Analytical strategies for the comprehensive profiling of histone post translational modifications by mass spectrometry and implications for functional analyses

Le long bio-polymère d'ADN est condensé à l’intérieur du noyau des cellules eukaryotes à l'aide de petites protéines appelées histones. En plus de leurs fonctions condensatrices,ces histones sont également la cible de nombreuses modifications post-traductionnelles(MPT), particulièrement au niveau de leur section N-terminale. Ces modifications réversibles font partie d’un code d’histones épi-génétique transmissible qui orchestre et module dynamiquement certains événements impliquant la chromatine, tels l’activation et la désactivation de gènes ainsi que la duplication et la réparation d’ADN. Ces modifications sont impliquées subséquemment dans la signalisation et la progression de cancers, tels que la leucémie. En conséquence, l'élucidation des modifications d’histones est importante pour comprendre leurs fonctions biologiques. Une méthodologie analytique a été mise au point en laboratoire pour isoler, détecter, et quantifier les MPT d’histones en utilisant une approche rapide à deux volets à l’aide d’outils bioinformatiques spécialisés. La méthodologie développée en laboratoire a été validée en utilisant des histones de souche sauvage ainsi que deux types d’histones mutants déficients en enzymes acétyltransferase. Des trois sources d’histones utilisées, la seule MPT qui a démontré un changement significatif est l’acétylation de l’histone H3 à lysine 56 (H3K56ac). L’expression et la stoechiométrie de cette MPT, issue de cellules de souche sauvage et de cellules mutantes, ont été déterminées avec précision et comparées. Les fonctions de balayage polyvalentes d'un instrument à trappe ionique quadrupôle linéaire hybride ont été utilisées pour améliorer la détection de protéines intactes. Le mode de balayage « enhanced multiply charged » (EMC) a été modifié pour contenir et détecter les ions de protéines intactes situées dans la trappe ionique linéaire. Ce mode de balayage nommé « targeted EMC » (tEMC) a permis de quadrupler le niveau de sensibilité (signal/interférence), et quintupler la résolution du mode de balayage conventionnel. De plus, la capacité de séparation des charges du tEMC a réduit de façon significative les effets de « space charge » dans la trappe ionique linéaire. La résolution supérieure du mode tEMC a permis de différencier plusieurs isoformes modifiées, particulièrement pour l’histone H3. L’analyse des peptides d’histones trypsiques à l’aide du mode de balayage « MRM » a permis le séquençage et la quantification de MPT avec un haut degré de précision. La seule MPT qui était sous-exprimée entre l’histone de souche sauvage et le mutant DOT1L fut la méthylation de l’histone H3 lysine 79(H3K79me1). Les effets de deux inhibiteurs d’enzymes HDAC (HDACi) sur l’expression de MPT d’histone ont été évalués en utilisant la méthodologie analytique mentionnée. Les histones extraites de cellules normales et cancéreuses ont été exposées à du Vorinostat(SAHA) ou du Entinostat (MS-275) pour une période de 24 à 72 heures. Deux histones furent principalement affectées, soit H3 et H4. Étonnamment, les mêmes effets n'ont pas été détectés lorsque les cellules normales ont été traitées avec le HDACi pour une période de 48 à 72 heures. Une méthode absolue de quantification avec une courbe d’étalonnage a été développée pour le peptide H3K56ac. Contrairement à certaines publications, nos résultats démontrent que cette MPT est présente dans les cellules mammifères avec une stoechiométrie très basse (< 0,1%) et n'est pas surexprimée de façon significative après le traitement au HDACi.

Essays in labor and public economics

Dans ma thèse, je me sers de modèles de recherche solides pour répondre à des questions importantes de politique publique. Mon premier chapitre évalue l’impact causal de l’allégeance partisane (républicain ou démocrate) des gouverneurs américains sur le marché du travail. Dans ce chapitre, je combine les élections des gouverneurs avec les données du March CPS pour les années fiscales 1977 à 2008. En utilisant un modèle de régression par discontinuité, je trouve que les gouverneurs démocrates sont associés à de plus faibles revenus individuels moyens. Je mets en évidence que cela est entrainée par un changement dans la composition de la main-d’oeuvre à la suite d’une augmentation de l’emploi des travailleurs à revenus faibles et moyens. Je trouve que les gouverneurs démocrates provoquent une augmentation de l’emploi des noirs et de leurs heures travaillées. Ces résultats conduisent à une réduction de l’écart salarial entre les travailleurs noir et blanc. Mon deuxième chapitre étudie l’impact causal des fusillades qui se produisent dans les écoles secondaires américaines sur les performances des éléves et les résultats des écoles tels que les effectifs et le nombre d’enseignants recruté, a l’aide d’une stratégie de différence-en-différence. Le chapitre est coécrit avec Dongwoo Kim. Nous constatons que les fusillades dans les écoles réduisent significativement l’effectif des élèves de 9e année, la proportion d’élèves ayant un niveau adéquat en anglais et en mathématiques. Nous examinons aussi l’effet hétérogene des tueries dans les écoles secondaires entre les crimes et les suicides. Nous trouvons que les fusillades de natures criminelles provoquent la diminution du nombre d’inscriptions et de la proportion d’élèves adéquats en anglais et mathématiques. En utilisant des données sur les élèves en Californie, nous confirmons qu’une partie de l’effet sur la performance des élèves provient des étudiants inscrits et ce n’est pas uniquement un effet de composition. Mon troisième chapitre étudie l’impact des cellulaires sur la performance scolaire des élèves. Le chapitre est coécrit avec Richard Murphy. Dans ce chapitre, nous combinons une base de données unique contenant les politiques de téléphonie mobile des écoles obtenues à partir d’une enquète auprès des écoles dans quatre villes en Angleterre avec des données administratives sur la performance scolaire des éleves. Nous étudions ainsi l’impact de l’introduction d’une interdiction de téléphonie mobile sur le rendement des éleves. Nos résultats indiquent qu’il y a une augmentation du rendement des éleves après l’instauration de l’interdiction des cellulaires à l’école, ce qui suggère que les téléphones mobiles sont sources de distraction pour l’apprentissage et l’introduction d’une interdiction à l’école limite ce problème.

A Synergistic Approach to Human Rights and Public Health Ethics: Effective or a Source of Conflict?

Commentaire / Commentary

Assessment of Tandem Mass Spectrometry and High Resolution Mass Spectrometry for the Analysis of Bupivacaine in Plasma

Triple quadrupole mass spectrometers coupled with high performance liquid chromatography are workhorses in quantitative bioanalyses. It provides substantial benefits including reproducibility, sensitivity and selectivity for trace analysis. Selected Reaction Monitoring allows targeted assay development but data sets generated contain very limited information. Data mining and analysis of non-targeted high-resolution mass spectrometry profiles of biological samples offer the opportunity to perform more exhaustive assessments, including quantitative and qualitative analysis. The objectives of this study was to test method precision and accuracy, statistically compare bupivacaine drug concentration in real study samples and verify if high resolution and accurate mass data collected in scan mode can actually permit retrospective data analysis, more specifically, extract metabolite related information. The precision and accuracy data presented using both instruments provided equivalent results. Overall, the accuracy was ranging from 106.2 to 113.2% and the precision observed was from 1.0 to 3.7%. Statistical comparisons using a linear regression between both methods reveal a coefficient of determination (R2) of 0.9996 and a slope of 1.02 demonstrating a very strong correlation between both methods. Individual sample comparison showed differences from -4.5% to 1.6% well within the accepted analytical error. Moreover, post acquisition extracted ion chromatograms at m/z 233.1648 ± 5 ppm (M-56) and m/z 305.2224 ± 5 ppm (M+16) revealed the presence of desbutyl-bupivacaine and three distinct hydroxylated bupivacaine metabolites. Post acquisition analysis allowed us to produce semiquantitative evaluations of the concentration-time profiles for bupicavaine metabolites.

Engaging Ethical Issues Associated with Research and Public Health Interventions During Humanitarian Crises: Review of a Dialogic Workshop

Compte-rendu / Review

Capital expenditure decision in private and public sector entreprises in India A comparative evaluation

This study is focussed on public and private sector The study is confined to industrial Public companies in the states of Kerala and Orissa along with companies in India. undertakings only. sector and private sector private sector companies in other states were studied. Even though the original plan of 190 companies as the sample size could not be accomplished, as the responses were very poor, but data could be collected frmn 6 public and 5 private sector companies in Kerala, 8 public and 8 private sector companies in Orissa along with 27 private sector companies in other states totalling to 54 companies. The number of years of data collected varies from 2 years to 6 years. Factors which are affecting capital expenditures and hence leading to the performance of private sector compared to public sector companies are studied. After the study and analysis, comparisons are made between public sector and private sector, and suitable recomendations are made so that public sector industries can also perform equally well as the private sector industries in India.

Economic Impact of Identity Theft in India: Lessons from Western Countries

In this computerized, globalised and internet world our computer collects various types of information’s about every human being and stores them in files secreted deep on its hard drive. Files like cache, browser history and other temporary Internet files can be used to store sensitive information like logins and passwords, names addresses, and even credit card numbers. Now, a hacker can get at this information by wrong means and share with someone else or can install some nasty software on your computer that will extract your sensitive and secret information. Identity Theft posses a very serious problem to everyone today. If you have a driver’s license, a bank account, a computer, ration card number, PAN card number, ATM card or simply a social security number you are more than at risk, you are a target. Whether you are new to the idea of ID Theft, or you have some unanswered questions, we’ve compiled a quick refresher list below that should bring you up to speed. Identity theft is a term used to refer to fraud that involves pretending to be someone else in order to steal money or get other benefits. Identity theft is a serious crime, which is increasing at tremendous rate all over the world after the Internet evolution. There is widespread agreement that identity theft causes financial damage to consumers, lending institutions, retail establishments, and the economy as a whole. Surprisingly, there is little good public information available about the scope of the crime and the actual damages it inflicts. Accounts of identity theft in recent mass media and in film or literature have centered on the exploits of 'hackers' - variously lauded or reviled - who are depicted as cleverly subverting corporate firewalls or other data protection defenses to gain unauthorized access to credit card details, personnel records and other information. Reality is more complicated, with electronic identity fraud taking a range of forms. The impact of those forms is not necessarily quantifiable as a financial loss; it can involve intangible damage to reputation, time spent dealing with disinformation and exclusion from particular services because a stolen name has been used improperly. Overall we can consider electronic networks as an enabler for identity theft, with the thief for example gaining information online for action offline and the basis for theft or other injury online. As Fisher pointed out "These new forms of hightech identity and securities fraud pose serious risks to investors and brokerage firms across the globe," I am a victim of identity theft. Being a victim of identity theft I felt the need for creating an awareness among the computer and internet users particularly youngsters in India. Nearly 70 per cent of Indian‘s population are living in villages. Government of India already started providing computer and internet facilities even to the remote villages through various rural development and rural upliftment programmes. Highly educated people, established companies, world famous financial institutions are becoming victim of identity theft. The question here is how vulnerable the illiterate and innocent rural people are if they suddenly exposed to a new device through which some one can extract and exploit their personal data without their knowledge? In this research work an attempt has been made to bring out the real problems associated with Identity theft in developed countries from an economist point of view.

TRIPS Agreement and Public Health: An Overview of International Issues

This paper deals with brief overview of the developments of international provisions on IPR related to public health. It discusses flexibilities before and after TRIPS Agreement and difficulties faced by developing countries in implementing TRIPS obligations and protecting public health. Also discussed are the reasons for the Doha Declaration and issues relating to implementation of Para 6 of the Declaration. Discusses the inadequacy in the compulsory licence based approach to solve public health crisis and argues for a more comprehensive approach to find a long term solution to the public health issues

From evolution to co-evolution – An empirical study of the role of innovation, policy and public research in industrial dynamics

In the past decades since Schumpeter’s influential writings economists have pursued research to examine the role of innovation in certain industries on firm as well as on industry level. Researchers describe innovations as the main trigger of industry dynamics, while policy makers argue that research and education are directly linked to economic growth and welfare. Thus, research and education are an important objective of public policy. Firms and public research are regarded as the main actors which are relevant for the creation of new knowledge. This knowledge is finally brought to the market through innovations. What is more, policy makers support innovations. Both actors, i.e. policy makers and researchers, agree that innovation plays a central role but researchers still neglect the role that public policy plays in the field of industrial dynamics. Therefore, the main objective of this work is to learn more about the interdependencies of innovation, policy and public research in industrial dynamics. The overarching research question of this dissertation asks whether it is possible to analyze patterns of industry evolution – from evolution to co-evolution – based on empirical studies of the role of innovation, policy and public research in industrial dynamics. This work starts with a hypothesis-based investigation of traditional approaches of industrial dynamics. Namely, the testing of a basic assumption of the core models of industrial dynamics and the analysis of the evolutionary patterns – though with an industry which is driven by public policy as example. Subsequently it moves to a more explorative approach, investigating co-evolutionary processes. The underlying questions of the research include the following: Do large firms have an advantage because of their size which is attributable to cost spreading? Do firms that plan to grow have more innovations? What role does public policy play for the evolutionary patterns of an industry? Are the same evolutionary patterns observable as those described in the ILC theories? And is it possible to observe regional co-evolutionary processes of science, innovation and industry evolution? Based on two different empirical contexts – namely the laser and the photovoltaic industry – this dissertation tries to answer these questions and combines an evolutionary approach with a co-evolutionary approach. The first chapter starts with an introduction of the topic and the fields this dissertation is based on. The second chapter provides a new test of the Cohen and Klepper (1996) model of cost spreading, which explains the relationship between innovation, firm size and R&D, at the example of the photovoltaic industry in Germany. First, it is analyzed whether the cost spreading mechanism serves as an explanation for size advantages in this industry. This is related to the assumption that the incentives to invest in R&D increase with the ex-ante output. Furthermore, it is investigated whether firms that plan to grow will have more innovative activities. The results indicate that cost spreading serves as an explanation for size advantages in this industry and, furthermore, growth plans lead to higher amount of innovative activities. What is more, the role public policy plays for industry evolution is not finally analyzed in the field of industrial dynamics. In the case of Germany, the introduction of demand inducing policy instruments stimulated market and industry growth. While this policy immediately accelerated market volume, the effect on industry evolution is more ambiguous. Thus, chapter three analyzes this relationship by considering a model of industry evolution, where demand-inducing policies will be discussed as a possible trigger of development. The findings suggest that these instruments can take the same effect as a technical advance to foster the growth of an industry and its shakeout. The fourth chapter explores the regional co-evolution of firm population size, private-sector patenting and public research in the empirical context of German laser research and manufacturing over more than 40 years from the emergence of the industry to the mid-2000s. The qualitative as well as quantitative evidence is suggestive of a co-evolutionary process of mutual interdependence rather than a unidirectional effect of public research on private-sector activities. Chapter five concludes with a summary, the contribution of this work as well as the implications and an outlook of further possible research.

Teoría y práctica de la diplomacia de celebridades : origen, desarrollo y límites de un concepto

Recientemente el mundo ha sido testigo de como las celebridades del espectáculo aparecen en los medios de comunicación comentando sobre asuntos políticos y humanitarios, el motivo de dichas apariciones es que cada día los famosos están interviniendo en temas internacionales de orden político y diplomático en un esfuerzo por solucionar algunas de las más graves crisis a las que se enfrenta el mundo en las últimas dos décadas, este fenómeno es conocido como Diplomacia de celebridades. El presente es un análisis acerca de este concepto en cuanto a su origen, desarrollo y alcance, apartir de la óptica del poder blando y ciertas condiciones que permiten la existencia del fenómeno, tales como el avance de las tecnologías digitales y de comunicación, fama de alcance global y nuevos espacios y actores en el ejercicio de las Relaciones Internacionales. La hipótesis sostiene que los famosos realizan una actividad no oficial y utilizan elementos como el poder blando, el dinero y los medios de comunicación para realizarla, tal y como lo haría una ONG. Ellos se identifican con una causa o un tema y utilizan los medios de comunicación como infraestructura para adentrarse en los escenarios de política internacional y promoverla a nivel de gobiernos y opinión pública. Usualmente los emprendimientos sociales y políticos de las celebridades suelen ser efectivos, sin embargo se enfrentan a campos donde su inexperiencia puede llevarlos al fracaso, la falta de comprensión de situaciones muy complejas puede considerarse otra limitante a la que se enfrentan los famosos, puesto que su mensaje o sus esfuerzos se distorsionan al no conocer los matices de una situación y de esta manera generar más perjuicios que beneficios.

Review: Social Media and Abstract Nouns

The revision slides for our Social Media course, contains major lessons learned throughout the course, and an example exam question (on trust).

Procesos de privatización del agua en América Latina: análisis y seguimiento de la interacción estratégica entre movimientos sociales versus multinacionales en Bolivia, Uruguay y México. Período 2000 – 2006

El presente escrito, constituye una aproximación a los procesos de privatización del agua en América Latina. Lo anterior, en el marco de la acción colectiva, específicamente de los movimientos sociales y su interacción estratégica con oponentes como las compañías multinacionales, las organizaciones financieras internacionales y el Estado. Tomando como referencia las luchas ocurridas en Bolivia, Uruguay y México, con especial énfasis en La Guerra del Agua en Cochabamba (Bolivia).

Influencia de las creencias religiosas cristianas en la decisión de realizar suicidio asistido al final de la vida

El suicidio asistido como una posible opción al final de la vida, es una idea que hasta ahora está siendo considerada, ya que existen argumentaciones a favor y en contra que han generado controvertidos debates a su alrededor. Algunos de los argumentos en contra están basados en los principios de las instituciones religiosas de orden cristiano, las cuales defienden el valor sagrado de la vida de las personas y la aceptación del sufrimiento como un acto de amor profundo y sumisión a los mandatos de Dios, el creador. Mientras del lado contrario, se encuentran quienes defienden el procedimiento, impulsando la autonomía y la autodeterminación que cada persona tiene sobre su vida. La revisión de la literatura realizada no sólo permite ampliar los argumentos de estas dos posiciones, sino que también permite conocer la historia del suicidio asistido, la posición que este procedimiento tiene en diferentes países del mundo, incluyendo a Colombia, y finalmente se presentan las contribuciones de la psicología entorno al procedimiento en discusión.

Maternal Mortality in Colombia: Some Thoughs on the Situation and Public Policy During Last Decade

In this article we make a critical analysis about the situation of maternal mortality in Colombia, considering some of the most current rates of this problem and the public policy rules adopted in the last years to decrease its prevalence. We think that the rate of maternal mortality in Colombia is too hight because the State does not guarantee the right health care to Colombian women. In this way, the structure and resources distribution established by the policies on sexual and reproductive health have not reflected the population necessities, specially in the case of young mothers left aside by the social security system.