (United States)

    Krivega, Ivan; Dean, Ann

    2015-03-05

    Pairing of homologous alleles is a phenomenon generally associated with imprinted and mono-allelically expressed loci. In this issue, Hogan et al. (2015) examine the earliest steps between pluripotency and lineage commitment in ESCs and find a critical role for transient pairing of Oct4 alleles in exiting the pluripotent state. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. A defined, feeder-free, serum-free system to generate in vitro hematopoietic progenitors and differentiated blood cells from hESCs and hiPSCs.

    Directory of Open Access Journals (Sweden)

    Giorgia Salvagiotto

    2011-03-01

    Full Text Available Human ESC and iPSC are an attractive source of cells of high quantity and purity to be used to elucidate early human development processes, for drug discovery, and in clinical cell therapy applications. To efficiently differentiate pluripotent cells into a pure population of hematopoietic progenitors we have developed a new 2-dimensional, defined and highly efficient protocol that avoids the use of feeder cells, serum or embryoid body formation. Here we showed that a single matrix protein in combination with growth factors and a hypoxic environment is sufficient to generate from pluripotent cells hematopoietic progenitors capable of differentiating further in mature cell types of different lineages of the blood system. We tested the differentiation method using hESCs and 9 iPSC lines generated from different tissues. These data indicate the robustness of the protocol providing a valuable tool for the generation of clinical-grade hematopoietic cells from pluripotent cells.

  9. The influence of gamma radiation on the ESC behaviour of a toughened PMMA through stress relaxation

    International Nuclear Information System (INIS)

    Sousa, Alexandre R.; Araujo, Elmo S.; Rabello, Marcelo S.

    2009-01-01

    On this work we studied the ESC degradation behaviour of a toughened PMMA irradiated with different gamma radiation doses. Tensile samples were obtained by injection moulding, and then irradiated using a 60 Co source. The samples irradiated on several doses were submitted to relaxation tests under air, ethanol and ethylene glycol. The results showed that the ESC action was intensified with the rising radiation doses when the relaxation tests were done under ethanol. On the tests under ethylene glycol the ESC effect was observed only to the irradiated polymer through the higher dose and under the higher relaxation load. The fracture surface analysis of tested relaxation samples, under ethanol, showed a dendritic pattern formed on fracture surfaces. (author)

  10. Derivation and characterisation of the human embryonic stem cell lines, NOTT1 and NOTT2.

    Science.gov (United States)

    Priddle, Helen; Allegrucci, Cinzia; Burridge, Paul; Munoz, Maria; Smith, Nigel M; Devlin, Lyndsey; Sjoblom, Cecilia; Chamberlain, Sarah; Watson, Sue; Young, Lorraine E; Denning, Chris

    2010-04-01

    The ability to maintain human embryonic stem cells (hESCs) during long-term culture and yet induce differentiation to multiple lineages potentially provides a novel approach to address various biomedical problems. Here, we describe derivation of hESC lines, NOTT1 and NOTT2, from human blastocysts graded as 3BC and 3CB, respectively. Both lines were successfully maintained as colonies by mechanical passaging on mouse embryonic feeder cells or as monolayers by trypsin-passaging in feeder-free conditions on Matrigel. Undifferentiated cells retained expression of pluripotency markers (OCT4, NANOG, SSEA-4, TRA-1-60 and TRA-1-81), a stable karyotype during long-term culture and could be transfected efficiently with plasmid DNA and short interfering RNA. Differentiation via formation of embryoid bodies resulted in expression of genes associated with early germ layers and terminal lineage specification. The electrophysiology of spontaneously beating NOTT1-derived cardiomyocytes was recorded and these cells were shown to be pharmacologically responsive. Histological examination of teratomas formed by in vivo differentiation of both lines in severe immunocompromised mice showed complex structures including cartilage or smooth muscle (mesoderm), luminal epithelium (endoderm) and neuroectoderm (ectoderm). These observations show that NOTT1 and NOTT2 display the accepted characteristics of hESC pluripotency.

  11. ESC resistance of commercial grade polycarbonates during exposure to butter and related chemicals

    DEFF Research Database (Denmark)

    Kjellander, Carina Koch; Nielsen, Tenna B; Ghanbari-Siahkali, Afshin

    2008-01-01

    differences in ESC resistance could be related to their structural or chemical properties. MALDI-TOF mass spectrometry revealed that Makrolon Rx:1805 contains a low molar mass material characterised as poly(propylene glycol)p, which was confirmed by ATR-FTIR and H-1 NMR. Some "non-absorbing" chemicals......, such as butter, cause the PCs to be less resistant to ESC under stress. The reason for this is that these chemicals and the PCs have sufficiently similar Hansen solubility parameters to allow surface conformational changes even though absorption is non-existent or extremely small. ATR-FTIR was used to detect...

  12. Derivation of Two New Human Embryonic Stem Cell Lines from Nonviable Human Embryos

    Directory of Open Access Journals (Sweden)

    Svetlana Gavrilov

    2011-01-01

    Full Text Available We report the derivation and characterization of two new human embryonic stem cells (hESC lines (CU1 and CU2 from embryos with an irreversible loss of integrated organismic function. In addition, we analyzed retrospective data of morphological progression from embryonic day (ED 5 to ED6 for 2480 embryos not suitable for clinical use to assess grading criteria indicative of loss of viability on ED5. Our analysis indicated that a large proportion of in vitro fertilization (IVF embryos not suitable for clinical use could be used for hESC derivation. Based on these combined findings, we propose that criteria commonly used in IVF clinics to determine optimal embryos for uterine transfer can be employed to predict the potential for hESC derivation from poor quality embryos without the destruction of vital human embryos.

  13. Icaritin enhances mESC self-renewal through upregulating core pluripotency transcription factors mediated by ER?

    OpenAIRE

    Tsang, Wing Pui; Zhang, Fengjie; He, Qiling; Cai, Waijiao; Huang, Jianhua; Chan, Wai Yee; Shen, Ziyin; Wan, Chao

    2017-01-01

    Utilization of small molecules in modulation of stem cell self-renewal is a promising approach to expand stem cells for regenerative therapy. Here, we identify Icaritin, a phytoestrogen molecule enhances self-renewal of mouse embryonic stem cells (mESCs). Icaritin increases mESCs proliferation while maintains their self-renewal capacity in vitro and pluripotency in vivo. This coincides with upregulation of key pluripotency transcription factors OCT4, NANOG, KLF4 and SOX2. The enhancement of m...

  14. How well are European Society of Cardiology (ESC) guidelines adhered to in patients with syncope?

    LENUS (Irish Health Repository)

    O'Dwyer, C

    2010-01-01

    The ESC guidelines on syncope were published in 2001 and updated in 2004. Adherence to the recommendations enables early stratification of low and high risk patients and prevents unnecessary investigations and admissions. Vasovagal syncope (VVS) is the commonest cause of syncope in all age groups and a low risk condition. The study objective was to determine whether the ESC guidelines were adhered to prior to referral to a syncope unit; 100 consecutive patients with unexplained syncope (52 +\\/- 23 (15-91) years); 53 female. Sixty-six patients had VVS. Forty nine (75%) of patients with VVS had undergone unnecessary investigations prior to diagnosis and 31 (47%) were admitted to hospital for investigation. Research from other countries confirms that adherence to the ESC guidelines expediates accurate diagnosis, improves resource utilization and reduces health care cost. Greater awareness amongst Irish practitioners of guidelines may improve syncope management and reduce costs.

  15. The Lyman Continuum Escape Fraction of Emission Line-selected z ∼ 2.5 Galaxies Is Less Than 15%

    Energy Technology Data Exchange (ETDEWEB)

    Rutkowski, Michael J.; Hayes, Matthew [Department of Astronomy, AlbaNova University Centre, Stockholm University, SE-10691 Stockholm (Sweden); Scarlata, Claudia; Mehta, Vihang [Minnesota Institute for Astrophysics, University of Minnesota, 116 Church Street SE, Minneapolis, MN 55455 (United States); Henry, Alaina; Hathi, Nimish; Koekemoer, Anton M. [Space Telescope Science Institute, Baltimore, MD 21218 (United States); Cohen, Seth; Windhorst, Rogier [School of Earth and Space Exploration, Arizona State University, Tempe, AZ 85281 (United States); Teplitz, Harry I. [Infrared Processing and Analysis Center, California Institute of Technology, Pasadena, CA 91125 (United States); Haardt, Francesco [DiSAT, Università dellInsubria, via Valleggio 11, I-22100 Como (Italy); Siana, Brian [Department of Physics, University of California, Riverside, CA 92521 (United States)

    2017-06-01

    Recent work suggests that strong emission line, star-forming galaxies (SFGs) may be significant Lyman continuum leakers. We combine archival Hubble Space Telescope broadband ultraviolet and optical imaging (F275W and F606W, respectively) with emission line catalogs derived from WFC3 IR G141 grism spectroscopy to search for escaping Lyman continuum (LyC) emission from homogeneously selected z ∼ 2.5 SFGs. We detect no escaping Lyman continuum from SFGs selected on [O ii] nebular emission ( N = 208) and, within a narrow redshift range, on [O iii]/[O ii]. We measure 1 σ upper limits to the LyC escape fraction relative to the non-ionizing UV continuum from [O ii] emitters, f {sub esc} ≲ 5.6%, and strong [O iii]/[O ii] > 5 ELGs, f {sub esc} ≲ 14.0%. Our observations are not deep enough to detect f {sub esc} ∼ 10% typical of low-redshift Lyman continuum emitters. However, we find that this population represents a small fraction of the star-forming galaxy population at z ∼ 2. Thus, unless the number of extreme emission line galaxies grows substantially to z ≳ 6, such galaxies may be insufficient for reionization. Deeper survey data in the rest-frame ionizing UV will be necessary to determine whether strong line ratios could be useful for pre-selecting LyC leakers at high redshift.

  16. The Lyman Continuum Escape Fraction of Emission Line-selected z ∼ 2.5 Galaxies Is Less Than 15%

    International Nuclear Information System (INIS)

    Rutkowski, Michael J.; Hayes, Matthew; Scarlata, Claudia; Mehta, Vihang; Henry, Alaina; Hathi, Nimish; Koekemoer, Anton M.; Cohen, Seth; Windhorst, Rogier; Teplitz, Harry I.; Haardt, Francesco; Siana, Brian

    2017-01-01

    Recent work suggests that strong emission line, star-forming galaxies (SFGs) may be significant Lyman continuum leakers. We combine archival Hubble Space Telescope broadband ultraviolet and optical imaging (F275W and F606W, respectively) with emission line catalogs derived from WFC3 IR G141 grism spectroscopy to search for escaping Lyman continuum (LyC) emission from homogeneously selected z ∼ 2.5 SFGs. We detect no escaping Lyman continuum from SFGs selected on [O ii] nebular emission ( N = 208) and, within a narrow redshift range, on [O iii]/[O ii]. We measure 1 σ upper limits to the LyC escape fraction relative to the non-ionizing UV continuum from [O ii] emitters, f _e_s_c ≲ 5.6%, and strong [O iii]/[O ii] > 5 ELGs, f _e_s_c ≲ 14.0%. Our observations are not deep enough to detect f _e_s_c ∼ 10% typical of low-redshift Lyman continuum emitters. However, we find that this population represents a small fraction of the star-forming galaxy population at z ∼ 2. Thus, unless the number of extreme emission line galaxies grows substantially to z ≳ 6, such galaxies may be insufficient for reionization. Deeper survey data in the rest-frame ionizing UV will be necessary to determine whether strong line ratios could be useful for pre-selecting LyC leakers at high redshift.

  17. Characterization of genetic determinants of extended-spectrum cephalosporinases (ESCs) in Escherichia coli isolates from Danish and imported poultry meat

    DEFF Research Database (Denmark)

    Bergenholz, Rikke; Jørgensen, Mikael Skaanning; Hansen, Lars H.

    2009-01-01

    Sir, The predominant cause of resistance towards cephalosporins in Escherichia coli is production of plasmid-encoded extended-spectrum ß-lactamases (ESBLs) and AmpC-type ß-lactamases, also referred to as extended-spectrum cephalosporinases (ESCs). Most studies have focused on description of ESCs...

  18. Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD) for monogenic disorder

    OpenAIRE

    Abdelkrim Hmadcha; Yolanda Aguilera; Maria Dolores Lozano-Arana; Nuria Mellado; Javier Sánchez; Cristina Moya; Luis Sánchez-Palazón; Jose Palacios; Guillermo Antiñolo; Bernat Soria

    2016-01-01

    From 106 human blastocyts donate for research after in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD) for monogenetic disorder, 3 human embryonic stem cells (hESCs) HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15)(q34.3;q14) detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were r...

  19. In Vitro Differentiation and Propagation of Urothelium from Pluripotent Stem Cell Lines.

    Science.gov (United States)

    Osborn, Stephanie L; Kurzrock, Eric A

    2018-01-01

    Bioengineering of bladder tissue, particularly for those patients who have advanced bladder disease, requires a source of urothelium that is healthy, capable of significant proliferation in vitro and immunologically tolerated upon transplant. As pluripotent stem cells have the potential to fulfill such criteria, they provide a critical cell source from which urothelium might be derived in vitro and used clinically. Herein, we describe the in vitro differentiation of urothelium from the H9 human embryonic stem cell (hESC) line through the definitive endoderm (DE) phase via selective culture techniques. The protocol can be used to derive urothelium from other hESCs or human-induced pluripotent stem cells.

  20. EURObservational Research Programme: the Heart Failure Pilot Survey (ESC-HF Pilot)

    DEFF Research Database (Denmark)

    Maggioni, Aldo P; Dahlström, Ulf; Filippatos, Gerasimos

    2010-01-01

    The primary objective of the new ESC-HF Pilot Survey was to describe the clinical epidemiology of outpatients and inpatients with heart failure (HF) and the diagnostic/therapeutic processes applied across 12 participating European countries. This pilot study was specifically aimed at validating...

  1. STS-48 ESC Earth observation of ice pack, Antarctic Ice Shelf

    Science.gov (United States)

    1991-01-01

    STS-48 Earth observation taken aboard Discovery, Orbiter Vehicle (OV) 103, is of the breakup of pack ice along the periphery of the Antarctic Ice Shelf. Strong offshore winds, probably associated with katabatic downdrafts from the interior of the continent, are seen peeling off the edges of the ice shelf into long filaments of sea ice, icebergs, bergy bits, and growlers to flow northward into the South Atlantic Ocean. These photos are used to study ocean wind, tide and current patterns. Similar views photographed during previous missions, when analyzed with these recent views may yield information about regional ice drift and breakup of ice packs. The image was captured using an electronic still camera (ESC), was stored on a removable hard disk or small optical disk, and was converted to a format suitable for downlink transmission. The ESC documentation was part of Development Test Objective (DTO) 648, Electronic Still Photography.

  2. Ansiedad escénica y flamenco

    Directory of Open Access Journals (Sweden)

    Mariola Lupiáñez Castillo

    2016-01-01

    Full Text Available Cuando el artista flamenco sube al escenario, además de expresar su arte y disfrutar con su ejecución, es también objeto de la valoración de su público. Cuando los pensamientos y la atención del artista se centran solo en este escrutinio de la audiencia se puede desarrollar miedo o incomodidad a la hora de actuar, padeciendo lo que se denomina ansiedad escénica. Esta fobia social aparece cuando la persona teme ser evaluada negativamente por los demás y mostrar síntomas de nerviosismo que puedan ser perceptibles por el público. Las manifestaciones de la ansiedad se pueden clasificar en tres categorías: Cognitivas, fisiológicas y conductuales. Estas manifestaciones afectan al estado de ánimo, dificultan la ejecución de la tarea y predisponen al artista a evitar enfrentarse a futuras actuaciones. No controlar estas manifestaciones de la forma adecuada puede suponer en el músico o bailaor el detrimento de su calidad artística y en numerosas ocasiones, el abandono de la carrera artística. Estudios anteriores revelan que la formación académica y el contexto socio cultural en el que se desenvuelve el artista pueden incidir en el desarrollo y permanencia de la ansiedad escénica. Teniendo en cuenta las peculiaridades históricas y sociales del Flamenco así como la evolución de su formación, que ha pasado de la transmisión oral de un ambiente informal al estudio dentro de los conservatorios, se considera necesario estudiar cómo afecta la ansiedad a los artistas de este género para poder prevenir e intervenir en los músicos que la padezcan.

  3. EURObservational Research Programme : The Heart Failure Pilot Survey (ESC-HF Pilot)

    NARCIS (Netherlands)

    Maggioni, Aldo P.; Dahlstrom, Ulf; Filippatos, Gerasimos; Chioncel, Ovidiu; Crespo Leiro, Marisa; Drozdz, Jaroslaw; Fruhwald, Friedrich; Gullestad, Lars; Logeart, Damien; Metra, Marco; Parissis, John; Persson, Hans; Ponikowski, Piotr; Rauchhaus, Mathias; Voors, Adriaan A.; Nielsen, Olav Wendelboe; Zannad, Faiez; Tavazzi, Luigi

    2010-01-01

    The primary objective of the new ESC-HF Pilot Survey was to describe the clinical epidemiology of outpatients and inpatients with heart failure (HF) and the diagnostic/therapeutic processes applied across 12 participating European countries. This pilot study was specifically aimed at validating the

  4. Ultrastructural characteristics of three undifferentiated mouse embryonic stem cell lines and their differentiated three-dimensional derivatives: a comparative study.

    Science.gov (United States)

    Alharbi, Suzan; Elsafadi, Mona; Mobarak, Mohammed; Alrwili, Ali; Vishnubalaji, Radhakrishnan; Manikandan, Muthurangan; Al-Qudsi, Fatma; Karim, Saleh; Al-Nabaheen, May; Aldahmash, Abdullah; Mahmood, Amer

    2014-04-01

    The fine structures of mouse embryonic stem cells (mESCs) grown as colonies and differentiated in three-dimensional (3D) culture as embryoid bodies (EBs) were analyzed by transmission electron microscopy. Undifferentiated mESCs expressed markers that proved their pluripotency. Differentiated EBs expressed different differentiation marker proteins from the three germ layers. The ultrastructure of mESCs revealed the presence of microvilli on the cell surfaces, large and deep infolded nuclei, low cytoplasm-to-nuclear ratios, frequent lipid droplets, nonprominent Golgi apparatus, and smooth endoplasmic reticulum. In addition, we found prominent juvenile mitochondria and free ribosomes-rich cytoplasm in mESCs. Ultrastructure of the differentiated mESCs as EBs showed different cell arrangements, which indicate the different stages of EB development and differentiation. The morphologies of BALB/c and 129 W9.5 EBs were very similar at day 4, whereas C57BL/6 EBs were distinct from the others at day 4. This finding suggested that differentiation of EBs from different cell lines occurs in the same pattern but not at the same rate. Conversely, the ultrastructure results of BALB/c and 129 W9.5 ESCs revealed differentiating features, such as the dilated profile of a rough endoplasmic reticulum. In addition, we found low expression levels of undifferentiated markers on the outer cells of BALB/c and 129 W9.5 mESC colonies, which suggests a faster differentiation potential.

  5. ELABELA Is an Endogenous Growth Factor that Sustains hESC Self-Renewal via the PI3K/AKT Pathway

    NARCIS (Netherlands)

    Ho, Lena; Tan, Shawn Y. X.; Wee, Sheena; Wu, Yixuan; Tan, Sam J. C.; Ramakrishna, Navin B.; Chng, Serene C.; Nama, Srikanth; Szczerbinska, Iwona; Sczerbinska, Iwona; Chan, Yun-Shen; Avery, Stuart; Tsuneyoshi, Norihiro; Ng, Huck Hui; Gunaratne, Jayantha; Dunn, N. Ray; Reversade, Bruno

    2015-01-01

    ELABELA (ELA) is a peptide hormone required for heart development that signals via the Apelin Receptor (APLNR, APJ). ELA is also abundantly secreted by human embryonic stem cells (hESCs), which do not express APLNR. Here we show that ELA signals in a paracrine fashion in hESCs to maintain

  6. Geological map of Uruguay Esc 1,100,000. Guayabos Sheet N-15

    International Nuclear Information System (INIS)

    Gancio, F.; Ford, I.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Guayabos) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils of Arapey, Guichon, Mercedes and Asencio formations in the Cretaceous period

  7. EseE of Edwardsiella tarda Augments Secretion of Translocon Protein EseC and Expression of the escC-eseE Operon.

    Science.gov (United States)

    Yi, Jia; Xiao, Shui Bing; Zeng, Zhi Xiong; Lu, Jin Fang; Liu, Lu Yi; Laghari, Zubair Ahmed; Nie, Pin; Yu, Hong Bing; Xie, Hai Xia

    2016-08-01

    Edwardsiella tarda is an important Gram-negative pathogen that employs a type III secretion system (T3SS) to deliver effectors into host cells to facilitate bacterial survival and replication. These effectors are translocated into host cells through a translocon complex composed of three secreted proteins, namely, EseB, EseC, and EseD. The secretion of EseB and EseD requires a chaperone protein called EscC, whereas the secretion of EseC requires the chaperone EscA. In this study, we identified a novel protein (EseE) that also regulates the secretion of EseC. An eseE deletion mutant secreted much less EseC into supernatants, accompanied by increased EseC levels within bacterial cells. We also demonstrated that EseE interacted directly with EseC in a pulldown assay. Interestingly, EseC, EseE, and EscA were able to form a ternary complex, as revealed by pulldown and gel filtration assays. Of particular importance, the deletion of eseE resulted in decreased levels of EseB and EseD proteins in both the bacterial pellet and supernatant fraction. Furthermore, real-time PCR assays showed that EseE positively regulated the transcription of the translocon operon escC-eseE, comprising escC, eseB, escA, eseC, eseD, and eseE These effects of EseE on the translocon components/operon appeared to have a functional consequence, since the ΔeseE strain was outcompeted by wild-type E. tarda in a mixed infection in blue gourami fish. Collectively, our results demonstrate that EseE not only functions as a chaperone for EseC but also acts as a positive regulator controlling the expression of the translocon operon escC-eseE, thus contributing to the pathogenesis of E. tarda in fish. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  8. Dynamic link between histone H3 acetylation and an increase in the functional characteristics of human ESC/iPSC-derived cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Tomomi G Otsuji

    Full Text Available Cardiomyocytes (CMs derived from human embryonic stem cells (hESCs or human induced pluripotent stem cells (hiPSCs are functionally heterogeneous, display insufficient biological efficacy and generally possess the electrophysiological properties seen in fetal CMs. However, a homogenous population of hESC/hiPSC-CMs, with properties similar to those of adult human ventricular cells, is required for use in drug cardiotoxicity screening. Unfortunately, despite the requirement for the functional characteristics of post-mitotic beating cell aggregates to mimic the behavior of mature cardiomyocytes in vitro, few technological improvements have been made in this field to date. Previously, we showed that culturing hESC-CMs under low-adhesion conditions with cyclic replating confers continuous contractility on the cells, leading to a functional increase in cardiac gene expression and electrophysiological properties over time. The current study reveals that culturing hESC/hiPSC-CMs under non-adhesive culture conditions enhances the electrophysiological properties of the CMs through an increase in the acetylation of histone H3 lysine residues, as confirmed by western blot analyses. Histone H3 acetylation was induced chemically by treating primitive hESC/hiPSC-CMs with Trichostatin A (TSA, a histone deacetylase (HDAC inhibitor, resulting in an immediate increase in global cardiac gene expression. In functional analyses using multi-electrode array (MEA recordings, TSA-treated hESC/hiPSC-CM colonies showed appropriate responses to particular concentrations of known potassium ion channel inhibitors. Thus, the combination of a cell-autonomous functional increase in response to non-adhesive culture and short-term TSA treatment of hESC/hiPSC-CM colonies cultured on MEA electrodes will help to make cardiac toxicity tests more accurate and reproducible via genome-wide chromatin activation.

  9. Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD for monogenic disorder

    Directory of Open Access Journals (Sweden)

    Abdelkrim Hmadcha

    2016-05-01

    Full Text Available From 106 human blastocyts donate for research after in vitro fertilization (IVF and preimplantation genetic diagnosis (PGD for monogenetic disorder, 3 human embryonic stem cells (hESCs HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15(q34.3;q14 detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were registered in the Spanish Stem Cell Bank.

  10. Geological map of Uruguay Esc 1,100,000. Cololo Sheet 0-19

    International Nuclear Information System (INIS)

    Morales, H.; Ford, I.; Montana, J.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Cololo) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils of Holocene, Oligocene and upper Cretaceous in Asencio and Fray Bentos formations

  11. Cost-effective master cell bank validation of multiple clinical-grade human pluripotent stem cell lines from a single donor.

    Science.gov (United States)

    Devito, Liani; Petrova, Anastasia; Miere, Cristian; Codognotto, Stefano; Blakely, Nicola; Lovatt, Archie; Ogilvie, Caroline; Khalaf, Yacoub; Ilic, Dusko

    2014-10-01

    Standardization guidelines for human pluripotent stem cells are still very broadly defined, despite ongoing clinical trials in the U.S., U.K., and Japan. The requirements for validation of human embryonic (hESCs) and induced pluripotent stem cells (iPSCs) in general follow the regulations for other clinically compliant biologics already in place but without addressing key differences between cell types or final products. In order to realize the full potential of stem cell therapy, validation criteria, methodology, and, most importantly, strategy, should address the shortfalls and efficiency of current approaches; without this, hESC- and, especially, iPSC-based therapy will not be able to compete with other technologies in a cost-efficient way. We addressed the protocols for testing cell lines for human viral pathogens and propose a novel strategy that would significantly reduce costs. It is highly unlikely that the multiple cell lines derived in parallel from a tissue sample taken from one donor would have different profiles of endogenous viral pathogens; we therefore argue that samples from the Master Cell Banks of sibling lines could be safely pooled for validation. We illustrate this approach with tiered validation of two sibling clinical-grade hESC lines, KCL033 and KCL034 (stage 1, sterility; stage 2, specific human pathogens; and stage 3, nonspecific human pathogens). The results of all tests were negative. This cost-effective strategy could also be applied for validation of Master Cell Banks of multiple clinical-grade iPSC lines derived from a single donor. ©AlphaMed Press.

  12. A comprehensive review of rollover accidents involving vehicles equipped with Electronic Stability Control (ESC) systems.

    Science.gov (United States)

    Padmanaban, Jeya; Shields, Leland E; Scheibe, Robert R; Eyges, Vitaly E

    2008-10-01

    This study investigated 478 police accident reports from 9 states to examine and characterize rollover crashes involving ESC-equipped vehicles. The focus was on the sequence of critical events leading to loss of control and rollover, and the interactions between the accident, driver, and environment. Results show that, while ESC is effective in reducing loss of control leading to certain rollover crashes, its effectiveness is diminished in others, particularly when the vehicle departs the roadway or when environmental factors such as slick road conditions or driver factors such as speeding, distraction, fatigue, impairment, or overcorrection are present.

  13. Introducción al libro La mujer en las artes visuales y escénicas

    OpenAIRE

    Barrios Herrero, Olga

    2010-01-01

    [ES]Este trabajo presenta el libro Women in the Visual and Performing Arts, haciendo hincapié en la variedad de estilos, alto grado de experimentación y géneros vanguardistas creados por mujeres artistas de todo el mundo, tanto en las artes visuales como en las escénicas, muchas de ellas combinando visual y escénico en sus expresiones artísticas. El libro incluye una panorámica histórica de las mujeres en las artes, presentando teoría al igual que testimonios de las propias artistas que nos c...

  14. Geological map of Uruguay Esc 1,100,000. Piriapolis Sheet G-29

    International Nuclear Information System (INIS)

    Preciozzi, F; Pena, S; Masquelin, E; Pias, J; Tabo, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Piriapolis) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils. Geomorphologically Piriapolis fotoplano is dominated by Las Animas and an important Cenozoic coverage

  15. Esc2 and Sgs1 act in functionally distinct branches of the homologous recombination repair pathway in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Mankouri, Hocine W; Ngo, Hien-Ping; Hickson, Ian D

    2009-01-01

    , the accumulation of these structures in esc2 (but not sgs1) mutants is entirely dependent on Mph1, a protein that shows structural similarity to the Fanconi anemia group M protein (FANCM). In the absence of both Esc2 and Sgs1, the intra-S-phase DNA damage checkpoint response is compromised after exposure to MMS...

  16. Eficiência de escórias de siderurgia na cultura do feijoeiro em sistema de semeadura direta

    Directory of Open Access Journals (Sweden)

    Angélica Cristina Fernandes Deus

    2013-10-01

    Full Text Available No sistema de semeadura direta (SSD, a prática da correção da acidez do solo se restringe ao uso do calcário, havendo poucas informações em relação às escórias de siderurgia. O trabalho objetivou avaliar as alterações em propriedades químicas do solo, rendimento e absorção de nutrientes do feijoeiro, em função das formas de aplicação de escórias de siderurgia, comparadas ao calcário, na implantação do sistema de semeadura direta. O experimento foi conduzido na área experimental da Faculdade de Ciências Agronômicas de Botucatu (SP, no período de dezembro de 2010 a maio de 2011. Os tratamentos foram duas formas de aplicação de sete corretivos da acidez do solo: escórias de aciaria, de alto forno, de forno de panela, do aço inox (agrosilício, wollastonita, calcário dolomítico e calcário dolomítico calcinado, além de uma testemunha sem aplicação de corretivo. A dose aplicada de cada corretivo foi calculada para elevar a saturação por bases do solo a 70. A acidez do solo foi neutralizada até a camada de 20cm com os calcários, enquanto a wollastonita e escória de forno de panela atingiram a camada de 10cm; para as escórias de aciaria, alto forno e o agrosilício, o efeito corretivo ficou restrito aos primeiros 5cm. O rendimento do feijoeiro aumentou com a aplicação de corretivos da acidez do solo, sem diferença entre o modo de aplicação.

  17. Comparison of Gene Expression in Human Embryonic Stem Cells, hESC-Derived Mesenchymal Stem Cells and Human Mesenchymal Stem Cells

    OpenAIRE

    Romain Barbet; Isabelle Peiffer; Antoinette Hatzfeld; Pierre Charbord; Jacques A. Hatzfeld

    2011-01-01

    We present a strategy to identify developmental/differentiation and plasma membrane marker genes of the most primitive human Mesenchymal Stem Cells (hMSCs). Using sensitive and quantitative TaqMan Low Density Arrays (TLDA) methodology, we compared the expression of 381 genes in human Embryonic Stem Cells (hESCs), hESC-derived MSCs ...

  18. Explicative memory of the geologic map of Uruguay. Esc. 1.500.000

    International Nuclear Information System (INIS)

    Preciozzi Porta, F.; Spoturno, J.; Rossi, P.; Heinzen, W.

    1985-01-01

    The Geological Map of Uruguay Esc. 1.500.000 is part of the Geological Map Programme developed by the Institute Geologic during the years 1977 - 1980. Its memory describe the geography, the lit ho stratigraphy and the crystalline area in the soil of Uruguay.

  19. Geological map of Uruguay Esc 1,100,000. Talita Sheet J-24

    International Nuclear Information System (INIS)

    Campal, N; Chulepin, H

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Talita) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils.The area belongs to the Rio de la Plata socket basin is associated with the orogenic cycle

  20. Geological map of Uruguay Esc 1,100,000. Molles Sheet K-19

    International Nuclear Information System (INIS)

    Ford, I

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Molles) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the Cretaceous, Tertiary and Quaternary period in Arapey, Mercedes, Asencio, Palmitas, Fray Bentos and Libertad formations

  1. Geological map of Uruguay Esc 1,100,000. Chafalote Sheet D-26

    International Nuclear Information System (INIS)

    Masquelin, H; Tabo, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Chafalote) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils. The area corresponding to Chafalote is located in central and eastern metamorphic formation constituted by Lavalleja-Rocha group

  2. Geological map of Uruguay Esc 1,100,000. Palmitas Sheet 0- 22

    International Nuclear Information System (INIS)

    Ford, I.; Gancio, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Molles) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the Proterozoic, Cretaceous, Tertiary and Quaternary period in Mercedes, Asencio, Palmitas and Fray Bentos formations

  3. Benefits and Disadvantages of Neoadjuvant Radiochemotherapy (RCT) in the Multimodal Therapy of Squamous Esophageal Cancer (ESC).

    Science.gov (United States)

    Hanna, Adrian; Birla, Rodica; Iosif, Cristina; Boeriu, Marius; Constantinoiu, Silviu

    2016-01-01

    The purpose of this paper is to present the advantages and disadvantages of neoadjuvant RCT in multimodal therapyof ESC. Between 1998-2014 221 patients were treated with ESC, 85 of whom received neoadjuvant RCT. For these we have made imaging and pathologic assessment of response using RECIST and MANDARD criteria and statistical data were interpreted in terms of the factors that influence the response. Also, they were evaluated statistical correlations between RCT and resectability, postoperative morbidity, mortality and long-term survival. 45 patients were imagistic responders and 34 underwent surgery, 40 non-responders of which 14 underwent surgery. Of the 48 surgical patients with preoperative RCT, histopathological evaluation showed that 32 were pathological responders and 16 non responders. There were performed statistical analyzes of correlations between RCT and resectability, stage, location of ESC, morbidity, mortality and survival. RCT increase resectability, improves survival and maximum duration of survival, more in responders than in nonresponders and does not affect postoperative complications and postoperative mortality, nor among the responders or nonresponders. Imaging evaluation result of the response to RCT overestimate responders. Celsius.

  4. Linee guida per la gestione della Volpe in Italia

    Directory of Open Access Journals (Sweden)

    Silvano Toso

    2003-10-01

    Full Text Available La plasticità ecologica della Volpe (Vulpes vulpes fa di questa specie il Carnivoro più diffuso e più adattabile al mondo. Proprio per la sua diffusione la Volpe è oggetto di un intenso prelievo sia venatorio, sia finalizzato a contenerne l'impatto predatorio ed allo sfruttamento commerciale della pelliccia. In Italia esiste una forte pressione dei cacciatori perché si realizzino programmi di controllo delle popolazioni di questo predatore, al fine di ridurne l'impatto sulle specie d'interesse cinegetico. Gli effetti del contenimento della Volpe sulla dinamica delle popolazioni di specie preda sono stati oggetto di numerosi studi, ma i risultati di tali ricerche non sono univoci; se in alcuni casi si è infatti verificato, in seguito a decrementi delle popolazioni di Volpe, un incremento di consistenza e successo riproduttivo delle specie preda, in altri casi non si è registrato alcun significativo effetto demografico. Ancora meno chiaro è quale sia la soglia di contrazione delle popolazioni di Volpe necessaria per ottenere un effetto significativo sulle popolazioni delle specie preda, considerato che questo Carnivoro ha un potenziale riproduttivo elevato ed un'organizzazione sociale che determina la rapida rioccupazione dei territori lasciati liberi. La legislazione italiana in materia di controllo è, rispetto ad altri paesi europei ed extraeuropei, particolarmente rigida: la legge n. 157/92 identifica infatti un obiettivo generale di mantenimento delle popolazioni dei predatori a livelli adeguati ad assicurare una funzione regolatrice delle biocenosi. La stessa norma impone inoltre che ogni intervento di abbattimento di predatori finalizzato al contenimento dei danni da essi arrecati sia autorizzato dalla Regione o dalla Provincia competente, sulla base di un parere tecnico dell'INFS che è chiamato a verificare l'inapplicabilità di metodi incruenti di prevenzione dei danni, la selettività delle tecniche di abbattimento e la correttezza tecnica del piano di prelievo proposto. Inoltre, la legge prescrive che gli interventi di contenimento debbano essere direttamente realizzati - o coordinati - da personale delle Amministrazioni provinciali, la cui operatività è spesso limitata dalla carenza degli organici. La situazione normativa e gestionale che caratterizza il nostro Paese determina tassi di prelievo sulle popolazioni di Volpe generalmente molto limitati, assolutamente inadeguati al raggiungimento, su ampia scala geografica, di significativi effetti demografici sulle popolazioni di specie preda d'interesse venatorio. In tale contesto l'INFS ritiene che la programmazione di azioni di controllo della Volpe debba prevedere: 1 la definizione di obiettivi chiari e coerenti con l'attuale quadro normativo, misurabili e realizzabili; 2 l'applicazione di misure incruente di contenimento del danno (in particolare la sospensione dei ripopolamenti con selvaggina allevata; 3 l'utilizzo di metodi di prelievo selettivi e che minimizzino gli effetti indesiderati; 4 la selezione delle aree di intervento sulla base della reale capacità operativa e delle finalità dei diversi istituti territoriali 5 la costante verifica dei risultati conseguiti in relazione agli obiettivi, in modo da permettere una periodica revisione dei programmi operativi attraverso un approccio costantemente adattativo.

  5. Methodological procedures and analytical instruments to evaluate an indicators integrated archive for urban management; Guida metodologica per la costruzione di un archivio integrato di indicatori urbani

    Energy Technology Data Exchange (ETDEWEB)

    Del Ciello, R; Napoleoni, S [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1998-07-01

    This guide provides the results of a research developed at ENEA (National Agency for new Technology, Energy and the Environment) Casaccia center (Rome, Italy) aimed to define methodological procedures and analytical instruments needed to carry out an indicators integrated archive for urban management. The guide also defines the scheme of a negotiation process aimed to reach and exchange data and information among governmental and local administrations, non-governmental organizations and scientific bodies. [Italian] Il lavoro presenta una sintesi dei risultati di una ricerca condotta presso il C.R. Casaccia dell'ENEA, relativia alla definizione di procedure metodologiche e strumenti di analisi ed elaborazione per realizzare un archivio integrato di indicatori per la gestione dei sistemi urbani. La guida, rivolta ai responsabili delle politiche urbane, deifinisce uno schema dei processi di condivisione degli indicatori urbani attraverso l'organizzazione di opportuni tavoli negoziali, costituiti da rappresentanti delle amministrazioni locali, dell'amministrazione centrale, delle categorie produttive e sociali e delle strutture tecniche operanti sul territorio.

  6. Methodological procedures and analytical instruments to evaluate an indicators integrated archive for urban management; Guida metodologica per la costruzione di un archivio integrato di indicatori urbani

    Energy Technology Data Exchange (ETDEWEB)

    Del Ciello, R.; Napoleoni, S. [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1998-07-01

    This guide provides the results of a research developed at ENEA (National Agency for new Technology, Energy and the Environment) Casaccia center (Rome, Italy) aimed to define methodological procedures and analytical instruments needed to carry out an indicators integrated archive for urban management. The guide also defines the scheme of a negotiation process aimed to reach and exchange data and information among governmental and local administrations, non-governmental organizations and scientific bodies. [Italian] Il lavoro presenta una sintesi dei risultati di una ricerca condotta presso il C.R. Casaccia dell'ENEA, relativia alla definizione di procedure metodologiche e strumenti di analisi ed elaborazione per realizzare un archivio integrato di indicatori per la gestione dei sistemi urbani. La guida, rivolta ai responsabili delle politiche urbane, deifinisce uno schema dei processi di condivisione degli indicatori urbani attraverso l'organizzazione di opportuni tavoli negoziali, costituiti da rappresentanti delle amministrazioni locali, dell'amministrazione centrale, delle categorie produttive e sociali e delle strutture tecniche operanti sul territorio.

  7. Murine transgenic embryonic stem cell lines for the investigation of sinoatrial node-related molecular pathways

    Directory of Open Access Journals (Sweden)

    Stefanie Schmitteckert

    2017-12-01

    Full Text Available The elucidation of molecular mechanisms that restrict the potential of pluripotent stem cells and promote cardiac lineage differentiation is of crucial relevance, since embryonic stem cells (ESCs hold great potential for cell based heart therapies. The homeodomain transcription factor Shox2 is essential for the development and proper function of the native cardiac pacemaker, the sinoatrial node. This prompted us to develop a cardiac differentiation model using ESC lines isolated from blastocysts of Shox2-deficient mice. The established cell model provides a fundamental basis for the investigation of molecular pathways under physiological and pathophysiological conditions for evaluating novel therapeutic approaches.

  8. The synergistic effect of 5Azadc and TSA on maintenance of pluripotency of chicken ESCs by overexpression of NANOG gene.

    Science.gov (United States)

    Wang, Xiaoyan; Wang, Yingjie; Zuo, Qisheng; Li, Dong; Zhang, Wenhui; Lian, Chao; Tang, Beibei; Xiao, Tianrong; Wang, Man; Wang, Kehua; Li, Bichun; Zhang, Yani

    2016-04-01

    NANOG is a transcription factor that functions in embryonic stem cells (ESCs) and a key factor in maintaining pluripotency. Here, we cloned the NANOG gene promoter from the Rugao yellow chicken and constructed a dual luciferase reporter vector to detect its transcriptional activity and analyze the effects of 5-aza-2'-deoxycytidine (5-Azadc) and trichostatin A (TSA) on NANOG promoter activity and ESC pluripotency maintenance in vitro. NANOG transcriptional activity was enhanced when 5-Azadc and TSA were used alone or together, suggesting the possibility of elevated methylation of the CpG island in the NANOG regulatory region. When ESCs were cultured in basic medium with 5-Azadc and TSA in vitro, significantly more cell colonies were maintained in the 5-Azadc + TSA group than in the control group, which had many differentiated cells and few cell colonies after 6 d of induction. On the tenth day of induction, the cells in the control group fully differentiated and no cell colonies remained, but many cell colonies were present in the 5-Azadc + TSA group. The expression of NANOG in the cell colonies was confirmed by indirect immunofluorescence. Furthermore, ESCs could be passaged to the 12th generation under 5-Azadc and TSA treatment and maintained their pluripotency. Thus, we showed that 5-Azadc and TSA can effectively maintain chicken ESC pluripotency in vitro by increasing NANOG gene expression.

  9. Dosage and cell line dependent inhibitory effect of bFGF supplement in human pluripotent stem cell culture on inactivated human mesenchymal stem cells.

    Science.gov (United States)

    Quang, Tara; Marquez, Maribel; Blanco, Giselle; Zhao, Yuanxiang

    2014-01-01

    Many different culture systems have been developed for expanding human pluripotent stem cells (hESCs and hiPSCs). In general, 4-10 ng/ml of bFGF is supplemented in culture media in feeder-dependent systems regardless of feeder cell types, whereas in feeder-free systems, up to 100 ng/ml of bFGF is required for maintaining long-term culture on various substrates. The amount of bFGF required in native hESCs growth niche is unclear. Here we report using inactivated adipose-derived human mesenchymal stem cells as feeder cells to examine long-term parallel cultures of two hESCs lines (H1 and H9) and one hiPSCs line (DF19-9-7T) in media supplemented with 0, 0.4 or 4 ng/ml of bFGF for up to 23 passages, as well as parallel cultures of H9 and DF19 in media supplemented with 4, 20 or 100 ng/ml bFGF for up to 13 passages for comparison. Across all cell lines tested, bFGF supplement demonstrated inhibitory effect over growth expansion, single cell colonization and recovery from freezing in a dosage dependent manner. In addition, bFGF exerted differential effects on different cell lines, inducing H1 and DF19 differentiation at 4 ng/ml or higher, while permitting long-term culture of H9 at the same concentrations with no apparent dosage effect. Pluripotency was confirmed for all cell lines cultured in 0, 0.4 or 4 ng/ml bFGF excluding H1-4 ng, as well as H9 cultured in 4, 20 and 100 ng/ml bFGF. However, DF19 demonstrated similar karyotypic abnormality in both 0 and 4 ng/ml bFGF media while H1 and H9 were karyotypically normal in 0 ng/ml bFGF after long-term culture. Our results indicate that exogenous bFGF exerts dosage and cell line dependent effect on human pluripotent stem cells cultured on mesenchymal stem cells, and implies optimal use of bFGF in hESCs/hiPSCs culture should be based on specific cell line and its culture system.

  10. [The characters and specific features of new human embryonic stem cells lines].

    Science.gov (United States)

    Krylova, T A; Kol'tsova, A M; Zenin, V V; Gordeeva, O F; Musorina, A S; Goriachaia, T S; Shlykova, S A; Kamenetskaia, Iu K; Pinaev, G P; Polianskaia, G G

    2009-01-01

    Four continuous human embryonic stem cell lines (SC1, SC2, SC3 and SC4), derived from the blastocysts has been described. The cell lines were cultivated on mitotically inactivated human feeder cells. The cell lines SC1 and SC2 have passed through 150 population doublings and the cell lines SC3 and SC4 -- near 120 populations doublings, which exceeds Hayflick limit sufficiently. These cell lines maintain high activity of alkaline phosphatase, expression of transcription factor OCT-4 and cell surface antigens (SSEA-4, TRA-1-60 and TRA-1-81), confirming their ESC status and human specificity. Immunofluorescent detection of antigens, characteristic of ectoderm, endoderm and mesoderm confirms the ability of these cells to retain their pluripotency under in vitro condition. PCR analysis revealed expression of six genes specific for pluripotent cells (OCT-4, NANOG, DPPA3/STELLA, TDGF/CRIPTO and LEFTYA). Correlation between the level of proliferative activity and the character of DNA-bound fluorescent staining was found. Fluorescent dyes, Hoechst 33342 and PI, produced diffuse staining of the nuclei in slowly proliferating cells of the SC1 and SC2 lines. In contrast, in actively proliferating cells of the SC3 and SC4 lines, the clear staining of the nuclei was observed. Upon changing the cultivation condition, proliferative activity of SC3 and SC4 lines decreased and became similar to that of SC1 and SC2 lines. The character of the fluorescent staining of all these lines was also shown to be similar. These results show that quality of the fluorescent staining with Hoechst 33342 and PI reflects the level of proliferation. Possible causes and mechanisms of this feature of human ESC are discussed.

  11. English for Scientific Purposes (EScP): Technology, Trends, and Future Challenges for Science Education

    Science.gov (United States)

    Liu, Gi-Zen; Chiu, Wan-Yu; Lin, Chih-Chung; Barrett, Neil E.

    2014-12-01

    To date, the concept of English for Specific Purposes has brought about a great impact on English language learning across various disciplines, including those in science education. Hence, this review paper aimed to address current English language learning in the science disciplines through the practice of computer-assisted language learning to identify the use of learning technologies in science-based literacy. In the literature review, the researchers found that science-based literacy instruction shares many pedagogical aims with English language teaching in terms of reading, writing, listening and speaking, allowing it to be classified as English for Scientific Purposes (EScP). To answer the research questions, the researchers conducted the survey by extracting related articles and teaching examples from the Web of Science. In the search procedure, the researchers used the keywords science OR scientific AND technolog* OR comput* in ten selected journals of social science citation index. Only articles which are specified as journal articles rather than other document types were included. After compiling the corpora, the researchers compared the trends, methodologies and results of EScP instruction in science education. The implications of this study include the opportunities, advantages and challenges for EScP instruction in science education to further develop better educational approaches, adopt new technologies, as well as offer some directions for researchers to conduct future studies.

  12. Posibilidades y límites del escándalo político como una forma de control social

    Directory of Open Access Journals (Sweden)

    FERNANDO JIMÉNEZ SÁNCHEZ

    1994-01-01

    Full Text Available El punto de partida del trabajo es una definición del escándalo político como un fenómeno distinto al de la corrupción. De este modo, se define el escándalo político como una reacción de la opinión pública contra un agente político al que se considera responsable de una conducta que es percibida como un abuso de poder o una traición de la confianza social sobre la que se basa la posición de la autoridad que mentiene o puede llegar a mantener tal agente. En este sentido, el escándalo puede ser considerado como una forma de control social (un medio a través del cuál una sociedad se regula a sí misma y coordina el comportamiento de sus miembros. Ahora bien, el escándalo político no es proceso automático de control social en el que a una violación de la confianza social le siga la sanción correspondiente, sino que es un proceso abierto cuyo resultado es incierto y depende del juego de unas élites sociales que se desarrolla sobre el fondo de un contexto cultural, institucional e histórico determinado.

  13. ESC-B: The Cryogenic Upper Stage for Europe's Heavy Lift Launcher Ariane 5ECB

    Science.gov (United States)

    Juhls, A.

    2002-01-01

    -A. Juhls, Astrium GmbH -M. Lepelletier, Snecma Moteurs -JM. Bahu, CNES -C. Poincheval, CNES. In the year 1998 the European ministerial council decided to initiate the Ariane 5 Plus programme in order to upgrade the European heavy lift launcher Ariane 5. The market was changing more rapidly than predicted showing steadily growing satellite mass and the demand for flexible missions while strong competitors were intensifying their preparations to enter the commercial business. The answer was to improve the Ariane 5 launcher by modifying the cryogenic first (or lower ?) stage and the solid boosters and by introducing two cryogenic upper stages in two steps: In order to cope with the short term need of a significant growth of GTO lift capacity up to 10 t the first denoted ESC-A shall enter commercial service in 2002. Four years later a more powerful second version shall take over enabling a GTO performance of 12 t and providing versatile mission capability. The paper will focus on this new cryogenic upper stage denoted ESC-B giving first a general description of main characteristics and constituents. The article will highlight different challenging aspects of the ESC-B development: Ambitious economical conditions regarding both limited development budgets and the strong need to reduce production cost require improved working methods and an adjustment of the conventional development logic, in particular regarding new verification methods. Furthermore Europe is now facing the complex combination of versatile mission capability together with a powerful cryogenic upper stage. The paper will present the approach to define reasonable mission scenarios in order to cover customer demands while avoiding too stringent system requirements. Along with VINCI, Europe's first expander cycle type engine featuring an extendable nozzle dedicated subsystems will be described which allow 4 re-ignitions and 6 hours of ballistic flight. The paper concludes with the summary of the

  14. Geological map of Uruguay Esc 1,100,000. Carmelo Sheet P-24

    International Nuclear Information System (INIS)

    Ferrando, L; Eugui, W; Cabrera, Z; Elias, R

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Carmelo) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils of the precambrian period in the Mercedes , Fray Bentos, Libertad and Raigon formations. The Precambrian rocks are found in large quarries in Carmelo hill

  15. Geological map of Uruguay Esc 1,100,000. Algorta Sheet N-16

    International Nuclear Information System (INIS)

    Ford, I; Gancio, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Algorta) and the explanatory memoranda which describes the geological , lithological and sedimentological soil characteristics of the upper cretaceous in Guichon, Mercedes and Asencio formaltions as well as the lower tertiary period of Palmitas and Fray Bentos formations

  16. Derivation of Huntington Disease affected Genea046 human embryonic stem cell line

    Directory of Open Access Journals (Sweden)

    Biljana Dumevska

    2016-03-01

    Full Text Available The Genea046 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying HTT gene CAG expansion of 45 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 85% of cells expressed Nanog, 92% Oct4, 75% Tra1–60 and 99% SSEA4 and demonstrated Alkaline Phosphatase activity. The cell line was negative for Mycoplasma and visible contamination.

  17. Variations in Humanized and Defined Culture Conditions Supporting Derivation of New Human Embryonic Stem Cell Lines

    DEFF Research Database (Denmark)

    Fletcher, Judy M; Ferrier, Patricia M; Gardner, John O

    2006-01-01

    matrix substrate of purified human laminin (Ln) with transitional reliance on mitotically inactivated human fibroblast (HDF) feeder cells. With this integrated system hESC lines were isolated using either HDF conditioned medium supplemented with a bovine-sourced serum replacement (bSRM), or a defined...

  18. Geological map of Uruguay Esc 1,100,000. Bequelo Sheet N0-20

    International Nuclear Information System (INIS)

    Montana, J.; Ford, I.; Morales, H.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Bequelo) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics of the precambrian period in the Bequelo stream as well as the Mercedes , Asencio and Fray Bentos formations

  19. Effects of Feeder Cells on Dopaminergic Differentiation of Human Embryonic Stem Cells

    Directory of Open Access Journals (Sweden)

    Zhenqiang Zhao

    2016-12-01

    Full Text Available Mouse embryonic fibroblasts (MEFs and human foreskin fibroblasts (HFFs are used for the culture of human embryonic stem cells (hESCs. MEFs and HFFs differed in their capacity to support the proliferation and pluripotency of hESCs and could affect cardiac differentiation potential of hESCs. The aim of this study was to evaluate the effect of MEFs and HFFs feeders on dopaminergic differentiation of hESCs lines. To minimize the impact of culture condition variation, two hESCs lines were cultured on mixed feeder cells (MFCs, MEFs: HFFs =1:1 and HFFs feeder respectively, and then were differentiated into DA neurons under the identical protocol. Dopaminergic differentiation was evaluated by immunocytochemistry, quantitative fluorescent real-time PCR (qRT-PCR, transmission and scanning electron microscopy, and patch clamp. Our results demonstrated that these hESCs-derived neurons were genuine and functional DA neurons. However, compared to hESCs line on MFCs feeder, hESCs line on HFFs feeder had a higher proportion of TH positive cells and expressed higher levels of FOXA2, PITX3, NURR1 and TH genes. In addition, the values of threshold intensity and threshold membrane potential of DA neurons from hESCs line on HFFs feeder were lower than those of DA neurons from hESCs line on the MFCs feeder. In conclusion, HFFs feeder not only facilitated the differentiation of hESCs cells into dopaminergic neurons, but also induced hESCs-derived DA neurons to express higher electrophysiological excitability. Therefore, feeder cells could affect not only dopaminergic differentiation potential of different hESCs lines, but also electrophysiological properties of hESCs-derived DA neurons.

  20. Geological map of Uruguay Esc 1,100,000. Zapican Sheet F-22

    International Nuclear Information System (INIS)

    Preciozzi, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Zapican) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils.The fotoplano Zapican is located in the north of Lavalleja town and geologically is formed by Lavalleja group and the west socket represented by granites and volcanic reefs

  1. Smad, PI3K/Akt, and Wnt-dependent signaling pathways are involved in BMP-4-induced ESC self-renewal.

    Science.gov (United States)

    Lee, Min Young; Lim, Hyun Woo; Lee, Sang Hun; Han, Ho Jae

    2009-08-01

    It is known that bone morphogenetic protein 4 (BMP-4) has a diverse effect on ESCs. However, its precise mechanism in mouse ESCs is not fully understood. We evaluated the effect of BMP-4 on ESC proliferation and its related signal cascades in this study. BMP-4 significantly increased the level of [(3)H]-thymidine incorporation in time- (> or =8 hours) and dose- (> or =10 ng/ml) dependent manners. Additionally, BMP-4 increased cyclin D1 and decreased p27(kip1) expression values in a time-dependent manner. The increases in BMP-4-induced [(3)H]-thymidine incorporation and cyclin D1 expression were inhibited by the BMP-4 receptor antagonist noggin. BMP-4 increased Wnt1 expression. Wnt1 expression was attenuated by Smad4 small interfering RNA (siRNA), and BMP-4-induced cyclin D1 expression was inhibited by Smad4 and Wnt1 siRNAs. BMP-4 also activated beta-catenin, which was blocked by Smad4 and Wnt1 siRNAs. In addition, BMP-4 induced Akt phosphorylation. BMP-4-induced beta-catenin activation and cyclin D1 expression were attenuated by phosphatidyl inositol 3-kinase (PI3K) siRNA and Akt inhibitor. Additionally, downregulation of Smad4, Wnt1, and PI3K expression by siRNA decreased the levels of pluripotency marker mRNAs of ESCs, including Oct4, Sox2, and FoxD3. Our results suggested that BMP-4-induced [(3)H]-thymidine incorporation was significantly attenuated by Smad4, Wnt1, and PI3K knockdown. In conclusion, BMP-4 contributed to the maintenance of cell proliferation and the pluripotent state by Smad, PI3K/Akt, and Wnt1/beta-catenin in mouse ESCs.

  2. Geological map of Uruguay Esc 1,100,000. El Ombu Sheet N-18

    International Nuclear Information System (INIS)

    Ford, I

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (El Ombu) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the Cretaceous, Tertiary and Quaternary period in Arapey, Mercedes, Asencio, Palmitas, Fray Bentos and Libertad formations

  3. Geological map of Uruguay Esc 1,100,000. Cerro Partido Sheet F-24

    International Nuclear Information System (INIS)

    Diaz, R; Albanell, H.; Bossi, J.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Cerro Partido) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils. The area corresponding to the Cerro Partido fotoplano is located in the orogenic belt or Don Feliciano belt

  4. Avaliação da escória de siderurgia e do calcário como corretivos para cultura cafeeira

    Directory of Open Access Journals (Sweden)

    Bruno Manoel Rezende de Melo

    2013-04-01

    Full Text Available O objetivo deste trabalho foi avaliar a eficiência de correção da escória de siderurgia comparativamente ao calcário em função do tempo e profundidade. O experimento foi instalado no Instituto Federal de Educação, Ciência e Tecnologia do Sul de Minas Gerais, campus Muzambinho, em lavoura de café arábica da cultivar Catuai, implantada em  solo Latossolo Vermelho Amarelo (LVA, sendo que os corretivos foram aplicados em função da elevação da saturação de bases para 60% (V%. O delineamento experimental foi o de blocos casualizados com parcelas subdivididas no tempo, com quatro repetições, considerando os seguintes tratamentos: testemunha (sem calcário ou escória, com calcário e escória, sendo as subparcelas amostradas nos respectivos tempos e profundidade: 30 dias (0 a 0,1m, 60 dias (0 a 0,1m e 0,1m a 0,2m e 90 dias (0 a 0,1m e 0,1m a 0,2m. Os resultados obtidos demonstraram que os corretivos não diferiram entre si. Conclui-se, portanto, que a escória de siderurgia apresentou potencial de uso na cafeicultura comparativamente ao calcário na profundidade de 0 a 10 cm, apresentando resultado significativo para o tempo 30 dias após aplicação dos corretivos, entretanto, a exemplo do calcário, a escória não corrigiu o solo na profundidade de 10 a 20 cm.

  5. Geological map of Uruguay Esc 1,100,000. Melo Sheet D-15

    International Nuclear Information System (INIS)

    Ferrando, L.; Andreis, R.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Melo) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the pre devonian period in Melo, Buena Vista Yaguari and Tres islas formations. These metamorphic rocks would be compared with the orogenic cycle of the east and southeast groups

  6. Maturation of human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in 3D collagen matrix: Effects of niche cell supplementation and mechanical stimulation.

    Science.gov (United States)

    Zhang, W; Kong, C W; Tong, M H; Chooi, W H; Huang, N; Li, R A; Chan, B P

    2017-02-01

    Cardiomyocytes derived from human embryonic stem cells (hESC-CMs) are regarded as a promising source for regenerative medicine, drug testing and disease modeling. Nevertheless, cardiomyocytes are immature in terms of their contractile structure, metabolism and electrophysiological properties. Here, we fabricate cardiac muscle strips by encapsulating hESC-CMs in collagen-based biomaterials. Supplementation of niche cells at 3% to the number of hESC-CMs enhance the maturation of the hESC-CMs in 3D tissue matrix. The benefits of adding mesenchymal stem cells (MSCs) are comparable to that of adding fibroblasts. These two cell types demonstrate similar effects in promoting the compaction and cell spreading, as well as expression of maturation markers at both gene and protein levels. Mechanical loading, particularly cyclic stretch, produces engineered cardiac tissues with higher maturity in terms of twitch force, elastic modulus, sarcomere length and molecular signature, when comparing to static stretch or non-stretched controls. The current study demonstrates that the application of niche cells and mechanical stretch both stimulate the maturation of hESC-CMs in 3D architecture. Our results therefore suggest that this 3D model can be used for in vitro cardiac maturation study. Cardiomyocytes derived from human embryonic stem cells (hESC-CMs) are regarded as being a promising source of cells for regenerative medicine, drug testing and disease modeling. Nevertheless, cardiomyocytes are immature in terms of their contractile structure, metabolism and electrophysiological properties. In the current study, we have fabricated cardiac muscle strips by encapsulating hESC-CMs in collagen-based biomaterials and demonstrated that supplementation of mesenchymal niche cells as well as provision of mechanical loading particularly stretching have significantly promoted the maturation of the cardiomyocytes and hence improved the mechanical functional characteristics of the tissue strips

  7. Geological map of Uruguay Esc 1,100,000. Guaycuru Sheet M-24

    International Nuclear Information System (INIS)

    Garat, I.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Guaycuru)) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the Cenozoic and Precambrian period. The Cuaycuru area is located in the West of the Rio de la Plata socket constituted by metamorphic belts and is associated with migmatitic and intrusive granitoids

  8. Geological map of Uruguay Esc 1,100,000. Piraraja Sheet F-23

    International Nuclear Information System (INIS)

    Preciozzi, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Piraraja) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to Cenozoic, Cretaceous, Jurassic and Precambrian period.The Pirafja fotoplano is located in Lavalleja and geomorphologically is constituted by the Lavalleja group, the Atlantic socket, Piraraja granite and the Barriga Negra group

  9. Geological map of Uruguay Esc 1,100,000. La Union Sheet J-29

    International Nuclear Information System (INIS)

    Goso, C.; Veroslavsky, G.; Oyantcabal, P.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (La Union) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to the Holocene Pleistocene, Pliocene, Oligocene and Precambrian period. This area is located in the eastern of Montevideo and the South West of Canelones

  10. Comparing ESC and iPSC?Based Models for Human Genetic Disorders

    OpenAIRE

    Halevy, Tomer; Urbach, Achia

    2014-01-01

    Traditionally, human disorders were studied using animal models or somatic cells taken from patients. Such studies enabled the analysis of the molecular mechanisms of numerous disorders, and led to the discovery of new treatments. Yet, these systems are limited or even irrelevant in modeling multiple genetic diseases. The isolation of human embryonic stem cells (ESCs) from diseased blastocysts, the derivation of induced pluripotent stem cells (iPSCs) from patients’ somatic cells, and the ne...

  11. Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    Directory of Open Access Journals (Sweden)

    Rao Mahendra S

    2009-06-01

    Full Text Available Abstract Background A unique and essential property of embryonic stem cells is the ability to self-renew and differentiate into multiple cell lineages. However, the possible differences in proliferation and differentiation capabilities among independently-derived human embryonic stem cells (hESCs are not well known because of insufficient characterization. To address this question, a side-by-side comparison of 1 the ability to maintain an undifferentiated state and to self-renew under standard conditions; 2 the ability to spontaneously differentiate into three primary embryonic germ lineages in differentiating embryoid bodies; and 3 the responses to directed neural differentiation was made between three NIH registered hES cell lines I3 (TE03, I6 (TE06 and BG01V. Lines I3 and I6 possess normal XX and a normal XY karyotype while BG01V is a variant cell line with an abnormal karyotype derived from the karyotypically normal cell line BG01. Results Using immunocytochemistry, flow cytometry, qRT-PCR and MPSS, we found that all three cell lines actively proliferated and expressed similar "stemness" markers including transcription factors POU5F1/Oct3/4 and NANOG, glycolipids SSEA4 and TRA-1-81, and alkaline phosphatase activity. All cell lines differentiated into three embryonic germ lineages in embryoid bodies and into neural cell lineages when cultured in neural differentiation medium. However, a profound variation in colony morphology, growth rate, BrdU incorporation, and relative abundance of gene expression in undifferentiated and differentiated states of the cell lines was observed. Undifferentiated I3 cells grew significantly slower but their differentiation potential was greater than I6 and BG01V. Under the same neural differentiation-promoting conditions, the ability of each cell line to differentiate into neural progenitors varied. Conclusion Our comparative analysis provides further evidence for similarities and differences between three hESC

  12. El presidente Mauricio Macri y los Panama Papers. Periodismo, justicia y política entre las denuncias y el escándalo

    Directory of Open Access Journals (Sweden)

    Jorge Andrés Stefoni

    2016-09-01

    Full Text Available El presente artículo aborda la magnitud y las limitaciones del “escándalo” producido alrededor del presidente argentino Mauricio Macri tras las revelaciones de los Panama Papers. Desde una perspectiva sociológico-pragmática, se recorrerán los marcos de situación generados por las denuncias, se evaluarán las intervenciones desde sus exigencias pragmáticas de justificación y se analizarán las legitimidades variables de los distintos actores involucrados en la dinámica denuncia-escándalo. Analizando dimensiones como I la articulación de la denuncia y la percepción del escándalo, II las características de la denuncia, III las particularidades del denunciado, IV el lugar de los medios de comunicación, V la movilización de la denuncia por parte de los distintos actores y VI las formas de legitimación de los denunciantes, se buscará explicar cuáles fueron las condiciones de felicidad del escándalo y por qué el develamiento estuvo lejos de corresponderse con las expectativas de sus impulsores. Como conclusión sostiene que fueron las características de la denuncia periodística y las condiciones contextuales en el ámbito periodístico-mediático, judicial y político las que atenuaron la denuncia, les imposibilitaron a varios actores acreditarse como intérpretes legítimos y terminaron por disminuir la magnitud del escándalo.

  13. Síndrome do aprisionamento fascial do nervo torácico longo: escápula alada

    Directory of Open Access Journals (Sweden)

    Jefferson Braga Silva

    2015-10-01

    Full Text Available resumo Objetivo: Analisar os resultados de cirurgia de intervenção precoce em pacientes com sín drome do aprisionamento fascial do nervo torácico longo e consequente escápula alada. Métodos: Acompanhamos seis pacientes com uma síndrome de aprisionamento sem restricões específicas de estiramento ao nervo. Resultados: Pacientes tiveram melhoria em seus sintomas seis a 20 meses após o proce dimento. Sintomas motores melhoraram completamente sem qualquer dor persistente. A deformidade medial da escápula alada melhorou em todos os casos sem distúrbios estéticos residuais. Conclusão: A abordagem de liberação cirúrgica precoce parece ser um melhor preditor na recuperação de paralisia não traumática do músculo serrátil anterior.

  14. Geological map of Uruguay Esc 1,100,000. Paso del Palmar Sheet N-19

    International Nuclear Information System (INIS)

    Ford, I.; Montana, J.; Morales, H.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Paso del Palmar) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils belong to Cretaceous, Tertiary and Quaternary period in San Gregorio, Arapey, Mercedes, Asencio and Fray Bentos formations

  15. Geological map of Uruguay Esc 1,100,000. Cuchilla del Ombu. Sheet H-12

    International Nuclear Information System (INIS)

    Montana, J.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Cuchilla del Ombu) and the explanatory memoranda which describes the geological, lithological and sedimentological characteristics soils. In crystalline rocks have been recognized four basic units: porphyritic granite, Cunapiru granite, Cunapiru subvolcanic microgranite and metamorfites

  16. A guide for estimating dynamic panel models: the macroeconomics models specifiness; Una guida per la stima dei modelli panel dinamici: le specificita dei modelli macroeconomici

    Energy Technology Data Exchange (ETDEWEB)

    Coletta, Gaetano [ENEA, Centro Ricerche Casaccia, Rome (Italy). Unita di Agenzia per lo Sviluppo Sostenibile Advisor

    2005-10-15

    The aim of this paper is to review estimators for dynamic panel data models, a topic in which the interest has grown recently. As a consequence 01 this late interest, different estimation techniques have been proposed in the last few years and, given the last development of the subject, there is still a lack 01 a comprehensive guide for panel data applications, and for macroeconomics panel data models in particular. Finally, we also provide some indications about the Stata software commands to estimate dynamic panel data models with the techniques illustrated in the paper. [Italian] L'obiettivo del presente lavoro e quello di effettuare una rassegna degli stimatori per modelli panel dinamici, una materia per cui l'interesse e cresciuto di recente. In conseguenza di questo crescente interesse, negli ultimi anni sono stati proposti una serie di differenti tecniche di stima ma, dato il rapido sviluppo della materia, non esiste, infatti, una guida esaustiva per le applicazioni su dati panel, in particolare nel caso di modelli macroeconomici. A conclusione del lavoro sono fornite, infine, alcune indicazioni relative ai comandi del software Stata per stimare i modelli panel dinamici con le tecniche illustrate nel testo.

  17. The Induction Effect of Am80 and TSA on ESC Differentiation via Regulation of Stra8 in Chicken.

    Science.gov (United States)

    Zhang, Yani; Zuo, Qisheng; Liu, Zhiyong; Li, Dong; Tang, Beibei; Xiao, Tian-Rong; Lian, Chao; Wang, Yingjie; Jin, Kai; Wang, Yilin; Zhang, Wenhui; Li, Bichun

    2015-01-01

    Stra8 encodes stimulated by retinoic acid gene 8, a protein that is important for initiation of meiosis in mammals and birds. This study was aimed at identifying the active control area of chicken STRA8 gene core promoter, to screen optimum inducers of the STRA8 gene, thus to enhance the differentiation of embryonic stem cells (ESCs) into spermatogonial stem cells. Fragments of chicken STRA8 gene promoter were cloned into fluorescent reporter plasmids and transfected into DF-1 cells. Then Dual-Luciferase® Reporter Assay System was used to identify the activity of the STRA8 gene under different inducers. Our studies showed that the promoter fragment -1055 bp to +54 bp of Suqin chicken Stra8 revealed the strongest activity. The dual-luciferase® reporter showed that Tamibarotene (Am80) and TrichostatinA (TSA) could significantly enhance STRA8 transcription. The in vitro inductive culture of chicken ESCs demonstrated that spermatogonial stem cells (SSC)-like cells appeared and Integrinβ1 protein was expressed on day 10, indicating that Am80 and TSA can promote ESCs differentiation into SSCs via regulation of Stra8.

  18. App per l’infanzia: linee guida per una progettazione efficace

    Directory of Open Access Journals (Sweden)

    Silvia Carbotti

    2015-04-01

    Full Text Available I bambini sono affascinati dai tablet e dai dispositivi mobili di ultima generazione. Allo stesso modo si trovano in una fase della vita in cui iniziano a formare preferenze e competenze così come ad esprimere le proprie emozioni e sensazioni attraverso la parola. Questo articolo descrive le strategie per la creazione di applicazioni (app per bambini che siano non solo divertenti ma anche efficaci per esperienze significative che li coinvolgano a livello cognitivo ed emotivo. Quando si disegnano app rivolte ai bambini occorre ricordare che i più piccoli non hanno ancora chiaro come dovrebbe funzionare una specifica tecnologia ma si stanno formando un’opinione a riguardo. Diventa dunque necessario considerare alcuni principi di interaction e visual design affinché l’esperienza vissuta con il tablet incontri bisogni e competenze.

  19. Derivation of NEM2 affected human embryonic stem cell line Genea079

    Directory of Open Access Journals (Sweden)

    Biljana Dumevska

    2016-03-01

    Full Text Available The Genea079 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying compound heterozygous mutations in the NEB gene, exon 55 deletion & c.15110dupA, indicative of Nemaline Myopathy Type 2 (NEM2. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male Allele pattern. The hESC line had pluripotent cell morphology, 86% of cells expressed Nanog, 95% Oct4, 54% Tra1-60 and 98% SSEA4 and gave a PluriTest Pluripotency score of 30.25, Novelty of 1.21. The cell line was negative for Mycoplasma and visible contamination.

  20. Simultaneous suppression of TGF-β and ERK signaling contributes to the highly efficient and reproducible generation of mouse embryonic stem cells from previously considered refractory and non-permissive strains.

    Science.gov (United States)

    Hassani, Seyedeh-Nafiseh; Totonchi, Mehdi; Farrokhi, Ali; Taei, Adeleh; Larijani, Mehran Rezaei; Gourabi, Hamid; Baharvand, Hossein

    2012-06-01

    Mouse embryonic stem cells (ESCs) are pluripotent stem cell lines derived from pre-implantation embryos. The efficiency of mESC generation is affected by genetic variation in mice; that is, some mouse strains are refractory or non-permissive to ESC establishment. Developing an efficient method to derive mESCs from strains of various genetic backgrounds should be valuable for establishment of ESCs in various mammalian species. In the present study, we identified dual inhibition of TGF-β and ERK1/2, by SB431542 and PD0325901, respectively led to the highly efficient and reproducible generation of mESC lines from NMRI, C57BL/6, BALB/c, DBA/2, and FVB/N strains, which previously considered refractory or non-permissive for ESC establishment. These mESCs expressed pluripotency markers and retained the capacity to differentiate into derivatives of all three germ layers. The evaluated lines exhibited high rates of chimerism when reintroduced into blastocysts. To our knowledge, this is the first report of efficient (100%) mESC lines generation from different genetic backgrounds. The application of these two inhibitors will not only solve the problems of mESC derivation but also clarifies new signaling pathways in pluripotent mESCs.

  1. Geological map of Uruguay Esc 1,100,000. Bizcocho Sheet N-0-21

    International Nuclear Information System (INIS)

    Ford, I; Gancio, F

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Bizcocho) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics of the lower Proterozoic in the socket of the La Plata river, Upper Cretaceous in Mercedes and Asencio formations, lower Tertiary in Palmitas and Fray Bentos formations and soils characteristics of the Quaternary period

  2. The effects of studded tires on fatal crashes with passenger cars and the benefits of electronic stability control (ESC) in Swedish winter driving.

    Science.gov (United States)

    Strandroth, Johan; Rizzi, Matteo; Olai, Maria; Lie, Anders; Tingvall, Claes

    2012-03-01

    This study set out to examine the effects of studded tires on fatal crashes on roads covered with ice or snow in Sweden and also to investigate the extra benefits of electronic stability control (ESC) during the winter months. Two different studies are presented in this paper. Both studies used an induced exposure approach. In the main study, 369 in-depth studies of fatal crashes with passenger cars were analyzed to determine whether loss-of-control (LOC) had been a major component or not. Only crashes involving cars without ESC and equipped with approved studded or non-studded winter tires were analyzed. The additional study used police-reported crashes that occurred during the winter seasons 2003-2010, involving passenger cars with and without ESC. While police records in Sweden do not include any tire information, it was assumed that most cars involved in crashes during the winter period would be equipped with studded tires. Findings in the main study showed that in 64% of the fatal crashes on roads covered with ice or snow LOC had been a major component. Furthermore, in 82% of LOC crashes, the passenger car over-steered prior to collision. Studded tires were found to have a statistically significant effect of 42% in terms of fatal crash reduction on roads covered with ice or snow, compared to non-studded winter tires. The effect on dry or wet roads in the winter was negative, although statistically non-significant. In the additional study, it was found that ESC further reduced crashes with injuries by 29%. The benefits on severe and fatal crashes were slightly greater (32%), although the lower 95% confidence limit was lower. Although studded tires were shown to reduce the risk of fatal crash involvement, compared to non-studded winter tires, the proportion of LOC and over-steering among cars with studded tires was large (59% and 49%, respectively). It was therefore concluded that studded tires do not prevent all LOC crashes, while ESC has benefits in those

  3. Correlation study of actual temperature profile and in-line metrology measurements for within-wafer uniformity improvement and wafer edge yield enhancement (Conference Presentation)

    Science.gov (United States)

    Fang, Fang; Vaid, Alok; Vinslava, Alina; Casselberry, Richard; Mishra, Shailendra; Dixit, Dhairya; Timoney, Padraig; Chu, Dinh; Porter, Candice; Song, Da; Ren, Zhou

    2018-03-01

    It is getting more important to monitor all aspects of influencing parameters in critical etch steps and utilize them as tuning knobs for within-wafer uniformity improvement and wafer edge yield enhancement. Meanwhile, we took a dive in pursuing "measuring what matters" and challenged ourselves for more aspects of signals acquired in actual process conditions. Among these factors which are considered subtle previously, we identified Temperature, especially electrostatic chuck (ESC) Temperature measurement in real etch process conditions have direct correlation to in-line measurements. In this work, we used SensArray technique (EtchTemp-SE wafer) to measure ESC temperature profile on a 300mm wafer with plasma turning on to reproduce actual temperature pattern on wafers in real production process conditions. In field applications, we observed substantial correlation between ESC temperature and in-line optical metrology measurements and since temperature is a process factor that can be tuning through set-temperature modulations, we have identified process knobs with known impact on physical profile variations. Furthermore, ESC temperature profile on a 300mm wafer is configured as multiple zones upon radius and SensArray measurements mechanism could catch such zonal distribution as well, which enables detailed temperature modulations targeting edge ring only where most of chips can be harvested and critical zone for yield enhancement. Last but not least, compared with control reference (ESC Temperature in static plasma-off status), we also get additional factors to investigate in chamber-to-chamber matching study and make process tool fleet match on the basis really matters in production. KLA-Tencor EtchTemp-SE wafer enables Plasma On wafer temperature monitoring of silicon etch process. This wafer is wireless and has 65 sensors with measurement range from 20 to 140°C. the wafer is designed to run in real production recipe plasma on condition with maximum RF power up

  4. Management of atrial fibrillation around the world: a comparison of current ACCF/AHA/HRS, CCS, and ESC guidelines.

    Science.gov (United States)

    Wasmer, Kristina; Eckardt, Lars

    2011-10-01

    New guidelines for the management of atrial fibrillation (AF) have recently been published by the American College of Cardiology Foundation/American Heart Association, and Heart Rhythm Society (ACCF/AHA/HRS) task force on practice guidelines, the Canadian Cardiovascular Society (CCS), and the European Society of Cardiology (ESC). Although they all refer to the same scientific data and agree in the majority of AF management, interpretation, and weighing of study results are quite different in some aspects. While recommendations for stroke risk assessment and prophylaxis are rather conservative in the ESC guidelines, the CCS guideline recommendations are more conservative with regard to lenient rate control and the ACCF/AHA/HRS recommendations are rather strict with regard to rhythm management.

  5. Efeito residual da escória de siderurgia como corretivo de acidez do solo na soqueira de cana-de-açúcar

    Directory of Open Access Journals (Sweden)

    Prado R. M.

    2003-01-01

    Full Text Available A escória de siderurgia, como material corretivo e efeito residual prolongado, pode beneficiar culturas de ciclo longo, a exemplo da cana-de-açúcar, minimizando a queda de produção ao longo do ciclo produtivo. Este trabalho objetivou avaliar diferentes níveis de saturação por bases, utilizando, como corretivo do solo, a escória de siderurgia, comparando-a com calcário calcítico, nas alterações de alguns atributos químicos do solo, bem como na resposta da soqueira da cana-de-açúcar. Para isto, realizou-se um experimento com a variedade SP 80-1842, durante o terceiro e o quarto corte, nos anos agrícolas 2000/01 e 2001/02. Os tratamentos, dispostos em blocos casualizados, em esquema fatorial com quatro repetições, constaram de duas fontes de corretivos, calcário calcítico e escória de siderurgia, e quatro níveis de correção, estimados pelo método da saturação por bases (V %: testemunha (sem correção e com correção para V % de 50; 75 e 100, tendo sido tais corretivos aplicados na época do plantio da cana-de-açúcar. O calcário calcítico e a escória de siderurgia promoveram efeito residual benéfico, após 48 meses da aplicação, na correção da acidez do solo e na elevação do valor da saturação por bases; a maior dose de calcário causou efeito depressivo no perfilhamento, no número de colmos industrializáveis e na produção da cana-de-açúcar, fato não observado com uso da escória de siderurgia; a aplicação da escória de siderurgia e do calcário, em pré-plantio, promoveu efeito residual positivo na produção da soqueira de cana-de-açúcar.

  6. A 3-dimensional human embryonic stem cell (hESC)-derived model to detect developmental neurotoxicity of nanoparticles.

    Science.gov (United States)

    Hoelting, Lisa; Scheinhardt, Benjamin; Bondarenko, Olesja; Schildknecht, Stefan; Kapitza, Marion; Tanavde, Vivek; Tan, Betty; Lee, Qian Yi; Mecking, Stefan; Leist, Marcel; Kadereit, Suzanne

    2013-04-01

    Nanoparticles (NPs) have been shown to accumulate in organs, cross the blood-brain barrier and placenta, and have the potential to elicit developmental neurotoxicity (DNT). Here, we developed a human embryonic stem cell (hESC)-derived 3-dimensional (3-D) in vitro model that allows for testing of potential developmental neurotoxicants. Early central nervous system PAX6(+) precursor cells were generated from hESCs and differentiated further within 3-D structures. The 3-D model was characterized for neural marker expression revealing robust differentiation toward neuronal precursor cells, and gene expression profiling suggested a predominantly forebrain-like development. Altered neural gene expression due to exposure to non-cytotoxic concentrations of the known developmental neurotoxicant, methylmercury, indicated that the 3-D model could detect DNT. To test for specific toxicity of NPs, chemically inert polyethylene NPs (PE-NPs) were chosen. They penetrated deep into the 3-D structures and impacted gene expression at non-cytotoxic concentrations. NOTCH pathway genes such as HES5 and NOTCH1 were reduced in expression, as well as downstream neuronal precursor genes such as NEUROD1 and ASCL1. FOXG1, a patterning marker, was also reduced. As loss of function of these genes results in severe nervous system impairments in mice, our data suggest that the 3-D hESC-derived model could be used to test for Nano-DNT.

  7. SOX2 co-occupies distal enhancer elements with distinct POU factors in ESCs and NPCs to specify cell state.

    Directory of Open Access Journals (Sweden)

    Michael A Lodato

    Full Text Available SOX2 is a master regulator of both pluripotent embryonic stem cells (ESCs and multipotent neural progenitor cells (NPCs; however, we currently lack a detailed understanding of how SOX2 controls these distinct stem cell populations. Here we show by genome-wide analysis that, while SOX2 bound to a distinct set of gene promoters in ESCs and NPCs, the majority of regions coincided with unique distal enhancer elements, important cis-acting regulators of tissue-specific gene expression programs. Notably, SOX2 bound the same consensus DNA motif in both cell types, suggesting that additional factors contribute to target specificity. We found that, similar to its association with OCT4 (Pou5f1 in ESCs, the related POU family member BRN2 (Pou3f2 co-occupied a large set of putative distal enhancers with SOX2 in NPCs. Forced expression of BRN2 in ESCs led to functional recruitment of SOX2 to a subset of NPC-specific targets and to precocious differentiation toward a neural-like state. Further analysis of the bound sequences revealed differences in the distances of SOX and POU peaks in the two cell types and identified motifs for additional transcription factors. Together, these data suggest that SOX2 controls a larger network of genes than previously anticipated through binding of distal enhancers and that transitions in POU partner factors may control tissue-specific transcriptional programs. Our findings have important implications for understanding lineage specification and somatic cell reprogramming, where SOX2, OCT4, and BRN2 have been shown to be key factors.

  8. A murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells

    NARCIS (Netherlands)

    C. Buecker (Christa); H.H. Chen; J.M. Polo (Jose); L. Daheron (Laurence); L. Bu (Lei); T.S. Barakat (Tahsin Stefan); P. Okwieka (Patricia); A. Porter (Andrew); J.H. Gribnau (Joost); K. Hochedlinger (Konrad); N. Geijsen (Niels)

    2010-01-01

    textabstractMurine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human

  9. Generation of iPSC line iPSC-FH2.1 in hypoxic conditions from human foreskin fibroblasts

    Directory of Open Access Journals (Sweden)

    María Questa

    2016-03-01

    Full Text Available Human foreskin fibroblasts were used to generate the iPSC line iPSC-FH2.1 using the EF1a-hSTEMCCA-loxP vector expressing OCT4, SOX2, c-MYC and KLF4, in 5% O2 culture conditions. Stemness was confirmed, as was pluripotency both in vivo and in vitro, in normoxia and hypoxia. Human Embryonic Stem Cell (hESC line WA-09 and reprogrammed fibroblast primary culture HFF-FM were used as controls.

  10. Survival and Functionality of hESC-Derived Retinal Pigment Epithelium Cells Cultured as a Monolayer on Polymer Substrates Transplanted in RCS Rats.

    Science.gov (United States)

    Thomas, Biju B; Zhu, Danhong; Zhang, Li; Thomas, Padmaja B; Hu, Yuntao; Nazari, Hossein; Stefanini, Francisco; Falabella, Paulo; Clegg, Dennis O; Hinton, David R; Humayun, Mark S

    2016-05-01

    To determine the safety, survival, and functionality of human embryonic stem cell-derived RPE (hESC-RPE) cells seeded on a polymeric substrate (rCPCB-RPE1 implant) and implanted into the subretinal (SR) space of Royal College of Surgeons (RCS) rats. Monolayers of hESC-RPE cells cultured on parylene membrane were transplanted into the SR space of 4-week-old RCS rats. Group 1 (n = 46) received vitronectin-coated parylene membrane without cells (rMSPM+VN), group 2 (n = 59) received rCPCB-RPE1 implants, and group 3 (n = 13) served as the control group. Animals that are selected based on optical coherence tomography screening were subjected to visual function assays using optokinetic (OKN) testing and superior colliculus (SC) electrophysiology. At approximately 25 weeks of age (21 weeks after surgery), the eyes were examined histologically for cell survival, phagocytosis, and local toxicity. Eighty-seven percent of the rCPCB-RPE1-implanted animals showed hESC-RPE survivability. Significant numbers of outer nuclear layer cells were rescued in both group 1 (rMSPM+VN) and group 2 (rCPCB-RPE1) animals. A significantly higher ratio of rod photoreceptor cells to cone photoreceptor cells was found in the rCPCB-RPE1-implanted group. Animals with rCPCB-RPE1 implant showed hESC-RPE cells containing rhodopsin-positive particles in immunohistochemistry, suggesting phagocytic function. Superior colliculus mapping data demonstrated that a significantly higher number of SC sites responded to light stimulus at a lower luminance threshold level in the rCPCB-RPE1-implanted group. Optokinetic data suggested both implantation groups showed improved visual acuity. These results demonstrate the safety, survival, and functionality of the hESC-RPE monolayer transplantation in an RPE dysfunction rat model.

  11. Aproveitamento de escória de siderurgia em substratos alternativos para produção de mudas de pimenteira Dedo-de-moça

    Directory of Open Access Journals (Sweden)

    Eduardo France Oza

    Full Text Available RESUMO Estudos relacionados com o aproveitamento de escória de siderurgia tornaram-se imprescindíveis, pela possibilidade de seu uso como componente de substratos na produção de mudas e por esse uso minimizar o risco de contaminação ambiental, decorrente do descarte inadequado. Assim, objetivou-se, com este trabalho, avaliar o efeito de níveis crescentes de escória de siderurgia em substratos, compostos por solo com esterco bovino curtido e areia, nas variáveis de crescimento e na qualidade de mudas de pimenteira dedo-de-moça (Capsicum baccatum L.. Utilizou-se o delineamento em blocos casualizados, tendo como tratamentos cinco concentrações de escória (0%; 2,5%; 5%; 10% e; 20% e quatro repetições. As avaliações ocorreram aos 55 dias após semeadura, consistindo na contagem do número de folhas, na medição da altura de planta e do diâmetro do coleto, na quantificação das massas secas de folhas e raízes da planta e na determinação do Índice de Qualidade de Dickson. Modelos de regressão foram ajustados (P < 0,05 aos tratamentos com níveis crescentes de escória de siderurgia. A adição em torno de 10% de escória ao substrato proporcionou os maiores valores das variáveis de crescimento, em mudas de pimenteira Dedo-de-moça.

  12. Risk prediction in stable cardiovascular disease using a high-sensitivity cardiac troponin T single biomarker strategy compared to the ESC-SCORE.

    Science.gov (United States)

    Biener, Moritz; Giannitsis, Evangelos; Kuhner, Manuel; Zelniker, Thomas; Mueller-Hennessen, Matthias; Vafaie, Mehrshad; Stoyanov, Kiril M; Neumann, Franz-Josef; Katus, Hugo A; Hochholzer, Willibald; Valina, Christian Marc

    2018-01-01

    To evaluate the prognostic performance of high-sensitivity cardiac troponin T (hs-cTnT) compared with the ESC-SCORE. We included low-risk outpatients with stable cardiovascular (CV) disease categorised into need for non-secondary and secondary prevention. The prognostication of hs-cTnT at index visit was compared with the European Society of Cardiology-Systematic COronary Risk Evaluation (ESC-SCORE) with respect to all-cause mortality (ACM) and two composite endpoints (ACM, acute myocardial infarction (AMI) and stroke and ACM, AMI, stroke and rehospitalisation for acute coronary syndrome (ACS) and decompensated heart failure (DHF)). Within a median follow-up of 796 days, a total of 16 deaths, 32 composite endpoints of ACM, AMI and stroke and 83 composite endpoints of ACM, AMI, stroke, rehospitalisation for ACS and DHF were observed among 693 stable low-risk outpatients. Using C-statistics, measurement of hs-cTnT alone outperformed the ESC-SCORE for the prediction of ACM in the entire study population (Δarea under the curve (AUC) 0.221, p=0.0039) and both prevention groups (non-secondary: ΔAUC 0.164, p=0.0208; secondary: ΔAUC 0.264, p=0.0134). For the prediction of all other secondary endpoints, hs-cTnT was at least as effective as the ESC-SCORE, both in secondary and non-secondary prevention. Using continuous and categorical net reclassification improvement and integrated discrimination improvement, hs-cTnT significantly improved reclassification regarding all endpoints in the entire population and in the secondary prevention cohort. In non-secondary prevention, hs-cTnT improved reclassification only for ACM. The results were confirmed in an independent external cohort on 2046 patients. Hs-cTnT is superior to the multivariable ESC-SCORE for the prediction of ACM and a composite endpoint in stable outpatients with and without relevant CV disease. NCT01954303; Pre-results.

  13. Notas sobre o escândalo e o cinismo de Manet

    Directory of Open Access Journals (Sweden)

    Tereza Calomeni

    2013-10-01

    Full Text Available Em 1971, Foucault profere, em Túnis, uma conferência sobre a pintura de Manet. Em 1984, em A coragem da verdade, último curso oferecido no Collège de France, analisando o cinismo como expressão da parresia, novamente se refere ao pintor ao relacionar arte moderna e cinismo. A partir dos dois textos, o artigo reflete sobre o escândalo de Olympia, uma das treze telas examinadas na conferência, e a relação, estabelecida no curso, entre arte moderna e cinismo.

  14. As gramáticas morais da corrupção: aportes para uma sociologia do escândalo

    Directory of Open Access Journals (Sweden)

    Edmilson Lopes Júnior

    2010-12-01

    Full Text Available Como analisar sociologicamente as práticas de corrupção? Quais aportes teóricos podem ser mobilizados para uma apreensão não essencialista dos escândalos de corrupção? Este artigo aborda estas questões combinando os aportes teóricos da sociologia econômica e da sociologia da moral. A ilustração empírica é dada pelo chamado "Escândalo do Mensalão", acontecimento que marcou a vida política brasileira na última década. Por fim, faço indicações para uma análise sociológica relacional das práticas de corrupção no Brasil.

  15. Generation of a Nrf2 homozygous knockout human embryonic stem cell line using CRISPR/Cas9

    Directory of Open Access Journals (Sweden)

    So-Jung Kim

    2017-03-01

    Full Text Available Nuclear factor erythroid 2-related factor 2 (NFE2L2 or Nrf2 is a well-known transcription factor that regulates the expression of a large number of anti-oxidant genes in mammalian cells (J.H. Kim et al., 2014. Here, we generated a homozygous Nrf2 knockout human embryonic stem cell (hESC line, H9Nrf2KO-A13, using the CRISPR/Cas9 genome editing method. The Nrf2 homozygous knockout H9 cell line maintains pluripotency, differentiation potential into three germ layers, and a normal karyotype.

  16. EZ spheres: a stable and expandable culture system for the generation of pre-rosette multipotent stem cells from human ESCs and iPSCs

    OpenAIRE

    Ebert, A.; Shelley, B.; Hurley, A.; Onorati, M.; Castiglioni, V.; Patitucci, T.; Svendsen, S.; Mattis, V.; Mcgivern, J.; Schwab, A.; Sareen, D.; Kim, H.; Cattaneo, E.; Svendsen, C.

    2013-01-01

    We have developed a simple method to generate and expand multipotent, self-renewing pre-rosette neural stem cells from both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs) without utilizing embryoid body formation, manual selection techniques, or complex combinations of small molecules. Human ESC and iPSC colonies were lifted and placed in a neural stem cell medium containing high concentrations of EGF and FGF-2. Cell aggregates (termed EZ spheres) could be...

  17. Comparison of Gene Expression in Human Embryonic Stem Cells, hESC-Derived Mesenchymal Stem Cells and Human Mesenchymal Stem Cells.

    Science.gov (United States)

    Barbet, Romain; Peiffer, Isabelle; Hatzfeld, Antoinette; Charbord, Pierre; Hatzfeld, Jacques A

    2011-01-01

    We present a strategy to identify developmental/differentiation and plasma membrane marker genes of the most primitive human Mesenchymal Stem Cells (hMSCs). Using sensitive and quantitative TaqMan Low Density Arrays (TLDA) methodology, we compared the expression of 381 genes in human Embryonic Stem Cells (hESCs), hESC-derived MSCs (hES-MSCs), and hMSCs. Analysis of differentiation genes indicated that hES-MSCs express the sarcomeric muscle lineage in addition to the classical mesenchymal lineages, suggesting they are more primitive than hMSCs. Transcript analysis of membrane antigens suggests that IL1R1(low), BMPR1B(low), FLT4(low), LRRC32(low), and CD34 may be good candidates for the detection and isolation of the most primitive hMSCs. The expression in hMSCs of cytokine genes, such as IL6, IL8, or FLT3LG, without expression of the corresponding receptor, suggests a role for these cytokines in the paracrine control of stem cell niches. Our database may be shared with other laboratories in order to explore the considerable clinical potential of hES-MSCs, which appear to represent an intermediate developmental stage between hESCs and hMSCs.

  18. Gene expression profile and functionality of ESC-derived Lin-ckit+Sca-1+ cells are distinct from Lin-ckit+Sca-1+ cells isolated from fetal liver or bone marrow.

    Directory of Open Access Journals (Sweden)

    Irina Fernandez

    Full Text Available In vitro bioreactor-based cultures are being extensively investigated for large-scale production of differentiated cells from embryonic stem cells (ESCs. However, it is unclear whether in vitro ESC-derived progenitors have similar gene expression profiles and functionalities as their in vivo counterparts. This is crucial in establishing the validity of ESC-derived cells as replacements for adult-isolated cells for clinical therapies. In this study, we compared the gene expression profiles of Lin-ckit+Sca-1+ (LKS cells generated in vitro from mouse ESCs using either static or bioreactor-based cultures, with that of native LKS cells isolated from mouse fetal liver (FL or bone marrow (BM. We found that in vitro-generated LKS cells were more similar to FL- than to BM LKS cells in gene expression. Further, when compared to cells derived from bioreactor cultures, static culture-derived LKS cells showed fewer differentially expressed genes relative to both in vivo LKS populations. Overall, the expression of hematopoietic genes was lower in ESC-derived LKS cells compared to cells from BM and FL, while the levels of non-hematopoietic genes were up-regulated. In order to determine if these molecular profiles correlated with functionality, we evaluated ESC-derived LKS cells for in vitro hematopoietic-differentiation and colony formation (CFU assay. Although static culture-generated cells failed to form any colonies, they did differentiate into CD11c+ and B220+ cells indicating some hematopoietic potential. In contrast, bioreactor-derived LKS cells, when differentiated under the same conditions failed to produce any B220+ or CD11c+ cells and did not form colonies, indicating that these cells are not hematopoietic progenitors. We conclude that in vitro culture conditions significantly affect the transcriptome and functionality of ESC-derived LKS cells and although in vitro differentiated LKS cells were lineage negative and expressed both ckit and Sca-1

  19. Ethanol negatively regulates hepatic differentiation of hESC by inhibition of the MAPK/ERK signaling pathway in vitro.

    Directory of Open Access Journals (Sweden)

    Wei Gao

    Full Text Available Alcohol insult triggers complex events in the liver, promoting fibrogenic/inflammatory signals and in more advanced cases, aberrant matrix deposition. It is well accepted that the regenerative capacity of the adult liver is impaired during alcohol injury. The liver progenitor/stem cells have been shown to play an important role in liver regeneration -in response to various chronic injuries; however, the effects of alcohol on stem cell differentiation in the liver are not well understood.We employed hepatic progenitor cells derived from hESCs to study the impact of ethanol on hepatocyte differentiation by exposure of these progenitor cells to ethanol during hepatocyte differentiation.We found that ethanol negatively regulated hepatic differentiation of hESC-derived hepatic progenitor cells in a dose-dependent manner. There was also a moderate cell cycle arrest at G1/S checkpoint in the ethanol treated cells, which is associated with a reduced level of cyclin D1 in these cells. Ethanol treatment specifically inhibited the activation of the ERK but not JNK nor the p38 MAP signaling pathway. At the same time, the WNT signaling pathway was also reduced in the cells exposed to ethanol. Upon evaluating the effects of the inhibitors of these two signaling pathways, we determined that the Erk inhibitor replicated the effects of ethanol on the hepatocyte differentiation and attenuated the WNT/β-catenin signaling, however, inhibitors of WNT only partially replicated the effects of ethanol on the hepatocyte differentiation.Our results demonstrated that ethanol negatively regulated hepatic differentiation of hESC-derived hepatic progenitors through inhibiting the MAPK/ERK signaling pathway, and subsequently attenuating the WNT signaling pathway. Thus, our finding provides a novel insight into the mechanism by which alcohol regulates cell fate selection of hESC-derived hepatic progenitor cells, and the identified pathways may provide therapeutic targets

  20. Derivation and characterisation of hESC lines from supernumerary embryos, experience from Odense, Denmark

    DEFF Research Database (Denmark)

    Harkness, Linda; Rasmussen, Iben Anne; Erb, Karin

    2010-01-01

    The derivation and characterisation of human embryonic stem cells provides a source of pluripotent stem cells with potential for clinical applications. Utilising locally sourced embryos from two IVF clinics, we derived and characterised five new cell lines for use in a non-clinical setting. Analy...

  1. Geological map of Uruguay Esc 1,100,000. Fuente del Puma Sheet G-27

    International Nuclear Information System (INIS)

    Preciozzi, F.; Pena, S.

    1990-01-01

    This work is about the geological map of Uruguay Esc.1.100.000 (Fuente del Puma) and the explanatory memoranda which describes the geological , lithological and sedimentological characteristics soils. The area corresponding to Fuente del Puma is located in the SW of Lavalleja and NW of Maldonado town and its stratigraphy belong to the Cretaceous and Cenozoic formations as well as the Cambrian and upper Precambrian

  2. CCL19/CCR7 contributes to the pathogenesis of endometriosis via PI3K/Akt pathway by regulating the proliferation and invasion of ESCs.

    Science.gov (United States)

    Diao, Ruiying; Wei, Weixia; Zhao, Jinghui; Tian, Fuying; Cai, Xueyong; Duan, Yong-Gang

    2017-11-01

    The level of CCL19 increased in the peritoneal fluid of women with endometriosis, but the precise mechanism of CCL19/CCR7 in the pathogenesis of endometriosis remains unknown. ELISA and immunohistochemistry were performed to analyze CCL19/CCR7 expressions in peritoneal fluid and endometrium from women with endometriosis (n = 38) and controls (n = 32). Cell proliferation and transwell invasion assays were applied to detect proliferation and invasion of human endometrial stromal cells (ESCs). Expressions of Bcl2, MMP2, MMP9, and p-AKT/AKT were analyzed by Western blot. Peritoneal fluid concentration of CCL19 in patients with endometriosis was higher than that in controls. Those patients with moderate/severe endometriosis had significantly higher peritoneal fluid concentrations of CCL19 compared to those with minimal/mild endometriosis. Higher CCL19 and CCR7 were found in the endometrium with endometriosis compared to control. CCL19 significantly enhanced ESC proliferation and invasion through CCR7 via activating PI3K/Akt signal pathways. CCL19/CCR7 interaction significantly enhanced phosphorylation of Akt, Bcl2, MMP2, and MMP9 in ESCs. These data indicate CCL19/CCR7 contributes to proliferation and invasion of ESCs, which are conducive to the pathogenesis of endometriosis through activating PI3K/Akt pathway. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. Striking similarity in the gene expression levels of individual Myc module members among ESCs, EpiSCs, and partial iPSCs.

    Directory of Open Access Journals (Sweden)

    Masataka Hirasaki

    Full Text Available Predominant transcriptional subnetworks called Core, Myc, and PRC modules have been shown to participate in preservation of the pluripotency and self-renewality of embryonic stem cells (ESCs. Epiblast stem cells (EpiSCs are another cell type that possesses pluripotency and self-renewality. However, the roles of these modules in EpiSCs have not been systematically examined to date. Here, we compared the average expression levels of Core, Myc, and PRC module genes between ESCs and EpiSCs. EpiSCs showed substantially higher and lower expression levels of PRC and Core module genes, respectively, compared with those in ESCs, while Myc module members showed almost equivalent levels of average gene expression. Subsequent analyses revealed that the similarity in gene expression levels of the Myc module between these two cell types was not just overall, but striking similarities were evident even when comparing the expression of individual genes. We also observed equivalent levels of similarity in the expression of individual Myc module genes between induced pluripotent stem cells (iPSCs and partial iPSCs that are an unwanted byproduct generated during iPSC induction. Moreover, our data demonstrate that partial iPSCs depend on a high level of c-Myc expression for their self-renewal properties.

  4. Single-Cell Analyses of ESCs Reveal Alternative Pluripotent Cell States and Molecular Mechanisms that Control Self-Renewal

    Directory of Open Access Journals (Sweden)

    Dmitri Papatsenko

    2015-08-01

    Full Text Available Analyses of gene expression in single mouse embryonic stem cells (mESCs cultured in serum and LIF revealed the presence of two distinct cell subpopulations with individual gene expression signatures. Comparisons with published data revealed that cells in the first subpopulation are phenotypically similar to cells isolated from the inner cell mass (ICM. In contrast, cells in the second subpopulation appear to be more mature. Pluripotency Gene Regulatory Network (PGRN reconstruction based on single-cell data and published data suggested antagonistic roles for Oct4 and Nanog in the maintenance of pluripotency states. Integrated analyses of published genomic binding (ChIP data strongly supported this observation. Certain target genes alternatively regulated by OCT4 and NANOG, such as Sall4 and Zscan10, feed back into the top hierarchical regulator Oct4. Analyses of such incoherent feedforward loops with feedback (iFFL-FB suggest a dynamic model for the maintenance of mESC pluripotency and self-renewal.

  5. Prevalence of extended-spectrum cephalosporinase (ESC)-producing Escherichia coli in Danish slaughter pigs and retail meat identified by selective enrichment and association with cephalosporin usage

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Aarestrup, Frank Møller; Pedersen, Karl

    2012-01-01

    OBJECTIVES: To investigate the prevalence of extended-spectrum cephalosporinase (ESC)-producing Escherichia coli in pigs at slaughter and retail meat, and possible associations with the consumption of third- and fourth-generation cephalosporins. METHODS: During 2009, faecal samples from Danish pigs...... (n = 786) were collected at slaughter, and 866 meat samples [Danish: pork (153), broiler meat (121) and beef (142); and imported: pork (173), broiler meat (193) and beef (84)] were randomly collected in retail stores and outlets. E. coli was isolated after enrichment in MacConkey broth.......7%–3.3% in other meat types. ESC E. coli from imported broiler meat (n = 69) contained blaCMY-2 (48%), blaCTX-M-1 (25%) and blaSHV-12 (16%). Without selective enrichment, no ESC E. coli from pigs and only 4.1% from imported broiler meat were found. CONCLUSIONS: The usage of cephalosporins for slaughter pigs may...

  6. Compósitos de cimento - borracha de pneus: efeito da escória nas propriedades Tire rubber-cement composites: effect of slag on properties

    Directory of Open Access Journals (Sweden)

    N. Segre

    2006-12-01

    Full Text Available Compósitos de cimento-borracha de pneus preparados com cimentos tipo I e contendo adições de três diferentes escórias foram estudados. Resistência à flexão, absorção de água e resistência ao ataque ácido foram investigadas. Um decréscimo no módulo de ruptura (MOR foi observado para todos os corpos de prova contendo borracha, quando comparados com os corpos de prova sem borracha (controles. Para as pastas controle e com borracha e argamassas controle, preparadas com cimento com escória, o MOR aumenta com o aumento da atividade hidráulica da escória utilizada, até valores indistinguíveis dos obtidos para as argamassas e pastas preparadas com cimento tipo I. Argamassas com borracha apresentaram valores de MOR independentes do cimento utilizado. Uma redução na sorção de água é observada para corpos de prova controle e contendo borracha, preparados com todos os cimentos contendo adição de escória, quando comparados com os corpos de prova preparados com cimento tipo I. Para corpos de prova de argamassa com borracha, quanto mais baixa é a basicidade da escória, menor é a sorção de água dos compósitos. Estes resultados indicam baixa porosidade e consequentemente uma melhor adesão entre a borracha e a matriz de cimento nestes corpos de prova. Além disso, uma velocidade menor de sorção de água é observada para os corpos de prova com borracha, com destaque para os corpos de prova com cimento que contém a escória menos básica, quando comparados com os controles. Resultados de ataque ácido aos corpos e prova de argamassas, preparadas com os cimentos contendo adição de escória, mostram que a susceptibilidade dos corpos de prova frente à degradação por ácido é governada não somente por aspectos microestruturais, como porosidade e permeabilidade, mas também por aspectos químicos, como diferenças na quantidade de álcalis presentes nas escórias ou quantidade de escória não reagida nos corpos de prova

  7. Resposta da cana-de-açúcar à aplicação da escória de siderurgia como corretivo de acidez do solo

    Directory of Open Access Journals (Sweden)

    R. M. Prado

    2001-03-01

    Full Text Available A acidez elevada dos solos da região sudeste e centro-oeste do Brasil predomina onde se concentra a maior área cultivada com cana-de-açúcar. Grandes depósitos de escória de siderurgia, que contêm nutrientes como cálcio e magnésio e apresentam ação corretiva de acidez, localizam-se também naquelas regiões. O presente trabalho objetivou avaliar diferentes níveis de saturação por bases, utilizando como corretivo do solo a escória de siderurgia, comparando-a com calcário calcítico, nas alterações de alguns atributos químicos do solo, bem como na resposta da cana-de-açúcar, durante os dois primeiros cortes. Para isto, foi realizado um experimento com a cana-de-açúcar, variedade SP 80-1842, nos anos agrícolas 98/99 e 99/00. Os tratamentos, dispostos em blocos casualizados em esquema fatorial com quatro repetições, constaram de duas fontes de corretivos, calcário calcítico e escória de siderurgia, e quatro níveis de correção, estimados pelo método da saturação por bases (V%: testemunha sem correção e correção para V% de 50, 75 e 100. Concluiu-se que os efeitos da aplicação do calcário e da escória de siderurgia foram semelhantes na correção da acidez do solo, na elevação da concentração de cálcio e magnésio do solo e na saturação por bases. As doses de calcário proporcionaram efeito quadrático sobre a produtividade e perfilhamento da cana-de-açúcar, enquanto as de escória resultaram em resposta linear.

  8. La Validez y la Eficacia de los Ejercicios Respiratorios para Reducir la Ansiedad Escénica en el Aula de Música

    Directory of Open Access Journals (Sweden)

    Pablo Ramos Ramos

    2013-06-01

    Full Text Available El presente texto repasa la literatura científica más relevante en el ámbito de la ansiedad escénica asociada a la interpretación musical, tanto en edad adulta como durante la infancia. Además, se exponen las conclusiones de aquellos estudios que han analizado este rasgo de la personalidad en el medio escolar y se destacan las causas que lo provocan: presión ante la presencia de padres y de otros alumnos, perfeccionismo, baja resistencia al estrés.Por otra parte, se relaciona el desarrollo de la ansiedad escénica con la pérdida de autoestima, lo que repercute en la educación del niño a un nivel general. Ante esta problemática se discuten las posibles terapias que podrían implementarse en el aula; de entre todas ellas se destacan –por su eficacia y por la facilidad a la hora de aplicarlas– las técnicas de respiración.Finalmente, se establece un decálogo a modo de guía de actuación para combatir la ansiedad escénica, que el maestro o profesor de música puede tener en cuenta de cara a una primera intervención en el medio escolar.

  9. Boa noite, e boa sorte: determinantes da demissão de ministros envolvidos em escândalos de corrupção no primeiro governo Dilma Rousseff

    Directory of Open Access Journals (Sweden)

    Cletiane Medeiros Araújo

    2016-04-01

    Full Text Available Resumo Quais os determinantes para demissão de ministros? Este artigo analisa a onda de escândalos de corrupção envolvendo ministros do primeiro governo Dilma Rousseff. O desenho de pesquisa combina estatística descritiva e análise multivariada para testar as hipóteses de que notícias sobre escândalos políticos, competição política e gerenciamento da coalizão impactam positivamente na probabilidade de o ministro denunciado deixar a pasta ministerial. O estudo foi realizado a partir da construção de uma base de dados, para a qual foram coletadas 677 notícias de três jornais de larga circulação nacional, a saber: Folha de S. Paulo, O Estado de S. Paulo e O Globo, ao longo dos 17 primeiros meses do governo. Os resultados apontam que quanto maior o número de matérias sobre os escândalos de corrupção do ministro denunciado maior a probabilidade de ele ser demitido. Espera-se, através deste estudo de caso, contribuir para a agenda de pesquisa sobre a relação escândalos políticos/instituições democráticas/accountability, buscando situar esse fenômeno na democracia brasileira recente.

  10. Thrombo-embolism and antithrombotic therapy for heart failure in sinus rhythm. A joint consensus document from the ESC Heart Failure Association and the ESC Working Group on Thrombosis.

    Science.gov (United States)

    Lip, Gregory Y H; Ponikowski, Piotr; Andreotti, Felicita; Anker, Stefan D; Filippatos, Gerasimos; Homma, Shunichi; Morais, Joao; Pullicino, Patrick; Rasmussen, Lars H; Marin, Francisco; Lane, Deirdre A

    2012-07-01

    Chronic heart failure (HF) with either reduced or preserved ejection fraction is common and remains an extremely serious disorder with a high mortality and morbidity. Many complications related to HF can be related to thrombosis. Epidemiological and pathophysiological data also link HF to an increased risk of thrombosis, leading to the clinical consequences of sudden death, stroke, systemic thrombo-embolism, and/or venous thrombo-embolism. This consensus document of the Heart Failure Association (EHFA) of the European Society of Cardiology (ESC) and the ESC Working Group on Thrombosis reviews the published evidence and summarizes 'best practice', and puts forward consensus statements that may help to define evidence gaps and assist management decisions in everyday clinical practice. In HF patients with atrial fibrillation, oral anticoagulation is recommended, and the CHA(2)DS(2)-VASc and HAS-BLED scores should be used to determine the likely risk-benefit ratio (thrombo-embolism prevention vs. risk of bleeding) of oral anticoagulation. In HF patients with reduced left ventricular ejection fraction who are in sinus rhythm there is no evidence of an overall benefit of vitamin K antagonists (e.g. warfarin) on mortality, with risk of major bleeding. Despite the potential for a reduction in ischaemic stroke, there is currently no compelling reason to use warfarin routinely for these patients. Risk factors associated with increased risk of thrombo-embolic events should be identified and decisions regarding use of anticoagulation individualized. Patient values and preferences are important determinants when balancing the risk of thrombo-embolism against bleeding risk. New oral anticoagulants that offer a different risk-benefit profile compared with warfarin may appear as an attractive therapeutic option, but this would need to be confirmed in clinical trials.

  11. Avaliação da escória de siderurgia e de calcários como corretivos da acidez do solo no cultivo da alface

    Directory of Open Access Journals (Sweden)

    Prado Renato de Mello

    2002-01-01

    Full Text Available O presente trabalho objetivou avaliar os efeitos da escória de siderurgia e dos calcários magnesiano e dolomítico, na produção de matéria seca da alface (Lactuca sativa L. e as alterações na sua nutrição e em alguns atributos químicos do solo. Os tratamentos consistiram dos corretivos calcário magnesiano e dolomítico e escória de siderurgia, em três doses: 2,36, 4,72 e 7,08 g por vaso. Utilizou-se o delineamento experimental inteiramente casualizado, com quatro repetições, em esquema fatorial 3x3+1 (três corretivos x três doses + testemunha. Todos os materiais corretivos aumentaram a produção de matéria seca da alface, porém destacaram-se os calcários, sendo que para a escória, a alta concentração do Mn na parte aérea limitou a produção. Na parte aérea da alface, os teores e quantidade acumulada de Ca, Mg e Cu aumentaram, e os teores de Mn e Zn não foram alterados com a aplicação dos corretivos. Todos os materiais corretivos foram eficientes na correção da acidez do solo, especialmente quando se utilizou calcário magnesiano. A escória de siderurgia não é o corretivo mais apropriado para a cultura da alface.

  12. Sourcing human embryos for embryonic stem cell lines: Problems & perspectives

    Directory of Open Access Journals (Sweden)

    Rajvi H Mehta

    2014-01-01

    Full Text Available The ability to successfully derive human embryonic stem cells (hESC lines from human embryos following in vitro fertilization (IVF opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been ′discarded′ or ′spare′ fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. In case a couple does not desire to ′cryopreserve′ their embryos then all the embryos remaining following embryo transfer can be considered ′spare′ or if a couple is no longer in need of the ′cryopreserved′ embryos then these also can be considered as ′spare′. But, the question raised by the ethicists is, "what about ′slightly′ over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to ′discarded′ embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of ′discarding′ embryos. What would be the criteria for discarding embryos and the potential ′use′ of ESC derived from the ′abnormal appearing′ embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material.

  13. Comparative study of human embryonic stem cells (hESC and human induced pluripotent stem cells (hiPSC as a treatment for retinal dystrophies

    Directory of Open Access Journals (Sweden)

    Marina Riera

    2016-01-01

    Full Text Available Retinal dystrophies (RD are major causes of familial blindness and are characterized by progressive dysfunction of photoreceptor and/or retinal pigment epithelium (RPE cells. In this study, we aimed to evaluate and compare the therapeutic effects of two pluripotent stem cell (PSC-based therapies. We differentiated RPE from human embryonic stem cells (hESCs or human-induced pluripotent stem cells (hiPSCs and transplanted them into the subretinal space of the Royal College of Surgeons (RCS rat. Once differentiated, cells from either source of PSC resembled mature RPE in their morphology and gene expression profile. Following transplantation, both hESC- and hiPSC-derived cells maintained the expression of specific RPE markers, lost their proliferative capacity, established tight junctions, and were able to perform phagocytosis of photoreceptor outer segments. Remarkably, grafted areas showed increased numbers of photoreceptor nuclei and outer segment disk membranes. Regardless of the cell source, human transplants protected retina from cell apoptosis, glial stress and accumulation of autofluorescence, and responded better to light stimuli. Altogether, our results show that hESC- and hiPSC-derived cells survived, migrated, integrated, and functioned as RPE in the RCS rat retina, providing preclinical evidence that either PSC source could be of potential benefit for treating RD.

  14. Comparative study of human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) as a treatment for retinal dystrophies

    Science.gov (United States)

    Riera, Marina; Fontrodona, Laura; Albert, Silvia; Ramirez, Diana Mora; Seriola, Anna; Salas, Anna; Muñoz, Yolanda; Ramos, David; Villegas-Perez, Maria Paz; Zapata, Miguel Angel; Raya, Angel; Ruberte, Jesus; Veiga, Anna; Garcia-Arumi, Jose

    2016-01-01

    Retinal dystrophies (RD) are major causes of familial blindness and are characterized by progressive dysfunction of photoreceptor and/or retinal pigment epithelium (RPE) cells. In this study, we aimed to evaluate and compare the therapeutic effects of two pluripotent stem cell (PSC)-based therapies. We differentiated RPE from human embryonic stem cells (hESCs) or human-induced pluripotent stem cells (hiPSCs) and transplanted them into the subretinal space of the Royal College of Surgeons (RCS) rat. Once differentiated, cells from either source of PSC resembled mature RPE in their morphology and gene expression profile. Following transplantation, both hESC- and hiPSC-derived cells maintained the expression of specific RPE markers, lost their proliferative capacity, established tight junctions, and were able to perform phagocytosis of photoreceptor outer segments. Remarkably, grafted areas showed increased numbers of photoreceptor nuclei and outer segment disk membranes. Regardless of the cell source, human transplants protected retina from cell apoptosis, glial stress and accumulation of autofluorescence, and responded better to light stimuli. Altogether, our results show that hESC- and hiPSC-derived cells survived, migrated, integrated, and functioned as RPE in the RCS rat retina, providing preclinical evidence that either PSC source could be of potential benefit for treating RD. PMID:27006969

  15. Susceptibility of Human Oral Squamous Cell Carcinoma (OSCC H103 and H376 cell lines to Retroviral OSKM mediated reprogramming

    Directory of Open Access Journals (Sweden)

    Nalini Devi Verusingam

    2017-04-01

    Full Text Available Although numbers of cancer cell lines have been shown to be successfully reprogrammed into induced pluripotent stem cells (iPSCs, reprogramming Oral Squamous Cell Carcinoma (OSCC to pluripotency in relation to its cancer cell type and the expression pattern of pluripotent genes under later passage remain unexplored. In our study, we reprogrammed and characterised H103 and H376 oral squamous carcinoma cells using retroviral OSKM mediated method. Reprogrammed cells were characterized for their embryonic stem cells (ESCs like morphology, pluripotent gene expression via quantitative real-time polymerase chain reaction (RT-qPCR, immunofluorescence staining, embryoid bodies (EB formation and directed differentiation capacity. Reprogrammed H103 (Rep-H103 exhibited similar ESCs morphologies with flatten cells and clear borders on feeder layer. Reprogrammed H376 (Rep-H376 did not show ESCs morphologies but grow with a disorganized morphology. Critical pluripotency genes Oct4, Sox2 and Nanog were expressed higher in Rep-H103 against the parental counterpart from passage 5 to passage 10. As for Rep-H376, Nanog expression against its parental counterpart showed a significant decrease at passage 5 and although increased in passage 10, the level of expression was similar to the parental cells. Rep-H103 exhibited pluripotent signals (Oct4, Sox2, Nanog and Tra-1-60 and could form EB with the presence of three germ layers markers. Rep-H103 displayed differentiation capacity into adipocytes and osteocytes. The OSCC cell line H103 which was able to be reprogrammed into an iPSC like state showed high expression of Oct4, Sox2 and Nanog at late passage and may provide a potential iPSC model to study multi-stage oncogenesis in OSCC.

  16. Naked regulators: moral pluralism, deliberative democracy and authoritative regulation of human embryonic stem cell (hESC) research.

    Science.gov (United States)

    Parker, Malcolm

    2009-02-01

    Bioethical issues pose challenges for pluralist, democratic societies due to the need to arbitrate between incompatible views over fundamental beliefs. The legitimacy of public policy is increasingly seen to depend on taking public consultation seriously, and subsequently regulating contested activities such as therapeutic cloning and hESC research. In December 2006, the Australian Federal Parliament lifted the ban on therapeutic cloning, following recommendations of the Legislation Review Committee (Lockhart Committee), which recently reported on its approach and methods in this journal. This column analyses recent accounts of democratic deliberative processes, authoritative regulation and the committee's own account. Authoritative regulation turns out to be largely an appeasement strategy, directed towards the losers of the contest, in this case the opponents of therapeutic cloning and hESC research. This is because regulation fails to minimise harm as perceived by the losers, and fails to meaningfully limit what it is the winners wish to do. Moreover, regulation adds an unnecessary layer of red tape to the work of the winners. Committees of inquiry in bioethical matters should be more open about their processes and their normative recommendations, at the risk of eroding trust in parts of their processes.

  17. FOXN1GFP/w Reporter hESCs Enable Identification of Integrin-β4, HLA-DR, and EpCAM as Markers of Human PSC-Derived FOXN1+ Thymic Epithelial Progenitors

    Directory of Open Access Journals (Sweden)

    Chew-Li Soh

    2014-06-01

    Full Text Available Thymic epithelial cells (TECs play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1GFP/w line as a readout, we developed a reproducible protocol for generating FOXN1-GFP+ thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-β4 (ITGB4, CD104 and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1+ TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1+ TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.

  18. Heat shock instructs hESCs to exit from the self-renewal program through negative regulation of OCT4 by SAPK/JNK and HSF1 pathway.

    Science.gov (United States)

    Byun, Kyunghee; Kim, Taek-Kyun; Oh, Jeehyun; Bayarsaikhan, Enkhjargal; Kim, Daesik; Lee, Min Young; Pack, Chan-Gi; Hwang, Daehee; Lee, Bonghee

    2013-11-01

    Environmental factors affect self-renewal of stem cells by modulating the components of self-renewal networks. Heat shock, an environmental factor, induces heat shock factors (HSFs), which up-regulate stress response-related genes. However, the link of heat shock to self-renewal of stem cells has not been elucidated yet. Here, we present the direct link of heat shock to a core stem cell regulator, OCT4, in the self-renewal network through SAPK/JNK and HSF1 pathway. We first showed that heat shock initiated differentiation of human embryonic stem cells (hESCs). Gene expression analysis revealed that heat shock increased the expression of many genes involved in cellular processes related to differentiation of stem cells. We then examined the effects of HSFs induced by heat shock on core self-renewal factors. Among HSFs, heat shock induced mainly HSF1 in hESCs. The HSF1 repressed the expression of OCT4, leading to the differentiation of hESCs and the above differentiation-related gene expression change. We further examined the effects of the upstream MAP (mitogen-activated protein) kinases of HSF1 on the repression of OCT4 expression by HSF1. Among the MAP kinases, SAPK/JNK controlled predominantly the repression of the OCT4 expression by HSF1. The direct link of heat shock to the core self-renewal regulator through SAPK/JNK and HSF1 provides a fundamental basis for understanding the effect of heat and other stresses involving activation of HSF1 on the self-renewal program and further controlling differentiation of hESCs in a broad spectrum of stem cell applications using these stresses. © 2013.

  19. Escória de siderurgia e calcário na correção da acidez do solo cultivado com cana-de-açúcar em vaso

    Directory of Open Access Journals (Sweden)

    Prado Renato de Mello

    2000-01-01

    Full Text Available A avaliação química da escória de siderurgia, como corretivo de acidez do solo, pode sofrer interferências em razão da presença da alta energia de ligação dos seus constituintes neutralizantes e a presença de diversos elementos metálicos. Tendo como objetivo avaliar se a recomendação de correção baseada no poder de neutralização adotado para o calcário é compatível para escória de siderurgia, em função das alterações do valor pH, teores de H+Al, Ca+Mg em solos ácidos da região dos cerrados cultivados com cana-de-açúcar, realizou-se o presente experimento, em condições de casa de vegetação, em vaso com 20 dm³ do Latossolo Vermelho e do Neossolo Quartzarênico em dois cultivos sucessivos da cana-de-açúcar (cana-planta e cana-soca por 210 dias após a incorporação dos corretivos, a cada cultivo. Os tratamentos, foram constituídos de dois corretivos calcário e escória em dois níveis de aplicação, como segue: nível 1= a dose para elevar V=50% e nível 2= dobro da dose necessária para elevar V=50%. Ao término de cada cultivo, os solos foram amostrados e analisados quimicamente. A reatividade da escória de siderurgia depende da classe de solo. A eficiência da escória de siderurgia baseado no poder de neutralização adotado para o calcário não apresentou comportamento satisfatório para estimar a necessidade de produto para a correção da acidez do solo, sugerindo a necessidade de mais estudos.

  20. Mivoq Evalita 2016 PosTwITA tagger

    OpenAIRE

    Paci, Giulio

    2017-01-01

    The POS tagger developed by Mivoq to tag tweets according to PosTwITA task guidelines as defined at Evalita 2016 is presented. The system obtained third position with 92.7% of accuracy. Si presenta il POS tagger sviluppato da Mivoq per etichettare i tweet secondo le linee guida del task PosT-wITA, cos come definite per Evalita 2016. Il sistema ha ottenuto la terza posizione con un accuratezza del 92.7%.

  1. Characterization of Tetraploid Somatic Cell Nuclear Transfer-Derived Human Embryonic Stem Cells.

    Science.gov (United States)

    Shin, Dong-Hyuk; Lee, Jeoung-Eun; Eum, Jin Hee; Chung, Young Gie; Lee, Hoon Taek; Lee, Dong Ryul

    2017-12-01

    Polyploidy is occurred by the process of endomitosis or cell fusion and usually represent terminally differentiated stage. Their effects on the developmental process were mainly investigated in the amphibian and fishes, and only observed in some rodents as mammalian model. Recently, we have established tetraploidy somatic cell nuclear transfer-derived human embryonic stem cells (SCNT-hESCs) and examined whether it could be available as a research model for the polyploidy cells existed in the human tissues. Two tetraploid hESC lines were artificially acquired by reintroduction of remained 1st polar body during the establishment of SCNT-hESC using MII oocytes obtained from female donors and dermal fibroblasts (DFB) from a 35-year-old adult male. These tetraploid SCNT-hESC lines (CHA-NT1 and CHA-NT3) were identified by the cytogenetic genotyping (91, XXXY,-6, t[2:6] / 92,XXXY,-12,+20) and have shown of indefinite proliferation, but slow speed when compared to euploid SCNT-hESCs. Using the eight Short Tendem Repeat (STR) markers, it was confirmed that both CHA-NT1 and CHA-NT3 lines contain both nuclear and oocyte donor genotypes. These hESCs expressed pluripotency markers and their embryoid bodies (EB) also expressed markers of the three embryonic germ layers and formed teratoma after transplantation into immune deficient mice. This study showed that tetraploidy does not affect the activities of proliferation and differentiation in SCNT-hESC. Therefore, tetraploid hESC lines established after SCNT procedure could be differentiated into various types of cells and could be an useful model for the study of the polyploidy cells in the tissues.

  2. Perspectives on the 2014 ESC/EACTS Guidelines on Myocardial Revascularization: Fifty Years of Revascularization: Where Are We and Where Are We Heading?

    NARCIS (Netherlands)

    F. Costa (Francesco); S. Ariotti (Sara); M. Valgimigli (Marco); P.H. Kolh (Philippe); S.W. Windecker (Stephan)

    2015-01-01

    textabstractThe joint European Society of Cardiology and European Association of Cardio-Thoracic Surgery (ESC/EACTS) guidelines on myocardial revascularization collect and summarize the evidence regarding decision-making, diagnostics, and therapeutics in various clinical scenarios of coronary artery

  3. Adherence to the European Society of Cardiology (ESC) guidelines for chronic heart failure - A national survey of the cardiologists in Pakistan

    Science.gov (United States)

    2011-01-01

    Background The aims of this study were to evaluate the awareness of and attitudes towards the 2005 European Society of Cardiology (ESC) guidelines for Heart Failure (HF) of the cardiologists in Pakistan and assess barriers to adherence to guidelines. Methods A cross-sectional survey was conducted in person from March to July 2009 to all cardiologists practicing in 4 major cities in Pakistan (Karachi, Lahore, Quetta and Peshawar). A validated, semi-structured questionnaire assessing ESC 2005 Guidelines for HF was used to obtain information from cardiologists. It included questions about awareness and relevance of HF guidelines (See Additional File 1). Respondents' management choices were compared with those of an expert panel based on the guidelines for three fictitious patient cases. Cardiologists were also asked about major barriers to adherence to guidelines. Results A total of 372 cardiologists were approached; 305 consented to participate (overall response rate, 82.0%). The survey showed a very high awareness of CHF guidelines; 97.4% aware of any guideline. About 13.8% considered ESC guidelines as relevant or very relevant for guiding treatment decisions while 92.8% chose AHA guidelines in relevance. 87.2% of respondents perceived that they adhered to the HF guidelines. For the patient cases, the proportions of respondents who made recommendations that completely matched those of the guidelines were 7% (Scenario 1), 0% (Scenario 2) and 20% (Scenario 3). Respondents considered patient compliance (59%) and cost/health economics (50%) as major barriers to guideline implementation. Conclusion We found important self reported departures from recommended HF management guidelines among cardiologists of Pakistan. PMID:22093082

  4. Alianzas y redes de colaboración entre las agrupaciones culturales de las Artes Escénicas en Andalucía

    Directory of Open Access Journals (Sweden)

    I. Ramos

    2013-01-01

    Full Text Available el análisis de redes sociales se ha utilizado de manera efectiva en la evalua- ción de programas (eisenberg & swanson, 1996; Fujimoto, valente & Pentz, 2009, la gestión de información y la difusión de innovaciones (valente, 1995; Kraatz, 1998; ahuja, 2000, el estudio de la evolución de las coaliciones comu- nitarias (Feinberg, Riggs & greenberg, 2005; Wendel et al., 2010 y las iniciati- vas para promover la capacidad de las organizaciones (nowel & Foster-Fishman, 2010. en este estudio se analizan 6 tipos de relaciones entre 32 organizaciones culturales del sector de las artes escénicas en andalucía. las agrupaciones de las artes escénicas en andalucía conforman una estructura centro-periferia. el es- pacio central se articula en torno a una tríada formada por tres entidades jóvenes que destacan, respectivamente, por (a gestionar recursos de formación en artes escénicas, (b coordinar la asociación de los profesionales del sector y (c tener un perfil activo en la producción de espectáculos en los últimos años. a través del análisis de clúster han sido identificados cuatro conglomerados integrados por organizaciones que han desplegado diferentes estrategias relacionales y cuentan con características diferenciales. los resultados se devolvieron a repre- sentantes del sector en un grupo de discusión. se presentan las reacciones de los participantes.

  5. O perfil da relação profissional entre as empresas de serviços contábeis (ESC) e a micro e a pequena empresa (MPE)

    OpenAIRE

    Palma, Daniel Azevedo

    2005-01-01

    Neste trabalho, além do referencial teórico, envolvendo conceitos e classificações da MPE, e também da Contabilidade, realizou-se uma pesquisa de campo na cidade de Unaí -MG, em que se procurou identificar as principais características existentes na relação profissional entre as Empresas de Serviços Contábeis (ESC) e a Micro e Pequena Empresa (MPE) e verificar como as informações geradas pelas ESCs podem ser mais bem utilizadas na gestão das MPEs. Obteve-se uma amostra corroborando a caracter...

  6. Thromboembolism and antithrombotic therapy for heart failure in sinus rhythm: an executive summary of a joint consensus document from the ESC Heart Failure Association and the ESC Working Group on Thrombosis.

    Science.gov (United States)

    Lip, Gregory Y H; Piotrponikowski, Piotr; Andreotti, Felicita; Anker, Stefan D; Filippatos, Gerasimos; Homma, Shunichi; Morais, Joao; Pullicino, Patrick; Rasmussen, Lars H; Marín, Francisco; Lane, Deirdre A

    2012-12-01

    Chronic heart failure (HF) with either reduced or preserved left ventricular (LV) ejection fraction is common and remains an extremely serious disorder with a high mortality and morbidity. Many complications related to heart failure can be related to thrombosis. Epidemiological and pathophysiological data also link HF to an increased risk of thrombosis, leading to the clinical consequences of sudden death, stroke, systemic thromboembolism and/or venous thromboembolism. This executive summary of a joint consensus document of the Heart Failure Association (EHFA) of the European Society of Cardiology (ESC) and the ESC Working Group on Thrombosis reviews the published evidence, summarises 'best practice', and puts forward consensus statements that may help to define evidence gaps and assist management decisions in everyday clinical practice. In HF patients with atrial fibrillation, oral anticoagulation is clearly recommended, and the CHA2DS2-VASc and HAS-BLED scores should be used to determine the likely risk-benefit ratio (thromboembolism prevention versus risk of bleeding) of oral anticoagulation. In HF patients with reduced LV ejection fraction who are in sinus rhythm there is no evidence of an overall benefit of vitamin K antagonists (e.g. warfarin) on mortality, with risk of major bleeding. Whilst there is the potential for a reduction in ischaemic stroke, there is currently no compelling reason to routinely use warfarin for these patients. Risk factors associated with increased risk of thromboembolic events should be identified and decisions regarding use of anticoagulation individualised. Patient values and preferences are important determinants when balancing the risk of thromboembolism against bleeding risk. Novel oral anticoagulants that offer a different risk-benefit profile compared with warfarin may appear as an attractive therapeutic option, but this would need to be confirmed in clinical trials.

  7. Comparison of a teratogenic transcriptome-based predictive test based on human embryonic versus inducible pluripotent stem cells.

    Science.gov (United States)

    Shinde, Vaibhav; Perumal Srinivasan, Sureshkumar; Henry, Margit; Rotshteyn, Tamara; Hescheler, Jürgen; Rahnenführer, Jörg; Grinberg, Marianna; Meisig, Johannes; Blüthgen, Nils; Waldmann, Tanja; Leist, Marcel; Hengstler, Jan Georg; Sachinidis, Agapios

    2016-12-30

    Human embryonic stem cells (hESCs) partially recapitulate early embryonic three germ layer development, allowing testing of potential teratogenic hazards. Because use of hESCs is ethically debated, we investigated the potential for human induced pluripotent stem cells (hiPSCs) to replace hESCs in such tests. Three cell lines, comprising hiPSCs (foreskin and IMR90) and hESCs (H9) were differentiated for 14 days. Their transcriptome profiles were obtained on day 0 and day 14 and analyzed by comprehensive bioinformatics tools. The transcriptomes on day 14 showed that more than 70% of the "developmental genes" (regulated genes with > 2-fold change on day 14 compared to day 0) exhibited variability among cell lines. The developmental genes belonging to all three cell lines captured biological processes and KEGG pathways related to all three germ layer embryonic development. In addition, transcriptome profiles were obtained after 14 days of exposure to teratogenic valproic acid (VPA) during differentiation. Although the differentially regulated genes between treated and untreated samples showed more than 90% variability among cell lines, VPA clearly antagonized the expression of developmental genes in all cell lines: suppressing upregulated developmental genes, while inducing downregulated ones. To quantify VPA-disturbed development based on developmental genes, we estimated the "developmental potency" (D p ) and "developmental index" (D i ). Despite differences in genes deregulated by VPA, uniform D i values were obtained for all three cell lines. Given that the D i values for VPA were similar for hESCs and hiPSCs, D i can be used for robust hazard identification, irrespective of whether hESCs or hiPSCs are used in the test systems.

  8. Dal CAD al GIS: il trasferimento di informazioni cartografiche

    Directory of Open Access Journals (Sweden)

    Andrea Scianna

    2005-10-01

    necessario verificare quali siano le informazioni che rimangono associate agli oggetti della classe al momento del caricamento dei dati, in formato DXF, all’interno di software GIS. Negli ultimi anni a livello internazionale sono state fissate norme tendenti a individuare linee guida per la definizione della struttura del contenuto semantico della cartografia numerica, delle caratteristiche geometriche e delle relazioni topologiche dell’informazione geografica.

  9. H3S10ph broadly marks early-replicating domains in interphase ESCs and shows reciprocal antagonism with H3K9me2.

    Science.gov (United States)

    Chen, Carol C L; Goyal, Preeti; Karimi, Mohammad M; Abildgaard, Marie H; Kimura, Hiroshi; Lorincz, Matthew C

    2018-01-01

    Phosphorylation of histone H3 at serine 10 (H3S10ph) by Aurora kinases plays an important role in mitosis; however, H3S10ph also marks regulatory regions of inducible genes in interphase mammalian cells, implicating mitosis-independent functions. Using the fluorescent ubiquitin-mediated cell cycle indicator (FUCCI), we found that 30% of the genome in interphase mouse embryonic stem cells (ESCs) is marked with H3S10ph. H3S10ph broadly demarcates gene-rich regions in G1 and is positively correlated with domains of early DNA replication timing (RT) but negatively correlated with H3K9me2 and lamin-associated domains (LADs). Consistent with mitosis-independent kinase activity, this pattern was preserved in ESCs treated with Hesperadin, a potent inhibitor of Aurora B/C kinases. Disruption of H3S10ph by expression of nonphosphorylatable H3.3S10A results in ectopic spreading of H3K9me2 into adjacent euchromatic regions, mimicking the phenotype observed in Drosophila JIL-1 kinase mutants . Conversely, interphase H3S10ph domains expand in Ehmt1 (also known as Glp ) null ESCs, revealing that H3S10ph deposition is restricted by H3K9me2. Strikingly, spreading of H3S10ph at RT transition regions (TTRs) is accompanied by aberrant transcription initiation of genes co-oriented with the replication fork in Ehmt1 -/- and Ehmt2 -/- ESCs, indicating that establishment of repressive chromatin on the leading strand following DNA synthesis may depend upon these lysine methyltransferases. H3S10ph is also anti-correlated with H3K9me2 in interphase murine embryonic fibroblasts (MEFs) and is restricted to intragenic regions of actively transcribing genes by EHMT2. Taken together, these observations reveal that H3S10ph may play a general role in restricting the spreading of repressive chromatin in interphase mammalian cells. © 2018 Chen et al.; Published by Cold Spring Harbor Laboratory Press.

  10. A new FACS approach isolates hESC derived endoderm using transcription factors.

    Directory of Open Access Journals (Sweden)

    Yuqiong Pan

    2011-03-01

    Full Text Available We show that high quality microarray gene expression profiles can be obtained following FACS sorting of cells using combinations of transcription factors. We use this transcription factor FACS (tfFACS methodology to perform a genomic analysis of hESC-derived endodermal lineages marked by combinations of SOX17, GATA4, and CXCR4, and find that triple positive cells have a much stronger definitive endoderm signature than other combinations of these markers. Additionally, SOX17(+ GATA4(+ cells can be obtained at a much earlier stage of differentiation, prior to expression of CXCR4(+ cells, providing an important new tool to isolate this earlier definitive endoderm subtype. Overall, tfFACS represents an advancement in FACS technology which broadly crosses multiple disciplines, most notably in regenerative medicine to redefine cellular populations.

  11. Adherence to the European Society of Cardiology (ESC guidelines for chronic heart failure - A national survey of the cardiologists in Pakistan

    Directory of Open Access Journals (Sweden)

    Shoukat Sana

    2011-11-01

    Full Text Available Abstract Background The aims of this study were to evaluate the awareness of and attitudes towards the 2005 European Society of Cardiology (ESC guidelines for Heart Failure (HF of the cardiologists in Pakistan and assess barriers to adherence to guidelines. Methods A cross-sectional survey was conducted in person from March to July 2009 to all cardiologists practicing in 4 major cities in Pakistan (Karachi, Lahore, Quetta and Peshawar. A validated, semi-structured questionnaire assessing ESC 2005 Guidelines for HF was used to obtain information from cardiologists. It included questions about awareness and relevance of HF guidelines (See Additional File 1. Respondents' management choices were compared with those of an expert panel based on the guidelines for three fictitious patient cases. Cardiologists were also asked about major barriers to adherence to guidelines. Additional file 1 Questionnaire. Description: Questionnaire that was administered to participants. Click here for file Results A total of 372 cardiologists were approached; 305 consented to participate (overall response rate, 82.0%. The survey showed a very high awareness of CHF guidelines; 97.4% aware of any guideline. About 13.8% considered ESC guidelines as relevant or very relevant for guiding treatment decisions while 92.8% chose AHA guidelines in relevance. 87.2% of respondents perceived that they adhered to the HF guidelines. For the patient cases, the proportions of respondents who made recommendations that completely matched those of the guidelines were 7% (Scenario 1, 0% (Scenario 2 and 20% (Scenario 3. Respondents considered patient compliance (59% and cost/health economics (50% as major barriers to guideline implementation. Conclusion We found important self reported departures from recommended HF management guidelines among cardiologists of Pakistan.

  12. An integrative analysis of reprogramming in human isogenic system identified a clone selection criterion.

    Science.gov (United States)

    Shutova, Maria V; Surdina, Anastasia V; Ischenko, Dmitry S; Naumov, Vladimir A; Bogomazova, Alexandra N; Vassina, Ekaterina M; Alekseev, Dmitry G; Lagarkova, Maria A; Kiselev, Sergey L

    2016-01-01

    The pluripotency of newly developed human induced pluripotent stem cells (iPSCs) is usually characterized by physiological parameters; i.e., by their ability to maintain the undifferentiated state and to differentiate into derivatives of the 3 germ layers. Nevertheless, a molecular comparison of physiologically normal iPSCs to the "gold standard" of pluripotency, embryonic stem cells (ESCs), often reveals a set of genes with different expression and/or methylation patterns in iPSCs and ESCs. To evaluate the contribution of the reprogramming process, parental cell type, and fortuity in the signature of human iPSCs, we developed a complete isogenic reprogramming system. We performed a genome-wide comparison of the transcriptome and the methylome of human isogenic ESCs, 3 types of ESC-derived somatic cells (fibroblasts, retinal pigment epithelium and neural cells), and 3 pairs of iPSC lines derived from these somatic cells. Our analysis revealed a high input of stochasticity in the iPSC signature that does not retain specific traces of the parental cell type and reprogramming process. We showed that 5 iPSC clones are sufficient to find with 95% confidence at least one iPSC clone indistinguishable from their hypothetical isogenic ESC line. Additionally, on the basis of a small set of genes that are characteristic of all iPSC lines and isogenic ESCs, we formulated an approach of "the best iPSC line" selection and confirmed it on an independent dataset.

  13. Efeito da escória de siderurgia na química do solo e na absorção de nutrientes e metais pesados pela cana-de-açúcar

    Directory of Open Access Journals (Sweden)

    Luiz Carlos Prezotti

    2012-08-01

    Full Text Available Com a redução das reservas naturais de nutrientes e o alto custo dos fertilizantes, tem aumentado a busca por fontes alternativas, sendo a escória de siderurgia uma opção, uma vez que possui compostos neutralizantes da acidez do solo e elementos químicos importantes para a nutrição das plantas. Este trabalho teve por objetivo avaliar as alterações, nas características químicas do solo e nos teores de nutrientes e de metais pesados, na cana-de-açúcar, em função de doses crescentes de escória de siderurgia. Os tratamentos foram compostos por oito doses de escória de aciaria: 0; 0,5; 1,0; 2,5; 5,0; 10; 20; 40 t ha-1, mais um tratamento adicional composto por uma dose de 2,5 t ha-1 de calcário, totalizando nove tratamentos, distribuídos em blocos ao acaso, com três repetições. Foram determinadas as características químicas de solo e planta, bem como a produtividade da cana-de-açúcar. A aplicação de escória no solo elevou o pH, a saturação por bases e os teores de P, Ca, B, Fe e Mn, não influenciando os teores de K e Mg. Mesmo aplicando-se doses mais elevadas, não houve alteração nos teores dos metais pesados Cd, Pb e Ni do solo. Houve elevação do teor de Cr. O teor de P na folha e no colmo da cana-de-açúcar apresentou tendência de aumento com as doses de escória, não havendo influência para os demais nutrientes, como, também, para os metais pesados. Houve elevação da produtividade tanto da cana-planta como da cana-soca.

  14. The tense the hostile and the distressed : Multidimensional psychosocial risk profiles based on the ESC interview in coronary artery disease patients. The THORESCI study

    NARCIS (Netherlands)

    van Montfort, E.M.J.; Denollet, J.; Vermunt, J.K.; Widdershoven, J.W.M.G.; Kupper, N.

    2017-01-01

    Background While single psychosocial factors have been associated with cardiovascular outcomes, it is still unclear how they cluster. Therefore, we examined whether latent multidimensional psychosocial risk profiles could be identified in the European Society of Cardiology (ESC) psychosocial

  15. Derivation of xeno-free and GMP-grade human embryonic stem cells--platforms for future clinical applications.

    Directory of Open Access Journals (Sweden)

    Shelly E Tannenbaum

    Full Text Available Clinically compliant human embryonic stem cells (hESCs should be developed in adherence to ethical standards, without risk of contamination by adventitious agents. Here we developed for the first time animal-component free and good manufacturing practice (GMP-compliant hESCs. After vendor and raw material qualification, we derived xeno-free, GMP-grade feeders from umbilical cord tissue, and utilized them within a novel, xeno-free hESC culture system. We derived and characterized three hESC lines in adherence to regulations for embryo procurement, and good tissue, manufacturing and laboratory practices. To minimize freezing and thawing, we continuously expanded the lines from initial outgrowths and samples were cryopreserved as early stocks and banks. Batch release criteria included DNA-fingerprinting and HLA-typing for identity, characterization of pluripotency-associated marker expression, proliferation, karyotyping and differentiation in-vitro and in-vivo. These hESCs may be valuable for regenerative therapy. The ethical, scientific and regulatory methodology presented here may serve for development of additional clinical-grade hESCs.

  16. Single-Cell Gene Expression Analysis of a Human ESC Model of Pancreatic Endocrine Development Reveals Different Paths to β-Cell Differentiation.

    Science.gov (United States)

    Petersen, Maja Borup Kjær; Azad, Ajuna; Ingvorsen, Camilla; Hess, Katja; Hansson, Mattias; Grapin-Botton, Anne; Honoré, Christian

    2017-10-10

    The production of insulin-producing β cells from human embryonic stem cells (hESCs) in vitro represents a promising strategy for a cell-based therapy for type 1 diabetes mellitus. To explore the cellular heterogeneity and temporal progression of endocrine progenitors and their progeny, we performed single-cell qPCR on more than 500 cells across several stages of in vitro differentiation of hESCs and compared them with human islets. We reveal distinct subpopulations along the endocrine differentiation path and an early lineage bifurcation toward either polyhormonal cells or β-like cells. We uncover several similarities and differences with mouse development and reveal that cells can take multiple paths to the same differentiation state, a principle that could be relevant to other systems. Notably, activation of the key β-cell transcription factor NKX6.1 can be initiated before or after endocrine commitment. The single-cell temporal resolution we provide can be used to improve the production of functional β cells. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  17. Healthy human CSF promotes glial differentiation of hESC-derived neural cells while retaining spontaneous activity in existing neuronal networks

    Directory of Open Access Journals (Sweden)

    Heikki Kiiski

    2013-05-01

    The possibilities of human pluripotent stem cell-derived neural cells from the basic research tool to a treatment option in regenerative medicine have been well recognized. These cells also offer an interesting tool for in vitro models of neuronal networks to be used for drug screening and neurotoxicological studies and for patient/disease specific in vitro models. Here, as aiming to develop a reductionistic in vitro human neuronal network model, we tested whether human embryonic stem cell (hESC-derived neural cells could be cultured in human cerebrospinal fluid (CSF in order to better mimic the in vivo conditions. Our results showed that CSF altered the differentiation of hESC-derived neural cells towards glial cells at the expense of neuronal differentiation. The proliferation rate was reduced in CSF cultures. However, even though the use of CSF as the culture medium altered the glial vs. neuronal differentiation rate, the pre-existing spontaneous activity of the neuronal networks persisted throughout the study. These results suggest that it is possible to develop fully human cell and culture-based environments that can further be modified for various in vitro modeling purposes.

  18. Non-canonical microRNAs miR-320 and miR-702 promote proliferation in Dgcr8-deficient embryonic stem cells

    International Nuclear Information System (INIS)

    Kim, Byeong-Moo; Choi, Michael Y.

    2012-01-01

    Highlights: ► Embryonic stem cells (ESCs) lacking non-canonical miRNAs proliferate slower. ► miR-320 and miR-702 are two non-canonical miRNAs expressed in ESCs. ► miR-320 and miR-702 promote proliferation of Dgcr8-deficient ESCs. ► miR-320 targets p57 and helps to release Dgcr8-deficient ESCs from G1 arrest. ► miR-702 targets p21 and helps to release Dgcr8-deficient ESCs from G1 arrest. -- Abstract: MicroRNAs are known to contribute significantly to stem cell phenotype by post-transcriptionally regulating gene expression. Most of our knowledge of microRNAs comes from the study of canonical microRNAs that require two sequential cleavages by the Drosha/Dgcr8 heterodimer and Dicer to generate mature products. In contrast, non-canonical microRNAs bypass the cleavage by the Drosha/Dgcr8 heterodimer within the nucleus but still require cytoplasmic cleavage by Dicer. The function of non-canonical microRNAs in embryonic stem cells (ESCs) remains obscure. It has been hypothesized that non-canonical microRNAs have important roles in ESCs based upon the phenotypes of ESC lines that lack these specific classes of microRNAs; Dicer-deficient ESCs lacking both canonical and non-canonical microRNAs have much more severe proliferation defect than Dgcr8-deficient ESCs lacking only canonical microRNAs. Using these cell lines, we identified two non-canonical microRNAs, miR-320 and miR-702, that promote proliferation of Dgcr8-deficient ESCs by releasing them from G1 arrest. This is accomplished by targeting the 3′-untranslated regions of the cell cycle inhibitors p57 and p21 and thereby inhibiting their expression. This is the first report of the crucial role of non-canonical microRNAs in ESCs.

  19. Regional differences in expression of specific markers for human embryonic stem cells

    DEFF Research Database (Denmark)

    Laursen, Steen B; Møllgård, Kjeld; Olesen, Christian

    2007-01-01

    Characterization of human embryonic stem cell (hESC) lines derived from the inner cell masses of blastocysts generally includes expression analysis of markers such as OCT4, NANOG, SSEA3, SSEA4, TRA-1-60 and TRA-1-81. Expression is usually detected by immunocytochemical staining of entire colonies...... of hESC, using one colony for each individual marker. Four newly established hESC lines showed the expected expression pattern and were capable of differentiating into the three germ layers in vitro. Neighbouring sections of entire colonies grown for 4, 11, 21 and 28 days respectively were stained...

  20. Enrichment of short interspersed transposable elements to embryonic stem cell-specific hypomethylated gene regions.

    Science.gov (United States)

    Muramoto, Hiroki; Yagi, Shintaro; Hirabayashi, Keiji; Sato, Shinya; Ohgane, Jun; Tanaka, Satoshi; Shiota, Kunio

    2010-08-01

    Embryonic stem cells (ESCs) have a distinctive epigenome, which includes their genome-wide DNA methylation modification status, as represented by the ESC-specific hypomethylation of tissue-dependent and differentially methylated regions (T-DMRs) of Pou5f1 and Nanog. Here, we conducted a genome-wide investigation of sequence characteristics associated with T-DMRs that were differentially methylated between ESCs and somatic cells, by focusing on transposable elements including short interspersed elements (SINEs), long interspersed elements (LINEs) and long terminal repeats (LTRs). We found that hypomethylated T-DMRs were predominantly present in SINE-rich/LINE-poor genomic loci. The enrichment for SINEs spread over 300 kb in cis and there existed SINE-rich genomic domains spreading continuously over 1 Mb, which contained multiple hypomethylated T-DMRs. The characterization of sequence information showed that the enriched SINEs were relatively CpG rich and belonged to specific subfamilies. A subset of the enriched SINEs were hypomethylated T-DMRs in ESCs at Dppa3 gene locus, although SINEs are overall methylated in both ESCs and the liver. In conclusion, we propose that SINE enrichment is the genomic property of regions harboring hypomethylated T-DMRs in ESCs, which is a novel aspect of the ESC-specific epigenomic information.

  1. Immunoflourescence and mRNA analysis of human embryonic stem cells (hESCs) grown under feeder-free conditions

    DEFF Research Database (Denmark)

    Awan, Aashir; Oliveri, Roberto S; Jensen, Pernille L

    2010-01-01

    onto 16-well glass chambers, and continuing with the general IF and qPCR steps will be provided. The techniques will be illustrated with new results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, and PDGF signaling pathways to primary cilia in stem cell......This chapter describes the procedures in order to do immunofluorescence (IF) microscopy and quantitative PCR (qPCR) analysis of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol outlining the steps from initially growing the cells, passaging...

  2. High-fidelity Glucagon-CreER mouse line generated by CRISPR-Cas9 assisted gene targeting

    Directory of Open Access Journals (Sweden)

    Amanda M. Ackermann

    2017-03-01

    Full Text Available Objective: α-cells are the second most prominent cell type in pancreatic islets and are responsible for producing glucagon to increase plasma glucose levels in times of fasting. α-cell dysfunction and inappropriate glucagon secretion occur in both type 1 and type 2 diabetes. Thus, there is growing interest in studying both normal function and pathophysiology of α-cells. However, tools to target gene ablation or activation specifically of α-cells have been limited, compared to those available for β-cells. Previous Glucagon-Cre and Glucagon-CreER transgenic mouse lines have suffered from transgene silencing, and the only available Glucagon-CreER “knock-in” mouse line results in glucagon haploinsufficiency, which can confound the interpretation of gene deletion analyses. Therefore, we sought to develop a Glucagon-CreERT2 mouse line that would maintain normal glucagon expression and would be less susceptible to transgene silencing. Methods: We utilized CRISPR-Cas9 technology to insert an IRES-CreERT2 sequence into the 3′ UTR of the Glucagon (Gcg locus in mouse embryonic stem cells (ESCs. Targeted ESC clones were then injected into mouse blastocysts to obtain Gcg-CreERT2 mice. Recombination efficiency in GCG+ pancreatic α-cells and glucagon-like peptide 1 positive (GLP1+ enteroendocrine L-cells was measured in Gcg-CreERT2;Rosa26-LSL-YFP mice injected with tamoxifen during fetal development and adulthood. Results: Tamoxifen injection of Gcg-CreERT2;Rosa26-LSL-YFP mice induced high recombination efficiency of the Rosa26-LSL-YFP locus in perinatal and adult α-cells (88% and 95%, respectively, as well as in first-wave fetal α-cells (36% and adult enteroendocrine L-cells (33%. Mice homozygous for the Gcg-CreERT2 allele were phenotypically normal. Conclusions: We successfully derived a Gcg-CreERT2 mouse line that expresses CreERT2 in pancreatic α-cells and enteroendocrine L-cells without disrupting preproglucagon gene expression. These mice

  3. Design the Cost Approach in Trade-Off's for Structural Components, Illustrated on the Baseline Selection of the Engine Thrust Frame of Ariane 5 ESC-B

    Science.gov (United States)

    Appolloni, L.; Juhls, A.; Rieck, U.

    2002-01-01

    Designing for value is one of the very actual upcoming methods for design optimization, which broke into the domain of aerospace engineering in the late 90's. In the frame of designing for value two main design philosophies exist: Design For Cost and Design To Cost. Design To Cost is the iterative redesign of a project until the content of the project meets a given budget. Designing For Cost is the conscious use of engineering process technology to reduce life cycle cost while satisfying, and hopefully exceeding, customer demands. The key to understanding cost, and hence to reducing cost, is the ability to measure cost accurately and to allocate it appropriately to products. Only then can intelligent decisions be made. Therefore the necessity of new methods as "Design For Value" or "Design For Competitiveness", set up with a generally multidisciplinary approach to find an optimized technical solution driven by many parameters, depending on the mission scenario and the customer/market needs. Very often three, but not more than five parametric drivers are sufficient. The more variable exist, the higher is in fact the risk to find just a sub-optimized local and not the global optimum, and the less robust is the found solution against change of input parameters. When the main parameters for optimization have been identified, the system engineer has to communicate them to all design engineers, who shall take care of these assessment variables during the entire design and decision process. The design process which has taken to the definition of the feasible structural concepts for the Engine Thrust Frame of the Ariane 5 Upper Cryogenic Stage ESC-B follows these most actual design philosophy methodologies, and combines a design for cost approach, to a design to cost optimization loop. Ariane 5 is the first member of a family of heavy-lift launchers. It aims to evolve into a family of launchers that responds to the space transportation challenges of the 21st century. New

  4. Experiment list: SRX590302 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available type/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of ...origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibo

  5. Experiment list: SRX590318 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available type/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of ...origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibo

  6. Experiment list: SRX590312 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available type/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of ...origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibo

  7. Experiment list: SRX590304 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available type/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of ...origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibo

  8. Experiment list: SRX590308 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available type/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of ...origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibo

  9. Experiment list: SRX590300 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available notype/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line o...f origin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip anti

  10. Experiment list: SRX590316 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available pe/variation=Foxd3 conditional knockout || cell type=embryonic stem cells (ESCs; R cells) || cell line of or...igin=Foxd3 conditional knockout C57BL/6 ESCs || treatment=Lif 2i || chip antibody

  11. Human therapeutic cloning (NTSC): applying research from mammalian reproductive cloning.

    Science.gov (United States)

    French, Andrew J; Wood, Samuel H; Trounson, Alan O

    2006-01-01

    Human therapeutic cloning or nuclear transfer stem cells (NTSC) to produce patient-specific stem cells, holds considerable promise in the field of regenerative medicine. The recent withdrawal of the only scientific publications claiming the successful generation of NTSC lines afford an opportunity to review the available research in mammalian reproductive somatic cell nuclear transfer (SCNT) with the goal of progressing human NTSC. The process of SCNT is prone to epigenetic abnormalities that contribute to very low success rates. Although there are high mortality rates in some species of cloned animals, most surviving clones have been shown to have normal phenotypic and physiological characteristics and to produce healthy offspring. This technology has been applied to an increasing number of mammals for utility in research, agriculture, conservation, and biomedicine. In contrast, attempts at SCNT to produce human embryonic stem cells (hESCs) have been disappointing. Only one group has published reliable evidence of success in deriving a cloned human blastocyst, using an undifferentiated hESC donor cell, and it failed to develop into a hESC line. When optimal conditions are present, it appears that in vitro development of cloned and parthenogenetic embryos, both of which may be utilized to produce hESCs, may be similar to in vitro fertilized embryos. The derivation of ESC lines from cloned embryos is substantially more efficient than the production of viable offspring. This review summarizes developments in mammalian reproductive cloning, cell-to-cell fusion alternatives, and strategies for oocyte procurement that may provide important clues facilitating progress in human therapeutic cloning leading to the successful application of cell-based therapies utilizing autologous hESC lines.

  12. Generation of functional neuromuscular junctions from human pluripotent stem cell lines

    Directory of Open Access Journals (Sweden)

    Katja ePuttonen

    2015-12-01

    Full Text Available Several neuromuscular diseases involve dysfunction of neuromuscular junctions (NMJs, yet there are no patient-specific human models for electrophysiological characterization of NMJ. We seeded cells of neurally-induced embryoid body-like spheres derived from induced pluripotent stem cell (iPSC or embryonic stem cell (ESC lines as monolayers without basic fibroblast factor (bFGF and observed differentiation of neuronal as well as spontaneously contracting, multinucleated skeletal myotubes. The myotubes showed striation, immunoreactivity for myosin heavy chain, actin bundles typical for myo-oriented cells, and generated spontaneous and evoked action potentials (APs. The myogenic differentiation was associated with expression of MyoD1, myogenin and type I ryanodine receptor. Neurons formed end plate like structures with strong binding of α-bungarotoxin, a marker of nicotinic acetylcholine receptors highly expressed in the postsynaptic membrane of NMJs, and expressed SMI-32, a motoneuron marker, as well as SV2, a marker for synapses. Pharmacological stimulation of cholinergic receptors resulted in strong depolarization of myotube membrane and raised Ca2+ concentration in sarcoplasm, while electrical stimulation evoked Ca2+ transients in myotubes. Stimulation of motoneurons with N-Methyl-D-aspartate resulted in reproducible APs in myotubes and end plates displayed typical MEPs and tonic activity depolarizing myotubes of about 10 mV. We conclude that simultaneous differentiation of neurons and myotubes from patient-specific iPSCs or ESCs results also in the development of functional NMJs. Our human model of NMJ may serve as an important tool to investigate normal development, mechanisms of diseases and novel drug targets involving NMJ dysfunction and degeneration.

  13. ESC Track Fusion Demonstration Tool for Distributed Environments

    Science.gov (United States)

    Cox, C.; Degraaf, E.; Perry, R.; Diaz, R.

    A key requirement of future net-centric Space Situational Awareness systems development and operations will be decentralized operations, including multi-level distributed data fusion. Raytheon has developed a demonstration for ESC 850 ELSG/NS that fuses sensor-supplied tracks in a dense resident space object (RSO) environment. The demonstration use the state vector and covariance input data from single pass orbit solutions and applies track-to-track correlation algorithms to fuse the individual tracks into composite orbits. Platform independent Java technology and an agent-based software design using asynchronous inter-process communications was used in the demonstration tool development. The tool has been tested against a simulated scenario corresponding to the future 100,000+ object catalog environment. Ten days of simulated data from Fylingdales, Shemya, Eglin, and a future Space Fence sensor were generated for a co-orbiting family of 122 sun-synchronous objects between 700 and 800 km altitude from the NASA simulated small debris for 2015. The selected set exceeds the average object densities for the 100,000+ RSO environment, and provides a scenario similar to an evolved breakup where the debris has had time to disperse. The demo produced very good results using fast and simple astrodynamic models. A total of 16678 input tracks were fused, with less than 1.6% being misassociated. Pure tracks were generated for 65% of the 122 truth objects, and 97% of the objects had a misassociation rate tool that can be used to assess current breakups such as the Chinese ASAT event.

  14. [Effectiveness of school-based interventions to prevent traffic accidents: an overview of secondary literature].

    Science.gov (United States)

    Guerriero, Chiara; Verdina, Federico; Ferri, Chiara; Pizzaguerra, Martina; Silvestri, Silvia; Tober, Nastassja; Ristagno, Quenya; Costa, Alessandro; Cerasuolo, Michele; Faggiano, Fabrizio; Allara, Elias

    2016-01-01

    INTRODUZIONE: gli incidenti stradali sono l'ottava causa di morte al mondo e la prima tra i giovani di 15-29 anni. In Italia il Piano nazionale sicurezza stradale raccomanda l'educazione scolastica per la prevenzione degli incidenti stradali; ad oggi non esistono documenti che raccolgano evidenze di efficacia sugli interventi educativi stradali e le rapportino al contesto italiano. OBIETTIVI: riassumere e discutere ciò che è noto in letteratura riguardo agli interventi scolastici per la prevenzione degli incidenti stradali. METODI: sono state ricercate linee guida e revisioni sistematiche usando i seguenti criteri di inclusione: popolazione di età inferiore ai 25 anni di entrambi i sessi; interventi scolastici di educazione stradale; effetti su indicatori primari di esito come riduzione degli incidenti stradali, astinenza dalla guida sotto l'effetto di alcol e dall'accettare passaggi in macchina da guidatori che sono sotto l'effetto di alcol; effetti su indicatori secondari di esito come conoscenze e competenze sui comportamenti di guida sicura. RISULTATI: sono state identificate due revisioni sistematiche. L'educazione stradale nelle scuole non mostra evidenza di efficacia (rischio relativo 1,03; IC95% 0,98-1,08) nel ridurre gli incidenti. Programmi scolastici più specifici mostrano risultati solo in parte convincenti per l'adozione di comportamenti sicuri come l'astinenza dal guidare sotto l'effetto di alcol e dall'accettare passaggi in macchina da guidatori che sono sotto l'effetto di alcol. DISCUSSIONE: le revisioni incluse non hanno trovato programmi efficaci nella riduzione degli incidenti stradali o dei fattori che possano determinarli. Nell'attesa di studi più recenti, appare opportuno promuovere l'implementazione di interventi misti, scolastici e di comunità, che hanno mostrato maggiori prove di efficacia.

  15. Improved genetic manipulation of human embryonic stem cells.

    NARCIS (Netherlands)

    Braam, S.R.; Denning, C.; van den Brink, S.; Kats, P.; Hochstenbach, R.; Passier, R.; Mummery, C.L.

    2008-01-01

    Low efficiency of transfection limits the ability to genetically manipulate human embryonic stem cells (hESCs), and differences in cell derivation and culture methods require optimization of transfection protocols. We transiently transferred multiple independent hESC lines with different growth

  16. Non-canonical microRNAs miR-320 and miR-702 promote proliferation in Dgcr8-deficient embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Byeong-Moo [Department of Medicine/GI Unit, Massachusetts General Hospital, Boston, MA 02114 (United States); Department of Medicine, Harvard Medical School, Boston, MA 02115 (United States); Choi, Michael Y., E-mail: mchoi@partners.org [Department of Medicine/GI Unit, Massachusetts General Hospital, Boston, MA 02114 (United States); Department of Medicine, Harvard Medical School, Boston, MA 02115 (United States); Harvard Stem Cell Institute, Boston, MA 02114 (United States)

    2012-09-21

    Highlights: Black-Right-Pointing-Pointer Embryonic stem cells (ESCs) lacking non-canonical miRNAs proliferate slower. Black-Right-Pointing-Pointer miR-320 and miR-702 are two non-canonical miRNAs expressed in ESCs. Black-Right-Pointing-Pointer miR-320 and miR-702 promote proliferation of Dgcr8-deficient ESCs. Black-Right-Pointing-Pointer miR-320 targets p57 and helps to release Dgcr8-deficient ESCs from G1 arrest. Black-Right-Pointing-Pointer miR-702 targets p21 and helps to release Dgcr8-deficient ESCs from G1 arrest. -- Abstract: MicroRNAs are known to contribute significantly to stem cell phenotype by post-transcriptionally regulating gene expression. Most of our knowledge of microRNAs comes from the study of canonical microRNAs that require two sequential cleavages by the Drosha/Dgcr8 heterodimer and Dicer to generate mature products. In contrast, non-canonical microRNAs bypass the cleavage by the Drosha/Dgcr8 heterodimer within the nucleus but still require cytoplasmic cleavage by Dicer. The function of non-canonical microRNAs in embryonic stem cells (ESCs) remains obscure. It has been hypothesized that non-canonical microRNAs have important roles in ESCs based upon the phenotypes of ESC lines that lack these specific classes of microRNAs; Dicer-deficient ESCs lacking both canonical and non-canonical microRNAs have much more severe proliferation defect than Dgcr8-deficient ESCs lacking only canonical microRNAs. Using these cell lines, we identified two non-canonical microRNAs, miR-320 and miR-702, that promote proliferation of Dgcr8-deficient ESCs by releasing them from G1 arrest. This is accomplished by targeting the 3 Prime -untranslated regions of the cell cycle inhibitors p57 and p21 and thereby inhibiting their expression. This is the first report of the crucial role of non-canonical microRNAs in ESCs.

  17. Defining differentially methylated regions specific for the acquisition of pluripotency and maintenance in human pluripotent stem cells via microarray.

    Directory of Open Access Journals (Sweden)

    WenYin He

    Full Text Available Epigenetic regulation is critical for the maintenance of human pluripotent stem cells. It has been shown that pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, appear to have a hypermethylated status compared with differentiated cells. However, the epigenetic differences in genes that maintain stemness and regulate reprogramming between embryonic stem cells and induced pluripotent stem cells remain unclear. Additionally, differential methylation patterns of induced pluripotent stem cells generated using diverse methods require further study.Here, we determined the DNA methylation profiles of 10 human cell lines, including 2 ESC lines, 4 virally derived iPSC lines, 2 episomally derived iPSC lines, and the 2 parental cell lines from which the iPSCs were derived using Illumina's Infinium HumanMethylation450 BeadChip. The iPSCs exhibited a hypermethylation status similar to that of ESCs but with distinct differences from the parental cells. Genes with a common methylation pattern between iPSCs and ESCs were classified as critical factors for stemness, whereas differences between iPSCs and ESCs suggested that iPSCs partly retained the parental characteristics and gained de novo methylation aberrances during cellular reprogramming. No significant differences were identified between virally and episomally derived iPSCs. This study determined in detail the de novo differential methylation signatures of particular stem cell lines.This study describes the DNA methylation profiles of human iPSCs generated using both viral and episomal methods, the corresponding somatic cells, and hESCs. Series of ss-DMRs and ES-iPS-DMRs were defined with high resolution. Knowledge of this type of epigenetic information could be used as a signature for stemness and self-renewal and provides a potential method for selecting optimal pluripotent stem cells for human regenerative medicine.

  18. DNA methylation dynamics in human induced pluripotent stem cells over time.

    Directory of Open Access Journals (Sweden)

    Koichiro Nishino

    2011-05-01

    Full Text Available Epigenetic reprogramming is a critical event in the generation of induced pluripotent stem cells (iPSCs. Here, we determined the DNA methylation profiles of 22 human iPSC lines derived from five different cell types (human endometrium, placental artery endothelium, amnion, fetal lung fibroblast, and menstrual blood cell and five human embryonic stem cell (ESC lines, and we followed the aberrant methylation sites in iPSCs for up to 42 weeks. The iPSCs exhibited distinct epigenetic differences from ESCs, which were caused by aberrant methylation at early passages. Multiple appearances and then disappearances of random aberrant methylation were detected throughout iPSC reprogramming. Continuous passaging of the iPSCs diminished the differences between iPSCs and ESCs, implying that iPSCs lose the characteristics inherited from the parent cells and adapt to very closely resemble ESCs over time. Human iPSCs were gradually reprogrammed through the "convergence" of aberrant hyper-methylation events that continuously appeared in a de novo manner. This iPS reprogramming consisted of stochastic de novo methylation and selection/fixation of methylation in an environment suitable for ESCs. Taken together, random methylation and convergence are driving forces for long-term reprogramming of iPSCs to ESCs.

  19. Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

    DEFF Research Database (Denmark)

    Vestergaard, Maj Linea; Awan, Aashir; Warzecha, Caroline Becker

    2016-01-01

    onto 16-well glass chambers, and continuing with the general IFM and qPCR anlysis. The techniques are illustrated with results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, PDGF, and TGFβ signaling pathways to primary cilia in stem cell maintenance......This chapter describes the procedures for immunofluorescence microscopy (IFM) and quantitative PCR (qPCR) analyses of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol is provided outlining the steps from initially growing the cells, passaging...

  20. Induced pluripotent stem cells derived from rabbits exhibit some characteristics of naïve pluripotency

    Directory of Open Access Journals (Sweden)

    Pierre Osteil

    2013-05-01

    Not much is known about the molecular and functional features of pluripotent stem cells (PSCs in rabbits. To address this, we derived and characterized 2 types of rabbit PSCs from the same breed of New Zealand White rabbits: 4 lines of embryonic stem cells (rbESCs, and 3 lines of induced PSCs (rbiPSCs that were obtained by reprogramming adult skin fibroblasts. All cell lines required fibroblast growth factor 2 for their growth and proliferation. All rbESC lines showed molecular and functional properties typically associated with primed pluripotency. The cell cycle of rbESCs had a prolonged G1 phase and a DNA damage checkpoint before entry into the S phase, which are the 2 features typically associated with the somatic cell cycle. In contrast, the rbiPSC lines exhibited some characteristics of naïve pluripotency, including resistance to single-cell dissociation by trypsin, robust activity of the distal enhancer of the mouse Oct4 gene, and expression of naïve pluripotency-specific genes, as defined in rodents. According to gene expression profiles, rbiPSCs were closer to the rabbit inner cell mass (ICM than rbESCs. Furthermore, rbiPSCs were capable of colonizing the ICM after aggregation with morulas. Therefore, we propose that rbiPSCs self-renew in an intermediate state between naïve and primed pluripotency, which represents a key step toward the generation of bona fide naïve PSC lines in rabbits.

  1. Induced pluripotent stem cells derived from rabbits exhibit some characteristics of naïve pluripotency

    Science.gov (United States)

    Osteil, Pierre; Tapponnier, Yann; Markossian, Suzy; Godet, Murielle; Schmaltz-Panneau, Barbara; Jouneau, Luc; Cabau, Cédric; Joly, Thierry; Blachère, Thierry; Gócza, Elen; Bernat, Agnieszka; Yerle, Martine; Acloque, Hervé; Hidot, Sullivan; Bosze, Zsuzsanna; Duranthon, Véronique; Savatier, Pierre; Afanassieff, Marielle

    2013-01-01

    Summary Not much is known about the molecular and functional features of pluripotent stem cells (PSCs) in rabbits. To address this, we derived and characterized 2 types of rabbit PSCs from the same breed of New Zealand White rabbits: 4 lines of embryonic stem cells (rbESCs), and 3 lines of induced PSCs (rbiPSCs) that were obtained by reprogramming adult skin fibroblasts. All cell lines required fibroblast growth factor 2 for their growth and proliferation. All rbESC lines showed molecular and functional properties typically associated with primed pluripotency. The cell cycle of rbESCs had a prolonged G1 phase and a DNA damage checkpoint before entry into the S phase, which are the 2 features typically associated with the somatic cell cycle. In contrast, the rbiPSC lines exhibited some characteristics of naïve pluripotency, including resistance to single-cell dissociation by trypsin, robust activity of the distal enhancer of the mouse Oct4 gene, and expression of naïve pluripotency-specific genes, as defined in rodents. According to gene expression profiles, rbiPSCs were closer to the rabbit inner cell mass (ICM) than rbESCs. Furthermore, rbiPSCs were capable of colonizing the ICM after aggregation with morulas. Therefore, we propose that rbiPSCs self-renew in an intermediate state between naïve and primed pluripotency, which represents a key step toward the generation of bona fide naïve PSC lines in rabbits. PMID:23789112

  2. Specialized mouse embryonic stem cells for studying vascular development.

    Science.gov (United States)

    Glaser, Drew E; Burns, Andrew B; Hatano, Rachel; Medrzycki, Magdalena; Fan, Yuhong; McCloskey, Kara E

    2014-01-01

    Vascular progenitor cells are desirable in a variety of therapeutic strategies; however, the lineage commitment of endothelial and smooth muscle cell from a common progenitor is not well-understood. Here, we report the generation of the first dual reporter mouse embryonic stem cell (mESC) lines designed to facilitate the study of vascular endothelial and smooth muscle development in vitro. These mESC lines express green fluorescent protein (GFP) under the endothelial promoter, Tie-2, and Discomsoma sp. red fluorescent protein (RFP) under the promoter for alpha-smooth muscle actin (α-SMA). The lines were then characterized for morphology, marker expression, and pluripotency. The mESC colonies were found to exhibit dome-shaped morphology, alkaline phosphotase activity, as well as expression of Oct 3/4 and stage-specific embryonic antigen-1. The mESC colonies were also found to display normal karyotypes and are able to generate cells from all three germ layers, verifying pluripotency. Tissue staining confirmed the coexpression of VE (vascular endothelial)-cadherin with the Tie-2 GFP+ expression on endothelial structures and smooth muscle myosin heavy chain with the α-SMA RFP+ smooth muscle cells. Lastly, it was verified that the developing mESC do express Tie-2 GFP+ and α-SMA RFP+ cells during differentiation and that the GFP+ cells colocalize with the vascular-like structures surrounded by α-SMA-RFP cells. These dual reporter vascular-specific mESC permit visualization and cell tracking of individual endothelial and smooth muscle cells over time and in multiple dimensions, a powerful new tool for studying vascular development in real time.

  3. Contacting co-culture of human retinal microvascular endothelial cells alters barrier function of human embryonic stem cell derived retinal pigment epithelial cells.

    Science.gov (United States)

    Skottman, H; Muranen, J; Lähdekorpi, H; Pajula, E; Mäkelä, K; Koivusalo, L; Koistinen, A; Uusitalo, H; Kaarniranta, K; Juuti-Uusitalo, K

    2017-10-01

    Here we evaluated the effects of human retinal microvascular endothelial cells (hREC) on mature human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells. The hESC-RPE cells (Regea08/017, Regea08/023 or Regea11/013) and hREC (ACBRI 181) were co-cultured on opposite sides of transparent membranes for up to six weeks. Thereafter barrier function, small molecule permeability, localization of RPE and endothelial cell marker proteins, cellular fine structure, and growth factor secretion of were evaluated. After co-culture, the RPE specific CRALBP and endothelial cell specific von Willebrand factor were appropriately localized. In addition, the general morphology, pigmentation, and fine structure of hESC-RPE cells were unaffected. Co-culture increased the barrier function of hESC-RPE cells, detected both with TEER measurements and cumulative permeability of FD4 - although the differences varied among the cell lines. Co-culturing significantly altered VEGF and PEDF secretion, but again the differences were cell line specific. The results of this study showed that co-culture with hREC affects hESC-RPE functionality. In addition, co-culture revealed drastic cell line specific differences, most notably in growth factor secretion. This model has the potential to be used as an in vitro outer blood-retinal barrier model for drug permeability testing. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  4. Insulin redirects differentiation from cardiogenic mesoderm and endoderm to neuroectoderm in differentiating human embryonic stem cells.

    NARCIS (Netherlands)

    Freund, C.M.A.H.; Ward-van Oostwaard, D.; Monshouwer-Kloots, J.; van den Brink, S.; van Rooijen, M.A.; Xu, X.; Zweigerdt, R.; Mummery, C.L.; Passier, R.

    2008-01-01

    Human embryonic stem cells (hESC) can proliferate indefinitely while retaining the capacity to form derivatives of all three germ layers. We have reported previously that hESC differentiate into cardiomyocytes when cocultured with a visceral endoderm-like cell line (END-2). Insulin/insulin-like

  5. Porcine embryonic stem cells

    DEFF Research Database (Denmark)

    Hall, Vanessa Jane

    2008-01-01

    The development of porcine embryonic stem cell lines (pESC) has received renewed interest given the advances being made in the production of immunocompatible transgenic pigs. However, difficulties are evident in the production of pESCs in-vitro. This may largely be attributable to differences...

  6. Unraveling the global microRNAome responses to ionizing radiation in human embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Mykyta V Sokolov

    Full Text Available MicroRNAs (miRNA comprise a group of short ribonucleic acid molecules implicated in regulation of key biological processes and functions at the post-transcriptional level. Ionizing radiation (IR causes DNA damage and generally triggers cellular stress response. However, the role of miRNAs in IR-induced response in human embryonic stem cells (hESC has not been defined yet. Here, by using system biology approaches, we show for the first time, that miRNAome undergoes global alterations in hESC (H1 and H9 lines after IR. Interrogation of expression levels of 1,090 miRNA species in irradiated hESC showed statistically significant changes in 54 genes following 1 Gy of X-ray exposures; global miRNAome alterations were found to be highly temporally and cell line--dependent in hESC. Time-course studies showed that the 16 hr miRNAome radiation response of hESC is much more robust compared to 2 hr-response signature (only eight genes, and may be involved in regulating the cell cycle. Quantitative real-time PCR performed on some miRNA species confirms the robustness of our miRNA microarray platform. Positive regulation of differentiation-, cell cycle-, ion transport- and endomembrane system-related processes were predicted to be negatively affected by miRNAome changes in irradiated hESC. Our findings reveal a fundamental role of miRNAome in modulating the radiation response, and identify novel molecular targets of radiation in hESC.

  7. EZ spheres: a stable and expandable culture system for the generation of pre-rosette multipotent stem cells from human ESCs and iPSCs.

    Science.gov (United States)

    Ebert, Allison D; Shelley, Brandon C; Hurley, Amanda M; Onorati, Marco; Castiglioni, Valentina; Patitucci, Teresa N; Svendsen, Soshana P; Mattis, Virginia B; McGivern, Jered V; Schwab, Andrew J; Sareen, Dhruv; Kim, Ho Won; Cattaneo, Elena; Svendsen, Clive N

    2013-05-01

    We have developed a simple method to generate and expand multipotent, self-renewing pre-rosette neural stem cells from both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs) without utilizing embryoid body formation, manual selection techniques, or complex combinations of small molecules. Human ESC and iPSC colonies were lifted and placed in a neural stem cell medium containing high concentrations of EGF and FGF-2. Cell aggregates (termed EZ spheres) could be expanded for long periods using a chopping method that maintained cell-cell contact. Early passage EZ spheres rapidly down-regulated OCT4 and up-regulated SOX2 and nestin expression. They retained the potential to form neural rosettes and consistently differentiated into a range of central and peripheral neural lineages. Thus, they represent a very early neural stem cell with greater differentiation flexibility than other previously described methods. As such, they will be useful for the rapidly expanding field of neurological development and disease modeling, high-content screening, and regenerative therapies based on pluripotent stem cell technology. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Differentiation of stem cells upon deprivation of exogenous FGF2

    DEFF Research Database (Denmark)

    Kjartansdóttir, Kristín Rós; Gabrielsen, Anette; Reda, Ahmed

    2012-01-01

    Establishing a model for in vitro differentiation of human embryonic stem cells (hESCs) towards the germ cell lineage could be used to identify molecular mechanisms behind germ cell differentiation that may help in understanding human infertility. Here, we evaluate whether a lack of exogenous...... fibroblast growth factor 2 (FGF2) is supporting spontaneous differentiation of hESCs cultured on human foreskin fibroblast (hFF) monolayers towards germ cell lineage. Additionally to depriving the hESCs of exogenous FGF2, cells were stimulated with all-trans retinoic acid (ATRA). To get a more comprehensive...... impression on effects of removal of FGF2 and stimulation with ATRA, we combined the results of three cell lines for each experimental setting. When combining gene expression profiles of three cell lines for 96 genes, only 6 genes showed a significant up-regulation in all cell lines, when no FGF2 was added...

  9. Culture of hESC-derived pancreatic progenitors in alginate-based scaffolds.

    Science.gov (United States)

    Formo, Kjetil; Cho, Candy H-H; Vallier, Ludovic; Strand, Berit L

    2015-12-01

    The effect of alginate-based scaffolds with added basement membrane proteins on the in vitro development of hESC-derived pancreatic progenitors was investigated. Cell clusters were encapsulated in scaffolds containing the basement membrane proteins collagen IV, laminin, fibronectin, or extracellular matrix-derived peptides, and maintained in culture for up to 46 days. The cells remained viable throughout the experiment with no signs of central necrosis. Whereas nonencapsulated cells aggregated into larger clusters, some of which showed signs of morphological changes and tissue organization, the alginate matrix stabilized the cluster size and displayed more homogeneous cell morphologies, allowing culture for long periods of time. For all conditions tested, a stable or declining expression of insulin and PDX1 and an increase in glucagon and somatostatin over time indicated a progressive reduction in beta cell-related gene expression. Alginate scaffolds can provide a chemically defined, xeno-free and easily scalable alternative for culture of pancreatic progenitors. Although no increase in insulin and PDX1 gene expression after alginate-immobilized cell culture was seen in this study, further optimization of the matrix physicochemical and biological properties and of the medium composition may still be a relevant strategy to promote the stabilization or maturation of stem cell-derived beta cells. © 2015 Wiley Periodicals, Inc.

  10. Actas del III Encuentro Platense de Investigadores/as sobre Cuerpo en las Artes Escénicas y Performáticas

    OpenAIRE

    Sáez, Mariana Lucía

    2016-01-01

    Los días 4, 5 y 6 de septiembre de 2013, en el Centro Cultural Islas Malvinas de la ciudad de La Plata, se desarrolló el III ECART, Encuentro Platense de Investigadores sobre Cuerpo en las Artes Escénicas y Performáticas, organizado por el Grupo de Estudio sobre Cuerpo-Centro Interdisciplinario Cuerpo, Educación y Sociedad y la Compañía Proyecto en Bruto. En el transcurso de las tres jornadas de este nuevo encuentro se sucedieron talleres, conferencias, mesas de ponencias, obras de danza y...

  11. Sílica gel obtida de escória de alto forno: Marabá, Pará

    Directory of Open Access Journals (Sweden)

    M. M. Rebelo

    2015-09-01

    Full Text Available ResumoSílica gel com propriedades similares à sílica comercial foi obtida a partir de escória de alto forno (EAF, utilizando digestão com ácido clorídrico. A EAF-sílica obtida foi caracterizada por diferentes técnicas, mostrando-se amorfa, com pureza 99,7% e área específica 282 m2/g. Apresentou caráter hidrofílico alto (12,27%, com água de constituição de ~ 6,18%, o que foi confirmado pela perda de massa durante a análise termogravimétrica. As partículas de EAF-sílica apresentaram tamanhos micrométricos (< 1 µm em forma de agregados, distribuição granulométrica unimodal e D50 7,0 µm.

  12. A homozygous Keap1-knockout human embryonic stem cell line generated using CRISPR/Cas9 mediates gene targeting

    Directory of Open Access Journals (Sweden)

    So-Jung Kim

    2017-03-01

    Full Text Available Kelch-like ECH-associated protein 1 (keap1 is a cysteine-rich protein that interacts with transcription factor Nrf2 in a redox-sensitive manner, leading to the degradation of Nrf2 (Kim et al., 2014a. Disruption of Keap1 results in the induction of Nrf2-related signaling pathways involving the expression of a set of anti-oxidant and anti-inflammatory genes. We generated biallelic mutants of the Keap1 gene using a CRISPR-Cas9 genome editing method in the H9 human embryonic stem cell (hESC. The Keap1 homozygous-knockout H9 cell line retained normal morphology, gene expression, and in vivo differentiation potential.

  13. NRF2 Mutation Confers Malignant Potential and Resistance to Chemoradiation Therapy in Advanced Esophageal Squamous Cancer

    Directory of Open Access Journals (Sweden)

    Tatsuhiro Shibata

    2011-09-01

    Full Text Available Esophageal squamous cancer (ESC is one of the most aggressive tumors of the gastrointestinal tract. A combination of chemotherapy and radiation therapy (CRT has improved the clinical outcome, but the molecular background determining the effectiveness of therapy remains unknown. NRF2 is a master transcriptional regulator of stress adaptation, and gain of-function mutation of NRF2 in cancer confers resistance to stressors including anticancer therapy. Direct resequencing analysis revealed that Nrf2 gain-of-function mutation occurred recurrently (18/82, 22% in advanced ESC tumors and ESC cell lines (3/10. The presence of Nrf2 mutation was associated with tumor recurrence and poor prognosis. Short hairpin RNA-mediated down-regulation of NRF2 in ESC cells that harbor only mutated Nrf2 allele revealed that themutant NRF2 conferred increased cell proliferation, attachment-independent survival, and resistance to 5-fluorouracil and γ-irradiation. Based on the Nrf2 mutation status, gene expression signatures associated with NRF2 mutation were extracted from ESC cell lines, and their potential utility for monitoring and prognosis was examined in a cohort of 33 pre-CRT cases of ESC. The molecular signatures of NRF2 mutation were significantly predictive and prognostic for CRT response. In conclusion, recurrent NRF2 mutation confers malignant potential and resistance to therapy in advanced ESC, resulting in a poorer outcome. Molecular signatures of NRF2 mutation can be applied as predictive markers of response to CRT, and efficient inhibition of aberrant NRF2 activation could be a promising approach in combination with CRT.

  14. (L. Benth. en bosques secundarios en Escárcega, Campeche, México

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    J. L. López-Torres

    2005-01-01

    Full Text Available El T´zalam Lysiloma latisiliquum (L. Benth. en la región de Escárcega, Campeche, México, es la especie más abundante y con mayor importancia económica en bosques secundarios derivados de selva mediana subperennifolia, después haber sido intervenida mediante el sistema rosa-tumba-quema. El objetivo de este trabajo es analizar el comportamiento del crecimiento en diámetro de esta especie, en un periodo de 20 años de desarrollo. A partir de la variable diámetro normal (d1.3, agrupada en categorías diamétricas, se estimaron los incrementos anuales medio, máximo y mínimo para cada categoría. Los resultados indican que en el escenario más ventajoso se requiere de 10.6 años para que los árboles pasen de categoría reserva a categoría aprovechable y, en promedio, es necesario de 21.5 años para alcanzar la misma condición. El incremento en diámetro fue, en promedio, de 0.511 cm y se clasifica como medio.

  15. Procedures for Derivation and Characterisation of Human Embryonic Stem Cells from Odense, Denmark

    DEFF Research Database (Denmark)

    Harkness, Linda; Kassem, Moustapha

    2012-01-01

    In 1998, a development occurred in stem cell biology with the fi rst report of the derivation of a human embryonic stem cell (hESC) line. Since then a number of techniques have been used to derive and characterise hESCs. Here, we describe the derivation methods used by our laboratory for isolatio...

  16. Emotion in political discourse: contrasting approaches to stem cell governance in the USA, UK, Israel and Germany.

    Science.gov (United States)

    Gottweis, Herbert; Prainsack, Barbara

    2006-11-01

    In August 2004, Stojkovic and Murdoch from the University of Newcastle upon Tyne, UK, were granted the UK's first license to create human embryonic stem cells (hESCs) using cell nuclear replacement. While this news made headlines around the globe, a spokesman for the German Ministry of Research warned scientists in his country of the illegality of advising their English colleagues on hESC research. Meanwhile, US Members of Congress had asked President Bush to revoke his decision to limit federal funding to research on a limited number of hESC lines created before 9 August, 2001 (a decision that he confirmed in July 2006, while nonfederally funded research on hESC continues to be unrestricted). In Israel, where hESC research is legal and has never been a contested political issue, a bioethicist argued that, in light of the potential to alleviate human suffering, "banning research is against human dignity". How can such striking differences in the regulation of hESC research be explained?

  17. Gene Expression Music Algorithm-Based Characterization of the Ewing Sarcoma Stem Cell Signature

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    Martin Sebastian Staege

    2016-01-01

    Full Text Available Gene Expression Music Algorithm (GEMusicA is a method for the transformation of DNA microarray data into melodies that can be used for the characterization of differentially expressed genes. Using this method we compared gene expression profiles from endothelial cells (EC, hematopoietic stem cells, neuronal stem cells, embryonic stem cells (ESC, and mesenchymal stem cells (MSC and defined a set of genes that can discriminate between the different stem cell types. We analyzed the behavior of public microarray data sets from Ewing sarcoma (“Ewing family tumors,” EFT cell lines and biopsies in GEMusicA after prefiltering DNA microarray data for the probe sets from the stem cell signature. Our results demonstrate that individual Ewing sarcoma cell lines have a high similarity to ESC or EC. Ewing sarcoma cell lines with inhibited Ewing sarcoma breakpoint region 1-Friend leukemia virus integration 1 (EWSR1-FLI1 oncogene retained the similarity to ESC and EC. However, correlation coefficients between GEMusicA-processed expression data between EFT and ESC decreased whereas correlation coefficients between EFT and EC as well as between EFT and MSC increased after knockdown of EWSR1-FLI1. Our data support the concept of EFT being derived from cells with features of embryonic and endothelial cells.

  18. TRANSGENIC GDNF POSITIVELY INFLUENCES PROLIFERATION, DIFFERENTIATION, MATURATION AND SURVIVAL OF MOTOR NEURONS PRODUCED FROM MOUSE EMBRYONIC STEM CELLS.

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    Daniel Édgar Cortés

    2016-09-01

    Full Text Available Embryonic stem cells (ESC are pluripotent and thus can differentiate into every cell type present in the body. Directed differentiation into motor neurons has been described for pluripotent cells. Although neurotrophic factors promote neuronal survival, their role in neuronal commitment is elusive. Here, we developed double-transgenic lines of mouse ESC that constitutively produce Glial cell-derived neurotrophic factor (GDNF and also contain a GFP reporter, driven by HB9, which is expressed only by postmitotic motor neurons. After lentiviral transduction, ESC lines integrated and expressed the human GDNF gene without altering pluripotency markers before differentiation. Further, GDNF-ESC showed significantly higher spontaneous release of this neurotrophin to the medium, when compared to controls. To study motor neuron induction, control and GDNF cell lines were grown as embryoid bodies and stimulated with retinoic acid and Sonic Hedgehog. In GDNF-overexpressing cells, a significant increase of proliferative Olig2+ precursors, which are specified as spinal motor neurons, was found. Accordingly, GDNF increases the yield of cells with the pan motor neuronal markers HB9, monitored by GFP expression, and Isl1. At terminal differentiation, almost all differentiated neurons express phenotypic markers of motor neurons in GDNF cultures, with lower proportions in control cells. To test if the effects of GDNF were present at early differentiation stages, exogenous recombinant human GDNF was added to control ESC, also resulting in enhanced motor neuron differentiation. This effect was abolished by the co-addition of neutralizing anti-GDNF antibodies, strongly suggesting that differentiating ESC are responsive to GDNF. Using the HB9::GFP reporter, motor neurons were selected for electrophysiological recordings. Motor neurons differentiated from GDNF-ESC, compared to control motor neurons, showed greater electrophysiological maturation, characterized by

  19. GMP scale-up and banking of pluripotent stem cells for cellular therapy applications.

    Science.gov (United States)

    Ausubel, Lara J; Lopez, Patricia M; Couture, Larry A

    2011-01-01

    Human pluripotent stem cells (PSCs), which include human embryonic stem cells (ESCs) as well as induced pluripotent stem cells (iPSCs), represent an important source of cellular therapies in regenerative medicine and the study of early human development. As such, it is becoming increasingly important to develop methods for the large-scale banking of human PSC lines. There are several well-established methods for the propagation of human PSCs. The key to development of a good manufacturing practice (GMP) bank is to determine a manufacturing method that is amenable to large-scale production using materials that are fully documented. We have developed several banks of hESCs using animal feeder cells, animal-based matrices, or animal-free matrices. Protocols for growing hESCs on mouse embryonic fibroblasts (MEFs) are well established and are very helpful for producing research grade banks of cells. As most human ESCs cultured by research laboratories have been exposed to xenogeneic reagents, it is not imperative that all materials used in the production of a master cell bank be animal-free in origin. Nevertheless, as the field develops, it will no doubt become increasingly important to produce a bank of cells for clinical use without xenogeneic reagents, particularly nonhuman feeder cells which might harbor viruses with potential risk to human health or cell product integrity. Thus, even for cell lines previously exposed to xenogeneic reagents, it is important to minimize any subsequent exposure of the cell lines to additional adventitious agents. We have specifically described procedures for the growth of hESCs on Matrigel, an animal-matrix, and CELLstart, an animal-free matrix, and these can be used to produce hESCs as part of a clinical manufacturing process.

  20. Clinical phenotypes and outcome of patients hospitalized for acute heart failure: the ESC Heart Failure Long-Term Registry.

    Science.gov (United States)

    Chioncel, Ovidiu; Mebazaa, Alexandre; Harjola, Veli-Pekka; Coats, Andrew J; Piepoli, Massimo Francesco; Crespo-Leiro, Maria G; Laroche, Cecile; Seferovic, Petar M; Anker, Stefan D; Ferrari, Roberto; Ruschitzka, Frank; Lopez-Fernandez, Silvia; Miani, Daniela; Filippatos, Gerasimos; Maggioni, Aldo P

    2017-10-01

    To identify differences in clinical epidemiology, in-hospital management and 1-year outcomes among patients hospitalized for acute heart failure (AHF) and enrolled in the European Society of Cardiology Heart Failure Long-Term (ESC-HF-LT) Registry, stratified by clinical profile at admission. The ESC-HF-LT Registry is a prospective, observational study collecting hospitalization and 1-year follow-up data from 6629 AHF patients. Among AHF patients enrolled in the registry, 13.2% presented with pulmonary oedema (PO), 2.9% with cardiogenic shock (CS), 61.1% with decompensated heart failure (DHF), 4.8% with hypertensive heart failure (HT-HF), 3.5% with right heart failure (RHF) and 14.4% with AHF and associated acute coronary syndromes (ACS-HF). The 1-year mortality rate was 28.1% in PO, 54.0% in CS, 27.2% in DHF, 12.8% in HT-HF, 34.0% in RHF and 20.6% in ACS-HF patients. When patients were classified by systolic blood pressure (SBP) at initial presentation, 1-year mortality was 34.8% in patients with SBP 140 mmHg. These differences tended to diminish in the months post-discharge, and 1-year mortality for the patients who survived at least 6 months post-discharge did not vary significantly by either clinical profile or SBP classification. Rates of adverse outcomes in AHF remain high, and substantial differences have been found when patients were stratified by clinical profile or SBP. However, patients who survived at least 6 months post-discharge represent a more homogeneous group and their 1-year outcome is less influenced by clinical profile or SBP at admission. © 2017 The Authors. European Journal of Heart Failure © 2017 European Society of Cardiology.

  1. Proliferation of mouse endometrial stromal cells in culture is highly sensitive to lysophosphatidic acid signaling

    International Nuclear Information System (INIS)

    Aikawa, Shizu; Kano, Kuniyuki; Inoue, Asuka; Aoki, Junken

    2017-01-01

    Endometrial stromal cells (ESCs) proliferate rapidly both in vivo and in vitro. Here we show that proliferation of ESCs in vitro is strongly dependent on lysophosphatidic acid (LPA) signaling. LPA is produced by autotaxin (ATX) and induces various kinds of cellular processes including migration, proliferation and inhibition of cell death possibly through six G protein-coupled receptors (LPA 1-6 ). We found that ESCs proliferated rapidly in vitro in an autocrine manner and that the proliferation was prominently suppressed by either an ATX inhibitor (ONO-8430506) or an LPA 1/3 antagonist (Ki16425). Among the cells lines tested, mouse ESCs were the most sensitive to these inhibitors. Proliferation of ESCs isolated from either LPA 1 - or LPA 3 -deficient mice was comparable to proliferation of ESCs isolated from control mice. An LPA receptor antagonist (AM095), which was revealed to be a dual LPA 1 /LPA 3 antagonist, also suppressed the proliferation of ESCs. The present results show that LPA signaling has a critical role in the proliferation of ESCs, and that this role is possibly mediated redundantly by LPA 1 and LPA 3 . - Highlights: • Uterine endometrial stromal cells (ESCs) proliferate rapidly both in vivo and in vitro. • ESCs proliferated in vitro in an autocrine fashion. • Proliferation of mouse ESCs was prominently suppressed by inhibitors of lysophosphatidic acid (LPA) signaling. • LPA receptors, LPA 1 and LPA 3 , had redundant role in supporting the proliferation of ESCs.

  2. Analysis of mitochondrial function and localisation during human embryonic stem cell differentiation in vitro.

    Directory of Open Access Journals (Sweden)

    Andrew B J Prowse

    Full Text Available Human embryonic stem cell (hESC derivatives show promise as viable cell therapy options for multiple disorders in different tissues. Recent advances in stem cell biology have lead to the reliable production and detailed molecular characterisation of a range of cell-types. However, the role of mitochondria during differentiation has yet to be fully elucidated. Mitochondria mediate a cells response to altered energy requirements (e.g. cardiomyocyte contraction and, as such, the mitochondrial phenotype is likely to change during the dynamic process of hESC differentiation. We demonstrate that manipulating mitochondrial biogenesis alters mesendoderm commitment. To investigate mitochondrial localisation during early lineage specification of hESCs we developed a mitochondrial reporter line, KMEL2, in which sequences encoding the green fluorescent protein (GFP are targeted to the mitochondria. Differentiation of KMEL2 lines into the three germ layers showed that the mitochondria in these differentiated progeny are GFP positive. Therefore, KMEL2 hESCs facilitate the study of mitochondria in a range of cell types and, importantly, permit real-time analysis of mitochondria via the GFP tag.

  3. The promise of human embryonic stem cells in aging-associated diseases

    Science.gov (United States)

    Yabut, Odessa; Bernstein, Harold S.

    2011-01-01

    Aging-associated diseases are often caused by progressive loss or dysfunction of cells that ultimately affect the overall function of tissues and organs. Successful treatment of these diseases could benefit from cell-based therapy that would regenerate lost cells or otherwise restore tissue function. Human embryonic stem cells (hESCs) promise to be an important therapeutic candidate in treating aging-associated diseases due to their unique capacity for self-renewal and pluripotency. To date, there are numerous hESC lines that have been developed and characterized. We will discuss how hESC lines are derived, their molecular and cellular properties, and how their ability to differentiate into all three embryonic germ layers is determined. We will also outline the methods currently employed to direct their differentiation into populations of tissue-specific, functional cells. Finally, we will highlight the general challenges that must be overcome and the strategies being developed to generate highly-purified hESC-derived cell populations that can safely be used for clinical applications. PMID:21566262

  4. Derivation of Rabbit Embryonic Stem Cells from Vitrified–Thawed Embryos

    Science.gov (United States)

    Chen, Chien-Hong; Li, Yi; Hu, Yeshu; An, Li-You; Yang, Lan; Zhang, Jifeng; Chen, Y. Eugene

    2015-01-01

    Abstract The rabbit is a useful animal model for regenerative medicine. We previously developed pluripotent rabbit embryonic stem cell (rbESC) lines using fresh embryos. We also successfully cryopreserved rabbit embryos by vitrification. In the present work, we combined these two technologies to derive rbESCs using vitrified–thawed (V/T) embryos. We demonstrate that V/T blastocysts (BLs) can be used to derive pluripotent rbESCs with efficiencies comparable to those using fresh BLs. These ESCs are undistinguishable from the ones derived from fresh embryos. We tested the developmental capacity of rbESCs derived from V/T embryos by BL injection experiments and produced chimeric kits. Our work adds cryopreservation to the toolbox of rabbit stem cell research and applications and will greatly expand the available research materials for regenerative medicine in a clinically relevant animal model. PMID:26579970

  5. Site-Directed Genome Knockout in Chicken Cell Line and Embryos Can Use CRISPR/Cas Gene Editing Technology

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    Qisheng Zuo

    2016-06-01

    Full Text Available The present study established an efficient genome editing approach for the construction of stable transgenic cell lines of the domestic chicken (Gallus gallus domesticus. Our objectives were to facilitate the breeding of high-yield, high-quality chicken strains, and to investigate gene function in chicken stem cells. Three guide RNA (gRNAs were designed to knockout the C2EIP gene, and knockout efficiency was evaluated in DF-1 chicken fibroblasts and chicken ESCs using the luciferase single-strand annealing (SSA recombination assay, T7 endonuclease I (T7EI assay, and TA clone sequencing. In addition, the polyethylenimine-encapsulated Cas9/gRNA plasmid was injected into fresh fertilized eggs. At 4.5 d later, frozen sections of the embryos were prepared, and knockout efficiency was evaluated by the T7EI assay. SSA assay results showed that luciferase activity of the vector expressing gRNA-3 was double that of the control. Results of the T7EI assay and TA clone sequencing indicated that Cas9/gRNA vector-mediated gene knockdown efficiency was approximately 27% in both DF-1 cells and ESCs. The CRISPR/Cas9 vector was also expressed in chicken embryos, resulting in gene knockdown in three of the 20 embryos (gene knockdown efficiency 15%. Taken together, our results indicate that the CRISPR/Cas9 system can mediate stable gene knockdown at the cell and embryo levels in domestic chickens.

  6. Avaliação das possibilidades de escórias de siderurgia como corretivos da acidez do solo Evaluation of blast furnace slags as correctives for soil acidity

    Directory of Open Access Journals (Sweden)

    Antônio Carlos Pimentel Wutke

    1962-01-01

    Full Text Available No presente trabalho são apresentados os resultados obtidos em um ensaio de competição entre materiais corretivos, conduzido em vasos de Mitscherlich. Objetivou-se, principalmente, avaliar as possibilidades de dois tipos de escórias de siderurgia para substituir os corretivos usuais na agricultura. Foram comparados um calcário dolomítico cristalino, um calcário altamente cálcico, um material cálcico proveniente dos sambaquis do Litoral Sul paulista, uma escória de altos fornos a coque, produzida em Volta Redonda, e uma escória originária da produção de aços pelo processo Martin-Siemens básico, obtida em São Caetano. Os materiais corretivos foram comparados sempre em presença de adubação com NPK, feita em solução e na forma de drogas puras. Constaram, ainda, da experiência, um tratamento testemunha e outro com NPK, sem calagem. A planta indicadora utilizada foi a soja, Glycine max (L. Merril, variedade "Abura". No final do ensaio, além da colheita das sementes de soja, retiraram-se amostras de terra de todos os vasos, para determinações de seus índices pH e dos teores de H+ + AL+3 trocável. Com relação ao poder de neutralização da acidez do solo, revelaram-se mais eficiente o calcário dolomítico, o cálcico e o sambaqui Com eficiência intermediária colocou-se a escória de São Caetano e. em último lugar, a escória de Volta Redonda. As produções das sementes de soja, todavia, não confirmaram êstes resultados, uma vez que as diferenças, entre os tratamentos com corretivos não foram estatisticamente significativas. Moagem mais severa do que a utilizada neste ensaio possivelmente elevará a eficiência das duas escórias, como corretivos da acidez do solo. Com relação às suas influências sôbre a produção, avaliadas nas condições do ensaio, não há necessidade de se apurar mais o seu grau de moagem, a nao ser para atender às exigências legais, que disciplinam o comércio de adubos e corretivos

  7. Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells.

    Science.gov (United States)

    Wang, Ruoxing; Guo, Yan-Lin

    2012-10-01

    Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Epigenomic Co-localization and Co-evolution Reveal a Key Role for 5hmC as a Communication Hub in the Chromatin Network of ESCs

    Directory of Open Access Journals (Sweden)

    David Juan

    2016-02-01

    Full Text Available Summary: Epigenetic communication through histone and cytosine modifications is essential for gene regulation and cell identity. Here, we propose a framework that is based on a chromatin communication model to get insight on the function of epigenetic modifications in ESCs. The epigenetic communication network was inferred from genome-wide location data plus extensive manual annotation. Notably, we found that 5-hydroxymethylcytosine (5hmC is the most-influential hub of this network, connecting DNA demethylation to nucleosome remodeling complexes and to key transcription factors of pluripotency. Moreover, an evolutionary analysis revealed a central role of 5hmC in the co-evolution of chromatin-related proteins. Further analysis of regions where 5hmC co-localizes with specific interactors shows that each interaction points to chromatin remodeling, stemness, differentiation, or metabolism. Our results highlight the importance of cytosine modifications in the epigenetic communication of ESCs. : 5-hydroxymethylcytosine (5hmC plays a key role in the epigenomic communication network of embryonic stem cells. Juan et al. build a communication network based in co-localization of epigenomic data and literature. The analysis of the network and its components reveals that proteins reading and editing 5hmC co-evolve and serve as links between diverse molecular processes.

  9. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation.

    Science.gov (United States)

    Sun, Wu-Sheng; Chun, Ju-Lan; Do, Jeong-Tae; Kim, Dong-Hwan; Ahn, Jin-Seop; Kim, Min-Kyu; Hwang, In-Sul; Kwon, Dae-Jin; Hwang, Seong-Soo; Lee, Jeong-Woong

    2016-01-01

    Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE) and proximal enhancer (PE), in the 5' upstream regulatory sequences (URSs) of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs). We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free- Oct4 -promoter-driven EGFP reporter cassette with a PE-free- Oct4 -promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9) and a mouse EpiSC-like cell line (P19) before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naïve pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naïve pluripotent stem cells from other metastable state cells in a heterogenous population.

  10. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation

    Directory of Open Access Journals (Sweden)

    Wu-Sheng Sun

    2016-01-01

    Full Text Available Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE and proximal enhancer (PE, in the 5′ upstream regulatory sequences (URSs of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs and epiblast stem cells (EpiSCs. We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free-Oct4-promoter-driven EGFP reporter cassette with a PE-free-Oct4-promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9 and a mouse EpiSC-like cell line (P19 before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naïve pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naïve pluripotent stem cells from other metastable state cells in a heterogenous population.

  11. Prevention of Lysosomal Storage Diseases and Derivation of Mutant Stem Cell Lines by Preimplantation Genetic Diagnosis

    Science.gov (United States)

    Altarescu, Gheona; Beeri, Rachel; Eiges, Rachel; Epsztejn-Litman, Silvina; Eldar-Geva, Talia; Elstein, Deborah; Zimran, Ari; Margalioth, Ehud J.; Levy-Lahad, Ephrat; Renbaum, Paul

    2012-01-01

    Preimplantation genetic diagnosis (PGD) allows birth of unaffected children for couples at risk for a genetic disorder. We present the strategy and outcome of PGD for four lysosomal storage disorders (LSD): Tay-Sachs disease (TSD), Gaucher disease (GD), Fabry disease (FD), and Hunter syndrome (HS), and subsequent development of stem cell lines. For each disease, we developed a family-specific fluorescent multiplex single-cell PCR protocol that included the familial mutation and informative markers surrounding the mutation. Embryo biopsy and PGD analysis were performed on either oocytes (polar bodies one and two) or on single blastomeres from a six-cell embryo. We treated twenty families carrying mutations in these lysosomal storage disorders, including 3 couples requiring simultaneous analysis for two disorders (TSD/GD, TSD/balanced Robertsonian translocation 45XYder(21;14), and HS/oculocutaneus albinism). These analyses led to an overall pregnancy rate/embryo transfer of 38% and the birth of 20 unaffected children from 17 families. We have found that PGD for lysosomal disorders is a safe and effective method to prevent birth of affected children. In addition, by using mutant embryos for the derivation of stem cell lines, we have successfully established GD and HS hESC lines for use as valuable models in LSD research. PMID:23320174

  12. Identification of Proteins Related to Epigenetic Regulation in the Malignant Transformation of Aberrant Karyotypic Human Embryonic Stem Cells by Quantitative Proteomics

    Science.gov (United States)

    Sun, Yi; Yang, Yixuan; Zeng, Sicong; Tan, Yueqiu; Lu, Guangxiu; Lin, Ge

    2014-01-01

    Previous reports have demonstrated that human embryonic stem cells (hESCs) tend to develop genomic alterations and progress to a malignant state during long-term in vitro culture. This raises concerns of the clinical safety in using cultured hESCs. However, transformed hESCs might serve as an excellent model to determine the process of embryonic stem cell transition. In this study, ITRAQ-based tandem mass spectrometry was used to quantify normal and aberrant karyotypic hESCs proteins from simple to more complex karyotypic abnormalities. We identified and quantified 2583 proteins, and found that the expression levels of 316 proteins that represented at least 23 functional molecular groups were significantly different in both normal and abnormal hESCs. Dysregulated protein expression in epigenetic regulation was further verified in six pairs of hESC lines in early and late passage. In summary, this study is the first large-scale quantitative proteomic analysis of the malignant transformation of aberrant karyotypic hESCs. The data generated should serve as a useful reference of stem cell-derived tumor progression. Increased expression of both HDAC2 and CTNNB1 are detected as early as the pre-neoplastic stage, and might serve as prognostic markers in the malignant transformation of hESCs. PMID:24465727

  13. Gobernanza, riesgo y sistema financiero: el escándalo de la LIBOR

    Directory of Open Access Journals (Sweden)

    García Fronti, Javier

    2013-06-01

    Full Text Available The current financial crisis has hit us, compelling to reflect about the global financial system and its social effects. Political awareness can be radically altered by catastrophic experiences, enabling us to think of a transformation of the status quo. In this paper, we propose to reflect on the regional and global financial relations in light of the recent scandal LIBOR. It is essential that intergovernmental organizations, NGOs, democratic states, companies and civil society engage in processes of financial responsible innovations in the context of a global sustainable governance.La actual crisis financiera nos ha impactado de tal forma que es imposible escapar a una reflexión sobre el sistema financiero global y sus efectos sociales. La conciencia política puede ser radicalmente alterada a través de experiencias catastróficas, dándonos la posibilidad de pensar en una transformación del orden establecido. En este trabajo, nos proponemos reflexionar sobre las relaciones financieras regionales y globales a la luz del reciente escándalo de la LIBOR. Es fundamental que las organizaciones intergubernamentales, las organizaciones no gubernamentales, los Estados democráticos, las empresas y la sociedad civil se involucren en procesos de innovaciones financieras responsables en un contexto de gobernanza sostenible global.

  14. Efficient production of platelets from mouse embryonic stem cells by enforced expression of Gata2 in late hemogenic endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Kawaguchi, Manami [Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 (Japan); Kitajima, Kenji [Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Kanokoda, Mai [Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 (Japan); Suzuki, Hidenori [Division of Morphological and Biomolecular Research, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo, 113-8602 (Japan); Miyashita, Kazuya; Nakajima, Marino [Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 (Japan); Nuriya, Hideko [Core Technology and Research Center, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Kasahara, Kohji [Laboratory of Biomembrane, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Hara, Takahiko, E-mail: hara-tk@igakuken.or.jp [Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506 (Japan); Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 (Japan)

    2016-06-03

    Platelets are essential for blood circulation and coagulation. Previous study indicated that overexpression of Gata2 in differentiated mouse embryonic stem cells (ESCs) resulted in robust induction of megakaryocytes (Mks). To evaluate platelet production capacity of the Gata2-induced ESC-derived Mks, we generated iGata2-ESC line carrying the doxycycline-inducible Gata2 expression cassette. When doxycycline was added to day 5 hemogenic endothelial cells in the in vitro differentiation culture of iGata2-ESCs, c-Kit{sup −}Tie2{sup −}CD41{sup +} Mks were predominantly generated. These iGata2-ESC-derived Mks efficiently produced CD41{sup +}CD42b{sup +}CD61{sup +} platelets and adhered to fibrinogen-coated glass coverslips in response to thrombin stimulation. Transmission electron microscopy analysis demonstrated that the iGata2-ESC-derived platelets were discoid-shaped with α-granules and an open canalicular system, but were larger than peripheral blood platelets in size. These results demonstrated that an enforced expression of Gata2 in late HECs of differentiated ESCs efficiently promotes megakaryopoiesis followed by platelet production. This study provides valuable information for ex vivo platelet production from human pluripotent stem cells in future. -- Highlights: •Megakaryocytes are efficiently induced by Gata2 from ESC-derived day 5 HECs. •Gata2-induced ESC-derived megakaryocytes are c-Kit{sup −}Tie2{sup −}CD41{sup +}. •Gata2-induced ESC-derived megakaryocytes produce larger discoid-shaped platelets. •Gata2-induced ESC-derived platelets bind fibrinogen upon thrombin stimulation.

  15. Efficient production of platelets from mouse embryonic stem cells by enforced expression of Gata2 in late hemogenic endothelial cells

    International Nuclear Information System (INIS)

    Kawaguchi, Manami; Kitajima, Kenji; Kanokoda, Mai; Suzuki, Hidenori; Miyashita, Kazuya; Nakajima, Marino; Nuriya, Hideko; Kasahara, Kohji; Hara, Takahiko

    2016-01-01

    Platelets are essential for blood circulation and coagulation. Previous study indicated that overexpression of Gata2 in differentiated mouse embryonic stem cells (ESCs) resulted in robust induction of megakaryocytes (Mks). To evaluate platelet production capacity of the Gata2-induced ESC-derived Mks, we generated iGata2-ESC line carrying the doxycycline-inducible Gata2 expression cassette. When doxycycline was added to day 5 hemogenic endothelial cells in the in vitro differentiation culture of iGata2-ESCs, c-Kit − Tie2 − CD41 + Mks were predominantly generated. These iGata2-ESC-derived Mks efficiently produced CD41 + CD42b + CD61 + platelets and adhered to fibrinogen-coated glass coverslips in response to thrombin stimulation. Transmission electron microscopy analysis demonstrated that the iGata2-ESC-derived platelets were discoid-shaped with α-granules and an open canalicular system, but were larger than peripheral blood platelets in size. These results demonstrated that an enforced expression of Gata2 in late HECs of differentiated ESCs efficiently promotes megakaryopoiesis followed by platelet production. This study provides valuable information for ex vivo platelet production from human pluripotent stem cells in future. -- Highlights: •Megakaryocytes are efficiently induced by Gata2 from ESC-derived day 5 HECs. •Gata2-induced ESC-derived megakaryocytes are c-Kit − Tie2 − CD41 + . •Gata2-induced ESC-derived megakaryocytes produce larger discoid-shaped platelets. •Gata2-induced ESC-derived platelets bind fibrinogen upon thrombin stimulation.

  16. Atributos químicos do solo, crescimento radicular e produtividade do arroz de acordo com a aplicação de escórias Soil chemical attributes, root growth and rice yield according to slag application

    Directory of Open Access Journals (Sweden)

    Juliana Garcia Carvalho-Pupatto

    2004-12-01

    Full Text Available Resíduos industriais são fontes alternativas de nutrientes para as plantas e sua utilização decorre da necessidade de diminuir o acúmulo dos resíduos nos centros de produção. O objetivo deste trabalho foi avaliar o efeito de escórias de siderurgia nos atributos químicos do solo, no desenvolvimento e na produtividade de grãos do arroz de terras altas irrigado por aspersão. O delineamento experimental foi em blocos casualizados, com três tratamentos constituídos de duas escórias, alto-forno (196 g kg-1 de Si e aciaria (56 g kg-1 de Si, e a testemunha sem aplicação, com oito repetições. As escórias podem ser usadas como corretivo de acidez do solo e como fonte de silício. As alterações nos atributos químicos do solo estão relacionadas com a composição química das escórias. A escória de alto-forno proporcionou maior crescimento radicular em profundidade e melhor distribuição no perfil do solo e, conseqüentemente, maior produção de massa de matéria seca da parte aérea e produtividade de grãos de arroz.Use of industrial residues as alternative sources of nutrients to the plants is due to the necessity of reducing their accumulation in the production sites. The objective of this work was to evaluate the effect of metallurgy slag in soil chemical attributes, in development and in grain yield of upland rice sprinkler irrigated. The experiment was carried out in a randomized block design, with three treatments constituted by two scums, blast furnace (196 g kg-1 of Si and steel slag (56 g kg-1 of Si, and a control without application, with eight replications. The scums can be used as corrective of soil acidity and silicon source. Alterations in the soil chemical attributes are related to the chemical composition of the scums. Blast furnace slag resulted in higher root growth in depth and better distribution in the profile soil provides higher shoot dry matter yield and grain yield.

  17. Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruoxing [Department of Biological Sciences, The University of Southern Mississippi, 118 College Drive 5018, Hattiesburg, MS 39406 (United States); Guo, Yan-Lin, E-mail: yanlin.guo@usm.edu [Department of Biological Sciences, The University of Southern Mississippi, 118 College Drive 5018, Hattiesburg, MS 39406 (United States)

    2012-10-01

    Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs. -- Highlights: Black-Right-Pointing-Pointer Inhibition of Cdks slows down mESCs proliferation. Black-Right-Pointing-Pointer mESCs display remarkable recovery capacity from short-term cell cycle interruption. Black-Right-Pointing-Pointer Short-term cell cycle interruption does not compromise mESC self-renewal. Black

  18. Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

    International Nuclear Information System (INIS)

    Wang, Ruoxing; Guo, Yan-Lin

    2012-01-01

    Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs. -- Highlights: ► Inhibition of Cdks slows down mESCs proliferation. ► mESCs display remarkable recovery capacity from short-term cell cycle interruption. ► Short-term cell cycle interruption does not compromise mESC self-renewal. ► Oct4 and Nanog are up-regulated via de novo synthesis by cell cycle interruption.

  19. Generation of Corneal Keratocytes from Human Embryonic Stem Cells.

    Science.gov (United States)

    Hertsenberg, Andrew J; Funderburgh, James L

    2016-01-01

    Human Embryonic Stem Cells (hESC) offer an important resource as a limitless supply of any differentiated cell type of the human body. Keratocytes, cells from the corneal stroma, may have the potential for restoration of vision in cell therapy and biomedical engineering applications, but these specialized cells are not readily expanded in vitro. Here we describe a two-part method to produce keratocytes from the H1 hESC cell line. The hESC cells, maintained and expanded in feeder-free culture medium are first differentiated to neural crest cells using the stromal-derived inducing activity (SDIA) of the PA6 mouse embryonic fibroblast cell line. The resulting neural crest cells are selected by their expression of cell-surface CD271 and subsequently cultured as 3D pellets in a defined differentiation medium to induce a keratocyte phenotype.

  20. Prion protein expression regulates embryonic stem cell pluripotency and differentiation.

    Directory of Open Access Journals (Sweden)

    Alberto Miranda

    2011-04-01

    Full Text Available Cellular prion protein (PRNP is a glycoprotein involved in the pathogenesis of transmissible spongiform encephalopathies (TSEs. Although the physiological function of PRNP is largely unknown, its key role in prion infection has been extensively documented. This study examines the functionality of PRNP during the course of embryoid body (EB differentiation in mouse Prnp-null (KO and WT embryonic stem cell (ESC lines. The first feature observed was a new population of EBs that only appeared in the KO line after 5 days of differentiation. These EBs were characterized by their expression of several primordial germ cell (PGC markers until Day 13. In a comparative mRNA expression analysis of genes playing an important developmental role during ESC differentiation to EBs, Prnp was found to participate in the transcription of a key pluripotency marker such as Nanog. A clear switching off of this gene on Day 5 was observed in the KO line as opposed to the WT line, in which maximum Prnp and Nanog mRNA levels appeared at this time. Using a specific antibody against PRNP to block PRNP pathways, reduced Nanog expression was confirmed in the WT line. In addition, antibody-mediated inhibition of ITGB5 (integrin αvβ5 in the KO line rescued the low expression of Nanog on Day 5, suggesting the regulation of Nanog transcription by Prnp via this Itgb5. mRNA expression analysis of the PRNP-related proteins PRND (Doppel and SPRN (Shadoo, whose PRNP function is known to be redundant, revealed their incapacity to compensate for the absence of PRNP during early ESC differentiation. Our findings provide strong evidence for a relationship between Prnp and several key pluripotency genes and attribute Prnp a crucial role in regulating self-renewal/differentiation status of ESC, confirming the participation of PRNP during early embryogenesis.

  1. Dynamics of the transcriptome response of cultured human embryonic stem cells to ionizing radiation exposure

    International Nuclear Information System (INIS)

    Sokolov, Mykyta V.; Panyutin, Irina V.; Panyutin, Igor G.; Neumann, Ronald D.

    2011-01-01

    One of the key consequences of exposure of human cells to genotoxic agents is the activation of DNA damage responses (DDR). While the mechanisms underpinning DDR in fully differentiated somatic human cells have been studied extensively, molecular signaling events and pathways involved in DDR in pluripotent human embryonic stem cells (hESC) remain largely unexplored. We studied changes in the human genome-wide transcriptome of H9 hESC line following exposures to 1 Gy of gamma-radiation at 2 h and 16 h post-irradiation. Quantitative real-time PCR was performed to verify the expression data for a subset of genes. In parallel, the cell growth, DDR kinetics, and expression of pluripotency markers in irradiated hESC were monitored. The changes in gene expression in hESC after exposure to ionizing radiation (IR) are substantially different from those observed in somatic human cell lines. Gene expression patterns at 2 h post-IR showed almost an exclusively p53-dependent, predominantly pro-apoptotic, signature with a total of only 30 up-regulated genes. In contrast, the gene expression patterns at 16 h post-IR showed 354 differentially expressed genes, mostly involved in pro-survival pathways, such as increased expression of metallothioneins, ubiquitin cycle, and general metabolism signaling. Cell growth data paralleled trends in gene expression changes. DDR in hESC followed the kinetics reported for human somatic differentiated cells. The expression of pluripotency markers characteristic of undifferentiated hESC was not affected by exposure to IR during the time course of our analysis. Our data on dynamics of transcriptome response of irradiated hESCs may provide a valuable tool to screen for markers of IR exposure of human cells in their most naive state; thus unmasking the key elements of DDR; at the same time, avoiding the complexity of interpreting distinct cell type-dependent genotoxic stress responses of terminally differentiated cells.

  2. Dynamics of the transcriptome response of cultured human embryonic stem cells to ionizing radiation exposure

    Energy Technology Data Exchange (ETDEWEB)

    Sokolov, Mykyta V., E-mail: sokolovm@mail.nih.gov [Nuclear Medicine Division, Department of Radiology and Imaging Sciences, Clinical Center, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892 (United States); Panyutin, Irina V., E-mail: ipanyutinv@mail.nih.gov [Nuclear Medicine Division, Department of Radiology and Imaging Sciences, Clinical Center, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892 (United States); Panyutin, Igor G., E-mail: igorp@helix.nih.gov [Nuclear Medicine Division, Department of Radiology and Imaging Sciences, Clinical Center, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892 (United States); Neumann, Ronald D., E-mail: rneumann@mail.nih.gov [Nuclear Medicine Division, Department of Radiology and Imaging Sciences, Clinical Center, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892 (United States)

    2011-05-10

    One of the key consequences of exposure of human cells to genotoxic agents is the activation of DNA damage responses (DDR). While the mechanisms underpinning DDR in fully differentiated somatic human cells have been studied extensively, molecular signaling events and pathways involved in DDR in pluripotent human embryonic stem cells (hESC) remain largely unexplored. We studied changes in the human genome-wide transcriptome of H9 hESC line following exposures to 1 Gy of gamma-radiation at 2 h and 16 h post-irradiation. Quantitative real-time PCR was performed to verify the expression data for a subset of genes. In parallel, the cell growth, DDR kinetics, and expression of pluripotency markers in irradiated hESC were monitored. The changes in gene expression in hESC after exposure to ionizing radiation (IR) are substantially different from those observed in somatic human cell lines. Gene expression patterns at 2 h post-IR showed almost an exclusively p53-dependent, predominantly pro-apoptotic, signature with a total of only 30 up-regulated genes. In contrast, the gene expression patterns at 16 h post-IR showed 354 differentially expressed genes, mostly involved in pro-survival pathways, such as increased expression of metallothioneins, ubiquitin cycle, and general metabolism signaling. Cell growth data paralleled trends in gene expression changes. DDR in hESC followed the kinetics reported for human somatic differentiated cells. The expression of pluripotency markers characteristic of undifferentiated hESC was not affected by exposure to IR during the time course of our analysis. Our data on dynamics of transcriptome response of irradiated hESCs may provide a valuable tool to screen for markers of IR exposure of human cells in their most naive state; thus unmasking the key elements of DDR; at the same time, avoiding the complexity of interpreting distinct cell type-dependent genotoxic stress responses of terminally differentiated cells.

  3. La documentación geométrica de elementos patrimoniales con láser escáner terrestre. La experiencia del Lapa en Galicia

    Directory of Open Access Journals (Sweden)

    Mañana Borrazás, Patricia

    2009-12-01

    Full Text Available In this text we show some geometric documentation of various elements of Cultural Heritage. We use a Terrestrial Laser Scanner, a Leica HDS3000, a survey instrument that allows a high density register with pinpoint accuracy. The basic aim of this article is to evaluate the documentation posibilities of this kind of Terrestrial Laser Scanner and the different problems that emerged. As an example, we present the work processing of the topographic survey of a hillfort, the record in full detail of an archaeological excavation, the survey of a building and a two site of Rock Art.

    En este texto se presentan una serie de trabajos de documentación geométrica de diversos elementos patrimoniales realizados con láser escáner 3D. En ellos hemos empleado el modelo de Leica HDS3000, un instrumento topográfico que permite una captura de datos densa y con una resolución milimétrica. El objetivo principal de este documento es evaluar las posibilidades de registro de este escáner en distintos tipos de sitios así como los problemas que se generaron. Como ejemplos, presentamos el proceso de trabajo para obtener la topografía de un castro, el registro de detalle de una excavación, el levantamiento de un edificio y de unos petroglifos.

  4. Efflux protein expression in human stem cell-derived retinal pigment epithelial cells.

    Directory of Open Access Journals (Sweden)

    Kati Juuti-Uusitalo

    Full Text Available Retinal pigment epithelial (RPE cells in the back of the eye nourish photoreceptor cells and form a selective barrier that influences drug transport from the blood to the photoreceptor cells. At the molecular level, ATP-dependent efflux transporters have a major role in drug delivery in human RPE. In this study, we assessed the relative expression of several ATP-dependent efflux transporter genes (MRP1, -2, -3, -4, -5, -6, p-gp, and BCRP, the protein expression and localization of MRP1, MRP4, and MRP5, and the functionality of MRP1 efflux pumps at different maturation stages of undifferentiated human embryonic stem cells (hESC and RPE derived from the hESC (hESC-RPE. Our findings revealed that the gene expression of ATP-dependent efflux transporters MRP1, -3, -4, -5, and p-gp fluctuated during hESC-RPE maturation from undifferentiated hESC to fusiform, epithelioid, and finally to cobblestone hESC-RPE. Epithelioid hESC-RPE had the highest expression of MRP1, -3, -4, and P-gp, whereas the most mature cobblestone hESC-RPE had the highest expression of MRP5 and MRP6. These findings indicate that a similar efflux protein profile is shared between hESC-RPE and the human RPE cell line, ARPE-19, and suggest that hESC-RPE cells are suitable in vitro RPE models for drug transport studies. Embryonic stem cell model might provide a novel tool to study retinal cell differentiation, mechanisms of RPE-derived diseases, drug testing and targeted drug therapy.

  5. Implementing the ESC/ERS pulmonary hypertension guidelines: real-life cases from a national referral centre

    Directory of Open Access Journals (Sweden)

    D. Montani

    2009-12-01

    Full Text Available Pulmonary hypertension (PH comprises a heterogeneous group of disorders characterised by increased pulmonary vascular resistance that results in progressive right ventricular failure. In order to translate current evidence into routine clinical practice, the European Society of Cardiology (ESC and the European Respiratory Society (ERS have recently jointly proposed evidence-based guidelines for the optimal management of different PH patient groups. This article describes a series of clinical cases of PH due to various aetiologies that were referred to a large national PH expert referral centre. In each case, the assessment and therapeutic approach undertaken is described in the context of the new ECS/ERS guidelines. The routine diagnostic work-up of suspected idiopathic pulmonary arterial hypertension (PAH and recommended treatments for patients with functional class II, III and IV disease is emphasised. Familial screening and management of heritable PAH is discussed. Appropriate investigation and therapeutic strategies for patients with chronic thromboembolic disease and PH that is associated with congenital heart disease, pulmonary veno-occlusive disease and systemic sclerosis are also highlighted.

  6. All-trans retinoic acid directs urothelial specification of murine embryonic stem cells via GATA4/6 signaling mechanisms.

    Directory of Open Access Journals (Sweden)

    Joshua R Mauney

    2010-07-01

    Full Text Available The urinary bladder and associated tract are lined by the urothelium, a transitional epithelium that acts as a specialized permeability barrier that protects the underlying tissue from urine via expression of a highly specific group of proteins known as the uroplakins (UP. To date, our understanding of the developmental processes responsible for urothelial differentiation has been hampered due to the lack of suitable models. In this study, we describe a novel in vitro cell culture system for derivation of urothelial cells from murine embryonic stem cells (ESCs following cultivation on collagen matrices in the presence all trans retinoic acid (RA. Upon stimulation with micromolar concentrations of RA, ESCs significantly downregulated the pluripotency factor OCT-4 but markedly upregulated UP1A, UP1B, UP2, UP3A, and UP3B mRNA levels in comparison to naïve ESCs and spontaneously differentiating controls. Pan-UP protein expression was associated with both p63- and cytokeratin 20-positive cells in discrete aggregating populations of ESCs following 9 and 14 days of RA stimulation. Analysis of endodermal transcription factors such as GATA4 and GATA6 revealed significant upregulation and nuclear enrichment in RA-treated UP2-GFP+ populations. GATA4-/- and GATA6-/- transgenic ESC lines revealed substantial attenuation of RA-mediated UP expression in comparison to wild type controls. In addition, EMSA analysis revealed that RA treatment induced formation of transcriptional complexes containing GATA4/6 on both UP1B and UP2 promoter fragments containing putative GATA factor binding sites. Collectively, these data suggest that RA mediates ESC specification toward a urothelial lineage via GATA4/6-dependent processes.

  7. A unique epigenetic signature is associated with active DNA replication loci in human embryonic stem cells.

    Science.gov (United States)

    Li, Bing; Su, Trent; Ferrari, Roberto; Li, Jing-Yu; Kurdistani, Siavash K

    2014-02-01

    The cellular epigenetic landscape changes as pluripotent stem cells differentiate to somatic cells or when differentiated cells transform to a cancerous state. These epigenetic changes are commonly correlated with differences in gene expression. Whether active DNA replication is also associated with distinct chromatin environments in these developmentally and phenotypically diverse cell types has not been known. Here, we used BrdU-seq to map active DNA replication loci in human embryonic stem cells (hESCs), normal primary fibroblasts and a cancer cell line, and correlated these maps to the epigenome. In all cell lines, the majority of BrdU peaks were enriched in euchromatin and at DNA repetitive elements, especially at microsatellite repeats, and coincided with previously determined replication origins. The most prominent BrdU peaks were shared between all cells but a sizable fraction of the peaks were specific to each cell type and associated with cell type-specific genes. Surprisingly, the BrdU peaks that were common to all cell lines were associated with H3K18ac, H3K56ac, and H4K20me1 histone marks only in hESCs but not in normal fibroblasts or cancer cells. Depletion of the histone acetyltransferases for H3K18 and H3K56 dramatically decreased the number and intensity of BrdU peaks in hESCs. Our data reveal a unique epigenetic signature that distinguishes active replication loci in hESCs from normal somatic or malignant cells.

  8. Sulla digitalizzazione della street art. Linee guida per un'analisi del museo virtuale Reggiane Urban Gallery

    Directory of Open Access Journals (Sweden)

    Damiano Razzoli

    2017-09-01

    Full Text Available Reggiane Urban Gallery is a virtual museum based on the abandoned industrial plant of ex-Officine Reggiane in Reggio Emilia, now in restoration. Along the years the place has become an open space for local and national street-artists. The aim of this paper is to analyze the use of virtual reality and navigable interface as a solution for the conservation of the Street Art, in relation to the fact that many pieces would have been lost due to the demolition of several walls. Also, the analysis proposes guidelines for a digitalization of Street Art that could prevent the displacement of situated Street Art pieces in museums and encourage the creation of street art locative media museum, supported on four concepts - dispositive/device, locativity, topographic database, interface - and one discoursive feature designing the mediated experience of urban crossing practices: the mimetical relation between the software cultural components and Street Art.

  9. Resposta de mudas de goiabeira à aplicação de escória de siderurgia como corretivo de acidez do solo Response of guava plants to basic slag application as corrective of soil acidity

    Directory of Open Access Journals (Sweden)

    Renato de Mello Prado

    2003-04-01

    Full Text Available A escória de siderurgia pode constituir-se em uma fonte alternativa de Ca e Mg, bem como corretivo de acidez do solo, melhorando o estado nutricional de mudas de goiabeira, podendo contribuir para o sucesso da implantação de um pomar. O presente trabalho objetivou avaliar os efeitos da escória de siderurgia nas alterações dos atributos químicos do solo, na nutrição das plantas e no crescimento de mudas de goiabeira. Para tanto, instalou-se um experimento em Taquaritinga-SP, em condições de vasos, com doses crescentes de escória de siderurgia: zero; metade; uma vez; uma vez e meia; duas vezes e duas vezes e meia a dose para elevar a saturação por bases para 70%. Após 90 dias da incubação da escória no solo, procedeu-se o plantio das mudas de goiabeira (cv. Paluma, propagadas vegetativamente por estaquia, em substrato de um Argissolo Vermelho-Amarelo ácido (vaso com 2,8 dm³, cultivando-as por 105 dias. A aplicação de escória de siderurgia elevou os valores de pH, SB, V%, e as concentrações de Ca, Mg e P, diminuindo H+Al do solo. Nas mudas de goiabeira, houve aumento significativo na altura, no número de folhas, na área foliar, nas concentrações de Ca, Mg e P da parte aérea e das raízes das plantas e, conseqüentemente, na matéria seca da parte aérea e das raízes. Portanto, a escória de siderurgia mostrou-se viável na produção de mudas de goiabeira como corretivo de acidez do solo e fonte de nutrientes (Ca e Mg .The basic slag can consist in alternative source of Ca and Mg, as well as conecting soil acidity improving the nutricional state of guava plants, and determining the success of the implantation of an orchard. The present work was carried out to evaluate the basic slag in chemical attributes in soil in the nutrition of plants and the production of guava plants. By the way, an experiment was conducted in Taquaritinga-SP, in pot conditions, with increasing doses of basic slag: zero; half; once, once and

  10. Progress in human embryonic stem cell research in the United States between 2001 and 2010.

    Directory of Open Access Journals (Sweden)

    Keyvan Vakili

    Full Text Available On August 9th, 2001, the federal government of the United States announced a policy restricting federal funds available for research on human embryonic stem cell (hESCs out of concern for the "vast ethical mine fields" associated with the creation of embryos for research purposes. Until the policy was repealed on March 9th, 2009, no U.S. federal funds were available for research on hESCs extracted after August 9, 2001, and only limited federal funds were available for research on a subset of hESC lines that had previously been extracted. This paper analyzes how the 2001 U.S. federal funding restrictions influenced the quantity and geography of peer-reviewed journal publications on hESC. The primary finding is that the 2001 policy did not have a significant aggregate effect on hESC research in the U.S. After a brief lag in early 2000s, U.S. hESC research maintained pace with other areas of stem cell and genetic research. The policy had several other consequences. First, it was tied to increased hESC research funding within the U.S. at the state level, leading to concentration of related activities in a relatively small number of states. Second, it stimulated increased collaborative research between US-based scientists and those in countries with flexible policies toward hESC research (including Canada, the U.K., Israel, China, Spain, and South Korea. Third, it encouraged independent hESC research in countries without restrictions.

  11. Efficient differentiation of steroidogenic and germ-like cells from epigenetically-related iPSCs derived from ovarian granulosa cells.

    Directory of Open Access Journals (Sweden)

    Raymond Anchan

    Full Text Available To explore restoration of ovarian function using epigenetically-related, induced pluripotent stem cells (iPSCs, we functionally evaluated the epigenetic memory of novel iPSC lines, derived from mouse and human ovarian granulosa cells (GCs using c-Myc, Klf4, Sox2 and Oct4 retroviral vectors. The stem cell identity of the mouse and human GC-derived iPSCs (mGriPSCs, hGriPSCs was verified by demonstrating embryonic stem cell (ESC antigen expression using immunocytochemistry and RT-PCR analysis, as well as formation of embryoid bodies (EBs and teratomas that are capable of differentiating into cells from all three germ layers. GriPSCs' gene expression profiles associate more closely with those of ESCs than of the originating GCs as demonstrated by genome-wide analysis of mRNA and microRNA. A comparative analysis of EBs generated from three different mouse cell lines (mGriPSCs; fibroblast-derived iPSC, mFiPSCs; G4 embryonic stem cells, G4 mESCs revealed that differentiated mGriPSC-EBs synthesize 10-fold more estradiol (E2 than either differentiated FiPSC- or mESC-EBs under identical culture conditions. By contrast, mESC-EBs primarily synthesize progesterone (P4 and FiPSC-EBs produce neither E2 nor P4. Differentiated mGriPSC-EBs also express ovarian markers (AMHR, FSHR, Cyp19a1, ER and Inha as well as markers of early gametogenesis (Mvh, Dazl, Gdf9, Boule and Zp1 more frequently than EBs of the other cell lines. These results provide evidence of preferential homotypic differentiation of mGriPSCs into ovarian cell types. Collectively, our data support the hypothesis that generating iPSCs from the desired tissue type may prove advantageous due to the iPSCs' epigenetic memory.

  12. Molecular Characterization of Down Syndrome Embryonic Stem Cells Reveals a Role for RUNX1 in Neural Differentiation

    Directory of Open Access Journals (Sweden)

    Tomer Halevy

    2016-10-01

    Full Text Available Down syndrome (DS is the leading genetic cause of mental retardation and is caused by a third copy of human chromosome 21. The different pathologies of DS involve many tissues with a distinct array of neural phenotypes. Here we characterize embryonic stem cell lines with DS (DS-ESCs, and focus on the neural aspects of the disease. Our results show that neural progenitor cells (NPCs differentiated from five independent DS-ESC lines display increased apoptosis and downregulation of forehead developmental genes. Analysis of differentially expressed genes suggested RUNX1 as a key transcription regulator in DS-NPCs. Using genome editing we were able to disrupt all three copies of RUNX1 in DS-ESCs, leading to downregulation of several RUNX1 target developmental genes accompanied by reduced apoptosis and neuron migration. Our work sheds light on the role of RUNX1 and the importance of dosage balance in the development of neural phenotypes in DS.

  13. TALEN/CRISPR-mediated eGFP knock-in add-on at the OCT4 locus does not impact differentiation of human embryonic stem cells towards endoderm.

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    Nicole A J Krentz

    Full Text Available Human embryonic stem cells (hESCs have great promise as a source of unlimited transplantable cells for regenerative medicine. However, current progress on producing the desired cell type for disease treatment has been limited due to an insufficient understanding of the developmental processes that govern their differentiation, as well as a paucity of tools to systematically study differentiation in the lab. In order to overcome these limitations, cell-type reporter hESC lines will be required. Here we outline two strategies using Transcription Activator Like Effector Nucleases (TALENs and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR-CRISPR-Associated protein (Cas to create OCT4-eGFP knock-in add-on hESC lines. Thirty-one and forty-seven percent of clones were correctly modified using the TALEN and CRISPR-Cas9 systems, respectively. Further analysis of three correctly targeted clones demonstrated that the insertion of eGFP in-frame with OCT4 neither significantly impacted expression from the wild type allele nor did the fusion protein have a dramatically different biological stability. Importantly, the OCT4-eGFP fusion was easily detected using microscopy, flow cytometry and western blotting. The OCT4 reporter lines remained equally competent at producing CXCR4+ definitive endoderm that expressed a panel of endodermal genes. Moreover, the genomic modification did not impact the formation of NKX6.1+/SOX9+ pancreatic progenitor cells following directed differentiation. In conclusion, these findings demonstrate for the first time that CRISPR-Cas9 can be used to modify OCT4 and highlight the feasibility of creating cell-type specific reporter hESC lines utilizing genome-editing tools that facilitate homologous recombination.

  14. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling

    Science.gov (United States)

    Wang, Linlin; Schulz, Thomas C.; Sherrer, Eric S.; Dauphin, Derek S.; Shin, Soojung; Nelson, Angelique M.; Ware, Carol B.; Zhan, Mei; Song, Chao-Zhong; Chen, Xiaoji; Brimble, Sandii N.; McLean, Amanda; Galeano, Maria J.; Uhl, Elizabeth W.; D'Amour, Kevin A.; Chesnut, Jonathan D.; Rao, Mahendra S.

    2007-01-01

    Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs following stimulation with mouse embryonic fibroblast (MEF) conditioned medium (CM) revealed rapid and prominent tyrosine phosphorylation of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R); less prominent tyrosine phosphorylation of epidermal growth factor receptor (EGFR) family members, including ERBB2 and ERBB3; and trace phosphorylation of fibroblast growth factor receptors. Intense IGF1R and IR phosphorylation occurred in the absence of MEF conditioning (NCM) and was attributable to high concentrations of insulin in the proprietary KnockOut Serum Replacer (KSR). Inhibition of IGF1R using a blocking antibody or lentivirus-delivered shRNA reduced hESC self-renewal and promoted differentiation, while disruption of ERBB2 signaling with the selective inhibitor AG825 severely inhibited hESC proliferation and promoted apoptosis. A simple defined medium containing an IGF1 analog, heregulin-1β (a ligand for ERBB2/ERBB3), fibroblast growth factor-2 (FGF2), and activin A supported long-term growth of multiple hESC lines. These studies identify previously unappreciated RTKs that support hESC proliferation and self-renewal, and provide a rationally designed medium for the growth and maintenance of pluripotent hESCs. PMID:17761519

  15. Characterization of Bovine 5′-flanking Region during Differentiation of Mouse Embryonic Stem Cells

    Directory of Open Access Journals (Sweden)

    Hye-Jeong Jang

    2015-12-01

    Full Text Available Embryonic stem cells (ESCs have been used as a powerful tool for research including gene manipulated animal models and the study of developmental gene regulation. Among the critical regulatory factors that maintain the pluripotency and self-renewal of undifferentiated ESCs, NANOG plays a very important role. Nevertheless, because pluripotency maintaining factors and specific markers for livestock ESCs have not yet been probed, few studies of the NANOG gene from domestic animals including bovine have been reported. Therefore, we chose mouse ESCs in order to understand and compare NANOG expression between bovine, human, and mouse during ESCs differentiation. We cloned a 600 bp (−420/+181 bovine NANOG 5′-flanking region, and tagged it with humanized recombinant green fluorescent protein (hrGFP as a tracing reporter. Very high GFP expression for bovine NANOG promoter was observed in the mouse ESC line. GFP expression was monitored upon ESC differentiation and was gradually reduced along with differentiation toward neurons and adipocyte cells. Activity of bovine NANOG (−420/+181 promoter was compared with already known mouse and human NANOG promoters in mouse ESC and they were likely to show a similar pattern of regulation. In conclusion, bovine NANOG 5-flanking region functions in mouse ES cells and has characteristics similar to those of mouse and human. These results suggest that bovine gene function studied in mouse ES cells should be evaluated and extrapolated for application to characterization of bovine ES cells.

  16. Efeito da escória de siderurgia e calcário na disponibilidade de fósforo de um Latossolo Vermelho-Amarelo cultivado com cana-de-açúcar Effect of slag and limestone on the availability of phosphorus of an Oxisol planted with sugarcane

    Directory of Open Access Journals (Sweden)

    Renato de Mello Prado

    2001-09-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da escória de siderurgia e calcário na concentração do P disponível em um Latossolo Vermelho-Amarelo, cultivado com cana-de-açúcar (Saccharum spp., em Ituverava, SP. Foram aplicados calcário calcítico e a escória de siderurgia, objetivando elevar a saturação por bases a 50%, 75% e 100%, e usou-se testemunha (sem correção. O efeito linear da escória de siderurgia no P disponível do solo, em contraste com a ausência de relação quando se aplicou calcário, sugere que o efeito positivo da escória de siderurgia se deve mais ao efeito do silicato, de saturar os sítios de adsorção de P, do que ao efeito de aumento do pH.The aim of this work was to evaluate the effect of slag and limestone, on the available P concentration of the soil planted with sugarcane. A field experiment was conducted in Ituverava, SP, Brazil, in a Red-Yellow Oxisol. Calcitic limestone and the slag were added in order to increase the base saturation to 50%, 75% and 100%, in addition to the control without correction. The linear effect of the slag on the soil available P, contrasting the lack of relationship when the limestone was applied, suggests that the positive effect of the slag is mainly due to the silicate effect than to the pH effect.

  17. Function of FEZF1 during early neural differentiation of human embryonic stem cells.

    Science.gov (United States)

    Liu, Xin; Su, Pei; Lu, Lisha; Feng, Zicen; Wang, Hongtao; Zhou, Jiaxi

    2018-01-01

    The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model for dissecting human development because of unlimited self-renewal and the capacity to differentiate into nearly all cell types in the human body. In this study, using a chemical defined neural induction protocol and molecular profiling, we identified Fez family zinc finger 1 (FEZF1) as a potential regulator of early human neural development. FEZF1 is rapidly up-regulated during neural differentiation in hESCs and expressed before PAX6, a well-established marker of early human neural induction. We generated FEZF1-knockout H1 hESC lines using CRISPR-CAS9 technology and found that depletion of FEZF1 abrogates neural differentiation of hESCs. Moreover, loss of FEZF1 impairs the pluripotency exit of hESCs during neural specification, which partially explains the neural induction defect caused by FEZF1 deletion. However, enforced expression of FEZF1 itself fails to drive neural differentiation in hESCs, suggesting that FEZF1 is necessary but not sufficient for neural differentiation from hESCs. Taken together, our findings identify one of the earliest regulators expressed upon neural induction and provide insight into early neural development in human.

  18. Liberação de micronutrientes de uma escória aplicada em um Argissolo Vermelho-Amarelo cultivado com mudas de goiabeira (Psidium guajava L. Micronutrients released from one basic slag applied a ultisol cultivated with guava plants (Psidium guajava L.

    Directory of Open Access Journals (Sweden)

    RENATO DE MELLO PRADO

    2002-08-01

    Full Text Available Os micronutrientes são importantes na nutrição das plantas, especialmente em solos tropicais com baixa concentração devido ao intemperismo. Como fonte alternativa de micronutrientes, tem-se a escória, resíduo da indústria de produção de ferro-gusa e aço. Assim, objetivou-se avaliar a escória como fonte de micronutrientes para mudas de goiabeira. O delineamento experimental foi em blocos ao acaso, com cinco repetições. As doses de escória foram aplicadas objetivando elevar em meia, uma vez, uma vez e meia, duas vezes e duas vezes e meia a saturação por bases do solo igual a 70%, correspondendo a 1,68; 3,36; 5,04; 6,72 e 8,40 g por vaso, além da testemunha sem aplicação. Após 90 dias de incubação da escória com o Argissolo Vermelho-Amarelo, cultivaram-se mudas de goiabeira (cv. Paluma por 110 dias em vasos com 2,8 dm-3 de substrato, em viveiro telado, em Taquaritinga-SP, no período de outubro de 2000 a abril de 2001. A escória promoveu um efeito favorável na reação do solo e na disponibilidades de Zn, Cu, Mn e B do solo. Houve efeitos quadráticos nas concentrações de Zn, Cu e Mn do solo, que, por sua vez, estiveram associadas às doses de escória superiores a 5,8; 6,3 e 7,5 g por vaso, respectivamente, enquanto, para o B, esse efeito foi linear. A saturação por bases do solo, entre 51 e 55%, resultou em maior disponibilidade dos micronutrientes Zn, Cu e Mn no solo, ao passo que, para o B, esse valor foi de 65%. Da mesma forma que ocorreu no solo, a aplicação da escória apresentou efeitos quadráticos nos teores de Zn, Cu e Mn da parte aérea e das raízes das mudas de goiabeira, enquanto, para o B, esse efeito foi linear. Concluiu-se, portanto, que a escória se comportou como material corretivo da acidez e como fonte de micronutrientes.The micronutrients are important in the nutrition of the plants, especially in tropical ground with low concentration of these nutrients. As alternative source of

  19. Epigenetic mechanisms regulate MHC and antigen processing molecules in human embryonic and induced pluripotent stem cells.

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    Beatriz Suárez-Alvarez

    2010-04-01

    Full Text Available Human embryonic stem cells (hESCs are an attractive resource for new therapeutic approaches that involve tissue regeneration. hESCs have exhibited low immunogenicity due to low levels of Mayor Histocompatibility Complex (MHC class-I and absence of MHC class-II expression. Nevertheless, the mechanisms regulating MHC expression in hESCs had not been explored.We analyzed the expression levels of classical and non-classical MHC class-I, MHC class-II molecules, antigen-processing machinery (APM components and NKG2D ligands (NKG2D-L in hESCs, induced pluripotent stem cells (iPSCs and NTera2 (NT2 teratocarcinoma cell line. Epigenetic mechanisms involved in the regulation of these genes were investigated by bisulfite sequencing and chromatin immunoprecipitation (ChIP assays. We showed that low levels of MHC class-I molecules were associated with absent or reduced expression of the transporter associated with antigen processing 1 (TAP-1 and tapasin (TPN components in hESCs and iPSCs, which are involved in the transport and load of peptides. Furthermore, lack of beta2-microglobulin (beta2m light chain in these cells limited the expression of MHC class I trimeric molecule on the cell surface. NKG2D ligands (MICA, MICB were observed in all pluripotent stem cells lines. Epigenetic analysis showed that H3K9me3 repressed the TPN gene in undifferentiated cells whilst HLA-B and beta2m acquired the H3K4me3 modification during the differentiation to embryoid bodies (EBs. Absence of HLA-DR and HLA-G expression was regulated by DNA methylation.Our data provide fundamental evidence for the epigenetic control of MHC in hESCs and iPSCs. Reduced MHC class I and class II expression in hESCs and iPSCs can limit their recognition by the immune response against these cells. The knowledge of these mechanisms will further allow the development of strategies to induce tolerance and improve stem cell allograft acceptance.

  20. Epigenetic Mechanisms Regulate MHC and Antigen Processing Molecules in Human Embryonic and Induced Pluripotent Stem Cells

    Science.gov (United States)

    Suárez-Álvarez, Beatriz; Rodriguez, Ramón M.; Calvanese, Vincenzo; Blanco-Gelaz, Miguel A.; Suhr, Steve T.; Ortega, Francisco; Otero, Jesus; Cibelli, Jose B.; Moore, Harry; Fraga, Mario F.; López-Larrea, Carlos

    2010-01-01

    Background Human embryonic stem cells (hESCs) are an attractive resource for new therapeutic approaches that involve tissue regeneration. hESCs have exhibited low immunogenicity due to low levels of Mayor Histocompatibility Complex (MHC) class-I and absence of MHC class-II expression. Nevertheless, the mechanisms regulating MHC expression in hESCs had not been explored. Methodology/Principal Findings We analyzed the expression levels of classical and non-classical MHC class-I, MHC class-II molecules, antigen-processing machinery (APM) components and NKG2D ligands (NKG2D-L) in hESCs, induced pluripotent stem cells (iPSCs) and NTera2 (NT2) teratocarcinoma cell line. Epigenetic mechanisms involved in the regulation of these genes were investigated by bisulfite sequencing and chromatin immunoprecipitation (ChIP) assays. We showed that low levels of MHC class-I molecules were associated with absent or reduced expression of the transporter associated with antigen processing 1 (TAP-1) and tapasin (TPN) components in hESCs and iPSCs, which are involved in the transport and load of peptides. Furthermore, lack of β2-microglobulin (β2m) light chain in these cells limited the expression of MHC class I trimeric molecule on the cell surface. NKG2D ligands (MICA, MICB) were observed in all pluripotent stem cells lines. Epigenetic analysis showed that H3K9me3 repressed the TPN gene in undifferentiated cells whilst HLA-B and β2m acquired the H3K4me3 modification during the differentiation to embryoid bodies (EBs). Absence of HLA-DR and HLA-G expression was regulated by DNA methylation. Conclusions/Significance Our data provide fundamental evidence for the epigenetic control of MHC in hESCs and iPSCs. Reduced MHC class I and class II expression in hESCs and iPSCs can limit their recognition by the immune response against these cells. The knowledge of these mechanisms will further allow the development of strategies to induce tolerance and improve stem cell allograft acceptance

  1. Aplicación de las geotecnologías láser escáner para la digitalización tridimensional de pinturas rupestres de abrigos

    OpenAIRE

    Santisteban Martínez, Raúl

    2013-01-01

    Este proyecto tiene como finalidad documentar las pinturas rupestres descubiertas en Ojos Albos, de una manera no destructiva aprovechando la tecnología disponible en la escuela Politécnica Superior de Ávila y los conocimientos en el campo de la fotografía del alumno. Este documento aborda las distintitas tareas realizadas durante la documentación del yacimiento, desde la generación de un plano partiendo de los datos obtenidos con el láser-escáner 3D, hasta documentación fotográfica de los...

  2. Human embryonic stem cells: preclinical perspectives

    Directory of Open Access Journals (Sweden)

    Sarda Kanchan

    2008-01-01

    Full Text Available Abstract Human embryonic stem cells (hESCs have been extensively discussed in public and scientific communities for their potential in treating diseases and injuries. However, not much has been achieved in turning them into safe therapeutic agents. The hurdles in transforming hESCs to therapies start right with the way these cells are derived and maintained in the laboratory, and goes up-to clinical complications related to need for patient specific cell lines, gender specific aspects, age of the cells, and several post transplantation uncertainties. The different types of cells derived through directed differentiation of hESC and used successfully in animal disease and injury models are described briefly. This review gives a brief outlook on the present and the future of hESC based therapies, and talks about the technological advances required for a safe transition from laboratory to clinic.

  3. LIF supports primitive endoderm expansion during pre-implantation development

    DEFF Research Database (Denmark)

    Morgani, Sophie M; Brickman, Joshua M

    2015-01-01

    Embryonic stem cells (ESCs) are pluripotent cell lines that can be maintained indefinitely in an early developmental state. ESC culture conditions almost always require the cytokine LIF to maintain self-renewal. As ESCs are not homogeneous but contain multiple populations reminiscent...... of the blastocyst, identifying the target cells of LIF is necessary to understand the propagation of pluripotency. We recently found that LIF acts under self-renewing conditions to stimulate the fraction of ESCs that express extraembryonic markers, but has little impact on pluripotent gene expression. Here, we...... derivatives, whereas the increase in PrE is mediated both by an increase in proliferation and inhibition of PrE apoptosis that is normally triggered in embryos with an excess of GATA6(+) cells. Thus, it appears that the relative size of the PrE is determined by the number of LIF-producing cells in the embryo...

  4. Challenges of using pluripotent stem cells for safety assessments of substances

    International Nuclear Information System (INIS)

    Vojnits, Kinga; Bremer, Susanne

    2010-01-01

    Various European Union (EU) legislations request the use of in vitro tests for toxicological evaluations in order to increase the safety of consumer and patients but also to reduce the number vertebrates. The review provides a brief overview on EU legislations in place but without further interpretation. At present several ongoing EU projects address the need of developing predictive in vitro tests including projects assessing the potential of human embryonic stem cell (hESC) lines as basis for a range of toxicity tests. Tests based on human cells would avoid interspecies variations and as such predict more precisely adverse effects to the human body. However, the ethical situation on the use of toxicity tests based on hESCs is still under debate since no harmonization within Europe on the use of hESC lines has been achieved yet. A mutual acceptance of toxicity tests based on hESCs for regulatory applications is therefore challenging. Recent reports on the establishment of induced pluripotent stem cells (iPSC) are pointing to a way out of this dilemma, since these cells have apparently very similar characteristics as hESCs and could serve as basis for the development of toxicity tests. A careful scientific comparison between pluripotent cells of different origin is now needed in order to make final judgments. In any case, the development of reliable and relevant in vitro toxicity tests based on human pluripotent cells requires additional quality assessments of critical parameter that are also summarized within the review.

  5. A quantitative system for discriminating induced pluripotent stem cells, embryonic stem cells and somatic cells.

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    Anyou Wang

    Full Text Available Induced pluripotent stem cells (iPSCs derived from somatic cells (SCs and embryonic stem cells (ESCs provide promising resources for regenerative medicine and medical research, leading to a daily identification of new cell lines. However, an efficient system to discriminate the different types of cell lines is lacking. Here, we develop a quantitative system to discriminate the three cell types, iPSCs, ESCs, and SCs. The system consists of DNA-methylation biomarkers and mathematical models, including an artificial neural network and support vector machines. All biomarkers were unbiasedly selected by calculating an eigengene score derived from analysis of genome-wide DNA methylations. With 30 biomarkers, or even with as few as 3 top biomarkers, this system can discriminate SCs from pluripotent cells (PCs, including ESCs and iPSCs with almost 100% accuracy. With approximately 100 biomarkers, the system can distinguish ESCs from iPSCs with an accuracy of 95%. This robust system performs precisely with raw data without normalization as well as with converted data in which the continuous methylation levels are accounted. Strikingly, this system can even accurately predict new samples generated from different microarray platforms and the next-generation sequencing. The subtypes of cells, such as female and male iPSCs and fetal and adult SCs, can also be discriminated with this method. Thus, this novel quantitative system works as an accurate framework for discriminating the three cell types, iPSCs, ESCs, and SCs. This strategy also supports the notion that DNA-methylation generally varies among the three cell types.

  6. Aplicação de escória de aciaria e calcário no solo e desenvolvimento do primeiro ciclo da grama bermuda

    OpenAIRE

    Oliveira, Mauricio Roberto de [UNESP

    2012-01-01

    O mercado brasileiro de grama cultivada tem expandido em decorrência a maior demanda em áreas paisagísticas e esportivas. Para maximizar a produção de grama, de maneira econômica, os tapetes devem ser produzidos com qualidade e em um tempo reduzido, sendo assim, a preparação do solo tem grande influência na produtividade dos gramados. Diante disso, tem se estudado diferentes corretivos da acidez do solo, e o mais novo é um subproduto da fabricação do aço, conhecido como escória de aciaria. Um...

  7. Vico and Literary Mannerism

    DEFF Research Database (Denmark)

    Catana, Leo

    Reviews: Scott Samuelson, New Vico studies, vol. 18 (2000), pp. 111-115; Maurizio Martirano, Sesto contribuito alla bibliografia vichiana (1996-2000); Alfredo Guida Editore: ?, (2002), p. 70 (Studi vichiani, vol. 37)...

  8. GROα regulates human embryonic stem cell self-renewal or adoption of a neuronal fate

    Science.gov (United States)

    Krtolica, Ana; Larocque, Nick; Genbacev, Olga; Ilic, Dusko; Coppe, Jean-Philippe; Patil, Christopher K.; Zdravkovic, Tamara; McMaster, Michael; Campisi, Judith; Fisher, Susan J.

    2012-01-01

    Previously we reported that feeders formed from human placental fibroblasts (hPFs) support derivation and long-term self-renewal of human embryonic stem cells (hESCs) under serum-free conditions. Here, we show, using antibody array and ELISA platforms, that hPFs secrete ~6-fold higher amounts of the CXC-type chemokine, GROα, than IMR 90, a human lung fibroblast line, which does not support hESC growth. Furthermore, immunocytochemistry and immunoblot approaches revealed that hESCs express CXCR, a GROα receptor. We used this information to develop defined culture medium for feeder-free propagation of hESCs in an undifferentiated state. Cells passaged as small aggregates and maintained in the GROα-containing medium had a normal karyotype, expressed pluripotency markers, and exhibited apical–basal polarity, i.e., had the defining features of pluripotent hESCs. They also differentiated into the three primary (embryonic) germ layers and formed teratomas in immunocompromised mice. hESCs cultured as single cells in the GROα-containing medium also had a normal karyotype, but they downregulated markers of pluripotency, lost apical–basal polarity, and expressed markers that are indicative of the early stages of neuronal differentiation—βIII tubulin, vimentin, radial glial protein, and nestin. These data support our hypothesis that establishing and maintaining cell polarity is essential for the long-term propagation of hESCs in an undifferentiated state and that disruption of cell–cell contacts can trigger adoption of a neuronal fate. PMID:21396766

  9. Switching of the core structures of glycosphingolipids from globo- and lacto- to ganglio-series upon human embryonic stem cell differentiation.

    Science.gov (United States)

    Liang, Yuh-Jin; Kuo, Huan-Hsien; Lin, Chi-Hung; Chen, Yen-Ying; Yang, Bei-Chia; Cheng, Yuan-Yuan; Yu, Alice L; Khoo, Kay-Hooi; Yu, John

    2010-12-28

    A systematic survey of expression profiles of glycosphingolipids (GSLs) in two hESC lines and their differentiated embryoid body (EB) outgrowth with three germ layers was carried out using immunofluorescence, flow cytometry, and MALDI-MS and MS/MS analyses. In addition to the well-known hESC-specific markers stage-specific embryonic antigen 3 (SSEA-3) and SSEA-4, we identified several globosides and lacto-series GSLs, previously unrevealed in hESCs, including Gb(4)Cer, Lc(4)Cer, fucosyl Lc(4)Cer, Globo H, and disialyl Gb(5)Cer. During hESC differentiation into EBs, MS analysis revealed a clear-cut switch in the core structures of GSLs from globo- and lacto- to ganglio-series, which was not as evident by immunostaining with antibodies against SSEA-3 and SSEA-4, owing to their cross-reactivities with various glycosphingolipids. Such a switch was attributable to altered expression of key glycosyltransferases (GTs) in the biosynthetic pathways by the up-regulation of ganglio-series-related GTs with simultaneous down-regulation of globo- and lacto-series-related GTs. Thus, these results provide insights into the unique stage-specific transition and mechanism for alterations of GSL core structures during hESC differentiation. In addition, unique glycan structures uncovered by MS analyses may serve as surface markers for further delineation of hESCs and help identify of their functional roles not only in hESCs but also in cancers.

  10. NANOG reporter cell lines generated by gene targeting in human embryonic stem cells

    DEFF Research Database (Denmark)

    Fischer, Yvonne; Ganic, Elvira; Ameri, Jacqueline

    2010-01-01

    Pluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role...

  11. Embryonic Stem Cell-Derived Cardiomyocyte Heterogeneity and the Isolation of Immature and Committed Cells for Cardiac Remodeling and Regeneration

    Directory of Open Access Journals (Sweden)

    Kenneth R. Boheler

    2011-01-01

    Full Text Available Pluripotent stem cells represent one promising source for cell replacement therapy in heart, but differentiating embryonic stem cell-derived cardiomyocytes (ESC-CMs are highly heterogeneous and show a variety of maturation states. In this study, we employed an ESC clonal line that contains a cardiac-restricted ncx1 promoter-driven puromycin resistance cassette together with a mass culture system to isolate ESC-CMs that display traits characteristic of very immature CMs. The cells display properties of proliferation, CM-restricted markers, reduced mitochondrial mass, and hypoxia-resistance. Following transplantation into rodent hearts, bioluminescence imaging revealed that immature cells, but not more mature CMs, survived for at least one month following injection. These data and comparisons with more mature cells lead us to conclude that immature hypoxia resistant ESC-CMs can be isolated in mass in vitro and, following injection into heart, form grafts that may mediate long-term recovery of global and regional myocardial contractile function following infarction.

  12. Expresividades de la imagen: régimen escópico, espacialidad y sensibilidades

    Directory of Open Access Journals (Sweden)

    Ileana Desirée Ibáñez

    2013-01-01

    Full Text Available En este trabajo ponemos en común reflexiones a partir de un taller de fotografía llevado a cabo con un grupo de jóvenes de sectores subalternos que habitan contextos de socio-segregación. El taller como metodología de investigación surge de la necesidad de posibilitar el despliegue de otras maneras de expresión, alternativas a la palabra, que permitan acercarnos a las dimensiones sensibles de las vivencias de los sujetos. Propusimos realizar un taller de fotografía considerando que la materialidad de las imágenes es una vía para acercarnos al "régimen escópico", a las maneras en que en el capitalismo contemporáneo se configura el sentido de la vista, entendiendo a los sentidos como objeto de regulación cotidiana, provisoria e incompleta. En primer lugar abordaremos algunas consideraciones teóricas acerca de la especificidad de la técnica y de su producto: la fotografía como objeto social en relación a las posibilidades de expresividad. Luego esbozaremos un esquema de lectura a partir de los aportes de Adrián SCRIBANO, Roland BARTHES y John BERGER. Finalmente expondremos algunos resultados a través de una lectura sobre las espacialidades, movilidades y sensibilidades planteadas en las imágenes. URN: http://nbn-resolving.de/urn:nbn:de:0114-fqs1301265

  13. Generation of human β-thalassemia induced pluripotent cell lines by reprogramming of bone marrow-derived mesenchymal stromal cells using modified mRNA.

    Science.gov (United States)

    Varela, Ioanna; Karagiannidou, Angeliki; Oikonomakis, Vasilis; Tzetis, Maria; Tzanoudaki, Marianna; Siapati, Elena-Konstantina; Vassilopoulos, George; Graphakos, Stelios; Kanavakis, Emmanuel; Goussetis, Evgenios

    2014-12-01

    Synthetic modified mRNA molecules encoding pluripotency transcription factors have been used successfully in reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs). We have applied this method on bone marrow-derived mesenchymal stromal cells (BM-MSCs) obtained from a patient with β-thalassemia (β-thal) with the aim to generate trangene-free β-thal-iPSCs. Transfection of 10(4) BM-MSCs by lipofection with mRNA encoding the reprogramming factors Oct4, Klf4, Sox2, cMyc, and Lin28 resulted in formation of five iPSC colonies, from which three were picked up and expanded in β-thal-iPSC lines. After 10 serial passages in vitro, β-thal-iPSCs maintain genetic stability as shown by array comparative genomic hybridization (aCGH) and are capable of forming embryoid bodies in vitro and teratomas in vivo. Their gene expression profile compared to human embryonic stem cells (ESCs) and BM-MSCs seems to be similar to that of ESCs, whereas it differs from the profile of the parental BM-MSCs. Differentiation cultures toward a hematopoietic lineage showed the generation of CD34(+) progenitors up to 10%, but with a decreased hematopoietic colony-forming capability. In conclusion, we report herein the generation of transgene-free β-thal-iPSCs that could be widely used for disease modeling and gene therapy applications. Moreover, it was demonstrated that the mRNA-based reprogramming method, used mainly in fibroblasts, is also suitable for reprogramming of human BM-MSCs.

  14. Effects of Low Doses of Ionizing Radiation Exposures on Stress-Responsive Gene Expression in Human Embryonic Stem Cells

    Directory of Open Access Journals (Sweden)

    Mykyta Sokolov

    2014-01-01

    Full Text Available There is a great deal of uncertainty on how low (≤0.1 Gy doses of ionizing radiation (IR affect human cells, partly due to a lack of suitable experimental model systems for such studies. The uncertainties arising from low-dose IR human data undermine practical societal needs to predict health risks emerging from diagnostic medical tests’ radiation, natural background radiation, and environmental radiological accidents. To eliminate a variability associated with remarkable differences in radioresponses of hundreds of differentiated cell types, we established a novel, human embryonic stem cell (hESC-based model to examine the radiobiological effects in human cells. Our aim is to comprehensively elucidate the gene expression changes in a panel of various hESC lines following low IR doses of 0.01; 0.05; 0.1 Gy; and, as a reference, relatively high dose of 1 Gy of IR. Here, we examined the dynamics of transcriptional changes of well-established IR-responsive set of genes, including CDKN1A, GADD45A, etc. at 2 and 16 h post-IR, representing “early” and “late” radioresponses of hESCs. Our findings suggest the temporal- and hESC line-dependence of stress gene radioresponses with no statistically significant evidence for a linear dose-response relationship within the lowest doses of IR exposures.

  15. Transplantation of rat embryonic stem cell-derived retinal progenitor cells preserves the retinal structure and function in rat retinal degeneration.

    Science.gov (United States)

    Qu, Zepeng; Guan, Yuan; Cui, Lu; Song, Jian; Gu, Junjie; Zhao, Hanzhi; Xu, Lei; Lu, Lixia; Jin, Ying; Xu, Guo-Tong

    2015-11-09

    Degenerative retinal diseases like age-related macular degeneration (AMD) are the leading cause of blindness. Cell transplantation showed promising therapeutic effect for such diseases, and embryonic stem cell (ESC) is one of the sources of such donor cells. Here, we aimed to generate retinal progenitor cells (RPCs) from rat ESCs (rESCs) and to test their therapeutic effects in rat model. The rESCs (DA8-16) were cultured in N2B27 medium with 2i, and differentiated to two types of RPCs following the SFEBq method with modifications. For rESC-RPC1, the cells were switched to adherent culture at D10, while for rESC-RPC2, the suspension culture was maintained to D14. Both RPCs were harvested at D16. Primary RPCs were obtained from P1 SD rats, and some of them were labeled with EGFP by infection with lentivirus. To generate Rax::EGFP knock-in rESC lines, TALENs were engineered to facilitate homologous recombination in rESCs, which were cotransfected with the targeting vector and TALEN vectors. The differentiated cells were analyzed with live image, immunofluorescence staining, flow cytometric analysis, gene expression microarray, etc. RCS rats were used to mimic the degeneration of retina and test the therapeutic effects of subretinally transplanted donor cells. The structure and function of retina were examined. We established two protocols through which two types of rESC-derived RPCs were obtained and both contained committed retina lineage cells and some neural progenitor cells (NPCs). These rESC-derived RPCs survived in the host retinas of RCS rats and protected the retinal structure and function in early stage following the transplantation. However, the glia enriched rESC-RPC1 obtained through early and longer adherent culture only increased the b-wave amplitude at 4 weeks, while the longer suspension culture gave rise to evidently neuronal differentiation in rESC-RPC2 which significantly improved the visual function of RCS rats. We have successfully differentiated

  16. Corrupção, fisiologismo, clientelismo e reacionarismo na morta-viva gestão presidencial temerária: anatomia de um escândalo antirrepublicano

    Directory of Open Access Journals (Sweden)

    Renato Nunes Bittencourt

    2017-08-01

    Full Text Available O artigo analisa a sobrevida do governo Temer, enredado em diversos escândalos que evidenciam seu descompromisso e sua ilegitimidade na condução da república brasileira, mais do que nunca comandada por estruturas plutocráticas desprovidas de qualquer projeto democrático de emancipação social. A sandice do projeto de reforma da previdência e da reforma trabalhista somente satisfaz aos interesses empresariais e aos espoliadores dos direitos sociais, perpetuando ainda mais a precariedade da vida pública brasileira, submetida ao crivo do reacionarismo autoritário.

  17. Tratamiento Manual Postcirugía en un caso Clínico de Transposición muscular del Romboides y Angular de la Escápula

    OpenAIRE

    Gómez García, Ana

    2009-01-01

    Descripción de un caso clínico en la que se secciona el nervio espinal derecho (neurotmesis) tras la exéresis de un quiste en el cuello. Debido a la falta de transmisión neuronal se produce una parálisis funcional en el músculo trapecio y en el músculo esternocleidomastoideo derecho y se suple sus funciones mediante una reinserción del músculo elevador de la escápula y del músculo romboides de dicho lado que recibe el nombre de transposición muscular. Se realiza tratamiento de fisioter...

  18. From "ES-like" cells to induced pluripotent stem cells: a historical perspective in domestic animals.

    Science.gov (United States)

    Koh, Sehwon; Piedrahita, Jorge A

    2014-01-01

    Pluripotent stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) provide great potential as cell sources for gene editing to generate genetically modified animals, as well as in the field of regenerative medicine. Stable, long-term ESCs have been established in laboratory mouse and rat; however, isolation of true pluripotent ESCs in domesticated animals such as pigs and dogs have been less successful. Initially, domesticated animal pluripotent cell lines were referred to as "embryonic stem-like" cells owing to their similar morphologic characteristics to mouse ESCs, but accompanied by a limited ability to proliferate in vitro in an undifferentiated state. That is, they shared some but not all the characteristics of true ESCs. More recently, advances in reprogramming using exogenous transcription factors, combined with the utilization of small chemical inhibitors of key biochemical pathways, have led to the isolation of iPSCs. In this review, we provide a historical perspective of the isolation of various types of pluripotent stem cells in domesticated animals. In addition, we summarize the latest progress and limitations in the derivation and application of iPSCs. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Minimal requirements for quality controls in radiotherapy with external beams; Controlli di qualita' essenziali in radioterapia con fasci esterni

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-07-01

    Physical dosimetric guidelines have been developed by the Italian National Institute of Health study group on quality assurance in radiotherapy to define protocols for quality controls in external beam radiotherapy. While the document does not determine strict rules or firm recommendations, it suggests minimal requirements for quality controls necessary to guarantee an adequate degree of accuracy in external beam radiotherapy. [Italian] Il gruppo di studio Assicurazione di qualita' in radioterapia dell'Istituto Superiore di Sanita' presenta le linee guida per la stesura dei protocolli di controllo di qualita' essenziali necessari a garantire un adeguato livello di accuratezza del trattamento radiante e rappresenta pertanto una parte essenziale del contributo fisico-dosimetrico globale di assicurazione di qualita' in radioterapia con fasci esterni.

  20. Minimal requirements for quality controls in radiotherapy with external beams; Controlli di qualita' essenziali in radioterapia con fasci esterni

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-07-01

    Physical dosimetric guidelines have been developed by the Italian National Institute of Health study group on quality assurance in radiotherapy to define protocols for quality controls in external beam radiotherapy. While the document does not determine strict rules or firm recommendations, it suggests minimal requirements for quality controls necessary to guarantee an adequate degree of accuracy in external beam radiotherapy. [Italian] Il gruppo di studio Assicurazione di qualita' in radioterapia dell'Istituto Superiore di Sanita' presenta le linee guida per la stesura dei protocolli di controllo di qualita' essenziali necessari a garantire un adeguato livello di accuratezza del trattamento radiante e rappresenta pertanto una parte essenziale del contributo fisico-dosimetrico globale di assicurazione di qualita' in radioterapia con fasci esterni.

  1. Construcción de un escáner tridimensional usando C++ y hardware libre

    Directory of Open Access Journals (Sweden)

    Cristian Enrique Gamboa

    2017-01-01

    Full Text Available En este trabajo, se desarrolló un sistema económico de escaneo tridimensional (3D que permite obtener información de la estructura física de un determinado objeto a partir de la cual, se genera un modelo digital. Este sistema opera proyectando un haz de luz láser en forma de línea sobre el objeto, mientras rota sobre su propio eje (eje z, el haz de luz crea un perfil del objeto, el cual es capturado mediante vídeo de alta definición (HD por una cámara. A través del procesamiento digital de las imágenes capturadas se obtienen los vértices del objeto y se construye el modelo tridimensional. Para llevar a cabo esta tarea, el escáner 3D está formado por un conjunto de elementos de hardware y software libre como lo son: la plataforma Arduino, que permite realizar el control del hardware de escaneo, y un software desarrollado en C++ que implementando las librerías OpenCv y OpenGl, permite la realización de las tareas de procesamiento y análisis de imágenes. Estos elementos hacen que este desarrollo, se  constituya como incursión económica en la tecnología de escaneo laser tridimensional.

  2. ISL1 protein transduction promotes cardiomyocyte differentiation from human embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Hananeh Fonoudi

    Full Text Available BACKGROUND: Human embryonic stem cells (hESCs have the potential to provide an unlimited source of cardiomyocytes, which are invaluable resources for drug or toxicology screening, medical research, and cell therapy. Currently a number of obstacles exist such as the insufficient efficiency of differentiation protocols, which should be overcome before hESC-derived cardiomyocytes can be used for clinical applications. Although the differentiation efficiency can be improved by the genetic manipulation of hESCs to over-express cardiac-specific transcription factors, these differentiated cells are not safe enough to be applied in cell therapy. Protein transduction has been demonstrated as an alternative approach for increasing the efficiency of hESCs differentiation toward cardiomyocytes. METHODS: We present an efficient protocol for the differentiation of hESCs in suspension by direct introduction of a LIM homeodomain transcription factor, Islet1 (ISL1 recombinant protein into the cells. RESULTS: We found that the highest beating clusters were derived by continuous treatment of hESCs with 40 µg/ml recombinant ISL1 protein during days 1-8 after the initiation of differentiation. The treatment resulted in up to a 3-fold increase in the number of beating areas. In addition, the number of cells that expressed cardiac specific markers (cTnT, CONNEXIN 43, ACTININ, and GATA4 doubled. This protocol was also reproducible for another hESC line. CONCLUSIONS: This study has presented a new, efficient, and reproducible procedure for cardiomyocytes differentiation. Our results will pave the way for scaled up and controlled differentiation of hESCs to be used for biomedical applications in a bioreactor culture system.

  3. Completely ES cell-derived mice produced by tetraploid complementation using inner cell mass (ICM deficient blastocysts.

    Directory of Open Access Journals (Sweden)

    Duancheng Wen

    Full Text Available Tetraploid complementation is often used to produce mice from embryonic stem cells (ESCs by injection of diploid (2n ESCs into tetraploid (4n blastocysts (ESC-derived mice. This method has also been adapted to mouse cloning and the derivation of mice from induced pluripotent stem (iPS cells. However, the underlying mechanism(s of the tetraploid complementation remains largely unclear. Whether this approach can give rise to completely ES cell-derived mice is an open question, and has not yet been unambiguously proven. Here, we show that mouse tetraploid blastocysts can be classified into two groups, according to the presence or absence of an inner cell mass (ICM. We designate these as type a (presence of ICM at blastocyst stage or type b (absence of ICM. ESC lines were readily derived from type a blastocysts, suggesting that these embryos retain a pluripotent epiblast compartment; whereas the type b blastocysts possessed very low potential to give rise to ESC lines, suggesting that they had lost the pluripotent epiblast. When the type a blastocysts were used for tetraploid complementation, some of the resulting mice were found to be 2n/4n chimeric; whereas when type b blastocysts were used as hosts, the resulting mice are all completely ES cell-derived, with the newborn pups displaying a high frequency of abdominal hernias. Our results demonstrate that completely ES cell-derived mice can be produced using ICM-deficient 4n blastocysts, and provide evidence that the exclusion of tetraploid cells from the fetus in 2n/4n chimeras can largely be attributed to the formation of ICM-deficient blastocysts.

  4. Medieval Studies in Austria: Research Infrastructure and Resources

    OpenAIRE

    Ertl, Thomas

    2014-01-01

    Questo testo intende fornire una veloce panoramica delle istituzioni e delle associazioni austriache che promuovono ricerche di storia medievale e una guida alle opportunità offerte dal mondo della ricerca scientifica e accademica austriaco.

  5. ON THE REDSHIFT EVOLUTION OF THE Lyα ESCAPE FRACTION AND THE DUST CONTENT OF GALAXIES

    International Nuclear Information System (INIS)

    Hayes, Matthew; Schaerer, Daniel; Oestlin, Goeran; Mas-Hesse, J. Miguel; Atek, Hakim; Kunth, Daniel

    2011-01-01

    The Lyα emission line has been proven to be a powerful tool for studying evolving galaxies at the highest redshift. However, in order to use Lyα as a physical probe of galaxies, it becomes vital to know the Lyα escape fraction (f Lyα esc ). Unfortunately, due to the resonant nature of Lyα, f Lyα esc may vary unpredictably and requires empirical measurement. Here, we compile Lyα luminosity functions (LFs) between redshifts z = 0 and 8 and, combined with Hα and ultraviolet data, assess how f Lyα esc evolves with redshift. We find a strong upward evolution in f Lyα esc over the range z = 0.3-6, which is well fit by the power law f Lyα esc ∝(1 + z) ξ with ξ = (2.57 +0.19 -0.12 ). This predicts that f Lyα esc should reach unity at z = 11.1. By comparing f Lyα esc and E B-V in individual galaxies we derive an empirical relationship between f Lyα esc and E B-V , which includes resonance scattering and can explain the redshift evolution of f Lyα esc between z = 0 and 6 purely as a function of the evolution in the dust content of galaxies. Beyond z ∼ 6.5, f Lyα esc drops more substantially, an effect attributed to either ionizing photon leakage, or an increase in the neutral gas fraction of the intergalactic medium. While distinguishing between these two scenarios may be extremely challenging, by framing the problem this way we remove the uncertainty of the halo mass from Lyα-based tests of reionization. We finally derive a new method by which to estimate the dust content of galaxies, based purely upon the observed Lyα and UV LFs. These data are characterized by an exponential with an e-folding scale of z EBV ∼ 3.4.

  6. Evaluating Electroporation and Lipofectamine Approaches for Transient and Stable Transgene Expressions in Human Fibroblasts and Embryonic Stem Cells

    Science.gov (United States)

    Sharifi Tabar, Mehdi; Hesaraki, Mahdi; Esfandiari, Fereshteh; Sahraneshin Samani, Fazel; Vakilian, Haghighat; Baharvand, Hossein

    2015-01-01

    Objective Genetic modification of human embryonic stem cells (hESCs) is critical for their extensive use as a fundamental tool for cell therapy and basic research. Despite the fact that various methods such as lipofection and electroporation have been applied to transfer the gene of interest (GOI) into the target cell line, however, there are few re- ports that compare all parameters, which influence transfection efficiency. In this study, we examine all parameters that affect the efficiency of electroporation and lipofection for transient and long-term gene expression in three different cell lines to introduce the best method and determinant factor. Materials and Methods In this experimental study, both electroporation and lipofection approaches were employed for genetic modification. pCAG-EGFP was applied for tran- sient expression of green fluorescent protein in two genetically different hESC lines, Roy- an H5 (XX) and Royan H6 (XY), as well as human foreskin fibroblasts (hFF). For long-term EGFP expression VASA and OLIG2 promoters (germ cell and motoneuron specific genes, respectively), were isolated and subsequently cloned into a pBluMAR5 plasmid backbone to drive EGFP expression. Flow cytometry analysis was performed two days after trans- fection to determine transient expression efficiency. Differentiation of drug resistant hESC colonies toward primordial germ cells (PGCs) was conducted to confirm stable integration of the transgene. Results Transient and stable expression suggested a variable potential for different cell lines against transfection. Analysis of parameters that influenced gene transformation ef- ficiency revealed that the vector concentrations from 20-60 μg and the density of the sub- jected cells (5×105and 1×106cells) were not as effective as the genetic background and voltage rate. The present data indicated that in contrast to the circular form, the linearized vector generated more distinctive drug resistant colonies. Conclusion

  7. Effects of Pulsed Electromagnetic Field on Differentiation of HUES-17 Human Embryonic Stem Cell Line

    Directory of Open Access Journals (Sweden)

    Yi-Lin Wu

    2014-08-01

    Full Text Available Electromagnetic fields are considered to potentially affect embryonic development, but the mechanism is still unknown. In this study, human embryonic stem cell (hESC line HUES-17 was applied to explore the mechanism of exposure on embryonic development to pulsed electromagnetic field (PEMF for 400 pulses at different electric field intensities and the differentiation of HUES-17 cells was observed after PEMF exposure. The expression of alkaline phosphatase (AP, stage-specific embryonic antigen-3 (SSEA-3, SSEA-4 and the mRNA level and protein level of Oct4, Sox2 and Nanog in HUES-17 cells remained unchanged after PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m. Four hundred pulses PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m did not affect the differentiation of HUES-17 cells. The reason why electromagnetic fields affect embryonic development may be due to other mechanisms rather than affecting the differentiation of embryonic stem cells.

  8. Avaliação agronômica da escória de siderurgia na cana-de-açúcar durante cinco ciclos de produção Agronomic evaluation of siderurgy slag in sugarcane during five cycles of production

    Directory of Open Access Journals (Sweden)

    Fabiano Barbiere Brassioli

    2009-01-01

    Full Text Available A escória de siderurgia, como material corretivo de acidez com reação mais lenta que o calcário, poderá beneficiar culturas de ciclo longo como a cana-de-açúcar. Assim, este trabalho teve como objetivo avaliar a resposta da cultura da cana-de-açúcar durante cinco ciclos de produção, decorrente da aplicação da escória de siderurgia. O trabalho foi realizado no município de Ituverava (SP, em Latossolo Vermelho-Amarelo (V=26%. Utilizou-se o delineamento experimental em blocos casualizados, em esquema fatorial 2 x 4, com quatro repetições. Os fatores constituíram-se de dois materiais corretivos (escória de siderurgia e calcário calcítico e quatro doses: 0; 1,23, 2,52 e 3,8 t ha-1 equivalente a CaCO3. Avaliou-se ao longo de cinco ciclos de cultivo, a produção de colmos (safras 1998/1999 até 2002/2003; nos quatro primeiros ciclos realizou-se a amostragem do solo (camada de 0-20 cm de profundidade e determinou-se a saturação por bases. A escória de siderurgia e o calcário calcítico foram semelhantes na correção da acidez do solo, ao longo dos ciclos de cultivo da cana-de-açúcar. A maior produção acumulada de colmos está associada à saturação por bases no solo próxima a 60% e 70% para uso do calcário e da escória de siderurgia, respectivamente.The siderurgy slag, as acidity correction material with reaction slower than the limestone, can benefit long-term crops such as sugar cane. This work had as objective to evaluate the response of the sugarcane during five cycles of production as function of the application of siderurgy slag. The work was carried out at Ituverava (SP, in a Red Yellow Latosol (V=26%. The experimental design was randomized blocks in factorial scheme 2 x 4, with four replications. The factors consisted of two corrective materials (slag of siderurgy and limestone at four levels: 0; 1.23, 2.52 and 3.8 t ha-1 equivalent of CaCO3. The cane stalk production was evaluated throughout five cycles of

  9. Effectiveness of electronic stability control on single-vehicle accidents

    DEFF Research Database (Denmark)

    Lyckegaard, Allan; Hels, Tove; Bernhoft, Inger Marie

    2015-01-01

    the injury severity categories (slight, severe, and fatal). Conclusions: In line with previous results, this study concludes that ESC reduces the risk for single-vehicle injury accidents by 31% when controlling for various confounding factors related to the driver, the car, and the accident surroundings......Objective: This study aims at evaluating the effectiveness of electronic stability control (ESC) on single-vehicle injury accidents while controlling for a number of confounders influencing the accident risk. Methods: Using police-registered injury accidents from 2004 to 2011 in Denmark with cars...... the following were significant. For the driver: Age, gender, driving experience, valid driving license, and seat belt use. For the vehicle: Year of registration, weight, and ESC. For the accident surroundings: Visibility, light, and location. Finally, for the road: Speed limit, surface, and section...

  10. Stable isotope labeling by amino acids in cell culture (SILAC) and quantitative comparison of the membrane proteomes of self-renewing and differentiating human embryonic stem cells

    DEFF Research Database (Denmark)

    Prokhorova, Tatyana A; Rigbolt, Kristoffer T G; Johansen, Pia T

    2009-01-01

    Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful quantitative proteomics platform for comprehensive characterization of complex biological systems. However, the potential of SILAC-based approaches has not been fully utilized in human embryonic stem cell (hESC) research...... embryonic stem cell lines. Of the 811 identified membrane proteins, six displayed significantly higher expression levels in the undifferentiated state compared with differentiating cells. This group includes the established marker CD133/Prominin-1 as well as novel candidates for hESC surface markers......: Glypican-4, Neuroligin-4, ErbB2, receptor-type tyrosine-protein phosphatase zeta (PTPRZ), and Glycoprotein M6B. Our study also revealed 17 potential markers of hESC differentiation as their corresponding protein expression levels displayed a dramatic increase in differentiated embryonic stem cell...

  11. ESH-ESC guidelines for the management of hypertension.

    Science.gov (United States)

    Erdine, Serap; Ari, Oben; Zanchetti, A; Cifkova, R; Fagard, R; Kjeldsen, S; Mancia, G; Poulter, N; Rahn, K H; Rodicio, J L; Ruilope, L M; Staessen, J; van Zwieten, P; Waeber, B; Williams, B

    2006-06-01

    The following is a brief statement of the 2003 European Society of Hypertension (ESH)-European Society of Cardiology (ESC) guidelines for the management of arterial hypertension. The continuous relationship between the level of blood pressure and cardiovascular risk makes the definition of hypertension arbitrary. Since risk factors cluster in hypertensive individuals, risk stratification should be made and decision about the management should not be based on blood pressure alone, but also according to the presence or absence of other risk factors, target organ damage, diabetes, and cardiovascular or renal damage, as well as on other aspects of the patient's personal, medical and social situation. Blood pressure values measured in the doctor's office or the clinic should commonly be used as reference. Ambulatory blood pressure monitoring may have clinical value, when considerable variability of office blood pressure is found over the same or different visits, high office blood pressure is measured in subjects otherwise at low global cardiovascular risk, there is marked discrepancy between blood pressure values measured in the office and at home, resistance to drug treatment is suspected, or research is involved. Secondary hypertension should always be investigated. The primary goal of treatment of patient with high blood pressure is to achieve the maximum reduction in long-term total risk of cardiovascular morbidity and mortality. This requires treatment of all the reversible factors identified, including smoking, dislipidemia, or diabetes, and the appropriate management of associated clinical conditions, as well as treatment of the raised blood pressure per se. On the basis of current evidence from trials, it can be recommended that blood pressure, both systolic and diastolic, be intensively lowered at least below 140/90 mmHg and to definitely lower values, if tolerated, in all hypertensive patients, and below 130/80 mmHg in diabetics. Lifestyle measures should be

  12. HLA engineering of human pluripotent stem cells.

    Science.gov (United States)

    Riolobos, Laura; Hirata, Roli K; Turtle, Cameron J; Wang, Pei-Rong; Gornalusse, German G; Zavajlevski, Maja; Riddell, Stanley R; Russell, David W

    2013-06-01

    The clinical use of human pluripotent stem cells and their derivatives is limited by the rejection of transplanted cells due to differences in their human leukocyte antigen (HLA) genes. This has led to the proposed use of histocompatible, patient-specific stem cells; however, the preparation of many different stem cell lines for clinical use is a daunting task. Here, we develop two distinct genetic engineering approaches that address this problem. First, we use a combination of gene targeting and mitotic recombination to derive HLA-homozygous embryonic stem cell (ESC) subclones from an HLA-heterozygous parental line. A small bank of HLA-homozygous stem cells with common haplotypes would match a significant proportion of the population. Second, we derive HLA class I-negative cells by targeted disruption of both alleles of the Beta-2 Microglobulin (B2M) gene in ESCs. Mixed leukocyte reactions and peptide-specific HLA-restricted CD8(+) T cell responses were reduced in class I-negative cells that had undergone differentiation in embryoid bodies. These B2M(-/-) ESCs could act as universal donor cells in applications where the transplanted cells do not express HLA class II genes. Both approaches used adeno-associated virus (AAV) vectors for efficient gene targeting in the absence of potentially genotoxic nucleases, and produced pluripotent, transgene-free cell lines.

  13. HLA Engineering of Human Pluripotent Stem Cells

    Science.gov (United States)

    Riolobos, Laura; Hirata, Roli K; Turtle, Cameron J; Wang, Pei-Rong; Gornalusse, German G; Zavajlevski, Maja; Riddell, Stanley R; Russell, David W

    2013-01-01

    The clinical use of human pluripotent stem cells and their derivatives is limited by the rejection of transplanted cells due to differences in their human leukocyte antigen (HLA) genes. This has led to the proposed use of histocompatible, patient-specific stem cells; however, the preparation of many different stem cell lines for clinical use is a daunting task. Here, we develop two distinct genetic engineering approaches that address this problem. First, we use a combination of gene targeting and mitotic recombination to derive HLA-homozygous embryonic stem cell (ESC) subclones from an HLA-heterozygous parental line. A small bank of HLA-homozygous stem cells with common haplotypes would match a significant proportion of the population. Second, we derive HLA class I–negative cells by targeted disruption of both alleles of the Beta-2 Microglobulin (B2M) gene in ESCs. Mixed leukocyte reactions and peptide-specific HLA-restricted CD8+ T cell responses were reduced in class I–negative cells that had undergone differentiation in embryoid bodies. These B2M−/− ESCs could act as universal donor cells in applications where the transplanted cells do not express HLA class II genes. Both approaches used adeno-associated virus (AAV) vectors for efficient gene targeting in the absence of potentially genotoxic nucleases, and produced pluripotent, transgene-free cell lines. PMID:23629003

  14. Precise gene modification mediated by TALEN and single-stranded oligodeoxynucleotides in human cells.

    Directory of Open Access Journals (Sweden)

    Xiaoling Wang

    Full Text Available The development of human embryonic stem cells (ESCs and induced pluripotent stem cells (iPSCs facilitates in vitro studies of human disease mechanisms, speeds up the process of drug screening, and raises the feasibility of using cell replacement therapy in clinics. However, the study of genotype-phenotype relationships in ESCs or iPSCs is hampered by the low efficiency of site-specific gene editing. Transcription activator-like effector nucleases (TALENs spurred interest due to the ease of assembly, high efficiency and faithful gene targeting. In this study, we optimized the TALEN design to maximize its genomic cutting efficiency. We showed that using optimized TALENs in conjunction with single-strand oligodeoxynucleotide (ssODN allowed efficient gene editing in human cells. Gene mutations and gene deletions for up to 7.8 kb can be accomplished at high efficiencies. We established human tumor cell lines and H9 ESC lines with homozygous deletion of the microRNA-21 (miR-21 gene and miR-9-2 gene. These cell lines provide a robust platform to dissect the roles these genes play during cell differentiation and tumorigenesis. We also observed that the endogenous homologous chromosome can serve as a donor template for gene editing. Overall, our studies demonstrate the versatility of using ssODN and TALEN to establish genetically modified cells for research and therapeutic application.

  15. EDA-containing fibronectin increases proliferation of embryonic stem cells.

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    Noelia Losino

    Full Text Available Embryonic stem cells (ESC need a set of specific factors to be propagated. They can also grow in conditioned medium (CM derived from a bovine granulosa cell line BGC (BGC-CM, a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA(+. Here, we investigated if the FN EDA(+ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA(-, and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC's proliferation rate. Here we showed for the first time that this FN isoform enhances ESC's proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.

  16. Moisture Control Guidance for Commercial and Public Buildings (EPA 402-F-13053)

    Science.gov (United States)

    This document provides guidance to designers, construction mangers, and building operation/maintenance managers to improve IEQ and reduce risks of encountering IEQ problems due to insufficient moisture control. EPA will be producing a document entitled "Moisture Control Guida...

  17. Accessibility-based evaluation of transportation and land-use planning : from laboratory to practice : USDOT Region V Regional University Transportation Center final report.

    Science.gov (United States)

    2016-12-16

    The concept of accessibility has made inroads into planning practice, largely at the system level. That is, accessibility is measured or modeled for current or future regional transportation and land-use scenarios for evaluation or broad policy guida...

  18. Efficient and scalable purification of cardiomyocytes from human embryonic and induced pluripotent stem cells by VCAM1 surface expression.

    Directory of Open Access Journals (Sweden)

    Hideki Uosaki

    Full Text Available RATIONALE: Human embryonic and induced pluripotent stem cells (hESCs/hiPSCs are promising cell sources for cardiac regenerative medicine. To realize hESC/hiPSC-based cardiac cell therapy, efficient induction, purification, and transplantation methods for cardiomyocytes are required. Though marker gene transduction or fluorescent-based purification methods have been reported, fast, efficient and scalable purification methods with no genetic modification are essential for clinical purpose but have not yet been established. In this study, we attempted to identify cell surface markers for cardiomyocytes derived from hESC/hiPSCs. METHOD AND RESULT: We adopted a previously reported differentiation protocol for hESCs based on high density monolayer culture to hiPSCs with some modification. Cardiac troponin-T (TNNT2-positive cardiomyocytes appeared robustly with 30-70% efficiency. Using this differentiation method, we screened 242 antibodies for human cell surface molecules to isolate cardiomyocytes derived from hiPSCs and identified anti-VCAM1 (Vascular cell adhesion molecule 1 antibody specifically marked cardiomyocytes. TNNT2-positive cells were detected at day 7-8 after induction and 80% of them became VCAM1-positive by day 11. Approximately 95-98% of VCAM1-positive cells at day 11 were positive for TNNT2. VCAM1 was exclusive with CD144 (endothelium, CD140b (pericytes and TRA-1-60 (undifferentiated hESCs/hiPSCs. 95% of MACS-purified cells were positive for TNNT2. MACS purification yielded 5-10×10(5 VCAM1-positive cells from a single well of a six-well culture plate. Purified VCAM1-positive cells displayed molecular and functional features of cardiomyocytes. VCAM1 also specifically marked cardiomyocytes derived from other hESC or hiPSC lines. CONCLUSION: We succeeded in efficiently inducing cardiomyocytes from hESCs/hiPSCs and identifying VCAM1 as a potent cell surface marker for robust, efficient and scalable purification of cardiomyocytes from hESC

  19. Evidence based medicine (EBM) and evidence based radiology (EBR) in the follow-up of the patients after surgery for lung and colon-rectal carcinoma; Medicina basata sulle evidenze (EMB) e radiologia basata sulle evidenze (EBR) nel follow-up dei pazienti operati per tumore del polmone e del colon

    Energy Technology Data Exchange (ETDEWEB)

    Giovagnoni, Andrea; Ottaviani, Letizia; Mensa' , Anna; Durastanti, Martina; Floriani, Irene; Cascinu, Stefano [Marche Univ., Ancona (Italy). Azienda ospedaliera Umberto I, Istituto di radiologia, oncologia clinica

    2005-04-01

    real costs of the follow-up programmes implemented in the 40 patients considered and the theoretical costs derived from the guide-line recommendations (an excess of 99.06% for lung cancer, and 93.6% for colon cancer). Conclusions: Our findings can serve as a basis to start a discussion within the scientific community about the role of radiological follow-up in cancer patients with the aim of defining a more rational use of resources. [Italian] Scopo: a) Utilizzare la metodologia di Evidence Based Medicine (EBM) nella definizione del ruolo della radiologia nel follow-up post chirurgico dei pazienti con neoplasia solida; b) valutare le eventuali discrepanze tra il modello teorico ed i protocolli clinici attualmente in uso; c) confrontare i costi degli esami radiologici ricavati da un campione di pazienti osservato con quelli ideali che si sarebbero sostenuti se i pazienti fossero stati seguiti secindo le raccomandazioni fornite dalle linee guida delle Societa Scientifiche Internazionali. Materiale e metodi: Sono stati ricercati (utilizzando banche dati tradizionali e di editoria secondaria) i lavori scientifici e le linee guida degli ultimi dieci anni inerenti l'impatto clinico dell'imaging nel follow-up dei pazienti con tumori solidi (polmone, colon). Utilizzando la metodologia delle revisioni sistematiche proprie della EBM. Sono stati scelti i lavori piu significativi per numero di pazienti inclusi e livello di affidabilita. Sono stati valutati i seguenti parametri: timing delle indagini, sopravvivenza, periodo libero di malattia, qualita della vita, morbilita indotta, costi, aspetti psicologici. In una seconda fase stato analizzato retrospettivamente il follow-up clinico strumentale di 40 pazienti consecutivi affetti da tumore del colon (20 pazienti) e polmone (20 pazienti) giunti alla nostra osservazione nel periodo 1998-2004, valutato il costo reale e confrontato con quello teorico. Risultati: La revisione sistematica della letteratura ha permesso di

  20. Rappresentare e gestire patrimoni immobiliari: il BIM per il Facility Management

    Directory of Open Access Journals (Sweden)

    Massimiliano Lo Turco

    2015-12-01

    Full Text Available Il presente contributo intende esplorare le potenzialità delle metodologie di Building Information Modeling (BIM applicabili alle diverse fasi del processo edilizio, con specifico riferimento a linee guida e best practice di respiro internazionale che avvalorano flussi operativi virtuosi: la componente informativa, prevalentemente grafica in fase di progetto, si integra e si rinnova in formato prevalentemente alfanumerico, divenendo parte integrante dell’apparato gestionale e manutentivo dei manufatti. I casi studio sviluppati in collaborazione con il Servizio Edilizia e Logistica del Politecnico di Torino sostanziano tali procedure che, seppur ancora poco diffuse, traguardano obiettivi di interoperabilità: si illustrano approcci integrati di condivisione delle informazioni atti a definire una corretta programmazione degli interventi, applicabili dalla grande alla piccola scala, definendo strategie progettate per limitare l’attuazione di errate soluzioni.

  1. Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases.

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    Anna Osiak

    Full Text Available Gene knockout in murine embryonic stem cells (ESCs has been an invaluable tool to study gene function in vitro or to generate animal models with altered phenotypes. Gene targeting using standard techniques, however, is rather inefficient and typically does not exceed frequencies of 10(-6. In consequence, the usage of complex positive/negative selection strategies to isolate targeted clones has been necessary. Here, we present a rapid single-step approach to generate a gene knockout in mouse ESCs using engineered zinc-finger nucleases (ZFNs. Upon transient expression of ZFNs, the target gene is cleaved by the designer nucleases and then repaired by non-homologous end-joining, an error-prone DNA repair process that introduces insertions/deletions at the break site and therefore leads to functional null mutations. To explore and quantify the potential of ZFNs to generate a gene knockout in pluripotent stem cells, we generated a mouse ESC line containing an X-chromosomally integrated EGFP marker gene. Applying optimized conditions, the EGFP locus was disrupted in up to 8% of ESCs after transfection of the ZFN expression vectors, thus obviating the need of selection markers to identify targeted cells, which may impede or complicate downstream applications. Both activity and ZFN-associated cytotoxicity was dependent on vector dose and the architecture of the nuclease domain. Importantly, teratoma formation assays of selected ESC clones confirmed that ZFN-treated ESCs maintained pluripotency. In conclusion, the described ZFN-based approach represents a fast strategy for generating gene knockouts in ESCs in a selection-independent fashion that should be easily transferrable to other pluripotent stem cells.

  2. Inhibition of Sirt1 promotes neural progenitors toward motoneuron differentiation from human embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yun; Wang, Jing [Department of Neurology, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Clinical Stem Cell Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Chen, Guian [Clinical Stem Cell Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Reproductive Medical Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Fan, Dongsheng, E-mail: dsfan@yahoo.cn [Department of Neurology, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Clinical Stem Cell Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Deng, Min, E-mail: dengmin1706@yahoo.com.cn [Department of Neurology, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China); Clinical Stem Cell Center, Peking University Third Hospital, 49 North Garden Road, Haidian District, Beijing 100191 (China)

    2011-01-14

    Research highlights: {yields} Nicotinamide inhibit Sirt1. {yields} MASH1 and Ngn2 activation. {yields} Increase the expression of HB9. {yields} Motoneurons formation increases significantly. -- Abstract: Several protocols direct human embryonic stem cells (hESCs) toward differentiation into functional motoneurons, but the efficiency of motoneuron generation varies based on the human ESC line used. We aimed to develop a novel protocol to increase the formation of motoneurons from human ESCs. In this study, we tested a nuclear histone deacetylase protein, Sirt1, to promote neural precursor cell (NPC) development during differentiation of human ESCs into motoneurons. A specific inhibitor of Sirt1, nicotinamide, dramatically increased motoneuron formation. We found that about 60% of the cells from the total NPCs expressed HB9 and {beta}III-tubulin, commonly used motoneuronal markers found in neurons derived from ESCs following nicotinamide treatment. Motoneurons derived from ESC expressed choline acetyltransferase (ChAT), a positive marker of mature motoneuron. Moreover, we also examined the transcript levels of Mash1, Ngn2, and HB9 mRNA in the differentiated NPCs treated with the Sirt1 activator resveratrol (50 {mu}M) or inhibitor nicotinamide (100 {mu}M). The levels of Mash1, Ngn2, and HB9 mRNA were significantly increased after nicotinamide treatment compared with control groups, which used the traditional protocol. These results suggested that increasing Mash1 and Ngn2 levels by inhibiting Sirt1 could elevate HB9 expression, which promotes motoneuron differentiation. This study provides an alternative method for the production of transplantable motoneurons, a key requirement in the development of hESC-based cell therapy in motoneuron disease.

  3. Inhibition of Sirt1 promotes neural progenitors toward motoneuron differentiation from human embryonic stem cells

    International Nuclear Information System (INIS)

    Zhang, Yun; Wang, Jing; Chen, Guian; Fan, Dongsheng; Deng, Min

    2011-01-01

    Research highlights: → Nicotinamide inhibit Sirt1. → MASH1 and Ngn2 activation. → Increase the expression of HB9. → Motoneurons formation increases significantly. -- Abstract: Several protocols direct human embryonic stem cells (hESCs) toward differentiation into functional motoneurons, but the efficiency of motoneuron generation varies based on the human ESC line used. We aimed to develop a novel protocol to increase the formation of motoneurons from human ESCs. In this study, we tested a nuclear histone deacetylase protein, Sirt1, to promote neural precursor cell (NPC) development during differentiation of human ESCs into motoneurons. A specific inhibitor of Sirt1, nicotinamide, dramatically increased motoneuron formation. We found that about 60% of the cells from the total NPCs expressed HB9 and βIII-tubulin, commonly used motoneuronal markers found in neurons derived from ESCs following nicotinamide treatment. Motoneurons derived from ESC expressed choline acetyltransferase (ChAT), a positive marker of mature motoneuron. Moreover, we also examined the transcript levels of Mash1, Ngn2, and HB9 mRNA in the differentiated NPCs treated with the Sirt1 activator resveratrol (50 μM) or inhibitor nicotinamide (100 μM). The levels of Mash1, Ngn2, and HB9 mRNA were significantly increased after nicotinamide treatment compared with control groups, which used the traditional protocol. These results suggested that increasing Mash1 and Ngn2 levels by inhibiting Sirt1 could elevate HB9 expression, which promotes motoneuron differentiation. This study provides an alternative method for the production of transplantable motoneurons, a key requirement in the development of hESC-based cell therapy in motoneuron disease.

  4. Human embryonic stem cell lines model experimental human cytomegalovirus latency.

    Science.gov (United States)

    Penkert, Rhiannon R; Kalejta, Robert F

    2013-05-28

    Herpesviruses are highly successful pathogens that persist for the lifetime of their hosts primarily because of their ability to establish and maintain latent infections from which the virus is capable of productively reactivating. Human cytomegalovirus (HCMV), a betaherpesvirus, establishes latency in CD34(+) hematopoietic progenitor cells during natural infections in the body. Experimental infection of CD34(+) cells ex vivo has demonstrated that expression of the viral gene products that drive productive infection is silenced by an intrinsic immune defense mediated by Daxx and histone deacetylases through heterochromatinization of the viral genome during the establishment of latency. Additional mechanistic details about the establishment, let alone maintenance and reactivation, of HCMV latency remain scarce. This is partly due to the technical challenges of CD34(+) cell culture, most notably, the difficulty in preventing spontaneous differentiation that drives reactivation and renders them permissive for productive infection. Here we demonstrate that HCMV can establish, maintain, and reactivate in vitro from experimental latency in cultures of human embryonic stem cells (ESCs), for which spurious differentiation can be prevented or controlled. Furthermore, we show that known molecular aspects of HCMV latency are faithfully recapitulated in these cells. In total, we present ESCs as a novel, tractable model for studies of HCMV latency.

  5. Giustizia, uguaglianza e differenza

    Directory of Open Access Journals (Sweden)

    Maria Chiara Pievatolo

    2013-02-01

    Full Text Available E’ una guida alla lettura della filosofia politica contemporanea, a cura di Brunella Casalini e Lorenzo Cini, pubblicata da Firenze University Press per la nostra collana ad accesso aperto Methexis. La versione digitalizzata del testo è a disposizione di tutti.

  6. Comparison of the glycosphingolipids of human-induced pluripotent stem cells and human embryonic stem cells.

    Science.gov (United States)

    Säljö, Karin; Barone, Angela; Vizlin-Hodzic, Dzeneta; Johansson, Bengt R; Breimer, Michael E; Funa, Keiko; Teneberg, Susann

    2017-04-01

    High expectations are held for human-induced pluripotent stem cells (hiPSC) since they are established from autologous tissues thus overcoming the risk of allogeneic immune rejection when used in regenerative medicine. However, little is known regarding the cell-surface carbohydrate antigen profile of hiPSC compared with human embryonic stem cells (hESC). Here, glycosphingolipids were isolated from an adipocyte-derived hiPSC line, and hiPSC and hESC glycosphingolipids were compared by concurrent characterization by binding assays with carbohydrate-recognizing ligands and mass spectrometry. A high similarity between the nonacid glycosphingolipids of hiPSC and hESC was found. The nonacid glycosphingolipids P1 pentaosylceramide, x2 pentaosylceramide and H type 1 heptaosylceramide, not previously described in human pluripotent stem cells (hPSC), were characterized in both hiPSC and hESC. The composition of acid glycosphingolipids differed, with increased levels of GM3 ganglioside, and reduced levels of GD1a/GD1b in hiPSC when compared with hESC. In addition, the hESC glycosphingolipids sulf-globopentaosylceramide and sialyl-globotetraosylceramide were lacking in hiPSC. Neural stem cells differentiating from hiPSC had a reduced expression of sialyl-lactotetra, whereas expression of the GD1a ganglioside was significantly increased. Thus, while sialyl-lactotetra is a marker of undifferentiated hPSC, GD1a is a novel marker of neural differentiation. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Embryonic stem cells derived from in vivo or in vitro-generated murine blastocysts display similar transcriptome and differentiation potential.

    Directory of Open Access Journals (Sweden)

    Rhodel K Simbulan

    Full Text Available The use of assisted reproductive technologies (ART such as in vitro fertilization (IVF has resulted in the birth of more than 5 million children. While children conceived by these technologies are generally healthy, there is conflicting evidence suggesting an increase in adult-onset complications like glucose intolerance and high blood pressure in IVF children. Animal models indicate similar potential risks. It remains unclear what molecular mechanisms may be operating during in vitro culture to predispose the embryo to these diseases. One of the limitations faced by investigators is the paucity of the material in the preimplantation embryo to test for molecular analysis. To address this problem, we generated mouse embryonic stem cells (mESC from blastocysts conceived after natural mating (mESCFB or after IVF, using optimal (KSOM + 5% O2; mESCKAA and suboptimal (Whitten's Medium, + 20% O2, mESCWM conditions. All three groups of embryos showed similar behavior during both derivation and differentiation into their respective mESC lines. Unsupervised hierarchical clustering of microarray data showed that blastocyst culture does not affect the transcriptome of derived mESCs. Transcriptomic changes previously observed in the inner cell mass (ICM of embryos derived in the same conditions were not present in mESCs, regardless of method of conception or culture medium, suggesting that mESC do not fully maintain a memory of the events occurring prior to their derivation. We conclude that the fertilization method or culture media used to generate blastocysts does not affect differentiation potential, morphology and transcriptome of mESCs.

  8. Effectiveness of electronic stability control on single-vehicle accidents.

    Science.gov (United States)

    Lyckegaard, Allan; Hels, Tove; Bernhoft, Inger Marie

    2015-01-01

    This study aims at evaluating the effectiveness of electronic stability control (ESC) on single-vehicle injury accidents while controlling for a number of confounders influencing the accident risk. Using police-registered injury accidents from 2004 to 2011 in Denmark with cars manufactured in the period 1998 to 2011 and the principle of induced exposure, 2 measures of the effectiveness of ESC were calculated: The crude odds ratio and the adjusted odds ratio, the latter by means of logistic regression. The logistic regression controlled for a number of confounding factors, of which the following were significant. For the driver: Age, gender, driving experience, valid driving license, and seat belt use. For the vehicle: Year of registration, weight, and ESC. For the accident surroundings: Visibility, light, and location. Finally, for the road: Speed limit, surface, and section characteristics. The present study calculated the crude odds ratio for ESC-equipped cars of getting in a single-vehicle injury accident as 0.40 (95% confidence interval [CI], 0.34-0.47) and the adjusted odds ratio as 0.69 (95% CI, 0.54-0.88). No difference was found in the effectiveness of ESC across the injury severity categories (slight, severe, and fatal). In line with previous results, this study concludes that ESC reduces the risk for single-vehicle injury accidents by 31% when controlling for various confounding factors related to the driver, the car, and the accident surroundings. Furthermore, it is concluded that it is important to control for human factors (at a minimum age and gender) in analyses where evaluations of this type are performed.

  9. YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers.

    Science.gov (United States)

    Brøchner, Christian B; Johansen, Julia S; Larsen, Lars A; Bak, Mads; Mikkelsen, Hanne B; Byskov, Anne Grete; Andersen, Claus Yding; Møllgård, Kjeld

    2012-03-01

    The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3β, PDX1, CD34, p63, nestin, PAX6) markers. Double-labeling showed YKL-40 expression in OCT4-positive hESCs, PAX6-positive neuroectodermal cells, and HNF-3β-positive endodermal cells. The differentiating progeny showed strong YKL-40 expression. Abrupt transition between YKL-40 and OCT4-positive hESCs and YKL-40-positive ecto- and neuroectodermal lineages was observed within the same epithelial-like layer. YKL-40-positive cells within deeper layers lacked contact with OCT4-positive cells. YKL-40 may be important in initial cell differentiation from hESCs toward ectoderm and neuroectoderm, with retained epithelial morphology, whereas later differentiation into endoderm and mesoderm involves a transition into the deeper layers of the colony.

  10. Regulatory variants of FOXG1 in the context of its topological domain organisation.

    Science.gov (United States)

    Mehrjouy, Mana M; Fonseca, Ana Carolina S; Ehmke, Nadja; Paskulin, Giorgio; Novelli, Antonio; Benedicenti, Francesco; Mencarelli, Maria Antonietta; Renieri, Alessandra; Busa, Tiffany; Missirian, Chantal; Hansen, Claus; Abe, Kikue Terada; Speck-Martins, Carlos Eduardo; Vianna-Morgante, Angela M; Bak, Mads; Tommerup, Niels

    2018-02-01

    FOXG1 syndrome is caused by FOXG1 intragenic point mutations, or by long-range position effects (LRPE) of intergenic structural variants. However, the size of the FOXG1 regulatory landscape is uncertain, because the associated topologically associating domain (TAD) in fibroblasts is split into two domains in embryonic stem cells (hESC). Indeed, it has been suggested that the pathogenetic mechanism of deletions that remove the stem-cell-specific TAD boundary may be enhancer adoption due to ectopic activity of enhancer(s) located in the distal hESC-TAD. Herein we map three de novo translocation breakpoints to the proximal regulatory domain of FOXG1. The classical FOXG1 syndrome in these and in other translocation patients, and in a patient with an intergenic deletion that removes the hESC-specific TAD boundary, do not support the hypothesised enhancer adoption as a main contributor to the FOXG1 syndrome. Also, virtual 4 C and HiC-interaction data suggest that the hESC-specific TAD boundary may not be critical for FOXG1 regulation in a majority of human cells and tissues, including brain tissues and a neuronal progenitor cell line. Our data support the importance of a critical regulatory region (SRO) proximal to the hESC-specific TAD boundary. We further narrow this critical region by a deletion distal to the hESC-specific boundary, associated with a milder clinical phenotype. The distance from FOXG1 to the SRO ( > 500 kb) highlight a limitation of ENCODE DNase hypersensitivity data for functional prediction of LRPE. Moreover, the SRO has little overlap with a cluster of frequently associating regions (FIREs) located in the proximal hESC-TAD.

  11. Proteomic Analysis of Human Blastocoel Fluid and Blastocyst Cells

    DEFF Research Database (Denmark)

    Linnert Jensen, Pernille; Beck, Hans Christian; Petersen, Jørgen

    The human blastocyst consists of 100-200 cells that are organized in an outer layer of differentiated trophectoderm (TE) cells lining the blastocyst cavity into which the undifferentiated inner cell mass (ICM) protrudes. The cavity of the blastocyst is filled with blastocoel fluid to which all...... the cells of the blastocyst are exposed. The ICM is the starting point for the development of undifferentiated human embryonic stem cells (hESCs), which posses the potential to develop into any cell type present in the adult human body [1,2]. This ability makes hESCs a potential source of cells...

  12. Using therapeutic cloning to fight human disease: a conundrum or reality?

    Science.gov (United States)

    Hall, Vanessa J; Stojkovic, Petra; Stojkovic, Miodrag

    2006-07-01

    The development and transplantation of autologous cells derived from nuclear transfer embryonic stem cell (NT-ESC) lines to treat patients suffering from disease has been termed therapeutic cloning. Human NT is still a developing field, with further research required to improve somatic cell NT and human embryonic stem cell differentiation to deliver safe and effective cell replacement therapies. Furthermore, the implications of transferring mitochondrial heteroplasmic cells, which may harbor aberrant epigenetic gene expression profiles, are of concern. The production of human NT-ESC lines also remains plagued by ethical dilemmas, societal concerns, and controversies. Recently, a number of alternate therapeutic strategies have been proposed to circumvent the moral implications surrounding human nuclear transfer. It will be critical to overcome these biological, legislative, and moral restraints to maximize the potential of this therapeutic strategy and to alleviate human disease.

  13. Guiado de un vehículo autónomo mediante la detección de los límites del carril usando un escáner láser

    OpenAIRE

    Jiménez Alonso, Felipe; Clavijo Jiménez, Miguel; Naranjo Hernández, José Eugenio; Gómez Casado, O.

    2016-01-01

    Los vehículos autónomos están cada vez cobrando mayor repercusión, tanto con funciones de semiautomatización como de automatización completa. Se han explorado diferentes herramientas y tecnologías para realizar el control automático. En cuanto a tecnologías, el escáner láser 3D está cobrando gran ventaja dadas sus prestaciones. Cuando se plantea la estrategia de control en el caso de un vehículo totalmente autónomo, se pueden distinguir diversas soluciones: seguimiento de una trayectoria dada...

  14. Gene function in early mouse embryonic stem cell differentiation

    Directory of Open Access Journals (Sweden)

    Campbell Pearl A

    2007-03-01

    Full Text Available Abstract Background Little is known about the genes that drive embryonic stem cell differentiation. However, such knowledge is necessary if we are to exploit the therapeutic potential of stem cells. To uncover the genetic determinants of mouse embryonic stem cell (mESC differentiation, we have generated and analyzed 11-point time-series of DNA microarray data for three biologically equivalent but genetically distinct mESC lines (R1, J1, and V6.5 undergoing undirected differentiation into embryoid bodies (EBs over a period of two weeks. Results We identified the initial 12 hour period as reflecting the early stages of mESC differentiation and studied probe sets showing consistent changes of gene expression in that period. Gene function analysis indicated significant up-regulation of genes related to regulation of transcription and mRNA splicing, and down-regulation of genes related to intracellular signaling. Phylogenetic analysis indicated that the genes showing the largest expression changes were more likely to have originated in metazoans. The probe sets with the most consistent gene changes in the three cell lines represented 24 down-regulated and 12 up-regulated genes, all with closely related human homologues. Whereas some of these genes are known to be involved in embryonic developmental processes (e.g. Klf4, Otx2, Smn1, Socs3, Tagln, Tdgf1, our analysis points to others (such as transcription factor Phf21a, extracellular matrix related Lama1 and Cyr61, or endoplasmic reticulum related Sc4mol and Scd2 that have not been previously related to mESC function. The majority of identified functions were related to transcriptional regulation, intracellular signaling, and cytoskeleton. Genes involved in other cellular functions important in ESC differentiation such as chromatin remodeling and transmembrane receptors were not observed in this set. Conclusion Our analysis profiles for the first time gene expression at a very early stage of mESC

  15. Aproveitamento de silício proveniente de escória siderúrgica por cultivares de cana-de-açúcar Recovery of silicon from metallurgy slag by sugarcane cultivars

    Directory of Open Access Journals (Sweden)

    Robson Thiago Xavier de Sousa

    2010-01-01

    Full Text Available A cana-de-açúcar é uma espécie acumuladora de silício (Si, capaz de responder à adubação silicatada, principalmente, em solos pobres desse elemento. As escórias de siderurgia constituem em uma interessante fonte de Si para aplicação em solos cultivados com essa cultura. Objetivou-se estudar o aproveitamento do Si proveniente da escória siderúrgica pela cana-de-açúcar. O experimento foi desenvolvido em Uberlândia (MG, entre 15 de agosto de 2007 e 2 de maio de 2008. O delineamento experimental utilizado foi o de blocos ao acaso em esquema fatorial 2x4, sendo duas cultivares de cana-de-açúcar (SP81 - 3250 e RB86 - 7515 e quatro doses de Si (0, 100, 200 e 400 kg ha-1 em quatro repetições. A cana foi cultivada em tambores plásticos (200 L. A aplicação de escória de siderurgia (silicato aumentou o teor de Si disponível no solo e o teor foliar da cana-de-açúcar. A quantidade de Si acumulada pela parte aérea da cana proveniente do silicato aplicado variou entre 23% e 56%, respectivamente, para as variedades RB86-7515 e SP81-3250. Em média, 39% do Si absorvido pela parte aérea da cana foram provenientes do fertilizante aplicado (silicato.Sugarcane is a species that accumulates silicon (Si and is capable of responding to Si applications, especially in soils lacking Si.. Metallurgy slag is an interesting source of Si for soils cultivated with this crop. The aim of this study was to evaluate the recovery of Si from metallurgy slag by sugarcane. The experiment was carried out at Uberlândia-MG, from August 15, 2007 to May 02, 2008. The experimental design was randomized in a 2 x 4 factorial arrangement being two cultivate of sugarcane (SP81 - 3250 and RB86 - 7515 and four Silicon doses (0, 100, 200 and 400 kg ha-1 with four replications. The sugarcane was cultivated in plastic drums (200 L. The application of slag (silicates increased the content of available silicon in soil and leaf of sugarcane. The amount of Si accumulated in

  16. Optimisation of active suspension control inputs for improved performance of active safety systems

    Science.gov (United States)

    Čorić, Mirko; Deur, Joško; Xu, Li; Tseng, H. Eric; Hrovat, Davor

    2018-01-01

    A collocation-type control variable optimisation method is used to investigate the extent to which the fully active suspension (FAS) can be applied to improve the vehicle electronic stability control (ESC) performance and reduce the braking distance. First, the optimisation approach is applied to the scenario of vehicle stabilisation during the sine-with-dwell manoeuvre. The results are used to provide insights into different FAS control mechanisms for vehicle performance improvements related to responsiveness and yaw rate error reduction indices. The FAS control performance is compared to performances of the standard ESC system, optimal active brake system and combined FAS and ESC configuration. Second, the optimisation approach is employed to the task of FAS-based braking distance reduction for straight-line vehicle motion. Here, the scenarios of uniform and longitudinally or laterally non-uniform tyre-road friction coefficient are considered. The influences of limited anti-lock braking system (ABS) actuator bandwidth and limit-cycle ABS behaviour are also analysed. The optimisation results indicate that the FAS can provide competitive stabilisation performance and improved agility when compared to the ESC system, and that it can reduce the braking distance by up to 5% for distinctively non-uniform friction conditions.

  17. Expansion and maintenance of human embryonic stem cell–derived endothelial cells by TGFβ inhibition is Id1 dependent

    Science.gov (United States)

    James, Daylon; Nam, Hyung-song; Seandel, Marco; Nolan, Daniel; Janovitz, Tyler; Tomishima, Mark; Studer, Lorenz; Lee, Gabsang; Lyden, David; Benezra, Robert; Zaninovic, Nikica; Rosenwaks, Zev; Rabbany, Sina Y; Rafii, Shahin

    2010-01-01

    Previous efforts to differentiate human embryonic stem cells (hESCs) into endothelial cells have not achieved sustained expansion and stability of vascular cells. To define vasculogenic developmental pathways and enhance differentiation, we used an endothelial cell–specific VE-cadherin promoter driving green fluorescent protein (GFP) (hVPr-GFP) to screen for factors that promote vascular commitment. In phase 1 of our method, inhibition of transforming growth factor (TGF)β at day 7 of differentiation increases hVPr-GFP+ cells by tenfold. In phase 2, TGFβ inhibition maintains the proliferation and vascular identity of purified endothelial cells, resulting in a net 36-fold expansion of endothelial cells in homogenous monolayers, which exhibited a transcriptional profile of Id1highVEGFR2highVE-cadherin+ ephrinB2+. Using an Id1-YFP hESC reporter line, we showed that TGFβ inhibition sustains Id1 expression in hESC-derived endothelial cells and that Id1 is required for increased proliferation and preservation of endothelial cell commitment. Our approach provides a serum-free method for differentiation and long-term maintenance of hESC-derived endothelial cells at a scale relevant to clinical application. PMID:20081865

  18. Derivation of porcine pluripotent stem cells for biomedical research.

    Science.gov (United States)

    Shiue, Yow-Ling; Yang, Jenn-Rong; Liao, Yu-Jing; Kuo, Ting-Yung; Liao, Chia-Hsin; Kang, Ching-Hsun; Tai, Chein; Anderson, Gary B; Chen, Lih-Ren

    2016-07-01

    Pluripotent stem cells including embryonic stem cells (ESCs), embryonic germ cells (EGCs), and induced pluripotent stem cells (iPSCs) are capable of self-renew and limitlessly proliferating in vitro with undifferentiated characteristics. They are able to differentiate in vitro, spontaneously or responding to suitable signals, into cells of all three primary germ layers. Consequently, these pluripotent stem cells will be valuable sources for cell replacement therapy in numerous disorders. However, the promise of human ESCs and EGCs is cramped by the ethical argument about destroying embryos and fetuses for cell line creation. Moreover, there are still carcinogenic risks existing toward the goal of clinical application for human ESCs, EGCs, and iPSCs. Therefore, a suitable animal model for stem cell research will benefit the further development of human stem cell technology. The pigs, on the basis of their similarity in anatomy, immunology, physiology, and biochemical properties, have been wide used as model animals in the study of various human diseases. The development of porcine pluripotent stem cell lines will hold the opportunity to provide an excellent material for human counterpart to the transplantation in biomedical research and further development of cell-based therapeutic strategy. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Pluripotent stem cell models of Shwachman-Diamond syndrome reveal a common mechanism for pancreatic and hematopoietic dysfunction

    Science.gov (United States)

    Tulpule, Asmin; Kelley, James M.; Lensch, M. William; McPherson, Jade; Park, In Hyun; Hartung, Odelya; Nakamura, Tomoka; Schlaeger, Thorsten M.; Shimamura, Akiko; Daley, George Q.

    2013-01-01

    Summary Shwachman-Diamond syndrome (SDS), a rare autosomal recessive disorder characterized by exocrine pancreatic insufficiency and hematopoietic dysfunction, is caused by mutations in the Shwachman-Bodian-Diamond syndrome (SBDS) gene. We created human pluripotent stem cell models of SDS by knock-down of SBDS in human embryonic stem cells (hESCs) and generation of induced pluripotent stem cell (iPSC) lines from two SDS patients. SBDS-deficient hESCs and iPSCs manifest deficits in exocrine pancreatic and hematopoietic differentiation in vitro, enhanced apoptosis and elevated protease levels in culture supernatants, which could be reversed by restoring SBDS protein expression through transgene rescue or by supplementing culture media with protease inhibitors. Protease-mediated auto-digestion provides a mechanistic link between the pancreatic and hematopoietic phenotypes in SDS, highlighting the utility of hESCs and iPSCs in obtaining novel insights into human disease. PMID:23602541

  20. YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers

    DEFF Research Database (Denmark)

    Brøchner, Christian B; Johansen, Julia S; Larsen, Lars A

    2012-01-01

    oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted......The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high...... YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3ß, PDX1, CD34, p63, nestin, PAX6) markers. Double...

  1. Fabricação de lã de rocha a partir da escória da produção de ligas FeSiMn Manufacture of mineral wool from slag of ferroalloy production FeSiMn

    Directory of Open Access Journals (Sweden)

    G. F. Rodrigues

    2012-12-01

    Full Text Available Este trabalho visou o estudo da produção de lã de rocha utilizando escória da produção de ligas de ferro silício manganês. O resíduo em questão possui características químicas que indicam a possibilidade de adição deste resíduo em formulações de lãs minerais. Procedeu-se a fusão da escória em um forno elétrico a arco, em escala de laboratório. A escória foi fundida e vazada na temperatura de 1450 ºC em um recipiente com água, visando um resfriamento brusco e com isso uma aproximação do método de produção das lãs de rocha. Amostras da lã de rocha produzida foram caracterizadas por técnicas de análise química, difração de raios X, microscopia eletrônica de varredura e análise térmica diferencial sendo que, os resultados obtidos foram comparados aos valores encontrados na literatura. Os resultados encontrados mostram que a reciclagem dos resíduos para a produção de lãs de rocha é promissora, pois o reaproveitamento da escória de ferro silício manganês permite a transformação destes resíduos em subprodutos através da incorporação dos resíduos na produção de lã de rocha, que possui amplo mercado como isolante térmico, acústico e inibidor de propagação de chamas.This study aimed to investigate the production of rock wool using slag from the production of ferrosilicon manganese alloys. The waste in question has chemical characteristics that indicate the possibility of adding this in formulations of mineral wool. There has been melting the slag in an electric arc furnace in laboratory scale. The slag was melted and poured at a temperature of 1450 ºC in a container with water, aiming at a rapid cooling and thus an approximation of the method of production of rock wool. Samples of rock wool produced were characterized by techniques of chemical analysis, X-ray diffraction, scanning electron microscopy and differential thermal analysis is that the results were compared to those found in the

  2. Universitarios y redes sociales informativas: Escépticos totales, moderados duales o pro-digitales

    Directory of Open Access Journals (Sweden)

    2017-10-01

    Full Text Available El objetivo del presente trabajo es analizar el uso de las redes sociales informativas como herramienta de empoderamiento social por los universitarios españoles, y su percepción de la universidad como institución que contribuye a la formación de una ciudadanía crítica y activa, al tiempo que les proporciona las pertinentes competencias digitales. La revisión bibliográfica evidencia posibles discrepancias respecto al efecto que tienen las nuevas formas de comunicación digital en el empoderamiento de los jóvenes y en particular de los universitarios, así como la existencia de numerosas cuestiones por aclarar en este escenario digital. A continuación se presenta un análisis tipológico de la percepción de los estudiantes universitarios respecto a las redes sociales informativas, empoderamiento social y el papel de la universidad. A partir de los datos recogidos mediante un cuestionario estructurado de una muestra de 236 estudiantes de Grados de Ciencias Sociales, se realiza un análisis de tipologías con el algoritmo K Medias. Se identifican tres tipos –etiquetados como «escépticos totales», «moderados duales» y «pro-digitales»– significativamente diferentes. Se explica su prevalencia, y su caracterización: perfiles de creencias y conducta relacionadas con dichas creencias. El trabajo concluye con diversas recomendaciones para futuras investigaciones en cuanto a la percepción del universitario sobre el uso de las redes sociales como herramienta de transformación social y el papel de la universidad.

  3. "Lo que se hablaba de él y la escandalosa vida que estaba dando al público". aproximación a la noción de escándalo a partir de las huellas del rumor en las prácticas de conciliación informal. El caso de Don José Tadeo Alquizar por "vivir escandalosamente"

    Directory of Open Access Journals (Sweden)

    Nicolás Celis Valderrama

    2016-01-01

    Full Text Available Este artículo quiere analizar, a través del impacto del rumor en las prácticas de conciliación informal, la noción de escándalo en el Chile de fines del siglo XVIII. El escándalo es abordado como un fenómeno constituido desde y en la esfera de lo público. Asimismo, se revelan las huellas del rumor y la relevancia de la conciliación informal en el control de las dinámicas escandalosas. Lo anterior se analizará desde el caso de Don José Tadeo Alquizar por vivir escandalosamente con una mulata desarrollado en Santiago de Chile 1795.

  4. Efeitos da aplicação da escória de siderurgia ferrocromo no solo, no estado nutricional e na produção de matéria seca de mudas de maracujazeiro Aplication of basic slag iron-chromium in soil in nutritional state and dry matter production of passion fruit seedlings

    Directory of Open Access Journals (Sweden)

    Renato de Mello Prado

    2004-04-01

    Full Text Available A escória de siderurgia ferrocromo pode-se constituir em uma fonte alternativa de Ca e Mg, bem como em corretivo da acidez do solo, melhorando a sua fertilidade e o estado nutricional de culturas. Assim, objetivou-se avaliar os efeitos da escória de siderurgia ferrocromo nas alterações dos atributos químicos do solo, na nutrição e no desenvolvimento de mudas de maracujazeiro. Para tanto, instalou-se um experimento em condições de casa de vegetação, empregando-se as seguintes doses crescentes da escória: zero; metade; uma vez; uma vez e meia e duas vezes a dose para elevar a saturação por bases do solo a 80%, correspondendo às doses de: 0; 0,375; 0,750; 1,125 e 1,500 g dm-3, respectivamente. O substrato utilizado foi um Latossolo Vermelho distrófico, ácido (vasos com 2,8 dm³, que foi incubado com a escória de siderurgia, por 30 dias, para posterior semeadura do maracujazeiro, cultivando-as por 85 dias. A aplicação da escória de siderurgia ferrocromo promoveu a neutralização da acidez do solo. Entretanto, mesmo em doses relativamente baixas (360 kg ha-1, houve diminuição no acúmulo de nutrientes e na produção de matéria seca das mudas de maracujazeiro.The basic slag can consist in an alternative source of Ca and Mg, as well as punishment of soil acidity improving the nutricional state of passion plants, and determining the success of the implantation of an orchard. Thus, it was objectified to evaluate the effect of siderurgy slag iron-chromium, in the alterations of the chemical attributes of the soil, in the nutrition of the plants and the development of passion seedlings. For this, it was installed an experiment in conditions of greenhouse, using the following increasing doses of the residue: zero; half; once; once and half and twice to raise V =80%. After 30 days of incubation of the slag in the soil, it was proceeded the plantation of the passion seedlings, in substratum of a distrofic, acid Red Latossol (pot with

  5. Derivation and characterization of human fetal MSCs: an alternative cell source for large-scale production of cardioprotective microparticles.

    Science.gov (United States)

    Lai, Ruenn Chai; Arslan, Fatih; Tan, Soon Sim; Tan, Betty; Choo, Andre; Lee, May May; Chen, Tian Sheng; Teh, Bao Ju; Eng, John Kun Long; Sidik, Harwin; Tanavde, Vivek; Hwang, Wei Sek; Lee, Chuen Neng; El Oakley, Reida Menshawe; Pasterkamp, Gerard; de Kleijn, Dominique P V; Tan, Kok Hian; Lim, Sai Kiang

    2010-06-01

    The therapeutic effects of mesenchymal stem cells (MSCs) transplantation are increasingly thought to be mediated by MSC secretion. We have previously demonstrated that human ESC-derived MSCs (hESC-MSCs) produce cardioprotective microparticles in pig model of myocardial ischemia/reperfusion (MI/R) injury. As the safety and availability of clinical grade human ESCs remain a concern, MSCs from fetal tissue sources were evaluated as alternatives. Here we derived five MSC cultures from limb, kidney and liver tissues of three first trimester aborted fetuses and like our previously described hESC-derived MSCs; they were highly expandable and had similar telomerase activities. Each line has the potential to generate at least 10(16-19) cells or 10(7-10) doses of cardioprotective secretion for a pig model of MI/R injury. Unlike previously described fetal MSCs, they did not express pluripotency-associated markers such as Oct4, Nanog or Tra1-60. They displayed a typical MSC surface antigen profile and differentiated into adipocytes, osteocytes and chondrocytes in vitro. Global gene expression analysis by microarray and qRT-PCR revealed a typical MSC gene expression profile that was highly correlated among the five fetal MSC cultures and with that of hESC-MSCs (r(2)>0.90). Like hESC-MSCs, they produced secretion that was cardioprotective in a mouse model of MI/R injury. HPLC analysis of the secretion revealed the presence of a population of microparticles with a hydrodynamic radius of 50-65 nm. This purified population of microparticles was cardioprotective at approximately 1/10 dosage of the crude secretion. (c) 2009 Elsevier Ltd. All rights reserved.

  6. A defined and xeno-free culture method enabling the establishment of clinical-grade human embryonic, induced pluripotent and adipose stem cells.

    Directory of Open Access Journals (Sweden)

    Kristiina Rajala

    2010-04-01

    Full Text Available The growth of stem cells in in vitro conditions requires optimal balance between signals mediating cell survival, proliferation, and self-renewal. For clinical application of stem cells, the use of completely defined conditions and elimination of all animal-derived materials from the establishment, culture, and differentiation processes is desirable.Here, we report the development of a fully defined xeno-free medium (RegES, capable of supporting the expansion of human embryonic stem cells (hESC, induced pluripotent stem cells (iPSC and adipose stem cells (ASC. We describe the use of the xeno-free medium in the derivation and long-term (>80 passages culture of three pluripotent karyotypically normal hESC lines: Regea 06/015, Regea 07/046, and Regea 08/013. Cardiomyocytes and neural cells differentiated from these cells exhibit features characteristic to these cell types. The same formulation of the xeno-free medium is capable of supporting the undifferentiated growth of iPSCs on human feeder cells. The characteristics of the pluripotent hESC and iPSC lines are comparable to lines derived and cultured in standard undefined culture conditions. In the culture of ASCs, the xeno-free medium provided significantly higher proliferation rates than ASCs cultured in medium containing allogeneic human serum (HS, while maintaining the differentiation potential and characteristic surface marker expression profile of ASCs, although significant differences in the surface marker expression of ASCs cultured in HS and RegES media were revealed.Our results demonstrate that human ESCs, iPSCs and ASCs can be maintained in the same defined xeno-free medium formulation for a prolonged period of time while maintaining their characteristics, demonstrating the applicability of the simplified xeno-free medium formulation for the production of clinical-grade stem cells. The basic xeno-free formulation described herein has the potential to be further optimized for specific

  7. Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro.

    Science.gov (United States)

    Visan, Anke; Hayess, Katrin; Sittner, Dana; Pohl, Elena E; Riebeling, Christian; Slawik, Birgitta; Gulich, Konrad; Oelgeschläger, Michael; Luch, Andreas; Seiler, Andrea E M

    2012-10-01

    Mouse embryonic stem cells (mESCs) represent an attractive cellular system for in vitro studies in developmental biology as well as toxicology because of their potential to differentiate into all fetal cell lineages. The present study aims to establish an in vitro system for developmental neurotoxicity testing employing mESCs. We developed a robust and reproducible protocol for fast and efficient differentiation of the mESC line D3 into neural cells, optimized with regard to chemical testing. Morphological examination and immunocytochemical staining confirmed the presence of different neural cell types, including neural progenitors, neurons, astrocytes, oligodendrocytes, and radial glial cells. Neurons derived from D3 cells expressed the synaptic proteins PSD95 and synaptophysin, and the neurotransmitters serotonin and γ-aminobutyric acid. Calcium ion imaging revealed the presence of functionally active glutamate and dopamine receptors. In addition, flow cytometry analysis of the neuron-specific marker protein MAP2 on day 12 after induction of differentiation demonstrated a concentration dependent effect of the neurodevelopmental toxicants methylmercury chloride, chlorpyrifos, and lead acetate on neuronal differentiation. The current study shows that D3 mESCs differentiate efficiently into neural cells involving a neurosphere-like state and that this system is suitable to detect adverse effects of neurodevelopmental toxicants. Therefore, we propose that the protocol for differentiation of mESCs into neural cells described here could constitute one component of an in vitro testing strategy for developmental neurotoxicity. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Hacia un teatro de cyborgs: Artes escénicas, tecnología/s y subjetividad/es Córdoba-Argentina (1997-2007

    Directory of Open Access Journals (Sweden)

    Valeria Cotaimich

    2012-04-01

    Full Text Available Este artículo presenta una serie de reflexiones respecto a la incidencia performativa del desarrollo tecnológico en la configuracón de las subjetividad/es contemporáneas, en el contexto del capitalismo postindustrial, a través de un análisis cualitativo en torno al proceso de transformación política y poética del teatro independiente en Córdoba-Argentina en la última década. Teórica y metodólogicamente se retoman aportes de la Teatrología, los Estudios de la Peformance, el Psicoanálisis y otros abordajes que indagan la relación entre tecnología/s y arte/s. Se describen ciertos efectos del uso de tecnologías numéricas en diversas áreas: dramaturgia, producción escénica y gestión cultural, proponiendo reemplazar las categorías teatro virtual y teatro tecnológico por teatro de cyborgs a los fines de dar cuenta de la diversidad y el carácter glocal de estos acontecimientos.

  9. Reducción del miedo escénico y mejora del afecto positivo, la autoeficacia y el desempeño ante la actuación en público: un cuasi=experimento aplicando PNL

    OpenAIRE

    Serrano Leiva, Damaris

    2015-01-01

    Treball final de Grau en Psicologia. Codi: PS1048. Curs acadèmic 2014-2015 En este estudio, se llevó a cabo un cuasi-experimento con el fin de evaluar la eficacia de dos técnicas en Programación Neuro Linguística (PNL): anclas y submodalidades en la mejora del afecto positivo, autoeficacia y desempeño, y la reducción del miedo escénico y del afecto negativo en alumnos de teatro de la Escuela Municipal de Villarreal. Para ello, se establecieron dos condiciones: Un grupo experime...

  10. Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

    International Nuclear Information System (INIS)

    Varga, Nóra; Veréb, Zoltán; Rajnavölgyi, Éva; Német, Katalin; Uher, Ferenc; Sarkadi, Balázs; Apáti, Ágota

    2011-01-01

    Highlights: ► MSC like cells were derived from hESC by a simple and reproducible method. ► Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. ► MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

  11. Safety paradigm: genetic evaluation of therapeutic grade human embryonic stem cells.

    Science.gov (United States)

    Stephenson, Emma; Ogilvie, Caroline Mackie; Patel, Heema; Cornwell, Glenda; Jacquet, Laureen; Kadeva, Neli; Braude, Peter; Ilic, Dusko

    2010-12-06

    The use of stem cells for regenerative medicine has captured the imagination of the public, with media attention contributing to rising expectations of clinical benefits. Human embryonic stem cells (hESCs) are the best model for capital investment in stem cell therapy and there is a clear need for their robust genetic characterization before scaling-up cell expansion for that purpose. We have to be certain that the genome of the starting material is stable and normal, but the limited resolution of conventional karyotyping is unable to give us such assurance. Advanced molecular cytogenetic technologies such as array comparative genomic hybridization for identifying chromosomal imbalances, and single nucleotide polymorphism analysis for identifying ethnic background and loss of heterozygosity should be introduced as obligatory diagnostic tests for each newly derived hESC line before it is deposited in national stem cell banks. If this new quality standard becomes a requirement, as we are proposing here, it would facilitate and accelerate the banking process, since end-users would be able to select the most appropriate line for their particular application, thus improving efficiency and streamlining the route to manufacturing therapeutics. The pharmaceutical industry, which may use hESC-derived cells for drug screening, should not ignore their genomic profile as this may risk misinterpretation of results and significant waste of resources.

  12. Il Cratilo di Platone

    Directory of Open Access Journals (Sweden)

    Maria Chiara Pievatolo

    2015-06-01

    Full Text Available La guida ipertestuale alla lettura del Cratilo di Platone composta per gli studenti della facoltà di Scienze politiche dell’università di Pisa è ora visibile a tutti qui. L’ipertesto ha tratto vantaggio dall’Introduzione alla linguistica generale del professor Manuel Barbera dell’università...

  13. Experiment list: SRX590292 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available -rep1; Mus musculus; ChIP-Seq source_name=R-Ctrl-Flag ChIP || strain background=C57BL/6 || genotype/variation=Foxd3 conditional knock...out || cell type=embryonic stem cells (ESCs; R cells) || cell line of origin=Foxd3 conditional knock

  14. Comparison of the early response of human embryonic stem cells and human induced pluripotent stem cells to ionizing radiation.

    Science.gov (United States)

    Suchorska, Wiktoria Maria; Augustyniak, Ewelina; Łukjanow, Magdalena

    2017-04-01

    Despite the well-demonstrated efficacy of stem cell (SC) therapy, this approach has a number of key drawbacks. One important concern is the response of pluripotent SCs to treatment with ionizing radiation (IR), given that SCs used in regenerative medicine will eventually be exposed to IR for diagnostic or treatment‑associated purposes. Therefore, the aim of the present study was to examine and compare early IR‑induced responses of pluripotent SCs to assess their radioresistance and radiosensitivity. In the present study, 3 cell lines; human embryonic SCs (hESCs), human induced pluripotent SCs (hiPSCs) and primary human dermal fibroblasts (PHDFs); were exposed to IR at doses ranging from 0 to 15 gray (Gy). Double strand breaks (DSBs), and the gene expression of the following DNA repair genes were analyzed: P53; RAD51; BRCA2; PRKDC; and XRCC4. hiPSCs demonstrated greater radioresistance, as fewer DSBs were identified, compared with hESCs. Both pluripotent SC lines exhibited distinct gene expression profiles in the most common DNA repair genes that are involved in homologous recombination, non‑homologous end‑joining and enhanced DNA damage response following IR exposure. Although hESCs and hiPSCs are equivalent in terms of capacity for pluripotency and differentiation into 3 germ layers, the results of the present study indicate that these 2 types of SCs differ in gene expression following exposure to IR. Consequently, further research is required to determine whether hiPSCs and hESCs are equally safe for application in clinical practice. The present study contributes to a greater understanding of DNA damage response (DDR) mechanisms activated in pluripotent SCs and may aid in the future development of safe SC‑based clinical protocols.

  15. Coculturing with endothelial cells promotes in vitro maturation and electrical coupling of human embryonic stem cell-derived cardiomyocytes.

    Science.gov (United States)

    Pasquier, Jennifer; Gupta, Renuka; Rioult, Damien; Hoarau-Véchot, Jessica; Courjaret, Raphael; Machaca, Khaled; Al Suwaidi, Jassim; Stanley, Edouard G; Rafii, Shahin; Elliott, David A; Abi Khalil, Charbel; Rafii, Arash

    2017-06-01

    Pluripotent human embryonic stem cells (hESC) are a promising source of repopulating cardiomyocytes. We hypothesized that we could improve maturation of cardiomyocytes and facilitate electrical interconnections by creating a model that more closely resembles heart tissue; that is, containing both endothelial cells (ECs) and cardiomyocytes. We induced cardiomyocyte differentiation in the coculture of an hESC line expressing the cardiac reporter NKX2.5-green fluorescent protein (GFP), and an Akt-activated EC line (E4 + ECs). We quantified spontaneous beating rates, synchrony, and coordination between different cardiomyocyte clusters using confocal imaging of Fura Red-detected calcium transients and computer-assisted image analysis. After 8 days in culture, 94% ± 6% of the NKX2-5GFP + cells were beating when hESCs embryonic bodies were plated on E4 + ECs compared with 34% ± 12.9% for controls consisting of hESCs cultured on BD Matrigel (BD Biosciences) without ECs at Day 11 in culture. The spatial organization of beating areas in cocultures was different. The GFP + cardiomyocytes were close to the E4 + ECs. The average beats/min of the cardiomyocytes in coculture was faster and closer to physiologic heart rates compared with controls (50 ± 14 [n = 13] vs 25 ± 9 [n = 8]; p < 0.05). The coculture with ECs led to synchronized beating relying on the endothelial network, as illustrated by the loss of synchronization upon the disruption of endothelial bridges. The coculturing of differentiating cardiomyocytes with Akt-activated ECs but not EC-conditioned media results in (1) improved efficiency of the cardiomyocyte differentiation protocol and (2) increased maturity leading to better intercellular coupling with improved chronotropy and synchrony. Copyright © 2017. Published by Elsevier Inc.

  16. Assessing the Effectiveness of a Far-Red Fluorescent Reporter for Tracking Stem Cells In Vivo

    Directory of Open Access Journals (Sweden)

    Jing Zhou

    2017-12-01

    Full Text Available Far-red fluorescent reporter genes can be used for tracking cells non-invasively in vivo using fluorescence imaging. Here, we investigate the effectiveness of the far-red fluorescent protein, E2-Crimson (E2C, for tracking mouse embryonic cells (mESCs in vivo following subcutaneous administration into mice. Using a knock-in strategy, we introduced E2C into the Rosa26 locus of an E14-Bra-GFP mESC line, and after confirming that the E2C had no obvious effect on the phenotype of the mESCs, we injected them into mice and imaged them over nine days. The results showed that fluorescence intensity was weak, and cells could only be detected when injected at high densities. Furthermore, intensity peaked on day 4 and then started to decrease, despite the fact that tumour volume continued to increase beyond day 4. Histopathological analysis showed that although E2C fluorescence could barely be detected in vivo at day 9, analysis of frozen sections indicated that all mESCs within the tumours continued to express E2C. We hypothesise that the decrease in fluorescence intensity in vivo was probably due to the fact that the mESC tumours became more vascular with time, thus leading to increased absorbance of E2C fluorescence by haemoglobin. We conclude that the E2C reporter has limited use for tracking cells in vivo, at least when introduced as a single copy into the Rosa26 locus.

  17. Research on Airborne SAR Imaging Based on Esc Algorithm

    Science.gov (United States)

    Dong, X. T.; Yue, X. J.; Zhao, Y. H.; Han, C. M.

    2017-09-01

    Due to the ability of flexible, accurate, and fast obtaining abundant information, airborne SAR is significant in the field of Earth Observation and many other applications. Optimally the flight paths are straight lines, but in reality it is not the case since some portion of deviation from the ideal path is impossible to avoid. A small disturbance from the ideal line will have a major effect on the signal phase, dramatically deteriorating the quality of SAR images and data. Therefore, to get accurate echo information and radar images, it is essential to measure and compensate for nonlinear motion of antenna trajectories. By means of compensating each flying trajectory to its reference track, MOCO method corrects linear phase error and quadratic phase error caused by nonlinear antenna trajectories. Position and Orientation System (POS) data is applied to acquiring accuracy motion attitudes and spatial positions of antenna phase centre (APC). In this paper, extend chirp scaling algorithm (ECS) is used to deal with echo data of airborne SAR. An experiment is done using VV-Polarization raw data of C-band airborne SAR. The quality evaluations of compensated SAR images and uncompensated SAR images are done in the experiment. The former always performs better than the latter. After MOCO processing, azimuth ambiguity is declined, peak side lobe ratio (PSLR) effectively improves and the resolution of images is improved obviously. The result shows the validity and operability of the imaging process for airborne SAR.

  18. RESEARCH ON AIRBORNE SAR IMAGING BASED ON ESC ALGORITHM

    Directory of Open Access Journals (Sweden)

    X. T. Dong

    2017-09-01

    Full Text Available Due to the ability of flexible, accurate, and fast obtaining abundant information, airborne SAR is significant in the field of Earth Observation and many other applications. Optimally the flight paths are straight lines, but in reality it is not the case since some portion of deviation from the ideal path is impossible to avoid. A small disturbance from the ideal line will have a major effect on the signal phase, dramatically deteriorating the quality of SAR images and data. Therefore, to get accurate echo information and radar images, it is essential to measure and compensate for nonlinear motion of antenna trajectories. By means of compensating each flying trajectory to its reference track, MOCO method corrects linear phase error and quadratic phase error caused by nonlinear antenna trajectories. Position and Orientation System (POS data is applied to acquiring accuracy motion attitudes and spatial positions of antenna phase centre (APC. In this paper, extend chirp scaling algorithm (ECS is used to deal with echo data of airborne SAR. An experiment is done using VV-Polarization raw data of C-band airborne SAR. The quality evaluations of compensated SAR images and uncompensated SAR images are done in the experiment. The former always performs better than the latter. After MOCO processing, azimuth ambiguity is declined, peak side lobe ratio (PSLR effectively improves and the resolution of images is improved obviously. The result shows the validity and operability of the imaging process for airborne SAR.

  19. Láser escáner y nubes de puntos. Un horizonte aplicado al análisis arqueológico de edificios

    Directory of Open Access Journals (Sweden)

    García-Gómez, Ismael

    2011-12-01

    Full Text Available This paper analyses the laser scanner, a powerful tool for graphic recording of cultural heritage sites. Although attention will be drawn to those technical and logistical aspects and methods related to its employment, this report will mainly focus on its applicability on Building Archaeology, highlighting those diagnosing capacities which may help the stratigrapher to establish an evolutionary sequence of the building. The basis of this study is our work on San Miguel church in Vitoria-Gasteiz.En el siguiente artículo analizamos uno de los más modernos instrumentos de documentación gráfica del patrimonio, el láser escáner. Aunque profundizaremos en todos aquellos aspectos técnicos, logísticos y de método relativos a su empleo, nos centraremos en su efectiva aplicabilidad dentro del campo de la Arqueología de la Arquitectura, incidiendo en aquellas capacidades diagnósticas que pueden ayudar al estratígrafo a determinar la secuencia evolutiva de un edificio. Tomaremos como base nuestra experiencia en la iglesia de San Miguel de Vitoria-Gasteiz.

  20. Report of an ESC-EAPCI Task Force on the evaluation and use of bioresorbable scaffolds for percutaneous coronary intervention: executive summary.

    Science.gov (United States)

    Byrne, Robert A; Stefanini, Giulio G; Capodanno, Davide; Onuma, Yoshinobu; Baumbach, Andreas; Escaned, Javier; Haude, Michael; James, Stefan; Joner, Michael; Jüni, Peter; Kastrati, Adnan; Oktay, Semih; Wijns, William; Serruys, Patrick W; Windecker, Stephan

    2018-05-07

    A previous Task Force of the European Society of Cardiology (ESC) and European Association of Percutaneous Cardiovascular Interventions (EAPCI) provided a report on recommendations for the non-clinical and clinical evaluation of coronary stents. Following dialogue with the European Commission, the Task Force was asked to prepare an additional report on the class of devices known as bioresorbable scaffolds (BRS). Five BRS have CE-mark approval for use in Europe. Only one device-the Absorb bioresorbable vascular scaffold-has published randomized clinical trial data and this data show inferior outcomes to conventional drug-eluting stents (DES) at 2-3 years. For this reason, at present BRS should not be preferred to conventional DES in clinical practice. The Task Force recommends that new BRS devices should undergo systematic non-clinical testing according to standardized criteria prior to evaluation in clinical studies. A clinical evaluation plan should include data from a medium sized, randomized trial against DES powered for a surrogate end point of clinical efficacy. Manufacturers of successful devices receive CE-mark approval for use and must have an approved plan for a large-scale randomized clinical trial with planned long-term follow-up.

  1. Celano’s Naples: Itineraries through a Material City (1692

    Directory of Open Access Journals (Sweden)

    Joris van Gastel

    2014-10-01

    Full Text Available La Napoli di Carlo Celano (1692: itinerari in una città materialeIl saggio propone una rilettura della nota guida di Napoli pubblicata nel 1692 da Carlo Celano, le Notitie del bello, dell’antico e del curioso della città di Napoli, ma con particolare attenzione per l’interesse continuo ivi dimostrato per la dimensione materiale della città. Soffermandosi sistematicamente sulla qualità della pietra utilizzata in molti edifici napoletani, Celano offre nella sua guida un’illustrazione dettagliata ed esperta della notevole varietà e ricchezza di materiali edili, che nella realtà napoletana del Seicento contraddistingue non solo il panorama urbano ma anche la stessa identità cittadina. La documentazione contenuta nel libro di Celano illustra pertanto un fenomeno caratteristico per l’urbanistica napoletana, e in un momento che precede di poco la sua distruzione, in seguito all’introduzione di stucco come materiale adatto per coprire quasi tutte le superfici, anche quelle in pietre solo recentemente elogiate per la loro propria qualità estetica.

  2. Huntingtin Protein is Essential for Mitochondrial Metabolism, Bioenergetics and Structure in Murine Embryonic Stem Cells

    Science.gov (United States)

    Ismailoglu, Ismail; Chen, Qiuying; Popowski, Melissa; Yang, Lili; Gross, Steven S.; Brivanlou, Ali H.

    2014-01-01

    Mutations in the Huntington locus (htt) have devastating consequences. Gain-of-poly-Q repeats in Htt protein causes Huntington's disease (HD), while htt-/- mutants display early embryonic lethality. Despite its importance, the function of Htt remains elusive. To address this, we compared more than 3,700 compounds in three syngeneic mouse embryonic stem cell (mESC) lines: htt-/-, extended poly-Q (Htt-Q140/7), and wildtype mESCs (Htt-Q7/7) using untargeted metabolite profiling. While Htt-Q140/7 cells, did not show major differences in cellular bioenergetics, we find extensive metabolic aberrations in htt-/- mESCs, including: (i) complete failure of ATP production despite preservation of the mitochondrial membrane potential; (ii) near-maximal glycolysis, with little or no glycolytic reserve; (iii) marked ketogenesis; (iv) depletion of intracellular NTPs; (v) accelerated purine biosynthesis and salvage; and (vi) loss of mitochondrial structural integrity. Together, our findings reveal that Htt is necessary for mitochondrial structure and function from the earliest stages of embryogenesis, providing a molecular explanation for htt-/- early embryonic lethality. PMID:24780625

  3. REST/NRSF Knockdown Alters Survival, Lineage Differentiation and Signaling in Human Embryonic Stem Cells.

    Directory of Open Access Journals (Sweden)

    Kaushali Thakore-Shah

    Full Text Available REST (RE1 silencing transcription factor, also known as NRSF (neuron-restrictive silencer factor, is a well-known transcriptional repressor of neural genes in non-neural tissues and stem cells. Dysregulation of REST activity is thought to play a role in diverse diseases including epilepsy, cancer, Down's syndrome and Huntington's disease. The role of REST/NRSF in control of human embryonic stem cell (hESC fate has never been examined. To evaluate the role of REST in hESCs we developed an inducible REST knockdown system and examined both growth and differentiation over short and long term culture. Interestingly, we have found that altering REST levels in multiple hESC lines does not result in loss of self-renewal but instead leads to increased survival. During differentiation, REST knockdown resulted in increased MAPK/ERK and WNT signaling and increased expression of mesendoderm differentiation markers. Therefore we have uncovered a new role for REST in regulation of growth and early differentiation decisions in human embryonic stem cells.

  4. ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.

    Science.gov (United States)

    Pettinato, Giuseppe; Vanden Berg-Foels, Wendy S; Zhang, Ning; Wen, Xuejun

    2014-01-01

    We report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCKi/+spin, and -ROCKi/-spin. Cell suspensions of BG01V/hOG and H9 hESC lines were pipetted into non-adherent hydrogel substrates containing defined microwell arrays. hEBs of consistent size and spherical geometry can be formed in each of the four conditions, including the -ROCKi/-spin condition. The hEBs formed under the -ROCKi/-spin condition differentiated to develop the three embryonic germ layers and tissues derived from each of the germ layers. This simplified hEB production technique offers homogeneity in hEB size and shape to support synchronous differentiation, elimination of the ROCKi xeno-factor and rate-limiting centrifugation treatment, and low-cost scalability, which will directly support automated, large-scale production of hEBs and hESC-derived cells needed for clinical, research, or therapeutic applications.

  5. Exploración escénica de la obra la tortura de E. Buenaventura

    Directory of Open Access Journals (Sweden)

    Isabel Cristina Flores

    2016-04-01

    Full Text Available omo cada año, comenzamos los preparativos del Encuentro de la Red Latinoamericana de Creación e Investigación Teatral Universitaria (RED CITU, organización académica que reúne a la comunidad estudiantil y docente de once universidades latinoamericanas en una exploración teatral única, realizar un proceso inédito de exploración escénica, sobre temas de interés académico común. En el año 2015 se celebró el Delta 6 a la Universidad de Antioquia, Medellín, Colombia; el evento estuvo planificado del 19 al 27 de septiembre. Cada equipo de las once universidades participantes tuvo una gran tarea que cumplir, indagar sobre la propuesta académica establecida por la universidad sede, en esta ocasión determinada como, el montaje de la obra La Tortura, de Enrique Buenaventura, observando los principios del Método de creación colectiva, legado del maestro Enrique Buenaventura al teatro universal. Como parte de la planificación del evento en la Benemérita Universidad Autónoma de Puebla, cada año iniciamos el proceso con la presentación del proyecto a la Dirección de la Escuela de Artes, para contar con el soporte económico y dar inicio a la recopilación de materiales, requerimientos del encuentro, investigación sobre el tema por tratar e integración del equipo que estará a cargo de la realización del proyecto. El encuentro anual de la Red CITU (en la modalidad Delta es esperado con mucho interés y entusiasmo por la comunidad estudiantil de la Licenciatura en Arte Dramático, Escuela de Artes de la BUAP. Los estudiantes preguntan, se informan, se preparan, ensayan para ser los elegidos en la audición, a la cual se convoca de manera abierta a toda la comunidad estudiantil de los últimos dos años de la carrera.

  6. The U-line line balancing problem

    NARCIS (Netherlands)

    Miltenburg, G.J.; Wijngaard, J.

    1994-01-01

    The traditional line balancing (LB) problem considers a production line in which stations are arranged consecutively in a line. A balance is determined by grouping tasks into stations while moving forward (or backward) through a precedence network. Recently many production lines are being arranged

  7. Equivalency of Buffalo (Bubalus Bubalis) Embryonic Stem Cells Derived From Fertilized, Parthenogenetic, and Hand-Made Cloned Embryos

    Science.gov (United States)

    Muzaffar, Musharifa; Selokar, Naresh L.; Singh, Karn P.; Zandi, Mohammad; Singh, Manoj K.; Shah, Riaz A.; Chauhan, Manmohan S.; Singla, Suresh K.; Palta, Prabhat

    2012-01-01

    Abstract This study was aimed at establishing buffalo embryonic stem cells (ESCs) from in vitro fertilized (IVF), parthenogenetic, and hand-made cloned (HMC) embryos and to check their equivalency in terms of stem cell marker expression, longevity, proliferation, and differentiation pattern. ESCs derived from all three sources were found by immunofluorescence to express the pluripotency markers SSEA-4, TRA-1-60, TRA-1-81, OCT4, and SOX2 and were able to form embryoid bodies containing cells expressing genes specific to endoderm (AFP, HNF4, and GATA4), mesoderm (MSX1, BMP4, and ASA), and ectoderm (cytokeratin 8 and NF68). Reverse transcriptase PCR (RT-PCR) showed cells from all sources to be positive for pluripotency markers OCT4, SOX2, NANOG, STAT3, REX1, FOXD3, NUCLEOSTEMIN, and TELOMERASE. Pluripotency markers OCT4, SOX2, NANOG, and c-MYC were also analyzed by real-time PCR. No significant differences were observed among ESCs from all three sources for all these genes except NANOG, whose expression was higher (pcells (1.603±0.315 and 1±0, respectively). Pluripotent, stable buffalo ESC lines derived from IVF, parthenogenesis, and HMC embryos may be genetically manipulated to provide a powerful tool for studies involving embryonic development, genomic imprinting, gene targeting, cloning, chimera formation, and transgenic animal production. PMID:22582863

  8. Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

    Energy Technology Data Exchange (ETDEWEB)

    Varga, Nora [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Vereb, Zoltan; Rajnavoelgyi, Eva [Department of Immunology, Medical and Health Science Centre, University of Debrecen, Debrecen (Hungary); Nemet, Katalin; Uher, Ferenc; Sarkadi, Balazs [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Apati, Agota, E-mail: apati@kkk.org.hu [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary)

    2011-10-28

    Highlights: Black-Right-Pointing-Pointer MSC like cells were derived from hESC by a simple and reproducible method. Black-Right-Pointing-Pointer Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. Black-Right-Pointing-Pointer MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

  9. Mesoderm Lineage 3D Tissue Constructs Are Produced at Large-Scale in a 3D Stem Cell Bioprocess.

    Science.gov (United States)

    Cha, Jae Min; Mantalaris, Athanasios; Jung, Sunyoung; Ji, Yurim; Bang, Oh Young; Bae, Hojae

    2017-09-01

    Various studies have presented different approaches to direct pluripotent stem cell differentiation such as applying defined sets of exogenous biochemical signals and genetic/epigenetic modifications. Although differentiation to target lineages can be successfully regulated, such conventional methods are often complicated, laborious, and not cost-effective to be employed to the large-scale production of 3D stem cell-based tissue constructs. A 3D-culture platform that could realize the large-scale production of mesoderm lineage tissue constructs from embryonic stem cells (ESCs) is developed. ESCs are cultured using our previously established 3D-bioprocess platform which is amenable to mass-production of 3D ESC-based tissue constructs. Hepatocarcinoma cell line conditioned medium is introduced to the large-scale 3D culture to provide a specific biomolecular microenvironment to mimic in vivo mesoderm formation process. After 5 days of spontaneous differentiation period, the resulting 3D tissue constructs are composed of multipotent mesodermal progenitor cells verified by gene and molecular expression profiles. Subsequently the optimal time points to trigger terminal differentiation towards cardiomyogenesis or osteogenesis from the mesodermal tissue constructs is found. A simple and affordable 3D ESC-bioprocess that can reach the scalable production of mesoderm origin tissues with significantly improved correspondent tissue properties is demonstrated. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. La matematica e il calcolatore l'avventura pisana

    CERN Document Server

    Capovani, Milvio

    2004-01-01

    La matematica ha invaso domini regolati dall'autorità e dall'abitudine e li ha radicalmente modificati; il calcolatore guida una profonda rivoluzione nel sistema mondo: la matematica, attraverso il calcolatore, si è diffusa in vari campi del sapere. Questi i temi trattati. La storia delle macchine per il calcolo a Pisa è incastonata in questo scenario.

  11. Focus sul digital marketing

    OpenAIRE

    S. Vianello; G. Randisi

    2011-01-01

    Il marketing impatta o meno sulle performance aziendali? L’investimento in competenze di Marketing & Sales genera ritorni per le imprese? La risposta che emerge da questo volume è certa. Il marketing conta davvero, perché guida le scelte strategiche delle imprese e perché impatta direttamente sui loro indicatori economico-finanziari, attraverso competenze e metriche adeguate.

  12. The Making of RDA

    Directory of Open Access Journals (Sweden)

    Tom Delsey

    2016-05-01

    Full Text Available L'autore ripercorre lo sviluppo di RDA dal principio, nel 2005, fino alla sua prima pubblicazione, nel 2010. L'impegno di sviluppo è inserito nel contesto di un ambiente digitale in evoluzione che trasforma sia la produzione sia la diffusione delle risorse informative e delle risorse utilizzate per creare, immagazzinare e accedere ai dati che descrivono tali risorse. L'autore esamina l'interazione tra l'impegno strategico ad allineare RDA con i nuovi modelli concettuali, le strutture di database emergenti e lo sviluppo dei metadati nelle comunità alleate, da una parte, e la compatibilità con l'eredità di AACR2 e dei database esistenti dall'altra. Gli aspetti esaminati comprendono la strutturazione di RDA come linguaggio di descrizione delle risorse, l'organizzazione del nuovo standard come uno strumento di lavoro e il raffinamento delle linee guida e delle istruzioni per la registrazione dei dati secondo RDA.

  13. La interpretación musical y la ansiedad escénica : validación de un instrumento de diagnóstico y su aplicación en los estudiantes españoles de Conservatorio Superior de Música

    OpenAIRE

    Arnáiz Rodríguez, Mateo

    2016-01-01

    [Resumen] La ansiedad que sufren los músicos, estudiantes de los Conservatorios Superiores de Música en España, es un proceso que se denomina “ansiedad escénica en la interpretación musical” y que afecta negativamente a la interpretación, su futura actividad profesional. Esta denominación está basada en la teoría desarrollada por Kenny (2011) y que a su vez se fundamenta en la teoría de la ansiedad sobre la triple vulnerabilidad de Barlow (2002). El fenómeno de la ansiedad e...

  14. Rejuvenation of MPTP-induced human neural precursor cell senescence by activating autophagy

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Liang [East Hospital, Tongji University School of Medicine, Shanghai (China); Dong, Chuanming [East Hospital, Tongji University School of Medicine, Shanghai (China); Department of Anatomy and Neurobiology, The Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong (China); Sun, Chenxi; Ma, Rongjie; Yang, Danjing [East Hospital, Tongji University School of Medicine, Shanghai (China); Zhu, Hongwen, E-mail: hongwen_zhu@hotmail.com [Tianjin Hospital, Tianjin Academy of Integrative Medicine, Tianjin (China); Xu, Jun, E-mail: xunymc2000@yahoo.com [East Hospital, Tongji University School of Medicine, Shanghai (China)

    2015-08-21

    Aging of neural stem cell, which can affect brain homeostasis, may be caused by many cellular mechanisms. Autophagy dysfunction was found in aged and neurodegenerative brains. However, little is known about the relationship between autophagy and human neural stem cell (hNSC) aging. The present study used 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) to treat neural precursor cells (NPCs) derived from human embryonic stem cell (hESC) line H9 and investigate related molecular mechanisms involved in this process. MPTP-treated NPCs were found to undergo premature senescence [determined by increased senescence-associated-β-galactosidase (SA-β-gal) activity, elevated intracellular reactive oxygen species level, and decreased proliferation] and were associated with impaired autophagy. Additionally, the cellular senescence phenotypes were manifested at the molecular level by a significant increase in p21 and p53 expression, a decrease in SOD2 expression, and a decrease in expression of some key autophagy-related genes such as Atg5, Atg7, Atg12, and Beclin 1. Furthermore, we found that the senescence-like phenotype of MPTP-treated hNPCs was rejuvenated through treatment with a well-known autophagy enhancer rapamycin, which was blocked by suppression of essential autophagy gene Beclin 1. Taken together, these findings reveal the critical role of autophagy in the process of hNSC aging, and this process can be reversed by activating autophagy. - Highlights: • We successfully establish hESC-derived neural precursor cells. • MPTP treatment induced senescence-like state in hESC-derived NPCs. • MPTP treatment induced impaired autophagy of hESC-derived NPCs. • MPTP-induced hESC-derived NPC senescence was rejuvenated by activating autophagy.

  15. Periodic harvesting of embryonic stem cells from a hollow-fiber membrane based four-compartment bioreactor.

    Science.gov (United States)

    Knöspel, Fanny; Freyer, Nora; Stecklum, Maria; Gerlach, Jörg C; Zeilinger, Katrin

    2016-01-01

    Different types of stem cells have been investigated for applications in drug screening and toxicity testing. In order to provide sufficient numbers of cells for such in vitro applications a scale-up of stem cell culture is necessary. Bioreactors for dynamic three-dimensional (3D) culture of growing cells offer the option for culturing large amounts of stem cells at high densities in a closed system. We describe a method for periodic harvesting of pluripotent stem cells (PSC) during expansion in a perfused 3D hollow-fiber membrane bioreactor, using mouse embryonic stem cells (mESC) as a model cell line. A number of 100 × 10(6) mESC were seeded in bioreactors in the presence of mouse embryonic fibroblasts (MEF) as feeder cells. Over a cultivation interval of nine days cells were harvested by trypsin perfusion and mechanical agitation every second to third culture day. A mean of 380 × 10(6) mESC could be removed with every harvest. Subsequent to harvesting, cells continued growing in the bioreactor, as determined by increasing glucose consumption and lactate production. Immunocytochemical staining and mRNA expression analysis of markers for pluripotency and the three germ layers showed a similar expression of most markers in the harvested cells and in mESC control cultures. In conclusion, successful expansion and harvesting of viable mESC from bioreactor cultures with preservation of sterility was shown. The present study is the first one showing the feasibility of periodic harvesting of adherent cells from a continuously perfused four-compartment bioreactor including further cultivation of remaining cells. © 2015 American Institute of Chemical Engineers.

  16. Calcário e escória de siderurgia avaliados por análise foliar, acúmulo, e exportação de macronutrientes em cana-de-açúcar Lime and slag evaluated by leaf analysis, macronutrient accumulation and export of sugarcane

    Directory of Open Access Journals (Sweden)

    Renato de Mello Prado

    2002-03-01

    Full Text Available A opção de uso de materiais corretivos alternativos como as escórias de siderurgia, pode influenciar na absorção de nutrientes pela planta e, consequentemente, na diagnose foliar, acúmulo e exportação de nutrientes. Com o objetivo de estudar o efeito da aplicação de doses de escória de siderurgia e de calcário na absorção de macronutrientes pela cana-de-açúcar (variedade SP80-1842, foi instalado em janeiro de 1998, em Ituverava-SP, um experimento de campo. Aplicou-se escória de siderurgia e calcário, em quatro doses equivalentes 0; 1,23; 2,52 e 3,80 t ha-1 de CaCO3, em um Latossolo Vermelho-Amarelo distrófico, em pré-plantio na cana-planta. Determinaram-se, além da produção de colmos, as concentrações de macronutrientes na folha +3 da cana-planta e da cana-soca (aos 120 dias após a emergência dos brotos e na parte aérea da cana-soca (aos 345 dias após a emergência dos brotos. Os materiais corretivos e as doses não alteraram a absorção de macronutrientes pela cana-de-açúcar. Houve maior acúmulo e exportação de macronutrientes de acordo com as doses de ambos corretivos. A escória de siderurgia comportou-se de modo semelhante ao calcário na nutrição da cana-de-açúcar quanto aos macronutrientes.Slag can be used as an alternative for lime as soil amendment, having influence on nutrient uptake by the plant and, as a result, on leaf analysis and nutrient accumulation. To study the effect of the application of increasing rates of slag and lime stone on the uptake of macronutrients by sugar cane (variety SP 80-1842 an experiment was installed under field conditions, in January 1998, at Ituverava-SP, Brazil. Slag and limestone were applied to the sugarcane crop at planting, in four rates: 0; 1.23; 2.52 and 3.80 t ha-1 of CaCO3, using a Typic Haplorthox. In addition to the cane stalk production a verification was performed for the first and second ratoons (120 days and 345 after the emergence of the shoots

  17. Computational Biology Methods for Characterization of Pluripotent Cells.

    Science.gov (United States)

    Araúzo-Bravo, Marcos J

    2016-01-01

    Pluripotent cells are a powerful tool for regenerative medicine and drug discovery. Several techniques have been developed to induce pluripotency, or to extract pluripotent cells from different tissues and biological fluids. However, the characterization of pluripotency requires tedious, expensive, time-consuming, and not always reliable wet-lab experiments; thus, an easy, standard quality-control protocol of pluripotency assessment remains to be established. Here to help comes the use of high-throughput techniques, and in particular, the employment of gene expression microarrays, which has become a complementary technique for cellular characterization. Research has shown that the transcriptomics comparison with an Embryonic Stem Cell (ESC) of reference is a good approach to assess the pluripotency. Under the premise that the best protocol is a computer software source code, here I propose and explain line by line a software protocol coded in R-Bioconductor for pluripotency assessment based on the comparison of transcriptomics data of pluripotent cells with an ESC of reference. I provide advice for experimental design, warning about possible pitfalls, and guides for results interpretation.

  18. Improved cell therapy protocols for Parkinson's disease based on differentiation efficiency and safety of hESC-, hiPSC-, and non-human primate iPSC-derived dopaminergic neurons

    DEFF Research Database (Denmark)

    Sundberg, Maria; Bogetofte, Helle; Lawson, Tristan

    2013-01-01

    of safety and efficacy of stem cell-derived DA neurons. The aim of this study was to improve the safety of human- and non-human primate iPSC (PiPSC)-derived DA neurons. According to our results, NCAM(+) /CD29(low) sorting enriched VM DA neurons from pluripotent stem cell-derived neural cell populations......The main motor symptoms of Parkinson's disease are due to the loss of dopaminergic (DA) neurons in the ventral midbrain (VM). For the future treatment of Parkinson's disease with cell transplantation it is important to develop efficient differentiation methods for production of human iPSCs and h......ESCs-derived midbrain-type DA neurons. Here we describe an efficient differentiation and sorting strategy for DA neurons from both human ES/iPS cells and non-human primate iPSCs. The use of non-human primate iPSCs for neuronal differentiation and autologous transplantation is important for preclinical evaluation...

  19. Influência da incorporação de resíduo de escória de fluxo de soldagem nas propriedades tecnológicas de argamassa de múltiplo uso e cerâmica vermelha para construção civil Influence of the incorporation of waste of slag of welding flux on the technological properties of multiple use mortar and red ceramic for civil construction

    Directory of Open Access Journals (Sweden)

    C. E Viana

    2010-03-01

    Full Text Available O processo de soldagem a arco submerso é empregado em grande escala na indústria metal-mecânica. No entanto, este processo gera enorme quantidade de resíduo de escória. Em geral, este resíduo vem sendo disposto em lixões. Nos últimos anos, a área de cerâmica tem atraído grande atenção para reciclagem de resíduos industriais. Neste trabalho foi feito um estudo objetivando avaliar a influência do resíduo de escória de fluxo de soldagem sobre as propriedades tecnológicas de argamassa de múltiplo uso e cerâmica vermelha para uso na construção civil. O resíduo de escória de fluxo de soldagem foi caracterizado em relação à composição química, difração de raios X, análise do tamanho de partículas e massa específica real. A argamassa foi preparada com a incorporação de resíduo de escória de soldagem em substituição total de areia natural (agregado miúdo tradicional. As argamassas foram caracterizadas em termos de consistência, densidade de massa no estado fresco, teor de ar incorporado e resistência à compressão. Análise microestrutural das argamassas foi feita via MEV. Foi preparada também uma série de misturas argila/resíduo contendo até 10% em peso de resíduo de escória de fluxo de soldagem. As peças cerâmicas foram preparadas por prensagem uniaxial e queimadas entre 850 ºC e 950 ºC. As peças queimadas foram caracterizadas quanto à retração linear, absorção de água, porosidade aparente, massa específica aparente e tensão de ruptura à flexão. Os resultados mostraram que o resíduo de escória de fluxo de soldagem pode substituir areia natural como agregado miúdo em argamassa de múltiplo uso. Além do mais, o resíduo de escória pode ser incorporado em cerâmica vermelha (tijolos e blocos cerâmicos como substituto parcial de argila natural.The submerged-arc welding process is widely used in the metal-mechanic industry. However, this process generates huge amounts of slag waste. In

  20. Localized decrease of β-catenin contributes to the differentiation of human embryonic stem cells

    International Nuclear Information System (INIS)

    Lam, Hayley; Patel, Shyam; Wong, Janelle; Chu, Julia; Li, Adrian; Li, Song

    2008-01-01

    Human embryonic stem cells (hESC) are pluripotent, and can be directed to differentiate into different cell types for therapeutic applications. To expand hESCs, it is desirable to maintain hESC growth without differentiation. As hESC colonies grow, differentiated cells are often found at the periphery of the colonies, but the underlying mechanism is not well understood. Here, we utilized micropatterning techniques to pattern circular islands or strips of matrix proteins, and examined the spatial pattern of hESC renewal and differentiation. We found that micropatterned matrix restricted hESC differentiation at colony periphery but allowed hESC growth into multiple layers in the central region, which decreased hESC proliferation and induced hESC differentiation. In undifferentiated hESCs, β-catenin primarily localized at cell-cell junctions but not in the nucleus. The amount of β-catenin in differentiating hESCs at the periphery of colonies or in multiple layers decreased significantly at cell-cell junctions. Consistently, knocking down β-catenin decreased Oct-4 expression in hESCs. These results indicate that localized decrease of β-catenin contributes to the spatial pattern of differentiation in hESC colonies

  1. Modelos urbanos tridimensionales generados a partir de nubes de puntos de un escáner láser terrestre

    Directory of Open Access Journals (Sweden)

    Hernán Porras Díaz

    2014-07-01

    Full Text Available En este artículo se presenta una revisión del estado del arte de las diferentes metodologías que se han desarrollado para el tratamiento digital de nubes de puntos tridimensionales, recolectadas mediante un escáner láser terrestre. Este instrumento tiene la capacidad de capturar información topográfica y geométrica de cualquier estructura con precisiones hasta el orden de los milímetros, y en algunos casos también captura la información radiométrica de los objetos escaneados. Esta información recolectada se representa en forma de nubes de puntos que se utilizan en una amplia variedad de aplicaciones como la planeación del crecimiento urbano, el análisis estructural de construcciones, el modelado de fenómenos de erosión y deforestación, la documentación de patrimonio histórico y la navegación virtual a través de los sitios turísticos mediante modelos tridimensionales de edificaciones. Por lo tanto, el propósito de este trabajo es proveer una revisión en el estado del conocimiento acerca de las técnicas desarrolladas en las diferentes fases que se llevan a cabo con el tratamiento de estas nubes de puntos, como el registro y georreferenciación, la segmentación, clasificación y modelado tridimensional.

  2. La ansiedad escénica en intérpretes musicales chilenos

    Directory of Open Access Journals (Sweden)

    Mimí Marinovic

    2006-06-01

    Full Text Available La ansiedad escénica (AE es uno de los mayores problemas del ejercicio profesional de los intérpretes musicales. Encuestas realizadas en países desarrollados revelan cifras de prevalencia entre 24% -70% y citan casos severos, causantes del abandono de la carrera. Objetivos: detectar la prevalencia de la AE en músicos chilenos que desempeñan su profesión en los principales conjuntos de música de concierto y determinar algunos de los factores que, a su juicio, influyen en su aparición. Método: 249 intérpretes (122 instrumentistas, 26 directores y 101 cantantes respondieron un cuestionario autoaplicado, incluido en una investigación más amplia sobre los intérpretes de arte. Este instrumento fue complementado con 36 entrevistas abiertas (13 instrumentistas, 6 directores y 11 cantantes. Resultados: el 78% de los músicos estudiados admitió haber sufrido AE, más mujeres que varones. El 64% consideró que ella se relaciona principalmente con la tarea a ejecutar (preparación insuficiente, dificultad de la obra, mientras que el 32% lo atribuyó a factores personales (ser nervioso, temor al fracaso y sólo el 4% a factores situacionales. Hubo diferencias significativas por género, especialidad y edad. Las entrevistas permitieron profundizar acerca del alcance de estos resultados. Conclusiones: se constató una alta prevalencia de AE, la cual debiera ser abordada interdisciplinariamente por especialistas de la salud mental y la música en beneficio de la educación general, la formación especializada y el desarrollo profesional de los intérpretes musicalesPerformance anxiety (PA is one of the major problems that musicians face in their careers. Surveys carried out in developed countries show prevalence figures ranging 24% -70% and quote severe cases leading to abandonment of the profession. Objectives: detect the prevalence of PA in Chilean musicians working in the main groups of concert music and determine some of the factors that

  3. Prefazione—aspetti relativi al benessere animale nel trasporto a lunga distanza

    Directory of Open Access Journals (Sweden)

    Gardner Murray

    2008-03-01

    veterinario e di sanità animale. Riconoscendone l'importanza, la World Organisation for Animal Health (Office International des Épizooties: OIE nel suo IV piano Strategico (punti a e b, ha identificato il benessere animale come tema chiave ed ha lanciato una serie di iniziative come lo sviluppo di linee guida per il benessere animale comuni per istruire i paesi, le industrie ed il pubblico in genere sulle pratiche più appropriate. Le risultanti linee guide dell'OIE per il trasporto di animali via mare e terra hanno fornito l'indispensabile punto di riferimento per ulteriori sviluppi futuri.Questo numero di Veterinaria Italiana si occupa di trasporto del bestiame. Non è una semplice collazione di articoli scientifici sull'argomento. Piuttosto, la sua unicità deriva dal fatto che fornisce un'indicazione e una guida sulle misure pratiche per migliorare l'approccio basato su dati scientifici. Politici, analisti del rischio, legislatori, manager della qualità, progettisti e formatori, avranno grandi vantaggi da questo numero, che servirà come documento di riferimento sull'argomento.Il ruolo dei veterinari e dei sevizi veterinari nel fornire guida ed indicazioni su benessere, sanità e allevamento di animali è fondamentale e non va sottovalutato. Essi hanno la grande responsabilità di occuparsi di questo argomento e grazie alle competenze acquisite, possono ‘fare la differenza’ nel supportare il continuo miglioramento del benessere e della sanità animale.

  4. Transposon-mediated BAC transgenesis in human ES cells

    Science.gov (United States)

    Rostovskaya, Maria; Fu, Jun; Obst, Mandy; Baer, Isabell; Weidlich, Stefanie; Wang, Hailong; Smith, Andrew J. H.; Anastassiadis, Konstantinos; Stewart, A. Francis

    2012-01-01

    Transgenesis is a cornerstone of molecular biology. The ability to integrate a specifically engineered piece of DNA into the genome of a living system is fundamental to our efforts to understand life and exploit its implications for medicine, nanotechnology and bioprospecting. However, transgenesis has been hampered by position effects and multi-copy integration problems, which are mainly due to the use of small, plasmid-based transgenes. Large transgenes based on native genomic regions cloned into bacterial artificial chromosomes (BACs) circumvent these problems but are prone to fragmentation. Herein, we report that contrary to widely held notions, large BAC-sized constructs do not prohibit transposition. We also report the first reliable method for BAC transgenesis in human embryonic stem cells (hESCs). The PiggyBac or Sleeping Beauty transposon inverted repeats were integrated into BAC vectors by recombineering, followed by co-lipofection with the corresponding transposase in hESCs to generate robust fluorescent protein reporter lines for OCT4, NANOG, GATA4 and PAX6. BAC transposition delivers several advantages, including increased frequencies of single-copy, full-length integration, which will be useful in all transgenic systems but especially in difficult venues like hESCs. PMID:22753106

  5. Alternative sources of pluripotency: science, ethics, and stem cells.

    Science.gov (United States)

    Kastenberg, Zachary J; Odorico, Jon S

    2008-07-01

    Despite many advances in human embryonic stem cell (hESC) technology the ethical dilemma involving the destruction of a human embryo is one factor that has limited the development of hESC based clinical therapies. Two recent reports describing the production of pluripotent stem cells following the in vitro reprogramming of human somatic cells with certain defined factors illustrate one potential method of bypassing the ethical debate surrounding hESCs (Yu J, Vodyanik MA, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007 Dec;318(5858):1917-1920; Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007 Nov;131(5): 861-872.). Other alternative methods include nuclear transfer, altered nuclear transfer, and parthenogenesis; each with its own set of advantages and disadvantages. This review discusses recent advances in these technologies with specific focus on the issues of embryo destruction, oocyte recovery, and the potential of each technology to produce large scale, patient specific cell transplantation therapies that would require little or no immunosuppression.

  6. Electrochemical Characterization of Ni/ScYSZ Electrodes as SOFC Anodes

    DEFF Research Database (Denmark)

    Ramos, Tania; Søgaard, Martin; Mogensen, Mogens Bjerg

    2014-01-01

    Investigations of Ni/ScYSZ cermets were performed by electrochemical impedance spectroscopy (EIS) using different symmetric designs: electrolyte supported (ESC) and anode supported (ASC) cells. The obtained spectra were analyzed using distribution of relaxation times (DRT), and complex non......-linear least squares fitting (CNLS). Depending on the cell design, one or two low frequency gas transport related processes have been identified, and fitted with generalized finite Warburg (GFW) elements. One was related to gas diffusion in a stagnant layer above the anode (ESC+ASC), and the other to gas...... diffusion in the anode support layer (ASC). A higher frequency process has also been identified, and correlated to the charge transfer (CT) combined with ionic conduction in the ceramic matrix. This has been fitted using a transmission line model (TML), which correlates the exhibited responses...

  7. Line facilities outline

    International Nuclear Information System (INIS)

    1998-08-01

    This book deals with line facilities. The contents of this book are outline line of wire telecommunication ; development of line, classification of section of line and theory of transmission of line, cable line ; structure of line, line of cable in town, line out of town, domestic cable and other lines, Optical communication ; line of optical cable, transmission method, measurement of optical communication and cable of the sea bottom, Equipment of telecommunication line ; telecommunication line facilities and telecommunication of public works, construction of cable line and maintenance and Regulation of line equipment ; regulation on technique, construction and maintenance.

  8. Instructions for learning Italian in two early modern Dutch travel guides: Delitiae Italiae (1602 and Delitiae Urbis Romae (1625

    Directory of Open Access Journals (Sweden)

    José van der Helm

    2014-10-01

    Full Text Available Istruzioni per imparare l'italiano in due guide di viaggio olandesi della prima età moderna: Delitiae Italiae (1602 e Delitiae Urbis Romae (1625I primi decenni del secolo XVII hanno visto la pubblicazione di due guide di viaggio specificamente destinate a un pubblico neerlandese che intende a viaggiare in Italia. La Delitiae Italiae è stata pubblicata per la prima volta nel 1602, ad Arnhem, essendo un adattamento di un’edizione tedesca pubblicata in occcasione dell’anno giubilare (1600; la Delitiae Urbis Romae appare nel 1625 ad Amsterdam ed offre una descrizione dei luoghi di interesse di Roma includendo una serie di illustrazioni. L’ultima guida risulta anch’essa un adattamento in neerlandese di un’edizione scritta originariamente in latino e pubblicata da Domenicus Custodi ad Augusta (Germania. Le due guide non stanno a sé; esse fanno parte di un corpus di testi di viaggio prodotto dalla metà del Cinquecento in poi: lavori dettagliati che miravano a fornire informazioni su tutti gli aspetti dell’Italia accanto a guide di viaggio compatte, di carattere ibrido, che non descrivono solo i luoghi da visitare, ma vogliono fornire anche informazioni pratiche. Sono guide piccole, pubblicate nel formato comodo in ottavo o in duodecimo, e da paragonare alle guide Lonely Planet, riferimento moderno per i viaggiatori indipendenti. Le due Delitiae descritte in questo articolo entrano chiaramente nellla categoria di guida pratica. Inoltre esse contengono − come parte del loro obiettivo di essere di uso pratico − delle istruzioni per imparare la lingua italiana: una serie di dialoghi in neerlandese-italiano si trova in appendice. Viene analizzato il contenuto di questi dialoghi, nonché il materiale linguistico stesso proveniente da circoli mercantili Quattro- e Cinquecenteschi di Anversa. Infine si ricerca la motivazione degli editori di ripubblicare il materiale mettendolo in un nuovo contesto secentesco: il viaggiatore

  9. Similarity law for Widom lines and coexistence lines.

    Science.gov (United States)

    Banuti, D T; Raju, M; Ihme, M

    2017-05-01

    The coexistence line of a fluid separates liquid and gaseous states at subcritical pressures, ending at the critical point. Only recently, it became clear that the supercritical state space can likewise be divided into regions with liquidlike and gaslike properties, separated by an extension to the coexistence line. This crossover line is commonly referred to as the Widom line, and is characterized by large changes in density or enthalpy, manifesting as maxima in the thermodynamic response functions. Thus, a reliable representation of the coexistence line and the Widom line is important for sub- and supercritical applications that depend on an accurate prediction of fluid properties. While it is known for subcritical pressures that nondimensionalization with the respective species critical pressures p_{cr} and temperatures T_{cr} only collapses coexistence line data for simple fluids, this approach is used for Widom lines of all fluids. However, we show here that the Widom line does not adhere to the corresponding states principle, but instead to the extended corresponding states principle. We resolve this problem in two steps. First, we propose a Widom line functional based on the Clapeyron equation and derive an analytical, species specific expression for the only parameter from the Soave-Redlich-Kwong equation of state. This parameter is a function of the acentric factor ω and compares well with experimental data. Second, we introduce the scaled reduced pressure p_{r}^{*} to replace the previously used reduced pressure p_{r}=p/p_{cr}. We show that p_{r}^{*} is a function of the acentric factor only and can thus be readily determined from fluid property tables. It collapses both subcritical coexistence line and supercritical Widom line data over a wide range of species with acentric factors ranging from -0.38 (helium) to 0.34 (water), including alkanes up to n-hexane. By using p_{r}^{*}, the extended corresponding states principle can be applied within

  10. Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells

    International Nuclear Information System (INIS)

    Miyazaki, Takamichi; Futaki, Sugiko; Hasegawa, Kouichi; Kawasaki, Miwa; Sanzen, Noriko; Hayashi, Maria; Kawase, Eihachiro; Sekiguchi, Kiyotoshi; Nakatsuji, Norio; Suemori, Hirofumi

    2008-01-01

    Human embryonic stem cells (hESCs) are thought to be a promising cell source for cell transplantation therapy. For such a clinical application, the hESCs should be manipulated using appropriate and qualified materials. In this study, we examined the efficacy of recombinant human laminin (rhLM) isoforms on the undifferentiated growth of hESCs. We first determined the major integrins expressed on the hESCs to reveal the preference of the hESCs for rhLMs, and found that the hESCs mainly expressed integrin α6β1, which binds predominantly to laminin-111, -332 and -511/-521. When the hESCs were seeded onto rhLMs, the cells indeed adhered markedly to rhLM-332, and to rhLM-511 and rhLM-111 to a lesser extent. The hESCs proliferated on these three rhLMs for several passages while preserving their pluripotency. These results show that rhLM-111, -332, and -511 are good substrates to expand undifferentiated hESCs due to their high affinity to integrin α6β1 expressed on hESCs

  11. [ESC guidelines on the management of valvular heart disease. What has changed and what is new?].

    Science.gov (United States)

    Mangner, N; Schuler, G

    2013-12-01

    In 2012 the new and collaborative "Guidelines on the management of valvular heart disease (version 2012)" were published by the European Society of Cardiology (ESC) and the European Association for Cardio-Thoracic Surgery (EACTS). These guidelines emphasize that decision-making in patients with valvular heart disease should ideally be carried out by a"heart team" with particular expertise in valvular heart disease. In aortic regurgitation pathologies of the aortic root are frequent and in patients with Marfan syndrome, surgery is indicated when the maximal ascending aortic diameter is ≥50 mm, while the threshold for intervention should be lower in patients with risk factors for progression. Regarding aortic stenosis, transcatheter aortic valve implantation (TAVI) should be performed only in hospitals with on-site cardiac surgery and with a"heart team" available to assess patient risks. The TAVI procedure is indicated in patients with severe symptomatic aortic stenosis who are judged by the"heart team" to be unsuitable for surgery but have sufficient life expectancy. It should be considered for high-risk patients with severe symptomatic aortic stenosis based on the individual risk profile assessed by the"heart team". Furthermore, low flow - low gradient aortic stenosis with normal ejection fraction and the difficult topic of asymptomatic severe aortic stenosis and the indications for aortic valve replacement are discussed. With respect to mitral regurgitation, valve repair should be the preferred technique when it is expected to be durable. The topics of asymptomatic mitral regurgitation as well as percutaneous mitral valve repair using the edge to edge technique as an alternative for high risk patients are discussed. Tricuspid disease should not be forgotten and during left-sided valve surgery, tricuspid valve surgery should be considered in the presence of mild to moderate secondary regurgitation if there is significant annular dilatation. Last but not least

  12. A new mathematical formulation of the line-by-line method in case of weak line overlapping

    Science.gov (United States)

    Ishov, Alexander G.; Krymova, Natalie V.

    1994-01-01

    A rigorous mathematical proof is presented for multiline representation on the equivalent width of a molecular band which consists in the general case of n overlapping spectral lines. The multiline representation includes a principal term and terms of minor significance. The principal term is the equivalent width of the molecular band consisting of the same n nonoverlapping spectral lines. The terms of minor significance take into consideration the overlapping of two, three and more spectral lines. They are small in case of the weak overlapping of spectral lines in the molecular band. The multiline representation can be easily generalized for optically inhomogeneous gas media and holds true for combinations of molecular bands. If the band lines overlap weakly the standard formulation of line-by-line method becomes too labor-consuming. In this case the multiline representation permits line-by-line calculations to be performed more effectively. Other useful properties of the multiline representation are pointed out.

  13. Stemcell Information: SKIP000944 [SKIP Stemcell Database[Archive

    Lifescience Database Archive (English)

    Full Text Available inson disease 168006 ... 66 60-69 Female ... No No iPS cell line iPS細胞 human ES-like Research Grade Retrov... SKIP000944 ... dermal fibroblast 表皮繊維芽細胞 Diseased SP08.3 SP08.3 ... パーキンソン病 G20 Park...irus SOX2, KLF4 and OCT3/4 Yes Yes MEFs Lines of patient-specific iPS cells were maintained by mechanical di...ssociation of colonies and splitting 1:3 onto feeder cells in hESC medium or by l

  14. Stemcell Information: SKIP000940 [SKIP Stemcell Database[Archive

    Lifescience Database Archive (English)

    Full Text Available inson disease 168006 ... 46 40-49 Male ... No No iPS cell line iPS細胞 human ES-like Research Grade Retrovir... SKIP000940 ... dermal fibroblast 表皮繊維芽細胞 Diseased SP04.1 SP04.1 ... パーキンソン病 G20 Park...us SOX2, KLF4 and OCT3/4 Yes Yes MEFs Lines of patient-specific iPS cells were maintained by mechanical diss...ociation of colonies and splitting 1:3 onto feeder cells in hESC medium or by lim

  15. Generation of human embryonic stem cells from abnormal blastocyst diagnosed with albinism.

    Science.gov (United States)

    Sun, Yi; Zhou, Xiaoying; Chen, Jing; Du, Juan; Lu, Guangxiu; Lin, Ge; Ouyang, Qi

    2016-11-01

    Human embryonic stem cell (hESC) line chHES-478 was derived from abnormal blastocyst diagnosed with albinism after preimplantation genetic diagnosis (PGD) treatment. DNA sequencing analysis confirmed that chHES-478 cell line carried a compound heterozygous mutation, c.896G>A(p.Arg299His) and c.929_930insC(p.Pro310Glnfs*9), of TYR gene. Characteristic tests proved that the chHES-478 cell line presented typical markers of pluripotency and had the capability to form the three germ layers both in vitro and in vivo. Copyright © 2016 Michael Boutros, German Cancer Research Center, Heidelberg, Germany. Published by Elsevier B.V. All rights reserved.

  16. 76 FR 51367 - China Shipping Container Lines Co., Ltd.; COSCO Container Lines Company Limited; Evergreen Line A...

    Science.gov (United States)

    2011-08-18

    ... FEDERAL MARITIME COMMISSION [Docket No. 11-12] China Shipping Container Lines Co., Ltd.; COSCO Container Lines Company Limited; Evergreen Line A Joint Service Agreement; Hanjin Shipping Co., Ltd... Maritime Commission (Commission) by China Shipping Container Lines Co., Ltd.; COSCO Container Lines Company...

  17. Locust bean gum as an alternative polymeric coating for embryonic stem cell culture

    Energy Technology Data Exchange (ETDEWEB)

    Perestrelo, Ana Rubina [Regenerative Medicine Program, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve (Portugal); IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve (Portugal); PhD Program in Biomedical Sciences, Universidade do Algarve (Portugal); Grenha, Ana [IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve (Portugal); Rosa da Costa, Ana M. [Centro de Investigação em Química do Algarve (CIQA) and Departamento de Química e Farmácia, Faculdade de Ciências e Tecnologia, Universidade do Algarve (Portugal); Belo, José António, E-mail: jose.belo@fcm.unl.pt [Regenerative Medicine Program, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve (Portugal); IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve (Portugal); Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo Mártires da Pátria 130, 1169-056 Lisboa (Portugal)

    2014-07-01

    Pluripotent embryonic stem cells (ESCs) have self-renewal capacity and the potential to differentiate into any cellular type depending on specific cues (pluripotency) and, therefore, have become a vibrant research area in the biomedical field. ESCs are usually cultured in gelatin or on top of a monolayer of feeder cells such as mitotically inactivated mouse embryonic fibroblasts (MEFsi). The latter is the gold standard support to maintain the ESCs in the pluripotent state. Examples of versatile, non-animal derived and inexpensive materials that are able to support pluripotent ESCs are limited. Therefore, our aim was to find a biomaterial able to support ESC growth in a pluripotent state avoiding laborious and time consuming parallel culture of MEFsi and as simple to handle as gelatin. Many of the new biomaterials used to develop stem cell microenvironments are using natural polymers adsorbed or covalently attached to the surface to improve the biocompatibility of synthetic polymers. Locust beam gum (LBG) is a natural, edible polymer, which has a wide range of potential applications in different fields, such as food and pharmaceutical industry, due to its biocompatibility, adhesiveness and thickening properties. The present work brings a natural system based on the use of LBG as a coating for ESC culture. Undifferentiated mouse ESCs were cultured on commercially available LBG to evaluate its potential in maintaining pluripotent ESCs. In terms of morphology, ESC colonies in LBG presented the regular dome shape with bright borders, similar to the colonies obtained in co-cultures with MEFsi and characteristic of pluripotent ESC colonies. In short-term cultures, ESC proliferation in LBG coating was similar to ESC cultured in gelatin and the cells maintained their viability. The activity of alkaline phosphatase and Nanog, Sox2 and Oct4 expression of mouse ESCs cultured in LBG were comparable or in some cases higher than in ESCs cultured in gelatin. An in vitro

  18. Locust bean gum as an alternative polymeric coating for embryonic stem cell culture

    International Nuclear Information System (INIS)

    Perestrelo, Ana Rubina; Grenha, Ana; Rosa da Costa, Ana M.; Belo, José António

    2014-01-01

    Pluripotent embryonic stem cells (ESCs) have self-renewal capacity and the potential to differentiate into any cellular type depending on specific cues (pluripotency) and, therefore, have become a vibrant research area in the biomedical field. ESCs are usually cultured in gelatin or on top of a monolayer of feeder cells such as mitotically inactivated mouse embryonic fibroblasts (MEFsi). The latter is the gold standard support to maintain the ESCs in the pluripotent state. Examples of versatile, non-animal derived and inexpensive materials that are able to support pluripotent ESCs are limited. Therefore, our aim was to find a biomaterial able to support ESC growth in a pluripotent state avoiding laborious and time consuming parallel culture of MEFsi and as simple to handle as gelatin. Many of the new biomaterials used to develop stem cell microenvironments are using natural polymers adsorbed or covalently attached to the surface to improve the biocompatibility of synthetic polymers. Locust beam gum (LBG) is a natural, edible polymer, which has a wide range of potential applications in different fields, such as food and pharmaceutical industry, due to its biocompatibility, adhesiveness and thickening properties. The present work brings a natural system based on the use of LBG as a coating for ESC culture. Undifferentiated mouse ESCs were cultured on commercially available LBG to evaluate its potential in maintaining pluripotent ESCs. In terms of morphology, ESC colonies in LBG presented the regular dome shape with bright borders, similar to the colonies obtained in co-cultures with MEFsi and characteristic of pluripotent ESC colonies. In short-term cultures, ESC proliferation in LBG coating was similar to ESC cultured in gelatin and the cells maintained their viability. The activity of alkaline phosphatase and Nanog, Sox2 and Oct4 expression of mouse ESCs cultured in LBG were comparable or in some cases higher than in ESCs cultured in gelatin. An in vitro

  19. In vivo control of CpG and non-CpG DNA methylation by DNA methyltransferases.

    Directory of Open Access Journals (Sweden)

    Julia Arand

    2012-06-01

    Full Text Available The enzymatic control of the setting and maintenance of symmetric and non-symmetric DNA methylation patterns in a particular genome context is not well understood. Here, we describe a comprehensive analysis of DNA methylation patterns generated by high resolution sequencing of hairpin-bisulfite amplicons of selected single copy genes and repetitive elements (LINE1, B1, IAP-LTR-retrotransposons, and major satellites. The analysis unambiguously identifies a substantial amount of regional incomplete methylation maintenance, i.e. hemimethylated CpG positions, with variant degrees among cell types. Moreover, non-CpG cytosine methylation is confined to ESCs and exclusively catalysed by Dnmt3a and Dnmt3b. This sequence position-, cell type-, and region-dependent non-CpG methylation is strongly linked to neighboring CpG methylation and requires the presence of Dnmt3L. The generation of a comprehensive data set of 146,000 CpG dyads was used to apply and develop parameter estimated hidden Markov models (HMM to calculate the relative contribution of DNA methyltransferases (Dnmts for de novo and maintenance DNA methylation. The comparative modelling included wild-type ESCs and mutant ESCs deficient for Dnmt1, Dnmt3a, Dnmt3b, or Dnmt3a/3b, respectively. The HMM analysis identifies a considerable de novo methylation activity for Dnmt1 at certain repetitive elements and single copy sequences. Dnmt3a and Dnmt3b contribute de novo function. However, both enzymes are also essential to maintain symmetrical CpG methylation at distinct repetitive and single copy sequences in ESCs.

  20. Bioprinting of human pluripotent stem cells and their directed differentiation into hepatocyte-like cells for the generation of mini-livers in 3D.

    Science.gov (United States)

    Faulkner-Jones, Alan; Fyfe, Catherine; Cornelissen, Dirk-Jan; Gardner, John; King, Jason; Courtney, Aidan; Shu, Wenmiao

    2015-10-21

    We report the first investigation into the bioprinting of human induced pluripotent stem cells (hiPSCs), their response to a valve-based printing process as well as their post-printing differentiation into hepatocyte-like cells (HLCs). HLCs differentiated from both hiPSCs and human embryonic stem cells (hESCs) sources were bioprinted and examined for the presence of hepatic markers to further validate the compatibility of the valve-based bioprinting process with fragile cell transfer. Examined cells were positive for nuclear factor 4 alpha and were demonstrated to secrete albumin and have morphology that was also found to be similar to that of hepatocytes. Both hESC and hiPSC lines were tested for post-printing viability and pluripotency and were found to have negligible difference in terms of viability and pluripotency between the printed and non-printed cells. hESC-derived HLCs were 3D printed using alginate hydrogel matrix and tested for viability and albumin secretion during the remaining differentiation and were found to be hepatic in nature. 3D printed with 40-layer of HLC-containing alginate structures reached peak albumin secretion at day 21 of the differentiation protocol. This work demonstrates that the valve-based printing process is gentle enough to print human pluripotent stem cells (hPSCs) (both hESCs and hiPSCs) while either maintaining their pluripotency or directing their differentiation into specific lineages. The ability to bioprint hPSCs will pave the way for producing organs or tissues on demand from patient specific cells which could be used for animal-free drug development and personalized medicine.

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