Implementation of non-invasive prenatal testing by semiconductor sequencing in a genetic laboratory

Objectives: To implement non-invasive prenatal testing (NIPT) for fetal aneuploidies with semiconductor sequencing in an academic cytogenomic laboratory and to evaluate the first 15-month experience on clinical samples. Methods: We validated a NIPT protocol for cell-free fetal DNA sequencing from maternal plasma for the detection of trisomy 13, 18 and 21 on a semiconductor sequencing instrument. Fetal DNA fraction calculation for all samples and several quality parameters were implemented in the workflow. One thousand eighty-one clinical NIPT samples were analysed, following the described protocol. Results: Non-invasive prenatal testing was successfully implemented and validated on 201 normal and 74 aneuploid samples. From 1081 clinical samples, 17 samples showed an abnormal result: 14 trisomy 21 samples, one trisomy 18 and one trisomy 16 were detected. Also a maternal copy number variation on chromosome 13 was observed, which could potentially lead to a false positive trisomy 13 result. One sex discordant result was reported, possibly attributable to a vanishing twin. Moreover, our combined fetal fraction calculation enabled a more reliable risk estimate for trisomy 13, 18 and 21. Conclusions: Non-invasive prenatal testing for trisomy 21, 18 and 13 has a very high specificity and sensitivity. Because of several biological phenomena, diagnostic invasive confirmation of abnormal results remains required

Mechanismen der transkriptionellen Regulation durch c-Myc

Das Proto-Onkogen c-MYC spielt eine entscheidende Rolle bei der Entstehung zahlreicher humaner Tumoren. Um zu verstehen, wie Myc in der Tumorigenese wirkt, sind Kenntnisse über Interaktionspartner und Wirkmechanismen von Myc sehr wichtig. Myc ist ein evolutionär konservierter Transkriptionsfaktor, der Gene aktivieren oder reprimieren kann, wobei diesen Funktionen verschiedene und zum Teil wenig verstandene Mechanismen zugrunde liegen. In der vorliegenden Arbeit wurden transkriptionelle Regulationsmechanismen von Myc sowie biochemische Interaktionen mit Kofaktoren an den genomischen Myc-Bindestellen eines aktivierten (Nukleolin) und eines reprimierten Zielgens (c/EBPα) untersucht. Es wurde gezeigt, dass MB2-abhängige Kofaktoren nicht diffentiell zu beiden Zielgenen rekrutiert werden. Myc-induzierte Histon-Acetylierung sowie Serin5-Phosphorylierung des RNAPII-Komplexes wurde ebenfalls an beiden Zielgenen nachgewiesen. Der Kofaktor p300 sowie der Mediatorkomplex werden hingegen selektiv nur zu dem Myc-aktivierten Zielgen rekrutiert. Durch in vitro- und in vivo-Interaktionsstudien wurde Med24 als Myc-interagierende Untereinheit des Mediatorkomplexes identifziert und durch RNA-Interferenz-basierte Gendepletion von Med24 in HeLa-Zellen konnte die Bedeutung von Med24 für die Transaktivierung von Myc nachgewiesen werden

Molecular characterization of genetic variation in somaclones of durum wheat (Triticum durum Desf) using SSR markers

This study is a continuation of a previous work; the main objective is to characterize the induced somaclonal variation in the first generation of durum wheat plants. In a previous experiment, in vitro plants of three genotypes of durum wheat were obtained following treatment with PEG6000 (0%, 10% and 20%). First generation offspring was produced. Five microsatellite markers were used for genotyping 26 durum wheat somaclones obtained under stressful and non-stressful conditions from 3 durum wheat cultivars. Amplifiable and reproducible alleles were obtained in three of the tested loci. These loci provided in total 78 monomorphic alleles of which 69 were detected in somaclones while the remaining 9 in the mother plants. High values of among-population genetic diversity were found, which accounted for 71 % of the total genetic variation. The number of alleles per locus varied from three to six. Estimated genetic distances varied from 0.83 to 1.67 between populations. The somaclonal variation was identified with 2 SSR markers. Five new alleles were identified in somaclonal variants DKR1-S1, DKR1-S2, OZR1-S2, WR1-C3 and WR1-S1at loci gmw131 and gwm427. Genetic variation rate was 21.74%. Eighty % of the genetic variation was identified in plants obtained from callus undergoing high osmotic pressure. The presence of selective agent in the medium could explain the observed genetic variations. Somaclonal variation cannot always be detected at the gross morphological level. The selected SSR markers could be used to study the uniformity of plants obtained from tissue culture and varietal identification

Support for Higher Education in the 1992-1993 Louisiana Legislature.

The public higher education system in Louisiana is facing a variety of problems. The Louisiana Legislature plays a vital role in the determination of policy regarding higher education in the state. Legislators have the responsibility of addressing current and emerging issues and finding solutions to problems. The purpose of the study was to determine the voting record of members of the 1992 Louisiana Legislature on issues facing higher education so as to infer legislative support for higher education. This purpose was accomplished by studying roll call votes on bills considered important to higher education in relation to personal characteristics of legislators. Data were collected on personal characteristics for all members of the 1992 legislature, including replacements. A panel of experts rated selected roll call bills voted on during the 1992-1993 regular and special sessions to determine the mean score for each bill. The mean score of each bill was applied to the appropriate yea or nay vote in determining a support for higher education score for each legislator. A model was developed for the senate that explained 33.1% of the variance in support for higher education. Seven variables in the stepwise regression model included legislative experience, business occupation, black race, number of bills introduced, education committee membership, legal occupation, and legislative committee leadership. A model developed for the house explained 21.4% of the variance in support for higher education. Five variables entered the stepwise regression model, including male gender, other occupation, democrat party affiliation, white race, legislative experience. The findings of the study indicated that there were more differences than similarities between the senate and the house research models in support for higher education. Legislative experience, race, and occupations, three variables common to both models, influence support for higher education differently in the two chambers. It is recommended that further study of the legislature include survey techniques to explore the influence of other factors contributing to the unexplained variance of legislative support for higher education

Génie des procédés d'agglomération de poudres alimentaires (éléments de phénoménologie des apports d'eau et d'énergie mécanique)

Le procédé d'agglomération humide trouve des applications dans des secteurs industriels d'importance (e.g. agroalimentaire, pharmaceutique, génie civil, etc.) et s'appuie encore fortement sur le savoir-faire des opérateurs. Ce travail porte sur l'étude des contributions des apports hydriques et mécaniques à l'agglomération humide d'une poudre réactive (transformation de la semoule de blé en grains de couscous). Une approche découplée des apports d'eau et d'énergie mécanique est proposée. L'apport d'eau est étudié par la modélisation de l'influence des paramètres formulation et procédé de l'atomisation liquide sur la taille des gouttes pulvérisées. Les apports d'énergie mécanique sont étudiés à l'aide d'un équipement modèle de malaxage. La répartition des contraintes verticales dans le milieu granulaire à l'état statique est identifiée via une "cartographie 2D des isocontraintes". Le comportement de la poudre au malaxage est étudié au cours du déplacement de la pale dans le malaxeur modèle, par l'analyse des champs de vitesses des particules (obtenus par vélocimétrie par images de particules), et par la mesure des contraintes verticales au niveau de la pale. Une étude de la sthénique et de la cinématique des écoulements granulaires permet d'identifier des longueurs caractéristiques impliquées dans le comportement du milieu granulaire à l'état statique et sous sollicitation mécanique. L'étude couplée des apports hydriques et mécaniques est réalisée par suivi des dynamiques d'agglomération humide dans un malaxeur à pale. Elles sont décrites par des mesures en continu de la consommation énergétique et des mesures in situ des spectres d'absorption proche infra-rouge.The wet agglomeration process presents large applications in different industrial fields (e.g. food, pharmaceutics, civil engineering, etc.) and is still mainly based on technical know-how and empiricism of operators. This thesis work investigates the contributions of water and mechanical energy inputs to the wet agglomeration of a reactive powder, in the particular case of the transformation of durum wheat semolina in couscous grains. A uncoupled approach of both water and mechanical energy inputs is carried out. The water addition is studied through a modelisation of the influence of operating and formulation parameters of the liquid atomization process on the droplet size. Mechanical energy inputs are studied using a model experimental mixing equipment. Vertical stress distribution in the granular bed in static conditions is identified thanks to the establishment of a "2D iso-stress cartography". The granular medium behaviour under mechanical solicitation is studied during the blade motion in the model mixing equipment thanks to the analysis of granular flows and velocity fields (obtained by particle image velocimetry) and to vertical stress measurements directly on the blade. A sthenic and kinematic study of granular flows allows to identify characteristic lengths involved in the granular medium behaviour under mechanical solicitation. The coupled study of water and mechanical inputs is conducted by following in-line the wet agglomeration dynamics in a pilot mixing device using energetical consumption measurements as well as in situ acquiring of near infrared absorption spectra.MONTPELLIER-SupAgro La Gaillarde (341722306) / SudocSudocFranceF